Preparation of recombinant proteins using yeast

1. Preparation of recombinant proteins using yeast expression system

Capacity: 5 10

Department: Molecular Biochemistry

Teaching staff: Tetsuya Okajima

Contact: Tel 2068 E-mail

Time and Date: September 1 and 4 (2 days)

Meeting place: Dept. of Biochemistry II

Outline:

For protein expression, various expression systems such as E. coli, yeastand insect cells are available. The yeast expression system offers severaladvantages over other systems since it allows to produce the large amount ofcorrectly folded disulfide-bonded recombinant proteins. In this trainingcourse, EGF domains of Notch receptors are expressed using K. lactisProteins Expression Kit (NEB), purified using Ni-affinity beads, anddetected 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. Yeast two hybrid system

Capacity: 6

Department: Molecular Mycology and Medicine

Teaching staff: Tomohiro Akashi

Contact: Tel 2460 E-mail [email protected]

Time and Date: June 10, 13, 17, 20 Afternoon. The first day (June 10) starts at 2:00 p.m.

Meeting place: Basic Research Building, Annex 5F

Outline:

Yeast two hybrid system is one of excellent methods to examine protein-protein interactions. Typically, this method is used to detect genes whosetranslated products can interact with protein of interest, from a collection ofgenes like cDNA libraries. This is also used to analyze the detail ofprotein-protein interaction. This technique is basically applicable to anygenes, and is highly universal. Furthermore, its experimental proceduresare highly organized, so people can use it without skilled experiences. In the course, we demonstrate the outline of all the experimentalprocedures. The demonstration also focuses on the handling of yeast cellsfor the two hybrid system. This course requires 4 afternoons indicatedabove. Applicants to this course should be careful to the schedule.Applicants also would be required to answer a questionnaire for the coursebefore approval.

3. Laser scanning cytometer (LSC)

Capacity: 8 (maximum: 16)

Department: Molecular Mycology and Medicine

Teaching staff: Tomohiro Akashi


Time and Date: October 7, afternoon (2:00 p.m. – 5:00 p.m.)

Meeting place: Basic Research Building, Annex 5F

Outline:

Laser scanning cytometer (LSC) is an epifluorescence microscopeequipped with a laser. This special microscope is designed to quantitate thefluorescence emitted from the sample mounted on slide glass excited by thelaser beam. LSC can quantitate any fluorescence signals from cells stainedwith fluorochrome. For an example, DNA content of a cell can bequantitated by cells stained with propidium iodide (PI). LSC is occasionallymisunderstood as flow cyotometer. Both can give us similar data, but LSCis based on microscopy. In the course, the merit and demerit of LSC are explained. Stainingprocedure of samples with propidium iodide and measurement of the DNAcontent of the sample are demonstrated. Attendees would also learn how tohandle it practically.

When the number of applicants are over the capacity, the schedule may bechanged. Change of schedule would be notified via e-mail in that case.

4. Introduction to the NCBI Gene Database

Capacity: 100 students

Department: Neurogenetics

Teaching staff: Kinji Ohno

Contact: Phone. ext. 2446 E-mail [email protected]

Time and Date: June 11, 2007 (Wed) or June 13, 2008 (Fri), 13:00 – 17:00I will repeat the identical seminar on two days, because themaximum capacity of the satellite laboratory is limited to50. You will be assigned to either day.

Meeting place: Satellite laboratory, 2nd floor of the Annex Building of theMedical 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 avariety of medical and biological researches. The seminar is intended forstudents who employ the molecular biological techniques in their researchprograms. The seminar covers the basic strategies and techniques of theNCBI (the National Center for Biotechnology Information) gene databasesearch. The specific topics to be covered include BLAST, uniGene, uniSTS,Entrez Gene, SNP, OMIM, and PubMed. The course also covers the OVIDMedline database, the RefScout service, and the CiteTrack service. Do notforget to bring your Nagoya Univeristy ID and its password, when youattend the seminar.

