Preparation of recombinant proteins using yeast

Page 1

1.ࠈPreparation of recombinant proteins using yeast expression system Capacity:ࠈࠈࠈࠈ5ࠤ10 Department:ࠈ ࠈMolecular Biochemistry Teaching staff:ࠈ Tetsuya Okajima Contact: ࠈࠈ ࠈTel 2068 ࠈࠈࠈ ࠈE-mail Time and Date:ࠈSeptember 1 and 4 (2 days) Meeting place:ࠈDept. of Biochemistry II

Outline: For protein expression, various expression systems such as E. coli, yeast and insect cells are available. The yeast expression system offers several advantages over other systems since it allows to produce the large amount of correctly folded disulfide-bonded recombinant proteins. In this training course, EGF domains of Notch receptors are expressed using K. lactis Proteins Expression Kit (NEB), purified using Ni-affinity beads, and detected by dot-blotting. Day 1 (Mon/PM) Overview, preparation of culture media, inoculation Day2 (Thr/PM) Purification of recombinant proteins and its detection by dot blotting


2.ࠈYeast two hybrid system Capacity:ࠈࠈࠈࠈ6 Department:ࠈࠈࠈMolecular Mycology and Medicine Teaching staff:ࠈ

Tomohiro Akashi

Contact:ࠈࠈࠈࠈࠈTel 2460 ࠈࠈࠈࠈࠈE-mail akashi@med.nagoya-u.ac.jp Time and Date:ࠈ June 10, 13, 17, 20 Afternoon. ࠈࠈThe first day (June 10) starts at 2:00 p.m. Meeting place:ࠈ

Basic Research Building, Annex 5F

Outline: Yeast two hybrid system is one of excellent methods to examine proteinprotein interactions. Typically, this method is used to detect genes whose translated products can interact with protein of interest, from a collection of genes like cDNA libraries. This is also used to analyze the detail of protein-protein interaction. This technique is basically applicable to any genes, and is highly universal. Furthermore, its experimental procedures are highly organized, so people can use it without skilled experiences. In the course, we demonstrate the outline of all the experimental procedures. The demonstration also focuses on the handling of yeast cells for the two hybrid system. This course requires 4 afternoons indicated above. Applicants to this course should be careful to the schedule. Applicants also would be required to answer a questionnaire for the course before approval.


3.ࠈLaser scanning cytometer (LSC) Capacity:ࠈࠈ ࠈࠈ8 (maximum: 16) Department:ࠈࠈ ࠈMolecular Mycology and Medicine Teaching staff:ࠈࠈ Tomohiro Akashi Contact: ࠈࠈࠈࠈTel 2460 ࠈࠈࠈࠈࠈE-mail akashi@med.nagoya-u.ac.jp Time and Date:ࠈ October 7, afternoon (2:00 p.m. – 5:00 p.m.) Meeting place:ࠈ

Basic Research Building, Annex 5F

Outline: Laser scanning cytometer (LSC) is an epifluorescence microscope equipped with a laser. This special microscope is designed to quantitate the fluorescence emitted from the sample mounted on slide glass excited by the laser beam. LSC can quantitate any fluorescence signals from cells stained with fluorochrome. For an example, DNA content of a cell can be quantitated by cells stained with propidium iodide (PI). LSC is occasionally misunderstood as flow cyotometer. Both can give us similar data, but LSC is based on microscopy. In the course, the merit and demerit of LSC are explained. Staining procedure of samples with propidium iodide and measurement of the DNA content of the sample are demonstrated. Attendees would also learn how to handle it practically. When the number of applicants are over the capacity, the schedule may be changed. Change of schedule would be notified via e-mail in that case.


4.ࠈIntroduction to the NCBI Gene Database Capacity:ࠈࠈࠈࠈ100 students Department: ࠈࠈNeurogenetics Teaching staff: ࠈKinji Ohno Contact: ࠈࠈࠈࠈPhone. ext. 2446 ࠈࠈࠈࠈ E-mail ohnok@med.nagoya-u.ac.jp Time and Date: ࠈJune 11, 2007 (Wed) or June 13, 2008 (Fri), 13:00 – 17:00 I will repeat the identical seminar on two days, because the maximum capacity of the satellite laboratory is limited to 50. You will be assigned to either day. Meeting place: ࠈSatellite laboratory, 2nd floor of the Annex Building of the Medical Research (the Kisobekkan).

Outline: The genome information that we can access is expanding day by day. Handling of the genome information is getting more and more essential in a variety of medical and biological researches. The seminar is intended for students who employ the molecular biological techniques in their research programs. The seminar covers the basic strategies and techniques of the NCBI (the National Center for Biotechnology Information) gene database search. The specific topics to be covered include BLAST, uniGene, uniSTS, Entrez Gene, SNP, OMIM, and PubMed. The course also covers the OVID Medline database, the RefScout service, and the CiteTrack service. Do not forget to bring your Nagoya Univeristy ID and its password, when you attend the seminar.


5.ࠈInduction and analysis of bone marrow derived dendritic cells Capacity:ࠈࠈ

ࠈ6 persons

Department:ࠈ ࠈDepartment of Aging Research, Program in Integrated Molecular Medicine Teaching staff:ࠈ Mitsuo MARUYAMA Contact: ࠈ ࠈTel : 0562-46-2311 (Ext. 5101) ࠈࠈ ࠈE-mail: michan@nils.go.jp Time and Date:ࠈtotal 3 days around late in September or early in October Meeting place:ࠈ Department of Mechanism of Aging, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology

Outline: The dendritic cell plays an important role with the antigen presentation in acquired immune system. It is well established that in vitro differentiated dendritic cell is induced from bone marrow cells by GM-CSF. In this basic training course, students will prepare the murine bone marrow derived dendritic cells and characterize them by FACS analysis. All procedures and techniques are generally plain and simple therefore the Master's course students are also welcomed.


6.ࠈAnalysis of Cellular Senescence Capacity: ࠈࠈ ࠈ6 Department:

Department of Aging Research, Program in Integrated Molecular Medicine

Teaching staff: ࠈNoboru Motoyama Contact: ࠈࠈࠈࠈTel 0562-46-2311 (Ext. 5554) ࠈࠈࠈࠈ E-mail: motoyama@nils.go.jp Time and Date:ࠈTBA (2days) Meeting place:

Department of Geriatric Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology

Outline: Cellular senescence, which is a stable state of proliferative arrest, is triggered by telomere shortening during replication. The cellular senescence also is induced by activated oncogenes, DNA damage, and oxidative stress. Recent studies have revealed that cellular senescence plays a critical role for tumor suppression and organism aging. In this basic training course, students will learn techniques to study cellular senescence. An important element will be the informal interaction between students and course faculty.


7.ࠈFunctional imaging of neuronal cells in brain slice cultures Capacity: ࠈࠈ ࠈ10 people Department: ࠈࠈDept. of Physiology II Teaching staff: ࠈDr. Tatsumi Contact: ࠈࠈࠈࠈTel ࠈ2054 ࠈࠈࠈࠈ E-mail ࠈtatsumi@med.nagoya-u.ac.jp ࠈࠈࠈࠈNeed contact with Dr.Tatsumi via E-mail as well as university office (Jimu, Kyomugakari). Time and Date: ࠈ㸪th and 㸫th June Meeting place: ࠈ403 room at 4th floor Basic Science Building

Outline: A short lecture for introducing the scientific importance of live cell imaging and demonstration of GFP neurons in a brain slice culture.


8.ࠈExperimental methods of Neuropsychopharmacology Capacity:ࠈࠈࠈࠈ20 Department:ࠈ

Department of Medical Pharmacy (Department of Neuropsychopharmacology and Hospital Pharmacy)

Teaching staff:ࠈKiyofumi Yamada Ph.D. Contact: ࠈࠈࠈTel 052-744-2674 ࠈࠈࠈࠈE-mail a-nitta@med.nagoya-u.ac.jp Time and Date:ࠈ23-24, June Meeting place:ࠈLaboratory of Medical Pharmacy, 1st floor of 1st building of clinical departments.

