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

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7. To study the expression of acetylcholine esterase, choline acetyltransferase, muscarinic M1, muscarinic M3, GABAAα1, GABAB, GAD, insulin receptor, superoxide dismutase, Bax protein and phospholipase C in the pancreas of diabetic and hypoglycemic rats using Real Time PCR. 8. To study the localisation and expression status of muscarinic M1, muscarinic M3, α7 nicotinic acetylcholine receptor and GABAAα1 receptors in the brain slices of cerebral cortex, cerebellum, brain stem, corpus striatum and hippocampus of diabetic and hypoglycemic rats using specific antibodies in confocal microscope. 9. Immunocytochemical studies to analyse the localisation and expression status of muscarinic M1, muscarinic M3 and GABAAα1 receptors in the pancreas of diabetic and hypoglycemic rats using specific antibodies in confocal microscope. 9
Literature Review Diabetes mellitus is a major global health problem that affects more than 246 million people in the world, but by 2025, that number is estimated to reach 380 million (Sicree et al., 2006; Zimmet et al., 2001). It is an increasingly prevalent metabolic disorder in humans and is characterised by hyperglycemia (Kumar & Clarke, 2002; Dunne et al., 2004). The symptoms of diabetes mellitus results from abnormal glucose metabolism. The lack of insulin activity results in failure of transfer of glucose from the plasma into the cells. This situation is so called “starvation in the midst of plenty”. The pancreatic hormones have an important role in the regulation of glucose metabolism. The secretion of insulin by ß-cells of the endocrine pancreas is regulated by glucose and other circulating nutrients. It is also modulated by several hormones and neurotransmitters, among which acetylcholine and GABA plays a prominent role. Hypoglycemia is a serious complication of diabetes and insulin therapy. Hypoglycemic brain injury occurs most frequently in patients attempting tight glucose control (Auer, 2004; Davis et al., 1998). Increased incidence of hypoglycemia occurs when attempts are made to institute tight glycemic control using currently available regimens of subcutaneous insulin administration in diabetic patients (Cryer, 1994). Insulin and sulfonylurea therapy for diabetes mellitus carries the risk of hypoglycemic brain injury and this risk is a major impediment to optimal glucose regulation in diabetic patients (Davis et al., 1998). Tight blood glucose control reduces the risk of diabetes complications but also increases the risk of hypoglycemic episodes. Symptomatic hypoglycemia occurs frequently in insulin-treated patients and 36% of patients were found in one study to have experienced hypoglycemic coma in their lifetime (Pramming et al., 1991). Upto 10% of patients practicing conventional insulin therapy and 25% of those practicing intensive therapy suffer at least one episode of severe, temporarily disabling hypoglycaemia, often with seizure or coma, in a
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 and 12: PFC Prefrontal cortex PLC Phospholi
Page 13 and 14: GABAC Receptors 34 Glutamic Acid De
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: OBJECTIVES OF THE PRESENT STUDY 1.
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
Page 63 and 64: 38 Literature Review which could be
Page 65 and 66: 40 Literature Review administration
Page 67 and 68: 42 Literature Review the hippocampu
Page 69 and 70: 44 Literature Review understanding
Page 71 and 72: Confocal Dyes Rat primary antibody
Page 73 and 74: antipyryl)-p-benzo quinoneimine. Th
Page 75 and 76: was analyzed. Data was expressed as
Page 77 and 78: ANALYSIS OF THE RECEPTOR BINDING DA
Page 79 and 80: Taqman probes of muscarinic M1, M3,
Page 81 and 82: Taylor, 1968). The islets were isol
Page 83 and 84: 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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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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