5. Induction and analysis of bone marrow derived dendritic cells

Capacity: 6 persons

Department: Department of Aging Research, Program in Integrated Molecular Medicine

Teaching staff: Mitsuo MARUYAMA

Contact: Tel : 0562-46-2311 (Ext. 5101) E-mail: [email protected]

Time and Date: total 3 days around late in September or early in October

Meeting place: Department of Mechanism of Aging, National Institute forLongevity Sciences, National Center for Geriatrics andGerontology

Outline:

The dendritic cell plays an important role with the antigen presentation inacquired immune system. It is well established that in vitro differentiateddendritic cell is induced from bone marrow cells by GM-CSF. In this basictraining course, students will prepare the murine bone marrow deriveddendritic cells and characterize them by FACS analysis. All procedures andtechniques are generally plain and simple therefore the Master's coursestudents are also welcomed.

6. Analysis of Cellular Senescence

Capacity: 6

Department: Department of Aging Research, Program in Integrated Molecular Medicine

Teaching staff: Noboru Motoyama

Contact: Tel 0562-46-2311 (Ext. 5554) E-mail: [email protected]

Time and Date: TBA (2days)

Meeting place: Department of Geriatric Medicine, National Institute forLongevity Sciences, National Center for Geriatrics andGerontology

Outline:

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

7. Functional imaging of neuronal cells in brain slice cultures

Capacity: 10 people

Department: Dept. of Physiology II

Teaching staff: Dr. Tatsumi


Need contact with Dr.Tatsumi via E-mail as well as university office(Jimu, Kyomugakari).

Time and Date: th and th June

Meeting place: 403 room at 4th floor Basic Science Building

Outline:

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

8. Experimental methods of Neuropsychopharmacology Capacity: 20

Department: Department of Medical Pharmacy (Department ofNeuropsychopharmacology and Hospital Pharmacy)

Teaching staff: Kiyofumi Yamada Ph.D.

Contact: Tel 052-744-2674 E-mail [email protected]

Time and Date: 23-24, June

Meeting place: Laboratory of Medical Pharmacy, 1st floor of 1st building ofclinical departments.

Outline:

Purpose; Learning how to do experimental methods ofneuropsychopharmacology, using rats or mice.1. Experiments to evaluate the ability of learning and memory1) Morris’s water maze task2) Eight arms radial maze3) Habituation task4) Passive avoidance task2. Experiments to evaluate depression and anxiety in mice

9. Whole-cell patch clamp recordings using brain slice preparations

Capacity: 5

Department: Visual Neuroscience, Research Institute of EnvironmentalMedicine

Teaching staff: Yumiko Yoshimura

Contact: Tel: 3879 E-mail: [email protected]

Time and Date: October 偐,

Meeting place: Room 409 of the forth floor, Research Institute ofEnvirionmental Medicine on Higashiyama campus

Outline:

The aim of this program is to introduce students to the basic proceduresfor preparing brain slices and analyzing synaptic transmission with whole-cell patch clamp techniques. Students are asked to prepare slices fromdeveloping rat visual cortex and record synaptic responses evoked byelectrical stimulation. Synaptic transmissions will be analyzed with apharmacological method.

10. Isolation and culture of glial cells.

Capacity: 8

Department: Neuroimmunology, Research Inst. Environmental Med.

Teaching staff: Akio Suzumura

Contact: Tel Ext. 3881(Higashiyama) E-mail [email protected]

Time and Date: Sept. 5 (Wed), 2007

Meeting place: Department of Neuroimmunology, RIEM

Outline:

Glial cells consist of oligodendrocytes, astrocytes and microglia. We haveestablished the isolation method to purify each glial cells from the primarymixed glial cell cultures prepared from neonatal rodents. The method enableus to use purified cells in various biochemical, molecular and immunologicalexperiments.