Outline: Purpose; Learning how to do experimental methods of neuropsychopharmacology, using rats or mice. 1. Experiments to evaluate the ability of learning and memory 1) Morris’s water maze task 2) Eight arms radial maze 3) Habituation task 4) Passive avoidance task 2. Experiments to evaluate depression and anxietyࠈin mice


9.ࠈWhole-cell patch clamp recordings using brain slice preparations Capacity: ࠈࠈࠈ5 Department: ࠈ Visual Neuroscience, Research Institute of Environmental Medicine Teaching staff: ࠈYumiko Yoshimura Contact: ࠈࠈࠈࠈTel: 3879 ࠈࠈࠈࠈ E-mail: yyumiko@riem.nagoya-u.ac.jp Time and Date:ࠈOctober 偐, 㸨 Meeting place:ࠈ Room 409 of the forth floor, Research Institute of Envirionmental Medicine on Higashiyama campus

Outline: The aim of this program is to introduce students to the basic procedures for preparing brain slices and analyzing synaptic transmission with wholecell patch clamp techniques. Students are asked to prepare slices from developing rat visual cortex and record synaptic responses evoked by electrical stimulation. Synaptic transmissions will be analyzed with a pharmacological method.


10.ࠈIsolation and culture of glial cells. Capacity: ࠈࠈࠈ 8 Department: ࠈࠈNeuroimmunology, Research Inst. Environmental Med. Teaching staff:

Akio Suzumura

Contact: ࠈ Tel ࠈExt. 3881(Higashiyama) ࠈࠈE-mail suzumura@riem.nagoya-u.ac.jp Time and Date:

Sept. 5 (Wed), 2007

Meeting place: ࠈDepartment of Neuroimmunology, RIEM

Outline: Glial cells consist of oligodendrocytes, astrocytes and microglia. We have established the isolation method to purify each glial cells from the primary mixed glial cell cultures prepared from neonatal rodents. The method enable us to use purified cells in various biochemical, molecular and immunological experiments.


11.ࠈPrimary culture of cortical neurons Capacity:

ࠈࠈ8

Department: ࠈ Department of Neuroimmunology, Research Institute of Environmental Medicine Teaching staff: ࠈTetsuya Mizuno Contact: ࠈࠈࠈࠈTel 㸯052-789-3883 ࠈࠈࠈࠈE-mail㸯tmizuno@riem.nagoya-u.ac.jp Time and Date:ࠈSeptember 4

2007

13㸯30ࠤ16㸯00

Meeting place:ࠈDepartment of Neuroimmunology, Research Institute of Environmental Medicine (4F)

Outline: Using primary neuronal culture, co-culture of microglia and neuron, and organotypic cortical slice cultures, the course introduces how to evaluate neuronal damage.


12.ࠈin vivo imaging technique for small animals Capacity:ࠈࠈ

ࠈ5 to 6

Department:ࠈ ࠈBrain Life Science Teaching staff:ࠈ

Kenji Ono

Contact: ࠈ ࠈࠈTelࠈ052-789-5002 ࠈࠈࠈ ࠈE-mailࠈ k_ono@riem.nagoya-u.ac.jp Time and Date:ࠈ 8/1 (Fri) A.M.10:00-P.M.5:00 Meeting place:ࠈ Seminar room in Research Institute of Environmental Medicine

Outline: The fate of drug substances administered externally to living animals is usually determined with body fluids and dissected tissues. ࠈࠈࠈࠈࠈ Recent improvements of in vivo imaging technology such as PET, MRI and even optical imaging, however, provide a new strategy for analyzing drug distribution, adsorption and metabolism. These imaging technologies also have advantages to determine the tagged substances such as drug vehicles and cells in a living animal. In this course, we provide a short lecture and a fundamental practice for a small animal MRI instrument and an in vivo optical imager.


13.ࠈBehavioral analyses in rodents Capacity:ࠈࠈࠈࠈ5ࠤ6 Department: ࠈ ࠈ Department of Brain Function, Research Institute of Environmental Medicine Teaching staff:ࠈ Hiroyuki Mizoguchi Ph.D. Contact: ࠈࠈࠈTelࠈ052-789-3929 ࠈࠈࠈࠈE-mailࠈh-mizo@riem.nagoya-u.ac.jp Time and Date:ࠈOne day for the period of summer vacation Meeting place:ࠈ Research Institute of Environmental Medicine

Outline: Purpose; Learning behavioral experimental methods using rodents. Experiments to evaluate the ability of learning and memory 1) Morris’s water maze task 2) Novel object recognition task 3) Elevated plus-maze task 4) Y-maze task


14.ࠈHow to measure pain in human and animal pain models Capacity:    5 students Department:   Futuristic Environmental Simulation Center, Research Institute of Environmental Medicine Teaching staff:  Jun SATO, MD, PhD. Contact:

Tel Higashiyama -3865 E-mail jun@riem.nagoya-u.ac.jp

Time and Date:  Tuesday, June 19, 2007 from 10:00 a.m. to 3:00 p.m. Meeting place:

Futuristic Environmental Simulation Center, Research Institute of Environmental Medicine (Higashiyama campus)

Outline: Persistent or chronic pain is the primary reason people seek medical care. Therefore It is very important for physicians to know how to measure pain sensitivity in patients. This course aims to provide the student with understanding and knowledge of pain mechanism and pain tests. In this lecture you will learn: 1. Pain in human (Lecture) 1) profile and physiological significance of various types of pain 2) pain tests (VAS scale, etc.) 2. Pain in experimental animals (Lecture and Exercise) 1) methods of making acute and chronic pain models 2) pain tests (von Frey hair test, Randall-Selitto test, etc.)


15.ࠈIntroduction to the electrical recording of biological signals Capacity: ࠈࠈࠈ6 persons Department: ࠈ

Department of Neuroscience II, Research Institute of Environmental Medicine

Teaching staff: ࠈKazue Mizumura Contact: ࠈࠈࠈࠈTel (extension) 85Ѹ 3861 ࠈࠈࠈࠈE-mailࠈmizu@riem.nagoya-u.ac.jp Time and Date:ࠈJuly 30ࠈ(Wen)

and 31 (Thu). 10:00-16:00

Meeting place: Room 215, Department of Neuroscience II, Research Institute of Environmental Medicine (Higashiyama-campus)

Outline: ࠈIn clinical practice it is quite common and essential to record biological signals with electrical methods (e.g. ECG). These methods are also widely used in basic medical research, for example, patch clamp recording, intra- and extra-cellular recordings etc. Therefore, it is important to have basic knowledge on mechanisms of electrical recording and instruments for using them properly and obtaining correct data. In this seminar following points will be overviewed and practiced. 1) Different kinds of biological signals and their electrical characteristics 2) Amplifier, filter, pen recorder, oscilloscope, magnetic tape recorder, electrical stimulator, various transducers 3) Characteristics of digital instruments and points to notice 4) Electrodes 5) Countermeasures against electronic noises 6) Practice using operational amplifiers


16.ࠈRecording of primary afferents with the single-fiber recording technique (dissection method). Capacity: ࠈࠈࠈ5 Department: ࠈ

Neural Regulation

Field: ࠈࠈࠈࠈ

Regulation of Organ Function

Program:ࠈࠈࠈ

Program in Cell Information Medicine

Teaching staff:ࠈ Toru TAGUCHI Contact:

