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

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CONTENTS INTRODUCTION 1 OBJECTIVES OF THE PRESENT STUDY 8 LITERATURE REVIEW 10 Hypoglycemia and brain 12 Glucose homeostasis and pancreas 14 Brain neurotransmitters and diabetes 17 Brain neurotransmitters and hypoglycemia 19 Cholinergic regulation of glucose homeostasis 20 Acetylcholine 20 Cholinergic Innervation 21 Muscarinic receptors 23 Classification of Muscarinic receptors 23 Muscarinic M1 receptor 24 Muscarinic M2 receptor 25 Muscarinic M3 receptor 26 Muscarinic M4 receptor 26 Muscarinic M5 receptor 27 Signal transduction by muscarinic activation 28 Cyclic adenosine monophosphate 28 Phospholipase C 29 Nicotinic Receptors 29 GABA regulation of glucose homeostasis 30 GABA Receptors 31 GABAA Receptors 32 GABAB Receptors 33
GABAC Receptors 34 Glutamic Acid Decarboxylase (GAD) 35 Insulin and Insulin receptors in the brain 39 Alterations of neuronal glucose transport during diabetes and hypoglycemia 40 Oxidative stress and neuronal damage during hypoglycemia and diabetes 42 The cAMP responsive element binding protein (CREB) 43 MATERIALS AND METHODS 45 Chemicals used in the study and their sources 45 Biochemicals 45 Radiochemicals 45 Molecular Biology Chemicals 45 Confocal Dyes 46 Animals 46 Diabetes induction and hypoglycemia 46 Determination of blood glucose 47 Sacrifice and tissue preparation 47 Estimation of blood glucose 47 Estimation of circulating insulin By Radioimmunoassay 48 Principle of the assay 48 Assay protocol 48 Behavioural Studies 49 Y-Maze Test 49 Grid Walk Test 50 Muscarinic and GABA receptor binding
Page 2: MUSCARINIC M1, M3, NICOTINIC, GABAA
Page 5 and 6: ACKNOWLEDGEMENT To the Almighty God
Page 7 and 8: Seema Chandran, Ms. Neema Ani Manga
Page 9 and 10: ABBREVIATIONS 5-HIAA 5 Hydroxy indo
Page 11: PFC Prefrontal cortex PLC Phospholi
Page 15 and 16: Behavioural response of control and
Page 17 and 18: the cerebral cortex of control and
Page 19 and 20: Muscarinic M1 receptor analysis Sca
Page 21 and 22: REAL TIME-PCR ANALYSIS 81 Real Time
Page 23 and 24: Real Time PCR analysis of GABAAα1
Page 25 and 26: pancreas of control and experimenta
Page 27 and 28: utilization is decreased in the bra
Page 29 and 30: impairment, coma or seizure (Cryer,
Page 31 and 32: achieving optimal blood sugar contr
Page 33 and 34: OBJECTIVES OF THE PRESENT STUDY 1.
Page 35 and 36: Literature Review Diabetes mellitus
Page 37 and 38: 12 Literature Review from the adren
Page 39 and 40: 14 Literature Review Hypoglycemic c
Page 41 and 42: 16 Literature Review respectively)
Page 43 and 44: 18 Literature Review in extracellul
Page 45 and 46: 20 Literature Review substrates in
Page 47 and 48: 22 Literature Review nucleus, altho
Page 49 and 50: 24 Literature Review al., 1990; Lev
Page 51 and 52: 26 Literature Review muscarinic M2
Page 53 and 54: Signal transduction by muscarinic a
Page 55 and 56: 30 Literature Review nicotinic acet
Page 57 and 58: GABAA Receptors 32 Literature Revie
Page 59 and 60: 34 Literature Review GABAB-mediated
Page 61 and 62: 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
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96 Results control. D + IIH and C +
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Discussion Glucose is the principal
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100 Discussion The ability to sense
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102 Discussion The Y-maze test is a
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104 Discussion CHOLINERGIC ENZYME A
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106 Discussion Pancreas Insulin sec
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108 Discussion 2003). Cholinergic m
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110 Discussion brain for learning,
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112 Discussion hypoglycemia on brai
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114 Discussion shows impaired expre
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116 Discussion glycemic control is
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118 Discussion release (Ilcol et al
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120 Discussion al., 1992), it has t
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122 Discussion metabolic cycles are
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124 Discussion al., 1988). Low bloo
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126 Discussion modulate the second
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128 Discussion between excitation a
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130 Discussion hypoglycemic seizure
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132 Discussion Insulin secretion fr
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134 Discussion hyper and hypoglycem
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136 Discussion receptors in glucose
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138 Discussion exacerbated by recur
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140 Discussion Haces et al., 2010).
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142 Discussion hypoglycemic and dia
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144 Discussion transcription factor
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Summary 1. Insulin induced hypoglyc
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148 Summary specific antibodies con
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150 Summary 16. Transcription facto
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References Abdelmalik PA, Shannon P
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Caulfield MP, Birdsall NJM. (1998).
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Cherubini E, Martina M, Sciancalepo
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Cryer PE. (2002b). The pathophysiol
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6. Anju T R, Pretty Mary Abraham, S
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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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