11. Primary culture of cortical neurons

Capacity: 8

Department: Department of Neuroimmunology, Research Institute ofEnvironmental Medicine

Teaching staff: Tetsuya Mizuno


Time and Date: September 4 2007 13 30 16 00

Meeting place: Department of Neuroimmunology, Research Institute ofEnvironmental Medicine (4F)

Outline:

Using primary neuronal culture, co-culture of microglia and neuron, andorganotypic cortical slice cultures, the course introduces how to evaluateneuronal damage.

12. in vivo imaging technique for small animals

Capacity: 5 to 6

Department: Brain Life Science

Teaching staff: Kenji Ono


Time and Date: 8/1 (Fri) A.M.10:00-P.M.5:00

Meeting place: Seminar room in Research Institute of EnvironmentalMedicine

Outline:

The fate of drug substances administered externally to living animals isusually determined with body fluids and dissected tissues.Recent improvements of in vivo imaging technology such as PET, MRI andeven optical imaging, however, provide a new strategy for analyzing drugdistribution, adsorption and metabolism. These imaging technologiesalso have advantages to determine the tagged substances such as drugvehicles and cells in a living animal. In this course, we provide a shortlecture and a fundamental practice for a small animal MRI instrument andan in vivo optical imager.

13. Behavioral analyses in rodents Capacity: 5 6

Department: Department of Brain Function, Research Institute ofEnvironmental Medicine

Teaching staff: Hiroyuki Mizoguchi Ph.D.


Time and Date: One day for the period of summer vacation

Meeting place: Research Institute of Environmental Medicine

Outline:

Purpose; Learning behavioral experimental methods using rodents.Experiments to evaluate the ability of learning and memory1) Morris’s water maze task2) Novel object recognition task3) Elevated plus-maze task4) Y-maze task

14. How to measure pain in human and animal pain models

Capacity: 　　　5 students

Department: 　 Futuristic Environmental Simulation Center, ResearchInstitute of Environmental Medicine

Teaching staff: 　Jun SATO, MD, PhD.

Contact: 　　　Tel Higashiyama -3865E-mail [email protected]

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

Meeting place: 　Futuristic Environmental Simulation Center, ResearchInstitute of Environmental Medicine (Higashiyamacampus)

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 painsensitivity in patients. This course aims to provide the student withunderstanding and knowledge of pain mechanism and pain tests. In thislecture you will learn:1. Pain in human (Lecture)

1) profile and physiological significance of various types of pain2) pain tests (VAS scale, etc.)

2. Pain in experimental animals (Lecture and Exercise)1) methods of making acute and chronic pain models2) pain tests (von Frey hair test, Randall-Selitto test, etc.)

15. Introduction to the electrical recording of biological signals

Capacity: 6 persons

Department: Department of Neuroscience II, Research Institute ofEnvironmental Medicine

Teaching staff: Kazue Mizumura

Contact: Tel (extension) 85 3861 E-mail [email protected]

Time and Date: July 30 (Wen) and 31 (Thu). 10:00-16:00

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

Outline:

In clinical practice it is quite common and essential to record biological signalswith electrical methods (e.g. ECG). These methods are also widely used in basicmedical research, for example, patch clamp recording, intra- and extra-cellularrecordings etc. Therefore, it is important to have basic knowledge onmechanisms of electrical recording and instruments for using them properly andobtaining correct data. In this seminar following points will be overviewed andpracticed.

1) Different kinds of biological signals and their electrical characteristics2) Amplifier, filter, pen recorder, oscilloscope, magnetic tape recorder,

electrical stimulator, various transducers3) Characteristics of digital instruments and points to notice4) Electrodes5) Countermeasures against electronic noises6) Practice using operational amplifiers

16. Recording of primary afferents with the single-fiber recording technique(dissection method).

Capacity: 5

Department: Neural Regulation

Field: Regulation of Organ Function

Program: Program in Cell Information Medicine

Teaching staff: Toru TAGUCHI

Contact: Tel: 052-789-3863E-mail: [email protected]

Time and Date: 10:00-17:00, Aug 5, 2008. (Aug 6 for applicants)

Meeting place: Main Bld. 2F (Room 215), Research Institute of EnvironmentalMedicine, Higashiyama-campus.