ࠈࠈTel: 052-789-3863 E-mail: tagu@riem.nagoya-u.ac.jp

Time and Date: ࠈ10:00-17:00, Aug 5, 2008. (Aug 6 for applicants) Meeting place:ࠈMain Bld. 2F (Room 215), Research Institute of Environmental Medicine, Higashiyama-campus. Outline: The single-fiber recording technique has been established in the latter 1960s. It is a kind of electrophysiology and enables to get action potentials of a single sensory receptor fiber in the peripheral tissues. The technique is widely applicable both in vivo and in vitro. In our lab, we have been studied peripheral mechanisms of nociception with skin-, muscle-, and testis-nerve preparations. In this course, activities of a single nociceptor (myelinated A_- or unmyelinated Cfiber) will be recorded in a rat extensor digitorum longus muscle-peroneal nerve preparation in vitro. Participants will learn below. Students who will take this course are advisable to take the course ‘Introduction to the electrical recording of biological signals’. 1)

A brief introduction to the single-fiber recording method electrophysiological instruments (lecture) 2) How to make the rat muscle-nerve preparation (practice) 3) Recording of nociceptor activities in the preparation made (practice)

and


17.ŕ ˆHow to use antibodies (Immunoprecipitation & Western blotting) Capacity: Department: Teaching staff:ŕ ˆ Contact: Tel E-mail Time and Date: Meeting place:

6 Dept. of Neural Regulation (RIEM, Higashiyama-campus) Katanosaka, K. ex. 85-3862/3864 katano@riem.nagoya-u.ac.jp Two days in September (10:00-17:30) The conference room in the Northern building 2F, Research Institute of Environmental Medicine (RIEM), Higashiyama-campus

Outline: Specific antibodies are powerful tools for medical and life science studies. This course is opened for the students who want to learn the basic information that is necessary to use antibodies and interpret the results of experiments using antibodies. In the course, everyone can get training in preparation of protein samples from tissues, handling of antibodies, and techniques for western blotting and immunoprecipitation. The course includes some short lectures as shown below: 1, Lecture 1, How to make antibodies. 2, Lecture 2, How to treat protein samples (its physicochemical properties). 3, Experiment 1, Preparation of protein samples and immunoprecipitation. 4, Experiment 2, Western blotting 5, Lecture 3, Troubleshooting in western blotting for beautiful results NOTICE! To participate this course, you need to apply to the school office and also email to the teaching staff of the course (katano@riem.nago-ya-u.ac.jp, Mail title: BT20, Deadline: June 30). If the number of applicants exceeds the capacity, the members will be selected by the staff and announced by e-mail and on the WEB site for Basic training until July 4. In your e-mail, please mention 1: Name, 2: ID, 3: e-mail address and the phone number of your office, 4: Date you prefer ( A, Tue. & Wed. ; B, Thu. & Fri. ; C, either) and 5: Any other things (e.g. your motive, experience, troubles on your use of antibodies, etc.).


18.ࠈCulture of cardiomyocytes Capacity:ࠈࠈࠈࠈ5 persons Department:ࠈࠈDivision of Cardiovascular Research (2), (Research Institute of Environmental Medicine) Teaching staff:ࠈMasahide Harada Contact: ࠈࠈࠈTel 052-789-3871 ࠈࠈࠈࠈE-mail mharada@riem.nagoya-u.ac.jp Time and Date: ࠈSeptember 11 (Thur.), 12 (Fri.) Meeting place: ࠈDepartment of Cardiovascular Research (2), 3rd floor of main building, Research Institute of Environmental Medicine. Outline: (Purpose) The main purpose of this course is to master the fundamental techniques on culturing cardiomyocites in cardiovascular research field. You can experience the primary culture of neonatal rat cardiomyocytes and get the basic skill needed. (Schedule) First day: Introduction of the primary culture of myocytes including the fundamental procedure, medium and equipments. The neonatal rat ventricles are carefully excised out of the chest. The ventricles are plated in the culture dish and are isolated into single myocytes by enzyme reaction. Second day: Culture of isolated myocytes. Isolated myocytes are incubated in the culture dish containing appropriate medium and their growth is microscopically observed.


19.ࠈInduction of cardiac cells from embryonic stem cells ~ Creating regenerative myocardium ~ Capacity:ࠈࠈࠈ Ten students Department:

Department of Cardiovascular Research (Circulation) Research Institute of Environmental Medicine

Teaching staff: ࠈLEE Jong-Kook Contact: ࠈࠈࠈࠈTel (052) 789-3871 ࠈࠈࠈࠈE-mail: jlee@riem.nagoya-u.ac.jp (Note) Please inform us by e-mail if you cannot participate in the course after you enrollment. Time and Date: Day 1: Monday in May or June (1:00PM- 3:00 PM.) Day 2: Wednesday in the following week of Day1 (1:00PM- 3:00 PM.) These schedules may be subject to change due to the condition of ES cells. Meeting place: ࠈRoom 312 Main building Third floor Department of Cardiovascular Research, Research Institute of Environmental Medicine, Nagoya University (Higashiyama campus) Outline: “Regenerative medicine” is a field of great concern in recent years, which restore the lost functions of failing organs through actively utilizing stem cells or stem cell-derived tissues. Especially, embryonic stem (ES) cell is attracting attention as a potential cell source because the cell possesses prulipotency to differentiate into almost all kinds of cells. In the field of cardiovascular research, ES cell-derived myocardial and vascular tissues are extensively studied. In this basic training course, our goal is to outline the methods of induction and isolation of cardiac cells from murine ES cells and try to obtain basic knowledge and technique. On Day 1 of the basic training, we start differentiation from undifferentiated embryonic stem cells to form embryoid bodies. On Day 2, about 7-10 days later, it is scheduled that students will observe spontaneously beating embryoid bodies.


20.ࠈOccytes Expression Method for Ion Channel Measurement Capacity: Department:

ࠈ5 persons ࠈࠈHumoral Regulation (Res Inst of Environ Med)

Teaching staff:

ࠈKaichiro Kamiya (Ryoko Niwa)

Contact: ࠈࠈࠈࠈ Tel ࠈ85-5006 ࠈࠈࠈࠈࠈE-mail kamiya@riem.nagoya-u.ac.jp Time and Date:

Jul 16 (Mon) AM 10:00 - PM 4:00

Meeting place: ࠈࠈResearch Institute of Environmental Medicine (Higashiyama Campus) Room N209 (North building) Outline: Oocytes expression method is one of the standard experimental methods for cellular electrophysiology. Ion channel cRNA was injected in the Xenopus Oocytes and two-three days later channel activities were measured by using double-microelectrodes voltage clamp methods. Activities of specific ion channels or detailed action of drugs on ion channels were widely studied by this method. Especially, this method is mostly effective in the studies to clarify pathological analysis of ion channel diseases due to genomic disorder or molecular action of antiarrhythmic drugs. In this basic training, a surgical procedure to remove oocytes from frog, an enzyme treatment and cRNA injection were performed by the participants. In addition, measurement of ion channel currents in pre-injected oocytes was experienced by using voltage clamp method.


21.ࠈAnimal model (rabbits) for cardiovascular research Capacity:ࠈࠈࠈ 10 Department:ࠈࠈDepartment of Cardiovascular Research ࠈResearch Institute of Environmental Medicine Teaching staff:ࠈ Yukiomi Tsuji, MD, PhD Contact: ࠈࠈࠈTel: 052-789-5006 ࠈࠈࠈࠈE-mail: ytsuji@riem.nagoya-u.ac.jp Time and Date:ࠈ AM10:00-PM1:00, September 4, 2008 Meeting place:ࠈ N201, Research Institute of Environmental Medicine

Outline: This course is for learning the basic skills to operate a small animal (rabbit) including anesthesia and intubation techniques.


