1998 Fiscal Year Final Research Report Summary
Functional analysis of GABA_B receptors using RT-PCR in snail neurons
| Project/Area Number |
09670045
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
General physiology
|
|---|
| Research Institution | HIROSHIMA UNIVERSITY |
|---|
Principal Investigator |
OGATA Nobukuni Hiroshima University, Faculty of Mdicine, Professor, 医学部, 教授 (80091255)
|
|---|
| Project Period (FY) |
1997 – 1998
|
| Keywords | GABA-B receptor / nerve cell / ion channel / single cell / gene expression / patch clamp / calcium cahnnel / second messenger |
|---|
| Research Abstract |
The neurotransmitter GABA (gamma -aminobutyric acid) is involved in the inhibitory regulation of a variety of functions in the central nervous system. The receptors for GAGA comprise mainly GABA-A and GABA-B receptors. The GABA-A receptors has long a subject of intensive investigation and thus their structural and functional properties are now well understood. On the contrary, the role of GABA-B receptors still remains be elucidated. The purpose of this project was 1, to examine a modulatory role of GA B receptors on the function of voltage-gated calcium channels, and 2, to develop a model system in which functional GABA-B receptors can be expressed for electrophysiological analysis of GABA-B receptors. Regarding the first purpose, I have investigated the actions of GABA-B agonist, baclofen, on the High voltage activated (HVA)-calcium channels using patch clamp technique. The experimental results show that an activation of GABA-B receptors causes not only well-known inhibitory effect on the calcium cha opening, but also a transient augmentation of the HVA-calcium channel currents. suggested that this effect was probably mediated by a change in the intracellular se messenger system. Regarding the second purpose, I have established the model system that can be used as the expression system of GABA-B receptors. Cultured neurons of superior cervical ganglion provided a number of advantages, e.g., these cells are devoid inherent GABA-B receptors. By the microinjection technique of the mutated genes bined with the use of fluorescent GFP (green-fluorescent protein), exogenous mutated genes could be expressed in an identified cells of the culture disc.
|