Outline:The single-fiber recording technique has been established in the latter 1960s. Itis a kind of electrophysiology and enables to get action potentials of a singlesensory receptor fiber in the peripheral tissues. The technique is widelyapplicable both in vivo and in vitro. In our lab, we have been studied peripheralmechanisms of nociception with skin-, muscle-, and testis-nerve preparations. Inthis course, activities of a single nociceptor (myelinated A_- or unmyelinated C-fiber) will be recorded in a rat extensor digitorum longus muscle-peroneal nervepreparation in vitro. Participants will learn below. Students who will take thiscourse are advisable to take the course ‘Introduction to the electrical recording ofbiological signals’.

1) A brief introduction to the single-fiber recording method andelectrophysiological instruments (lecture)

2) How to make the rat muscle-nerve preparation (practice)3) Recording of nociceptor activities in the preparation made (practice)

17. How to use antibodies (Immunoprecipitation & Western blotting)

Capacity: 6Department: Dept. of Neural Regulation (RIEM, Higashiyama-campus)Teaching staff: Katanosaka, K.Contact: Tel ex. 85-3862/3864 E-mail [email protected] and Date: Two days in September (10:00-17:30)Meeting place: 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 informationthat is necessary to use antibodies and interpret the results of experiments usingantibodies. In the course, everyone can get training in preparation of proteinsamples from tissues, handling of antibodies, and techniques for westernblotting and immunoprecipitation. The course includes some short lectures asshown 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 e-mail to the teaching staff of the course ([email protected], Mailtitle: BT20, Deadline: June 30). If the number of applicants exceeds thecapacity, the members will be selected by the staff and announced by e-mail andon 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 youprefer ( 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. Culture of cardiomyocytes

Capacity: 5 persons

Department: Division of Cardiovascular Research (2),(Research Institute of Environmental Medicine)

Teaching staff: Masahide Harada


Time and Date: September 11 (Thur.), 12 (Fri.)

Meeting place: Department 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 onculturing cardiomyocites in cardiovascular research field. You can experiencethe primary culture of neonatal rat cardiomyocytes and get the basic skillneeded.

(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 ventriclesare plated in the culture dish and are isolated into single myocytesby enzyme reaction.

Second day: Culture of isolated myocytes.Isolated myocytes are incubated in the culture dish containingappropriate medium and their growth is microscopically observed.

19. Induction of cardiac cells from embryonic stem cells~ Creating regenerative myocardium ~

Capacity: Ten students

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

Teaching staff: LEE Jong-Kook

Contact: Tel (052) 789-3871 E-mail: [email protected]

(Note) Please inform us by e-mail if you cannot participate in thecourse 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: Room 312 Main building Third floorDepartment of Cardiovascular Research, Research Institute ofEnvironmental Medicine, Nagoya University (Higashiyama campus)

Outline:

“Regenerative medicine” is a field of great concern in recent years, which restore thelost functions of failing organs through actively utilizing stem cells or stem cell-derivedtissues. Especially, embryonic stem (ES) cell is attracting attention as a potential cellsource because the cell possesses prulipotency to differentiate into almost all kinds ofcells. In the field of cardiovascular research, ES cell-derived myocardial and vasculartissues are extensively studied. In this basic training course, our goal is to outline themethods of induction and isolation of cardiac cells from murine ES cells and try toobtain basic knowledge and technique. On Day 1 of the basic training, we startdifferentiation from undifferentiated embryonic stem cells to form embryoid bodies. OnDay 2, about 7-10 days later, it is scheduled that students will observe spontaneouslybeating embryoid bodies.