22.ࠈOptical recording of membrane potential Capacity:ࠈࠈࠈ 8 Department:ࠈ

Cardiovascular Research (Humoral Regulation) Research Institute of Environmental Medicine

Teaching staff:ࠈ Haruo Honjo Contact: ࠈࠈࠈTel ࠈ789-5007 ࠈࠈࠈࠈE-mailࠈhonjo@riem.nagoya-u.ac.jp Time and Date:ࠈ16-17 September, 2008 Meeting place:

Cardiovascular Research (Humoral Regulation) Research Institute of Environmental Medicine

Outline: Optical measurements of transmembrane potential using fluorescent voltage-sensitive dyes have made important contributions to our understanding of electrophysiological properties of excitable tissues. These methods are non-invasive compared with conventional microelectrode techniques, capable of simultaneous recording with spatiotemporal resolution, and measure membrane potential signals without artifacts caused by electrical stimulation. Optical mapping of cardiac excitation waves has been widely used to investigate mechanisms underlying cardiac arrhythmias, cardioversion and electrical activity in the heart. This program includes lecture to lean theoretical backgrounds and experimental technology of voltage-sensitive dye mapping and practical laboratory training using Langendorf-perfused whole heart preparations.


23.ࠈWhole-mount immunohistochemistry Capacity:

ࠈࠈ 6

Department: ࠈDepartment of Genetics, Research Institute of Environmental Medicine Teaching staff: Yasuhiko Kanou Contact: ࠈࠈࠈTel: 85 (Higashiyama) -3875 ࠈࠈࠈ E-mail; kanou@riem.nagoya-u.ac.jp Time and Date:ࠈJuly 24 – 25 Meeting place:

Department of Genetics, Research Institute of Environmental Medicine

Outline: Whole-mount immunohistochemistry is the localization of antigens in unsectioned tissue, such as embryos, using specific antibodies. Although it is similar in outline to immunohistochemistry of sectioned material and shares some problems, it has several advantages. In this training course, mouse embryos will be stained with specific antibodies for nervous tissue by wholemount immunohistochemistry. Outline: 1. Dissection 2. Fixation and storage 3. Whole-mount immunohistochemistry 4. Color development, clearing and observation with the stereomicroscope


24.ࠈImmunohistocyotchemistry by confocal laser scanning microscopy Capacity: ࠈࠈ

10

Department:

Department of Genetics, Division of Stress Adaptation and Protection, Research Institute of Environmental Medicine (RIEM)

Teaching staff: ࠈYoshiko Takagishi Contact: ࠈࠈࠈࠈTel: 052-789-3874 or Ex 85-3874 ࠈࠈࠈࠈ E-mail taka@rime.naogya-u.ac.jp Time and Date:ࠈJuly 21(in the afternoon) and 22 (in the morning or afternoon) Meeting place: ࠈA meeting venue at the north wing of RIEM

Outline: Confocal laser scanning microscopy offers several advantages over conventional optical microscopy, including obtaining of high-resolution images from relatively thick sections, simultaneous collection of multiple images, and collecting serial optical sections and their three dimensional representation. Application of confocal laser scanning microscopy has been gaining popularity in immunohistocytochemistry to visualize localization of specific proteins in cells and tissues. Practice 1: Immunofluorescence labeling using tissue sections 2: Image collections by confocal laser scanning microscopy ձSingle and double optical sections ղz-series and three-dimensional reconstructions


25.ࠈPrinciple and application of RT-PCR Capacity: ࠈࠈࠈaround 10 students per day (Total 20 students) Department: ࠈࠈDepartment of Endocrinology, Research Institute of Environmental Medicine (RIEM), Nagoya University Teaching staff: ࠈFukushi Kambe, Xia Cao Contact: ࠈࠈࠈࠈTel 85-3867 ࠈࠈࠈࠈE-mail kambe@riem.nagoya-u.ac.jp Time and Date: ࠈ2008.6.18 (Wed) and 19 (Thu) ࠈFrom 10:00 am - 5:00 pm ࠈEach day includes the same lecture and practice so that ࠈapplicant attends either day. Meeting place: ࠈMain Bldg. Room No.203, Department of Endocrinology, RIEM

Outline: The course includes the lecture on the principle of RT-PCR and its applications such as quantitative PCR, cloning of PCR product (TA cloning), in vitro transcription and translation of PCR product, PCR-based cDNA subtraction (representational difference analysis, RDA), rapid amplification of cDNA ends (RACE), and site-directed mutagenesis, etc. It also includes the practice of RTPCR.


26.ࠈBioimaging (Transmission Electron Microscopy) Capacity:ࠈࠈࠈࠈ5 Department: ࠈ ࠈ Anatomy and Cell Biology, Center for Research of Laboratory Animals and Medical Research engineering Teaching staff:ࠈࠈToyoshi Fujimoto, Yoshikazu Fujita, Ikuyo Mizuguchi Contact: ࠈࠈࠈࠈTelࠈ2404 ࠈࠈࠈࠈࠈE-mailࠈyfujita@med.nagoya-u.ac.jp Time and Date:ࠈࠈJuly 22-25, 2008 Meeting place:ࠈࠈto be announced

Outline: Hands-on training course for transmission electron microscopy, including fixation embedding, ultrathin sectioning, EM observation, and data acquisition.


27.ࠈFundamental experimental technology for Cell Biology / Molecular Biology: Observation paraspeckles of RNA binding motif protein (RBM) and glucose transport of human normal cells and tumor cells. Capacity:ࠈࠈ ࠈIt is around㸫 people from capacity 㸨 Department:ࠈࠈAnatomy and Functional Histology (Anatomy ϩ)

Teaching staff:ࠈKoji Nishio PhD. Contact: ࠈࠈࠈTelࠈ An extension number:2021 ࠈࠈࠈࠈE-mailࠈknishio@med.nagoya-u.ac.jp Time and Date:ࠈFive days of May 26 (Monday) to May 30 (Friday), 2008. ࠈ ࠈࠈ ࠈ Four days of extra day (optional): June 2 to 5, 2008. ࠈࠈ ࠈ You can participate on the way of the training course. Meeting place:ࠈFunctional Histology (Room# 329, Cell culture room (# 327) and laboratory (# 339) ( the third floor/ basic research build.).

Outline: Observation of and human normal cells and tumor cells with functional fluorescence probes for the nuclei and nuclear bodies. With the expression vectors of GFP or RFP-fused protein and a several fluorescent pigments which are specific for the sub-cellular organelles, we observe the remark changes such as nuclear morphology, nuclear body assembly and disassembly, glucose uptake in human normal cells and tumor cells. We learn a remark method and applications of GFP, RFP, fluorescent glucose. Through, this training, we learn about technique of RNA interference with the RBM siRNA that inhibits expression of the nuclear RNA binding motif protein. The training course contents: we do cell culture operation, immunefluorescence techniques and transfection of GFP or RFP-expression vectors, total RNA isolation & RTPCR. We observe the assembly of the PML nuclear bodies or RBM paraspeckles.


28.ࠈAnimal Models for Pain Research Capacity: Department:

ࠈ5 persons ࠈDepartment of Functional Anatomy and Neuroscience

Teaching staff: ࠈOzaki, N Contact: ࠈࠈࠈࠈTel: ࠈext. 2019 ࠈࠈࠈࠈ E-mail: noozaki@med.nagoya-u.ac.jp Time and Date: ࠈMay 20-21, 2008 ࠈࠈ10:00 a.m Meeting place: ࠈ Room # 329, Basic Science Building 3F

Outline: Animal models are fundamental resources in pain research. Studies using animal models are essential if new and clinically relevant knowledge about the mechanisms of pain is to be acquired. Participants will learn the basic mechanisms of pain sensation and how animal models can be used to analyze its mechanisms. Aim: Lecture: Laboratory:

Understand basic mechanisms of pain sensation. Understand animal models used for pain research. Basic mechanisms of pain sensation. Animal models used for pain research. 1. Models of visceral pain. Colorectal distension, writhing test, etc.. 2. Models of somatic pain. Von Frey test, Hargreaves test, etc.. 3. Models of chronic pain.


