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

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function, learning and memory in the diabetic, D + IIH and C + IIH rats compared to control. Behavioural deficit was more prominent in hypoglycemic rats. 6. AChE expression level has been used as a marker for cholinergic activity. AChE expression was analysed in the brain regions and pancreas. During diabetes, the expression was increased in the cerebral cortex, cerebellum, brainstem, hippocampus and pancreas while hypoglycemia caused further increase in expression. In corpus striatum, AChE expression decreased significantly in hypoglycemic and diabetic rats. 7. ChAT expression level has been used as a marker for acetylcholine synthesis. ChAT showed decreased expression in the cerebral cortex, cerebellum, corpus striatum, hippocampus, pancreas while in brainstem it was increased in diabetic rats. D + IIH and C + IIH group showed significant decreased expression in all the brain regions and the pancreas. 8. Total muscarinic receptor was analysed in the brain regions and pancreas of control and experimental rats. The receptor binding was decreased in cerebral cortex, corpus striatum and hippocampus while cerebellum and brainstem showed increased expression in diabetic, D + IIH and C + IIH rats. The Scatchard analysis and gene expression studies of muscarinic M1 receptor revealed a down regulation in cerebral cortex, brainstem and hippocampus whereas in cerebellum and corpus striatum it was up regulated. Muscarinic M3 receptor binding and expression in cerebral cortex, cerebellum, brain stem and hippocampus were increased and in corpus striatum there was a decrease in diabetic, D + IIH and C + IIH rats compared to control. In pancreas, total muscarinic, muscarinic M1 and muscarinic M3 receptors were down regulated in hypoglycemia and diabetic condition. Immunohistochemical studies using 147
148 Summary specific antibodies confirmed the Scatchard analysis and Real Time PCR analysis of muscarinic receptor expression at protein level in control and experimental rats. 9. α7 nicotinic acetylcholine receptor gene expression was studied in brain regions of experimental rats. In hypoglycemic and diabetic condition, the receptor was increased in cerebral cortex, cerebellum, brain stem, corpus striatum and decreased in hippocampus when compared to control. Immunohistochemical studies using specific antibody confirmed the gene expression of α7 nicotinic acetylcholine receptor expression at protein level in control and experimental rats. 10. Total GABA receptor binding and GAD expression was analysed in the brain regions and pancreas of control and experimental rats. The receptor binding decreased in cerebral cortex, cerebellum, brain stem, corpus striatum and hippocampus of hypoglycemic and diabetic rats. The gene expression studies of GABAAα1 and GABAB receptor showed down regulation in cerebral cortex, cerebellum, brain stem, corpus striatum and hippocampus. GAD mRNA expression significantly decreased in all the brain regions studied. In pancreas, GABA receptor binding along with GABAAα1, GABAB and GAD significantly decreased during hypoglycemia and diabetes. Immunohistochemical studies using specific antibody confirmed the binding analysis and Real Time PCR analysis of GABAAα1 receptor expression in control and experimental rats. 11. Decreased GLUT3 in brain regions- cerebral cortex, cerebellum, brainstem, corpus striatum and hippocampus of diabetic rats were decreased further in hypoglycemic group compared to diabetic and control.
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MUSCARINIC M1, M3, NICOTINIC, GABAA
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ACKNOWLEDGEMENT To the Almighty God
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Seema Chandran, Ms. Neema Ani Manga
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ABBREVIATIONS 5-HIAA 5 Hydroxy indo
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PFC Prefrontal cortex PLC Phospholi
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GABAC Receptors 34 Glutamic Acid De
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Behavioural response of control and
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the cerebral cortex of control and
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Muscarinic M1 receptor analysis Sca
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REAL TIME-PCR ANALYSIS 81 Real Time
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Real Time PCR analysis of GABAAα1
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pancreas of control and experimenta
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utilization is decreased in the bra
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impairment, coma or seizure (Cryer,
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achieving optimal blood sugar contr
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OBJECTIVES OF THE PRESENT STUDY 1.