20. Occytes Expression Method for Ion Channel Measurement

Capacity: 5 persons

Department: Humoral Regulation (Res Inst of Environ Med)

Teaching staff: Kaichiro Kamiya (Ryoko Niwa)

Contact: Tel 85-5006 E-mail [email protected]

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

Meeting place: Research 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 wasinjected in the Xenopus Oocytes and two-three days later channelactivities were measured by using double-microelectrodes voltageclamp methods. Activities of specific ion channels or detailed actionof drugs on ion channels were widely studied by this method.Especially, this method is mostly effective in the studies to clarifypathological analysis of ion channel diseases due to genomicdisorder or molecular action of antiarrhythmic drugs.

In this basic training, a surgical procedure to remove oocytes fromfrog, an enzyme treatment and cRNA injection were performed bythe participants. In addition, measurement of ion channel currentsin pre-injected oocytes was experienced by using voltage clampmethod.

21. Animal model (rabbits) for cardiovascular research

Capacity: 10

Department: Department of Cardiovascular ResearchResearch Institute of Environmental Medicine

Teaching staff: Yukiomi Tsuji, MD, PhD

Contact: Tel: 052-789-5006

E-mail: [email protected]

Time and Date: AM10:00-PM1:00, September 4, 2008

Meeting place: 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. Optical recording of membrane potential

Capacity: 8

Department: Cardiovascular Research (Humoral Regulation)Research Institute of Environmental Medicine

Teaching staff: Haruo Honjo


Time and Date: 16-17 September, 2008

Meeting place: Cardiovascular Research (Humoral Regulation)Research Institute of Environmental Medicine

Outline:

Optical measurements of transmembrane potential using fluorescentvoltage-sensitive dyes have made important contributions to ourunderstanding of electrophysiological properties of excitable tissues. Thesemethods are non-invasive compared with conventional microelectrodetechniques, capable of simultaneous recording with spatiotemporalresolution, and measure membrane potential signals without artifactscaused by electrical stimulation. Optical mapping of cardiac excitation waveshas been widely used to investigate mechanisms underlying cardiacarrhythmias, cardioversion and electrical activity in the heart. This programincludes lecture to lean theoretical backgrounds and experimentaltechnology of voltage-sensitive dye mapping and practical laboratorytraining using Langendorf-perfused whole heart preparations.

23. Whole-mount immunohistochemistry

Capacity: 6

Department: Department of Genetics, Research Institute ofEnvironmental Medicine

Teaching staff: Yasuhiko Kanou

Contact: Tel: 85 (Higashiyama) -3875 E-mail; [email protected]

Time and Date: July 24 – 25

Meeting place: Department of Genetics, Research Institute ofEnvironmental Medicine

Outline:

Whole-mount immunohistochemistry is the localization of antigens inunsectioned tissue, such as embryos, using specific antibodies. Although it issimilar in outline to immunohistochemistry of sectioned material and sharessome problems, it has several advantages. In this training course, mouseembryos will be stained with specific antibodies for nervous tissue by whole-mount immunohistochemistry.

Outline: 1. Dissection2. Fixation and storage3. Whole-mount immunohistochemistry4. Color development, clearing and observation withthe stereomicroscope

24. Immunohistocyotchemistry by confocal laser scanning microscopy

Capacity: 10

Department: Department of Genetics, Division of Stress Adaptation andProtection, Research Institute of Environmental Medicine (RIEM)

Teaching staff: Yoshiko TakagishiContact: Tel: 052-789-3874 or Ex 85-3874 E-mail [email protected]

Time and Date: July 21(in the afternoon) and 22 (in the morning orafternoon)

Meeting place: A meeting venue at the north wing of RIEM

Outline:

Confocal laser scanning microscopy offers several advantages over conventionaloptical microscopy, including obtaining of high-resolution images from relatively thicksections, simultaneous collection of multiple images, and collecting serial opticalsections and their three dimensional representation. Application of confocal laserscanning microscopy has been gaining popularity in immunohistocytochemistry tovisualize localization of specific proteins in cells and tissues.Practice1: Immunofluorescence labeling using tissue sections2: Image collections by confocal laser scanning microscopy