29.ࠈGene introduction into developing brain tissues by “electroporation” methods followed by slice culture Capacity: ࠈࠈࠈ 10-15 students Department: ࠈ Anatomy and Cell Biology Teaching staff: ࠈTakaki Miyata (Prof.) Contact: ࠈࠈࠈࠈTel 2030 ࠈࠈࠈࠈ E-mail tmiyata@med.nagoya-u.ac.jp, Time and Date:ࠈ September 12, 2008,

10:00 a.m. ~

Meeting place: ࠈNew Building for Basic Medical Science, 1F

Outline: “Electroporation”, which was originally developed for introducing foreign genes of interest into suspended cells, can now be used for gene delivery into three-dimensional in vivo tissues. In this training course, a standard protocol of gene delivery into developing mouse brains through “in utero electroporation” will be shown. First, plasmid vectors encoding a green fluorescent protein (GFP) gene will be micro-injected into lateral ventricles of embryonic mice by in utero surgery. Then, electric stimulations will be given to the embryos so that cells facing the lateral ventricle will be shocked and subsequently “eat” the plasmids. Since it will take about 10-12 hr until GFP proteins starts to be detected in the electroporated brain tissue, another litter of embryos previously electroporated will be used for visualization of GFP-labeled brain walls. Brains will be sliced, and mounted into collagen gels for time-lapse monitoring, by which GFP-labeled single cells can be seen.


30.ࠈImmunohistochemistry (Immunostaining) Capacity:ࠈࠈࠈࠈ30 persons Department:ࠈࠈࠈDepartment of Pathology of Biological Response Teaching staff: ࠈࠈYoriko Yamashita, MD, PhD Contact: ࠈࠈࠈࠈTel㸯2087 ࠈࠈࠈࠈ E-mail㸯yoriko@med.nagoya-u.ac.jp Time and Date:ࠈࠈ10:00 AM ̿ࠈࠈ3:00PM Meeting place: ࠈPractical Exercise Room for Anatomy and Pathology, 2F, Annex, Medical Research Building

Outline: Immunohistochemistry, also called immunostaining is a method to visualize intracellular or extracellular molecule using either a light microscope or, most recently, a laser microsope. Although this method is simple and mostly reliable, there are several limitations for application; 1. Target molecule must have antigenicity. 2. A specific antibody is required. 3. Target molecule must retain its antigenicity throughout the process of sample preparation. 4. Relatively large amount of antigen is necessary for detection. In this course, we are planning to briefly explain the principles and procedure of immunohistochemistry first, and then the students can actually perform immunohistochemistry using ready-to use formalin-fixed, paraffin-embedded samples. We will also show some important points for performing high-quality immunostaining. Furthermore, we will introduce a simple example of RNA in situ hybridization.


31.ࠈPreparation of mouse tissue lysate for SDS-PAGE Capacity : ࠈ ࠈࠈ10 people Department : ࠈࠈTumor pathology/Neuropathology Teaching staff : ࠈMayumi Jijiwa Contact : ࠈࠈࠈࠈTel Ext. 2093 ࠈ ࠈࠈࠈE-mail j2w@med.nagoya-u.ac.jp Time and Date : ࠈ15/08/08 10:00-12:00 Meeting place :

Building for Medical Research, Department of Pathology 2

Outline : In this course, we prepare general tissue lysate of mouse for SDS-PAGE. Practice includes dissection of mouse and measurement of protein concentration. It takes about two hours for entire process.


32.ࠈClinical application of indirect immunofluorescence methods - anti-nuclear antibody assays Capacity:

ࠈࠈࠈ5~7 persons

Department: ࠈConnective Tissue Disease & Autoimmunity Teaching staff: ࠈYoshinao Muro, M.D. and Kazumitsu Sugiura,M.D. Contact: ࠈࠈࠈ Tel Ext.2314 ࠈࠈࠈࠈE-mail ymuro@med.nagoya-u.ac.jp Time and Date: A Friday on middle of or late June Meeting place:

Staff Room of Department of Dermatology

Outline: Almost sixty years have passed since the discovery of LE cell phenomenon by Hargraves, and the measurement of anti-nuclear antibodies (ANA) has played important roles in diagnosis of autoimmune connective diseases, as well as in elucidation of pathogenesis of the diseases. ANA has clinically been used for screening test to detect particular autoantibodies with following specific examinations. The autoantibodies told us important clinical information including disease subsets, and prognosis of the disease. In order to interpret the immunofluorescence results of ANA exactly, it is essential to know this experimental method itself. ANA is also very useful tool for basic researches on cellular and molecular biology. In this course, you can examine immunofluorescence assays with patients sera and identify staining patterns. We could also introduce enzyme-linked immunosorbent assay (ELISA) method used for anti-extractable nuclear antigens (ENA) antibody detection in hospitals recently.


33.ࠈIntroduction for microsurgery (microvascular anastomosis) Capacity㸯ࠈ ࠈࠈ5 participants for each group (A,B) Department㸯ࠈࠈPlastic and Reconstructive Surgery Teaching stuff㸯ࠈShuhei torii, Keisuke Takanari Contact㸯ࠈࠈࠈࠈTelࠈ2525 ࠈࠈࠈࠈࠈࠈࠈࠈ E-mail

takanari@na.rim.or.jp (Keisuke Takanari)

Time and date㸯ࠈUndecided ( Thursday of Mid November) Meeting place㸯ࠈPlastic and Reconstructive Surgery Office (13F)

Outline㸯 This course is planned to introduce the basic knowledge and technique of the microsurgery. It will include the lecture of the microsurgical technique with slides, videos, and practical skills in the morning, animal practice in the evening. It will be held on Thursday of the middle of September. We will welcome the participant whoever is interested in microsurgery.


34.ࠈObservation of environment-derived nanoparticles and nanofibers in human lung Capacity: ࠈࠈࠈࠈ3 Department: ࠈࠈOccupational and Environmental Health Teaching staff: ࠈGaku Ichihara Contact: ࠈࠈࠈࠈTelࠈ2123 ࠈࠈࠈࠈ E-mailࠈgak@med.nagoya-u.ac.jp Time and Date: ࠈJune 30, Saturday, from 14:00 Meeting place: ࠈ Library of Dept. Occupational and Environmental Health

Outline: ES&H (Environmental, safety and health) problem of engineered nanomaterials has become of great concern as a consequence of growing nanotechnology. Especially behavior and kinetics of nanoparticles in humans is one of the greatest issues to be solved in the field of nanotoxicology. The present course will introduce the method of identification and analysis of environment-derived nanoparticles and nanofibers in human lung using electron microscope.


35.ࠈ Introductory course on statistical procedures using SPSS for Windows Capacity:

150 students Only e-learning course is available, however, office-hour is scheduled for your questions.

Department: ࠈࠈࠈDepartment of Public Health/Health Information Dynamics and Department of Preventive Medicine Teaching staff: ࠈࠈDr. H Yatsuya, M Naito, K Wakai, and N Hamajima Contact: ࠈࠈࠈࠈࠈTelࠈ(ext) 2128 (744-2128) ࠈࠈࠈࠈࠈ E-mailࠈh828@med.nagoya-u.ac.jp Time and Date:ࠈࠈIf you are interested in, please be registered at the graduate students’ affair section (Daigakuin-kakari), and then contact Dr. Yatsuya directly by an e-mail in advance. Office hour

ࠈࠈFrom 1:30pm to 4:30pm May 16th (Friday), 2008 ࠈࠈFrom 5:30pm to 7:30pm May 20th (Tuesday), 2008 Meeting place: ࠈࠈ2nd Floor, Annex, Medical Research building (Satellite Information Laboratory)

Outline: Creation of SPSS dataset from Microsoft Excel spread sheet Variable format/ data definition/ value label Changing, rearranging, or consolidating the values of an existing variable. Creation of new variable/ Ranking of the existing variable to create new one Frequency table/ Descriptive statistics Scatter plot/ correlation analysis Contingency table/ chi-squared test Calculation of means/ t-test/ ANOVA Multiple linear regression


36.

