| Project/Area Number |
05558093
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Nerve anatomy/Neuropathology
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
KOSAKA Toshio KYUSHU UNIVERSITY,FACULTY OF MEDICINE,DEPARTMENT OF ANATOMY,PROFESSOR, 医学部, 教授 (00126054)
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| Co-Investigator(Kenkyū-buntansha) |
AIKA Yusuke KYUSHU UNIVERSITY,FACULTY OF MEDICINE,DEPARTMENT OF ANATOMY,ASSOCIATE PROFESSOR, 医学部, 講師 (50021415)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥15,200,000 (Direct Cost: ¥15,200,000)
Fiscal Year 1994: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1993: ¥13,300,000 (Direct Cost: ¥13,300,000)
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| Keywords | QUANTITATIVE ANALYSIS / STEREOLOGY / DISECTOR / IMMUNOCYTOCHEMISTRY / CONFOCAL LIGHT MICROSCOPE / IMAGE ANALYSIS / NEURON / BRAIN / フォトCD / 中枢神経系 / disector / システマティク・サンプリング |
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| Research Abstract |
Detailed structural analyzes, especially quantitative analyzes, are essential to understanding the neuronal organization of the central nervous system and its function. In the present research project we have tried 1) to establish the bases for the stereological analyzes, particularly optical disector analysis, in the central nervous system and 2) to establish the methods of image analyzes for evaluating the immunoreaction intensities of individual somata and synaptic terminals in brain samples at the light microscopic level.
For the optical disector analysis, we stained 50 mm thick sections fluorescent immunocytochemically, and examined by means of a confocal laser scanning light microscope (CLSM). The penetration of immunostaining from the section surfaces was carefully evaluated and the conditions of samples which can be used for the optical disector analysis were established. Then the established methods allowed us to analyze various cell groups defined chemically by means of conventional light microscope as well as CLSM.
The methods for evaluating the intensities of immunoreactivities in individual elements such as neuronal somata and synaptic boutons were developed and applied on determining the distributions of two isozymes of GABA synthesizing enzymes, GAD 67 and GAD 65, in hippocampal GABAergic neurons. Images were acquired by TV camera attached to conventional light microscope for the analyzes of somata and by CLSM for the analyzes of synaptic boutons. Relative reaction intensities on individual elements were analyzed by personal computer using an image analysis software NIH Image.
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