Single and double optical sectionsz-series and three-dimensional reconstructions

25. Principle and application of RT-PCR

Capacity: around 10 students per day (Total 20 students)

Department: Department of Endocrinology, Research Institute ofEnvironmental Medicine (RIEM), Nagoya University

Teaching staff: Fukushi Kambe, Xia Cao


Time and Date: 2008.6.18 (Wed) and 19 (Thu) From 10:00 am - 5:00 pm Each day includes the same lecture and practice so that applicant attends either day.

Meeting place: Main Bldg. Room No.203, Department of Endocrinology,RIEM

Outline:

The course includes the lecture on the principle of RT-PCR and itsapplications such as quantitative PCR, cloning of PCR product (TA cloning), invitro 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 RT-PCR.

26. Bioimaging (Transmission Electron Microscopy)

Capacity: 5

Department: Anatomy and Cell Biology, Center for Research ofLaboratory Animals and Medical Research engineering

Teaching staff: Toyoshi Fujimoto, Yoshikazu Fujita, Ikuyo Mizuguchi


Time and Date: July 22-25, 2008

Meeting place: to be announced

Outline:

Hands-on training course for transmission electron microscopy, includingfixation embedding, ultrathin sectioning, EM observation, and dataacquisition.

27. Fundamental experimental technology for Cell Biology / MolecularBiology: Observation paraspeckles of RNA binding motif protein (RBM) andglucose transport of human normal cells and tumor cells.

Capacity: It is around people from capacity

Department: Anatomy and Functional Histology (Anatomy )

Teaching staff: Koji Nishio PhD.

Contact: Tel An extension number:2021 E-mail [email protected]

Time and Date: Five days of May 26 (Monday) to May 30 (Friday), 2008. Four days of extra day (optional): June 2 to 5, 2008. You can participate on the way of the training course.

Meeting place: Functional Histology (Room# 329, Cell culture room (# 327) andlaboratory (# 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 expressionvectors of GFP or RFP-fused protein and a several fluorescent pigmentswhich are specific for the sub-cellular organelles, we observe the remarkchanges such as nuclear morphology, nuclear body assembly anddisassembly, glucose uptake in human normal cells and tumor cells. Welearn a remark method and applications of GFP, RFP, fluorescent glucose.Through, this training, we learn about technique of RNA interference withthe RBM siRNA that inhibits expression of the nuclear RNA binding motifprotein. The training course contents: we do cell culture operation, immune-fluorescence techniques and transfection of GFP or RFP-expression vectors,total RNA isolation & RTPCR. We observe the assembly of the PML nuclearbodies or RBM paraspeckles.

28. Animal Models for Pain Research

Capacity: 5 persons

Department: Department of Functional Anatomy and Neuroscience

Teaching staff: Ozaki, N

Contact: Tel: ext. 2019 E-mail: [email protected]

Time and Date: May 20-21, 200810:00 a.m -

Meeting place: Room # 329, Basic Science Building 3F

Outline:

Animal models are fundamental resources in pain research. Studies usinganimal models are essential if new and clinically relevant knowledge aboutthe mechanisms of pain is to be acquired. Participants will learn the basicmechanisms of pain sensation and how animal models can be used to analyzeits mechanisms.

Aim: Understand basic mechanisms of pain sensation.Understand animal models used for pain research.

Lecture: Basic mechanisms of pain sensation.Animal models used for pain research.

Laboratory: 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. Gene introduction into developing brain tissues by “electroporation”methods followed by slice culture

Capacity: 10-15 students

Department: Anatomy and Cell Biology

Teaching staff: Takaki Miyata (Prof.)