Advanced course on statistical procedures using SPSS for Windows

Capacity:

50 students. Only e-learning course is provided

Department:

Department of Public Health/Health Information Dynamics and Department of Preventive Medicine

Teaching staff:

Dr. H Yatsuya, M Naito, K Wakai, and N Hamajima

Contact:

Tel (ext) 2128 (744-2128) E-mail h828@med.nagoya-u.ac.jp

If you are interested in and think that you are eligible to take the course, please be registered at the graduate students’ affair section (Daigakuin-kakari), and then contact Dr. Yatsuya directly by an e-mail in advance. To be eligible to take the advanced course, participants need in prior to finish the introductory course or have equivalent skills. Outline: 1st: one-way ANOVA, ANCOVA, two-way ANOVA estimation of adjusted mean, trend test nd Multiple linear regression analysis, Logistic regression analysis 2 : 3rd: Survival analysis, Cox proportional hazard analysis, th 4 nonparametric methods The contents may be subject to change. Participants are required to complete assignments (details will be informed later).


37.

Organizing and Delivering Presentations

Capacity:

6 Participants must attend on all 3 days. All lectures will be in Japanese. Each participant must make a presentation in Japanese.

Department:

International Health

Teaching staff: Contact:

Prof. Atsuko Aoyama Tel 2109 E-mail intnl-h@med.nagoya-u.ac.jp

Time and Date: May 22 (Thu), May 23 (Fri), 2008 May 26 (Mon), 2008 Meeting place:

1:00 pm – 2:30 pm 1:00 pm – 3:30 pm

Seminar Room, Department of International Health (4th Floor, Medical Research Building)

Outline: Making a presentation is a communication skill to orally deliver messages to audience. This skill is used at various occasions, such as making lectures, showing a business plan, and reporting a research achievement. For the International health specialists, making a presentation is one of the most important communication skills to deliver accurate messages to people with various social and cultural backgrounds, and to change their behaviors. The same message will or will not be delivered effectively, depending on the skill. Thus, it is important for everyone to learn how to organize and deliver a presentation. During the course, participants will learn the outline of the skill. Then, each participant will organize and deliver a presentation. The presentation will be videotaped and evaluated one another.


38.

Measurement of ion transport and intracellular Ca2+ in epithelia

Capacity:

~8

Department:

Human Nutrition (Research Center of Health, Physical Fitness, and Sports)

Teaching staff:

Takaharu Kondo, Hiroshi Ishiguro, Akiko Yamamoto

Contact:

Tel: 2183 E-mail: ishiguro@htc.nagoya-u.ac.jp

Time and Date:

September

Meeting place: Laboratory of Human Nutrition, Building for Medical Research (west wing, 2nd floor)

Outline: Concentrations of various ions (Ca2+, Na+, Cl-, pH, etc.) in intra- and extra-cellular microenvironment can be measured by microfluorometry uing ion-sensitive fluorescent dyes. Pancreatic juice contains unusually high concentrations of HCO3-, which is produced by epithelial cells lining pancreatic duct system. Our laboratory investigates cellular and molecular mechanisms of HCO3- transport by using interlobular duct segments (diameter: ~100 micron) isolated from mouse, rat, or guinea-pig. The isolated duct is monolayer that retains the polarity and function (HCO3- secretion) of epithelium. The aim of this laboratory training is to master the basic techniques for measuring intracellular pH and Ca2+ concentration in epithelial cells, which is important for studying vectorial transport of HCO3- across epithelia.


39.

Estimation of in vivo and in vitro insulin action in rats

Capacity:

~10 persons

Department:

Health and Sports Medicine

Teaching staff: Yoshiharu Oshida, Teruhiko Koike Contact:

Telďźš 789-3961 E-mailďźšoshida@htc.nagoya-u.ac.jp

Time and Date: 3rd July/2008 Meeting place:

from 2:00 PM

Meeting Room (2nd Floor Research Center of Health, Physical Fitness and Sports, Higashiyama Campus)

Outline: This course will be lectured in Japanese. You can study the principle of in vivo and in vitro insulin action measurement by using of the hyperinsulinemic euglycemic clamp technique and muscle glucose uptake stimulated by insulin in rats, respectively.


40.

Psychoanalytic, Understanding of

Capacity:

Doctor-Patient Relationship

High level of Japanese language ability

Department:

Psychopathology & Psychotherapy

Teaching staff:

Prof. Toyoaki Ogawa, Prof. Toshinori

Contact:

Tel 052-789-5836 E-mail ogawa@htc.nagoya-u.ac.jp

Time and Date:

10/9/,17/9,24/9/2008, 5:30

Meeting place:

seminar room 202,203

Outline:


41.

Document retrieval

Capacity:

100 persons

Department:

Director, Medical Library

Teaching staff:

Junki, TAKAMATSU

Contact:

Tel : ex.2509 E-mail: med@nul.nagoya-u.ac.jp

Time and Date:

The end of June and the beginning of July 13:30 – 17:00 To get credit, you need to attend the series of two classes and submit a report.

Meeting place:

Satellite Laboratory (Annex, Medical Research 2F)

Outline:

Lecture and practice on PubMed, Web of Science, and Electronic Journal for beginners. (Lecture is in Japanese.)

Notice:

Nagoya University ID and password are required to take practice course.


42. Downregulation of target gene expression in mammalian cells by RNA interference Capacity:

10

Department:

Molecular biology

Teaching staff:

Yoshifumi Takei

Contact:

Tel 2064 E-mail secret@med.nagoya-u.ac.jp

Time and Date:

Nov 10, and Nov 12

Meeting place:

Seminar room of Biochemistry

9:30-12:00

Outline: RNA interference (RNAi) methodologies are rapidly being established and hold promise to specifically inhibit gene expression in mammals. RNAi is the sequence-specific, posttranscriptional gene silencing method initiated by double-stranded RNAs, which are homologous to the gene being suppressed. RNAi technology, especially chemically synthesized siRNA, is currently being evaluated not only as an extremely powerful instrument for functional genomic analyses but also as a potentially useful method to develop highly specific gene-silencing therapeutics. In the basic training program, you will actually learn and experience how to introduce synthetic siRNA molecule into mammalian cells, and how to estimate its RNAi effect.


43. Extraction and analysis of glycolipids from tissues and cells by thin layer chromatography Capacity:

5-10 students

Department:

Department of Biochemistry II

Teaching staff: Koichi Furukawa Contact:

Tel: 052-744-2070 E-mail: koichi@med.nagoya-u.ac.jp

Time and Date: November 2008 Meeting place:

New Research Building, 3rd Floor

Outline: After extracting lipids from mouse brain and liver or cultured cancer cells using chroloform and methanol, acidic and neutral fractions will be separated using DEAE-Sephadex ion exchange column chromatography. Then, the extracts will be analyzed by thin layer chromatography (TLC) and subsequent spray with primulin or resorcinol.


44.

cholesterol and phospholipid analyses

Capacity: Department:

10 Biochemistry

Teaching staff:

Yoshio Yamauchi

Contact:

Tel 2068 E-mail yyoshio@med.nagoya-u.ac.jp

Time and Date:

the second semester

Meeting place:

Department of Biochemistry II

Outline: Cholesterol and phospholipid are important components of cells. Impaired metabolism of these lipids is often involved in various disorders. The purpose of this training course is to learn basic methods for extraction and measurement of cholesterol and phospholipid in cultured cells and/or animal tissues. 1. Extract lipid from cultured cells and/or tissues. 2. Measure cholesterol and phospholipids contents by enzymatic assays.


