Muscarinic M1, M3, Nicotinic,GABAA and GABAB Receptor ...

was analyzed. Data was expressed as percentage of performance in all three arms
during the five minutes of test (Akwa et al., 2001, Jobin, et al., 2010).
Grid Walk Test
Deficits in descending motor control were examined by assessing the
ability to navigate across a 1 m long runway with irregularly assigned gaps (0.5–5
cm) between round metal bars. Crossing this runway requires that animals
accurately place their limbs on the bars. In baseline training and postoperative
testing, every animal had to cross the grid for at least three times. The number of
footfalls (errors) was counted in each crossing for 3 minute and a mean error rate
was calculated (Z’Graggen et al., 1998).
MUSCARINIC AND GABA RECEPTOR BINDING STUDIES USING [ 3 H]
RADIOLIGANDS
Binding studies in the Brain regions
Total muscarinic, muscarinic M1 and M3 receptor binding studies
[ 3 H] QNB and [ 3 H] DAMP binding assay in cerebral cortex, cerebellum,
brain stem, hippocampus, corpus striatum and pancreas were done according to
the modified procedure of Yamamura & Snyder (1981). Brain tissues were
homogenised in a polytron homogeniser with 20 volumes of cold 50mM Tris-HCl
buffer, p H 7.4, containing 1mM EDTA. The supernatant was then centrifuged at
30,000xg for 30 minutes and the pellets were resuspended in appropriate volume
of the buffer.
Total muscarinic and muscarinic M1 receptor binding parameter assays
were done using 0.1-2.5nM of [ 3 H] QNB and M3 receptor using 0.01-5nM of
[ 3 H] DAMP in the incubation buffer, p H 7.4 in a total incubation volume of 250µl.
The non-specific binding was determined using 100µM atropine for total
muscarinic, pirenzepine for muscarinic M1 and 4-DAMP mustard for muscarinic
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