Contact: Tel 2030 E-mail [email protected],

Time and Date: September 12, 2008, 10:00 a.m. ~

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

Outline:

“Electroporation”, which was originally developed for introducing foreigngenes of interest into suspended cells, can now be used for gene delivery intothree-dimensional in vivo tissues. In this training course, a standard protocolof gene delivery into developing mouse brains through “in uteroelectroporation” will be shown. First, plasmid vectors encoding a greenfluorescent protein (GFP) gene will be micro-injected into lateral ventriclesof embryonic mice by in utero surgery. Then, electric stimulations will begiven to the embryos so that cells facing the lateral ventricle will be shockedand subsequently “eat” the plasmids. Since it will take about 10-12 hr untilGFP proteins starts to be detected in the electroporated brain tissue, anotherlitter of embryos previously electroporated will be used for visualization ofGFP-labeled brain walls. Brains will be sliced, and mounted into collagengels for time-lapse monitoring, by which GFP-labeled single cells can beseen.

30. Immunohistochemistry (Immunostaining)

Capacity: 30 persons

Department: Department of Pathology of Biological Response

Teaching staff: Yoriko Yamashita, MD, PhD


Time and Date: 10:00 AM 3:00PM

Meeting place: Practical Exercise Room for Anatomy and Pathology, 2F,Annex, Medical Research Building

Outline:

Immunohistochemistry, also called immunostaining is a method to visualizeintracellular or extracellular molecule using either a light microscope or,most recently, a laser microsope. Although this method is simple andmostly 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 andprocedure of immunohistochemistry first, and then the students canactually perform immunohistochemistry using ready-to use formalin-fixed,paraffin-embedded samples. We will also show some important points forperforming high-quality immunostaining. Furthermore, we will introduce asimple example of RNA in situ hybridization.

31. Preparation of mouse tissue lysate for SDS-PAGE

Capacity : 10 people

Department : Tumor pathology/Neuropathology

Teaching staff : Mayumi Jijiwa

Contact : Tel Ext. 2093 E-mail [email protected]

Time and Date : 15/08/08 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 proteinconcentration. It takes about two hours for entire process.

32. Clinical application of indirect immunofluorescence methods - anti-nuclear antibody assays -

Capacity: 5~7 persons

Department: Connective Tissue Disease & Autoimmunity

Teaching staff: Yoshinao Muro, M.D. and Kazumitsu Sugiura,M.D.

Contact: Tel Ext.2314 E-mail [email protected]

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 phenomenonby Hargraves, and the measurement of anti-nuclear antibodies (ANA) hasplayed important roles in diagnosis of autoimmune connective diseases, aswell as in elucidation of pathogenesis of the diseases. ANA has clinicallybeen used for screening test to detect particular autoantibodies withfollowing specific examinations. The autoantibodies told us importantclinical information including disease subsets, and prognosis of the disease.In order to interpret the immunofluorescence results of ANA exactly, it isessential to know this experimental method itself. ANA is also very usefultool for basic researches on cellular and molecular biology. In this course,you can examine immunofluorescence assays with patients sera and identifystaining patterns. We could also introduce enzyme-linked immunosorbentassay (ELISA) method used for anti-extractable nuclear antigens (ENA)antibody detection in hospitals recently.

33. Introduction for microsurgery (microvascular anastomosis)

Capacity 5 participants for each group (A,B)

Department Plastic and Reconstructive Surgery

Teaching stuff Shuhei torii, Keisuke Takanari

Contact Tel 2525 E-mail [email protected] (Keisuke Takanari)

Time and date Undecided ( Thursday of Mid November)

Meeting place Plastic and Reconstructive Surgery Office (13F)

Outline

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

34. Observation of environment-derived nanoparticles and nanofibers inhuman lung

Capacity: 3

Department: Occupational and Environmental Health

Teaching staff: Gaku Ichihara


Time and Date: June 30, Saturday, from 14:00

Meeting place: Library of Dept. Occupational and Environmental Health

Outline:

ES&H (Environmental, safety and health) problem of engineerednanomaterials has become of great concern as a consequence of growingnanotechnology. Especially behavior and kinetics of nanoparticles in humansis one of the greatest issues to be solved in the field of nanotoxicology. Thepresent course will introduce the method of identification and analysis ofenvironment-derived nanoparticles and nanofibers in human lung usingelectron microscope.