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Literature Review Diabetes mellitus
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12 Literature Review from the adren
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14 Literature Review Hypoglycemic c
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16 Literature Review respectively)
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18 Literature Review in extracellul
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20 Literature Review substrates in
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22 Literature Review nucleus, altho
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24 Literature Review al., 1990; Lev
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26 Literature Review muscarinic M2
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Signal transduction by muscarinic a
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30 Literature Review nicotinic acet
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GABAA Receptors 32 Literature Revie
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34 Literature Review GABAB-mediated
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36 Literature Review (Erlander et a
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38 Literature Review which could be
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40 Literature Review administration
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42 Literature Review the hippocampu
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44 Literature Review understanding
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Confocal Dyes Rat primary antibody
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antipyryl)-p-benzo quinoneimine. Th
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was analyzed. Data was expressed as
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ANALYSIS OF THE RECEPTOR BINDING DA
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Taqman probes of muscarinic M1, M3,
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Taylor, 1968). The islets were isol
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Results BODY WEIGHT AND BLOOD GLUCO
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60 Results IIH and C + IIH rats sho
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Real Time-PCR analysis of muscarini
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Real Time PCR analysis of SOD 64 Re
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66 Results GABAAα1 receptor antibo
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68 Results compared to diabetic gro
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70 Results compared to diabetic rat
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72 Results Muscarinic M3 receptor a
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Muscarinic M3 receptor analysis 74
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76 Results group, α7 nAChR mRNA si
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78 Results diabetic rats. In C + II
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Muscarinic M1 receptor analysis 80
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Real Time-PCR analysis of α7 nAChR
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Real Time PCR analysis of phospholi
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HIPPOCAMPUS Total muscarinic recept
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88 Results IIH and C + IIH group sh
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90 Results group, GLUT3 mRNA signif
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92 Results α7 nACh receptor antibo
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GABA receptor analysis 94 Results S
Page 121 and 122: 96 Results control. D + IIH and C +
Page 123 and 124: Discussion Glucose is the principal
Page 125 and 126: 100 Discussion The ability to sense
Page 127 and 128: 102 Discussion The Y-maze test is a
Page 129 and 130: 104 Discussion CHOLINERGIC ENZYME A
Page 131 and 132: 106 Discussion Pancreas Insulin sec
Page 133 and 134: 108 Discussion 2003). Cholinergic m
Page 135 and 136: 110 Discussion brain for learning,
Page 137 and 138: 112 Discussion hypoglycemia on brai
Page 139 and 140: 114 Discussion shows impaired expre
Page 141 and 142: 116 Discussion glycemic control is
Page 143 and 144: 118 Discussion release (Ilcol et al
Page 145 and 146: 120 Discussion al., 1992), it has t
Page 147 and 148: 122 Discussion metabolic cycles are
Page 149 and 150: 124 Discussion al., 1988). Low bloo
Page 151 and 152: 126 Discussion modulate the second
Page 153 and 154: 128 Discussion between excitation a
Page 155 and 156: 130 Discussion hypoglycemic seizure
Page 157 and 158: 132 Discussion Insulin secretion fr
Page 159 and 160: 134 Discussion hyper and hypoglycem
Page 161 and 162: 136 Discussion receptors in glucose
Page 163 and 164: 138 Discussion exacerbated by recur
Page 165 and 166: 140 Discussion Haces et al., 2010).
Page 167 and 168: 142 Discussion hypoglycemic and dia
Page 169 and 170: 144 Discussion transcription factor
Page 171: Summary 1. Insulin induced hypoglyc
Page 175 and 176: 150 Summary 16. Transcription facto
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Body weight (gm) 300 250 200 150 10
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Figure-3 Blood glucose levels at di
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Figure-5 Behavioural response of co
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Figure-7 Scatchard analysis of [ 3
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Figure-11 Real Time PCR amplificati
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C D + IIH Figure--25 Muscarinic M1
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Figure--27 α 7 nACh receptor expre
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Figure--47 Muscarinic M1 receptor e
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Figure--49 α7 nicotinic acetylchol
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Figure--71 α7 nicotinic acetylchol
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C Figure--93 GABAAα1 receptor expr
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Figure-101 Real Time PCR amplificat
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Figure—113 Muscarinic M3 receptor
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Figure--115 GABAAα1 receptor expre
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Figure-117 Scatchard analysis of [
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Figure-119 Scatchard analysis of [
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Figure-131 Muscarinic M1 receptor e
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C D + IIH Figure-133 GABAAα1 recep
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