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

Confocal Dyes
Rat primary antibody for muscarinic M1 (Cat. No. 087k1395), M3 (Cat.
No. 126k1205), α7 nicotinic acetylcholine receptor (Cat. No. 018k4811) and
GABAAα1 (Cat. No. 077k4838) and FITC coated secondary antibody (Cat. No.
No-AP307R) were purchased from Sigma Aldrich and Chemicon, USA.
ANIMALS
Adult male Wistar rats of 180-240g body weight purchased from Amrita
Institue of Medical Sciences, Cochin and Kerala Agriculture Unviersity,
Mannuthy were used for all experiments. They were housed in separate cages
under 12 hour light and 12 hour dark periods and were maintained on standard
food pellets and water ad libitum.
DIABETES INDUCTION AND HYPOGLYCEMIA
Experimental rats were divided into the following groups as i) Control ii)
Diabetic iii) Insulin induced hypoglycemia in diabetic rats (D+IIH) and iv) Insulin
induced hypoglycemia in control rats (C+IIH). Each group consisted of 6-8
animals.
Diabetes was induced by a single intrafemoral dose, 55 mg/kg body
weight, of STZ prepared in citrate buffer, p H 4.5 (Arison et al., 1967; Hohenegger
& Rudas, 1971). The D+IIH group received daily 2 doses of 10 Unit/Kg body
weight of regular human insulin (Actrapid) and C+IIH received daily 2 doses of
1.5 Unit/Kg body weight of regular human insulin (Flanagan et al., 2003). D+IIH
and C+IIH group had daily two episodes of insulin-induced hypoglycemia for 10
days. Control rats were injected with citrate buffer.
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