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

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DECLARATION I hereby declare that the thesis entitled “Muscarinic M1, M3, Nicotinic, GABAA and GABAB Receptor Subtypes Gene Expression in Insulin Induced Hypoglycemic Rat Brain Regions: Functional Regulation through Phospholipase C and CREB Protein” is the authentic record of research work carried out by me for my doctoral degree, under the supervision and guidance of Dr. C. S. Paulose, Professor & Head, Department of Biotechnology, Director, Centre for Neuroscience, Cochin University of Science and Technology and that no part thereof has previously formed the basis for the award of any degree or diploma, associateship or other similar titles or recognition. Cochin – 682022 Sherin Antony 26-05-2010 Reg. No. 3169 Department of Biotechnology Cochin University of Science and Technology
ACKNOWLEDGEMENT To the Almighty God; source of all life and goodness from whom everyone in heaven and on earth takes its name. His blessings and His power that work within me and also with the people behind my life for inspiring, guiding and accompanying; I bow before you with a sense of gratitude. It is a pleasant moment to express my heartfelt gratitude to all. I would like to extent my gratitude to Prof. C. S. Paulose, Director, Centre for Neuroscience and Head, Dept. of Biotechnology, Cochin University of Science & Technology for his supervision, advices, motivation and exemplary guidance throughout the course of my doctoral research. I extent my profound thankfulness, for his constant encouragement and timely advice. I am deeply grateful to him for providing me necessary facilities and giving me exposure to organizing skills which exceptionally inspired and enriched my growth as a student and a researcher as well. I offer my sincere thanks to Dr. E. Vijayan, ICMR Emeritus Professor, Centre for Neuroscience, Dept. of Biotechnology, Cochin University of Science and Technology and Dr. Oommen V Oommen, Department of Zoology, Kerala University and for their help and encouragement during the period of my research. Also, I wish to extend my gratitude to Dr. Babu Philip, Dept. of Marine Biology, Microbiology and Biochemistry, CUSAT for his help, encouragement and advice. I would like to acknowledge Prof. M. Chandrasekaran, Dr. Sarita G. Bhat, Dr. Padma Nambisan, faculties of the Department of Biotechnology, CUSAT, and Dr. Elyas K. K, former lecturer of our teachers of the Department for their advice and support throughout my work. I take this opportunity to thank Dr. K. K. Mohammed Yusuf, Dean and Faculty of Science, Cochin University of Science & Technology for his help extended to me.
Page 2: MUSCARINIC M1, M3, NICOTINIC, GABAA
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 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
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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
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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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Akwa Y, Ladurelle N, Covey F, Bauli
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Arison RN, Ciaccio EI, Glitzer MS,
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Balters E, Francesco M. (2002). New
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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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Golding DW, Pow DV. (1990). Neurose
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Haddad GG, Jiang C. (1993). Mechani
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Robinson R, Krishnakumar A, Paulose
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Salkovic M, Lackovic Z. (1992). Bra
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Scheucher A. Alvarez AL, Torres N,
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(2002). Sulfonylurea receptor type
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Weiner DM, Levey AI, Brann MR. (199
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Zhao W, Chen H, Xu H, Moore E, Meir
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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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