45.

Proteome analysis with two-dimensional gel electrophoresis

Capacity:

15 persons

Department:

Drug Resistance and Pathogenesis

Teaching staff: Contact:

Time and Date: Meeting place:

Keiko YAMADA Tel E-mail

052-744-2102 yakeiko@med.nagoya-u.ac.jp

Nov. 26 – 27, 2008 On 10:00, Nov 26 at Molecular Bacteriology Lab. 4F and east side of new 2nd Research building

Outline: Along with the advantages of genomic research, genome sequencing analyses of a numbers of various species have been accomplished. From the enormous genetic information to understanding of life phenomenon, two-dimensional gel electrophoresis (2DE) in which proteins are separated based on an isoelectric point and molecular weight is powerful tool for the systematic and exhaustive analysis for proteins in the Post-Genome research. At the beginning of training, I will provide a fundamental lecture of principle of 2DE analysis, and then you can practice the procedure of 2DE with proteins secreted by bacteria. Fundamental principle is applicable for mammalian cell science including human, mouse, and etc. Attendee who could participate both of days (10:00-17:00) is preferable.


46.

Quantification of viral genes by the real-time PCR assay

Capacity:

10 persons

Department:

Virology

Teaching staff:

Hiroshi Kimura

Contact:

Tel: ext 2207 E-mail: hkimura@med.nagoya-u.ac.jp

Time and Date:

January or February 2008

Meeting place:

Department of Virology,

Outline: The real-time PCR assay is rapid and convenient method to quantify specific sequences of nucleic acids and widely used for the quantification of viral loads. Training contents are as follows: 1)principle of real-time PCR, 2)handling of materials,3)extraction of DNA or RNA,4)quantification of viral genes by real-time PCR using fluorescent probes,5)application for basic and clinical research.


47.

Introduction to Diagnostic Pathology

Capacity: 5 persons/year, One person in each term Department in charge: Depertment of Clinical Pathophysiology and Clinical Pathology Stuff: Yoshiko Murakami, MD, PhD(Hospital Research Associate) Satoko Shimada, MD, PhD (Assistant professor) Tetsuro Nagasaka, MD, PhD (Associate professor) Shigeo Nakamura, MD, PhD (Professor, Course director) Application: Contact to Dr. Shigeo Nakamura, (Professor,Ccourse director) Ex:2582, e-mail:snakamur@med.nagoya-u.ac.jp Duration : the latter term, one week to one month Location: The division of Pathology which is located on the second floor of the Central Clinical Facilities Building Introduction: Surgical pathology, cytopathology, intra-operative consultation on frozen sections, which provide patients with the proper diagnosis for treatment, and autopsy are our routine work. Modern techniques such as immunostaining, and in situ hybridization are now employed to ascertain various diagnostic and prognostic markers for the benefit of the patients. Our main interest is to solve the problems encountered in the practice of these examinations and develop new techniques to help in diagnostic pathology. All trainees participate directly in handling, processing, and examination of pathology specimens. Trainees are expected to follow their cases through all stages of processing, including macroscopic description, blocking, and review the microscopy and sign out the case with the stuff pathologist. This allows trainees to lean an analytical approach to deriving a differential diagnosis and then the adjunctive techniques available to arrive at correct diagnosis. The importance of providing accurate, timely feedback to clinical staff, the role of intra-operative consultation, and the importance of Diagnostic Pathology in patient care, is stressed. Trainees must attend the daily pathology conference and autopsy conference which held in every two weeks.


48.

Controlling of genetic background of genetically engineered mice

Capacity:

5∼10

Department:

Laboratory Animal Sciences

Teaching staff:

Tamio Ohno

Contact:

Tel 2468 E-mail ohno@med.nagoya-u.ac.jp

Time and Date: two consecutive days in October or November Meeting place:

1F Room Number 110, Division of Experimental Animals, Center for Promotion of Medical Research and Education

Outline: Mice carrying the same genetic defects on different genetic backgrounds often exhibited phenotypic variations. In these cases, to control the genetic backgrounds is necessary for more strict analysis. The purpose of this course is to learn the theory and methods for controlling the genetic backgrounds of mouse strains. ・breeding methods of congenic strains ・extraction of genome DNA from mouse tail ・genotyping for the PCR-SSLP method


49.

Protein expression profiling with MALDI-TOF-MS

Capacity:

5 persons

Department:

IAR / Molecular Biology

Teaching staff: Kiyoshi Yanagisawa Contact:

Tel ext. 2454 E-mail kyana@med.nagoya-u.ac.jp

Time and Date: Nov. 19 (Wed) 10 am to 3 pm Meeting place:

Meeting room at department of Molecular Biology

Outline: MALDI-TOF-MS can analyze molecular weight of peptides, small molecules, proteins , etc. in samples. In this course, we will learn the theory how to analyze molecular weight of samples with MALDI-TOF-MS, methods of sample preparation (cell line) and analysis with MALDI-TOF-MS.


50.

Cell fractionation by using a centrifugal elutriation

Capacity: Department:

10 Molecular Carcinogenesis

Teaching staff: Motoshi Suzuki Contact:

Tel E-mail

2456 msuzuki@med.nagoya-u.ac.jp

Time and Date: 11/12/2008 Meeting place:

Molecular Carcinogenesis Office

Outline: Centrifugal elutriation is a popular method that has been developed for cell fractionation. One may enrich a unique cell population from mixed one, or collect a certain phase of cells from a randomized cell culture. In this course, G1 cells will be collected using culture cells.


51.

Purification of GST fused protein

Capacity:

10

Department:

Cancer Biology

Teaching staff:

Ito Satoko

Contact:

Tel 2463 E-mail s-ito@med.nagoya-u.ac.jp

Time and Date: December Meeting place:

Conference room of Dept. of Cancer Biology

Outline: GST protein is widely used for purification of recombinant protein. With this technique, it is possible to produce large amount of proteins in a short period of time. We will show how to produce GST fusion protein in bacteria and how to purify it. The technique is simple and easy to learn and basic knowledge for molecular biology is not required.


52.

How to use protein database

Capacity:

ďź’ďź?

Department:

Cancer Biology

Teaching staff:

Senga Takeshi

Contact:

Tel 2463 E-mail tsenga@med.nagoya-u.ac.jp

Time and Date: December Meeting place:

Conference Room of Dept. of Cancer Biology

Outline: There are various database for biology and you can speed up you experiments by using them. I will talk about mainly about database for analyzing proteins. Attendants are advised to have some basic knowledge about molecular biology.


53.

Electrophysiology using brain slice preparations

Capacity:

5

Department: Medicine

Neuroscience 1, Research Institute of Environmental

Teaching staff:

Takuro Maruyama

Contact:

Tel: 3878 E-mail: takuro@riem.nagoya-u.ac.jp

Time and Date:

October 23, 24

Meeting place:

main building the fourth floor Room 409 of Research Institute of Envirionmental Medicine

Outline: The aim of this program is to introduce students to the basic procedures for preparing brain slices and plasticity electrophysiologically. Students are asked to prepare slices from developing rat visual cortex and record synaptic responses evoked by electrical stimulation. Using these slices, long-term potentiation (LTP) will be induced by high-frequency conditioning stimulation and the molecular mechanisms of LTP induction will be analyzed with a pharmacological method.