35. Introductory course on statistical procedures using SPSS for Windows

Capacity: 150 studentsOnly e-learning course is available, however, office-hour isscheduled for your questions.

Department: Department of Public Health/Health InformationDynamics 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 [email protected]

Time and Date: If you are interested in, please be registered at thegraduate students’ affair section (Daigakuin-kakari),and then contact Dr. Yatsuya directly by an e-mail inadvance.

Office hour From 1:30pm to 4:30pm May 16th (Friday), 2008From 5:30pm to 7:30pm May 20th (Tuesday), 2008

Meeting place: 2nd Floor, Annex, Medical Research building(Satellite Information Laboratory)

Outline:

Creation of SPSS dataset from Microsoft Excel spread sheetVariable format/ data definition/ value labelChanging, rearranging, or consolidating the values of an existing variable.Creation of new variable/ Ranking of the existing variable to create new oneFrequency table/ Descriptive statisticsScatter plot/ correlation analysisContingency table/ chi-squared testCalculation of means/ t-test/ ANOVAMultiple 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 [email protected] 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 2nd: Multiple linear regression analysis, Logistic regression analysis 3rd: Survival analysis, Cox proportional hazard analysis, 4th 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: Prof. Atsuko Aoyama Contact: Tel 2109 E-mail [email protected] Time and Date: May 22 (Thu), May 23 (Fri), 2008 1:00 pm – 2:30 pm May 26 (Mon), 2008 1:00 pm – 3:30 pm Meeting place: 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: [email protected] 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: Tel： 789-3961 E-mail：[email protected] Time and Date: 3rd July/2008 from 2:00 PM Meeting place: 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 Doctor-Patient Relationship Capacity: High level of Japanese language ability Department: Psychopathology & Psychotherapy Teaching staff: Prof. Toyoaki Ogawa, Prof. Toshinori Contact: Tel 052-789-5836 E-mail [email protected] 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: [email protected] 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 [email protected] Time and Date: Nov 10, and Nov 12 9:30-12:00 Meeting place: Seminar room of Biochemistry 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: [email protected] 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: 10 Department: Biochemistry Teaching staff: Yoshio Yamauchi Contact: Tel 2068 E-mail [email protected] 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: Keiko YAMADA Contact: Tel 052-744-2102 E-mail [email protected] Time and Date: Nov. 26 – 27, 2008 Meeting place: 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: [email protected] 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:[email protected] 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 [email protected] 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 [email protected] 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: 10 Department: Molecular Carcinogenesis Teaching staff: Motoshi Suzuki Contact: Tel 2456 E-mail [email protected] 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 [email protected] 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: ２０ Department: Cancer Biology Teaching staff: Senga Takeshi Contact: Tel 2463 E-mail [email protected] 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: Neuroscience 1, Research Institute of Environmental Medicine Teaching staff: Takuro Maruyama Contact: Tel: 3878 E-mail: [email protected] 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 Institute of

Environmental Medicine (RIEM) Teaching staff: MURATA Yoshiharu and ODA Sen-ichi Contact: Tel 052-789-4179 E-mail [email protected] Time and Date: Nov. 4, 2008 9:30~17:00 Meeting place: 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: [email protected] 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 (Associate Professor) Rieko Uchihi (Research Associate) Contact: Tel: 2119 (or 2117) E-mail: [email protected] 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 [email protected] 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 [email protected] 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 [email protected] 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 [email protected] 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: [email protected] 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: [email protected] 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 [email protected] 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.

Preparation of recombinant proteins using yeast

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