54. Basic training for breeding and usage of laboratory animals, and for a comparative dissection of animals Capacity:

12

Department:

Department of Genetics, Research Environmental Medicine (RIEM)

Teaching staff:

MURATA Yoshiharu and ODA Sen-ichi

Contact:

Tel 052-789-4179 E-mail oda@agr.nagoya-u.ac.jp

Time and Date:

Nov. 4, 2008 9:30~17:00

Meeting place:

Institute

of

Conference Hall, South Building of RIEM

Outline: This basic training course will provide some techniques and knowledge of: 1)breeding methods and genetic control of laboratory animals, such as inbred strain, closed colony, and mutant strains, and also nomenclature of gene, chromosome, specific strain names such as congenic strain, recombinant inbred strain and so on. 2)characterization of species and strains as an animal model for human disease and their establishment, maintenance and usage. 3)comparative anatomy of laboratory rodent and insectivore animals Carrying with you: Dissection tools for a small laboratory animal (dissection scissors, gold crown scissors, tweezers)


55. Detection of mRNA expression in cells. - From RNA extraction to RT-PCR

Capacity:

about 15 students

Department:

Gastroenterological surgery

Teaching staff:

Shuji Nomoto M.D. Ph.D.

Contact:

Tel: 2239 E-mail: snomoto@med.nagoya-u.ac.jp

Time and Date:

toward the end of December

Meeting place:

Office of Gastroenterological Surgery (2nd floor)

Outline: There are two methods to study a gene expression. One is to check mRNA level, and the other is to measure protein level. The method by RT-PCR is an easy way to check mRNA expression, and is widely applicable to various genes. The steps of this course are 1, RNA extraction from cultured cells. 2, c-DNA synthesis. 3, Designing the primers of RT-PCR for the gene of interest. 4, RT-PCR. 5, Run the PCR product on Gel. You can learn above five steps from RNA extraction to get the result of gene expression


56.

DNA typing by fragment analysis

Capacity:

20

Department:

Legal Medicine and Bioethics

Teaching staff:

Toshimichi Yamamoto Rieko Uchihi

Contact:

Tel: 2119 (or 2117) E-mail: yamachan@med.nagoya-u.ac.jp

(Associate Professor) (Research Associate)

Time and Date: Twice for 2 days around the middle of November in 2007 About 10 persons at each 2 days Meeting place:

2F (Room 217) in the Building of the Basic Medicine

Outline: In this course, the participants learn how to genotype by DNA fragment analysis from PCR products for STR (short tandem repeat) (microsatellite) markers, which are widely applied in forensic field for personal identification and kinship analyses. This technique will make it possible for the participants to perform linkage analyses using microsatellite markers in clinical medicine. To be concrete, the participants extract DNA from their own buccal swabs by a convenient commercial kit. Those DNA samples are amplified using a commercially released PCR multiplex kit with multi-colored fluorescently labeled primer sets for personal identification. The PCR products are loaded, sized and genotyped by capillary electrophoresis with a Genetic Analyzer 310. The participants calculate the probabilities for personal identification.


57. Drug/toxicant metabolism and a measurement of metabolites with a chromatograph Capacity:

10 students

Department:

Department of Occupational and Environmental Health

Teaching staff:

Michihiro Kamijima and Tamie Nasu-Nakajima

Contact:

Tel 2125 E-mail kamijima@med.nagoya-u.ac.jp

Time and Date: December 5 (Fri), 2008 Meeting place:

Department of Occupational and Environmental Health (7th floor of 2nd research building)

Outline: In this course students have a lecture about metabolism of environmental chemicals, especially pesticides, and measure urinary pesticide metabolites with a high performance liquid chromatograph or a gas chromatograph-mass spectrometry. The aim of this course is to understand 1) concept and applications of chromatograph and 2) metabolism of drug/toxicant and biological monitoring.


58.

MRI Volume measurement of hippocampus

Capacity:

5 Students

Department:

Pediatrics

Teaching staff:

Jun Natsume

Contact:

Tel ex2294 E-mail junnatsu@med.nagoya-u.ac.jp

Time and Date:

February, 2009

Meeting place:

5th floor, Medical Science Research Building 1

Outline: MRI-based measurement of hippocampal volume has been used as a useful tool in the study of neuropsychiatric disorders, such as temporal lobe epilepsy. The accuracy and reproducibility of the volume measurement depend on both image processing technique and anatomical assessment. In this course, we learn how to deal with the imaging data and how to assess anatomical boundaries of the hippocampus with using software developed by Montreal Neurological Institute.


59.

FACS analysis and cell sorting

Capacity:

5 person in one week course and 10 person in one day preliminary course

Department:

Immunology

Teaching staff:

Ken-ichi Isobe, Masataka Haneda, Haruhiko Suzuki, Sachiko Ito, Minoru Tanaka

Contact:

Tel 052-744-2134 E-mail kisobe@med.nagoya-u.ac.jp

Time and Date: 7th July~ 11th July Meeting place:

Immunological Department, meeting room

Outline: training about FACS analysis and cell sorting


60. Live cell imaging Capacity:

four persons

Department:

Cell Physiology

Teaching staff: Kenzo Hirose Contact:

Tel 2042 E-mail kenzo@med.nagoya-u.ac.jp

Time and Date:

14-18 July

Meeting place:

Cell Physiology (Physiology I)

Outline: Live cell imaging technique is a major research maneuver for understanding intracellular molecular events in a context of time and space. In this course, students will learn two representative live cell imaging techniques: imaging of GFP-fused proteins and intracellular calcium imaging.


61.

Immuno-electron microscopy (Post embedding method)

Capacity:

10 persons

Department:

Molecular Cell Biology

Teaching staff:

Akikazu Fujita

Contact:

Tel 2001 E-mail: afujita@med.nagoya-u.ac.jp

Time and Date:

July 24, and 25

Meeting place:

room# 345

Outline: We will focus on the post embedding technique of the immuno-electron microscopy method. This technique has the advantage for quantifying immuno-labeling, and therefore the relative concentration of a protein in various subcellular compartments can estimate with high resolution. On this course, students will actually embed fixed cells with lowicryl resin and labeled ultrathin sections with antibodies.


62. Realtime PCR Capacity:

5

Department:

Biochemistry

Teaching staff: Kenji Kadomatsu, Satoshi Kishida, Yasutomo Ito Contact:

Tel: 2064 E-mail: kishida@med.nagoya-u.ac.jp

Time and Date: July 28 (Mon)-Aug. 1 (Fri), 2008 13:00-17:00 Meeting place:

Annex, Medical Research 4F (Experimental Room for Biochemistry)

Outline: The purpose of this course is to learn the principle and skill for realtime PCR. Participants are supposed to determine the genotype of transgenic mice by utilizing realtime PCR. Although the canonical method to distinguish hemizygote and homozygote is southern blot, the application of realtime PCR can save a lot of trouble. The genomic DNA from mouse tail is used as a template for realtime PCR. For comparative evaluation, genome DNA from hemizygote is used as a “calibrator� and the ratio toward that is calculated in each examined mouse. The ratios of hemizygote and homozygote should be 1.0 and 2.0 respectively. Because the difference to be detected is very subtle, we must be careful in order to get accurate results.


63.

Proteome analysis

Capacity:

Five participants are acceptable

Department:

Microbiology

Teaching staff:

Michio Ohta and Akira Okamoto

Contact:

Tel ext. 2102 E-mail yoh@med.nagoya-u.ac.jp

Time and Date: Aug 6 to 10, 10:00am Meeting place:

Microbiology department

Outline: The protein is the fundamental component for all living organisms, and its identification becomes one of the most basic and essential techniques for all biochemical scientist. This practice course mainly aims at the technical learning; protein separation by electrophoresis, handling of micro amount protein, peptide recovery technique, LC-MS/MS analysis, and mass-tag search protein identification by program MASCOT.


Issuu converts static files into: digital portfolios, online yearbooks, online catalogs, digital photo albums and more. Sign up and create your flipbook.