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Publié par | profil-zyak-2012 |
Nombre de lectures | 133 |
Langue | Français |
Poids de l'ouvrage | 2 Mo |
Extrait
Thèse présentée pour obtenir le grade de
Docteur de l’Université Louis Pasteur
Strasbourg I
Discipline :
par ANGELONE TOMMASO
Titre Titre de la thèse
“The emerging role of Vasostatins and Hippocampal
Cholinergic Neurostimulating Peptide: new putative
antiadrenergic factors”
thSoutenue publiquement le 21 NOVEMBER 2005
Membres du jury
Directeur de Thèse : Madame Marie-Hélène Metz-Buotigue, Dr, Institut National de la Santè
et de la Recherche Médical INSERM U575
5, rue Blaise Pascal 67084 Strasbourg Cedex France
Rapporteur Interne : M. Francis Schnejder, Pr, Service de Rèanimation CHU Hautepierre
Hòpital de Hautepierre , Avenue Molière 67098 Strasbourg Cedex
Rapporteur Externe : M. Youssef Anouar , Dr, Institut National de la Santè et de la Recherche
Médical INSERM U413
UFR de Sciences et Technique 76821 Mont-Saint Aignau
Rapporteur Externe : M. Giuseppe Alloatti , Pr, Dip.di Biologia Animale e dell’Uomo Facoltà di
Scienze Matematiche Fisiche e Naturali
Via Accademia Albertina, 13, Università di Torino 10100, Torino Italy
Examinateur : M. Giovanni Zummo, Pr, Facoltà di Medicina Dip. di Medicina Sperimentale
sezione di Anatomia Umana- Università di Palermo Via Del Vespro 129, Policlinico 90127 Palermo
Italy
University of Calabria, Cosenza - Italy
Inserm U575, Universitè “Louis Pasteur” Strasbourg - France
Doctorate Thesis on co-tutorship in “Animal Biology” and in
“Aspects Moléculaires et Cellulaires de la Biologie”
Discipline: Cardiovascular Physiology and Biochemistry
Autor: Dr. Tommaso Angelone
Title:
“The emerging role of Vasostatins and Hippocampal “The emerging role of Vasostatins and Hippocampal
Cholinergic Neurostimulating Peptide: new putative
antiadrenergic factors”
co-TUTOR TUTOR
Dr. M-H Metz-Boutigue Pr. Bruno Tota
stDate of discussion: 21 November 2005
Academic Year 2004/2005 Academic Year 2004/2005
INDEX
Resumé de la thèse ……………...…………...……………………………………...…..…............ 1
Backgroud ..…...………………………………………………...…………………………........... 13
Backgroud in French…...…………………………………………………….……………........... 22
Introduction ………..…………………………………………………....………….……...…....... 31
1. The chromaffin cells ……………………………………………………………….......….. 32
2. affin cells as source of cardioregulatory peptides….………………..….......….. 34
Materials and Methods ………………………………………...…………...……………….. …...41
1. Isolated and perfused rat Langendorff heart …………………………………………......... 42
1.1 Animals …………………………………………………………………………........... 42
1.2 Isolated heart preparation ………………………………………………………........... 42
2. Experimental protocols ………………………………………………………………......... 44
2.1 Basal Conditions …………………………………………………………………......... 44
2.2 Drug application …………………………………………………………………......... 44
2.2.1 VS1 and VS2 stimulated preparations …………………………………………............ 44
2.2.2 Isoproterenol stimulated preparations …………………………………………….........45
2.2.3 Ouabain stimulated preparation ………………………………………………….......... 45
2.2.4 Mechanism of action of VS1 ……………………………………………………...........46
2.2.5 Cholinergic stimulated preparations ………………………………………………........46
2.2.6 Gi/0 Protein involvement ………………………………………………………........... 47
2.3 Myocardial Protective Effects ………………………………………………….............47
2.4 Statistics ………………………………………………………………………….......... 48
3. Cell Culture and Immunochemistry …………………………………………………..........49
4. Mass Spectra Analysis ………………………………………………………………...........50
Results………………………………………………….…….…………..…...…….........................51
1. Isolated heart preparations ………...……..…………………………………………............52
1.1 Basal Conditions …………………………………………………………………......... 52
1.2 VS1 and VS2 stimulated preparations …………………………………………............ 52
1.3 Inotropic effect …………………………………………………………………........... 52
1.4 Lusitropic effect …………………………………………………………………...........54
1.5 Isoproterenol-stimulated preparations ……………………………………………....... .56
1.6 Mechanism of action of VS1 …………………………………………………….......... 61
1.6.1 Membrane receptors ……………………………………………………………............61
1.6.2 Gi/0 Protein ……………………………………………………………………….........61
1.6.3 NO-cGMP-PKG System …………………………………………………………........ 61
1.7 Myocardial Protective Effects ………………………………………………….............65
2. Cell Culture and Immunochemistry ………………………………………………....... 66
3. Western Blot Analysis of CGA N-terminal Peptides Present in Rat heart Extracts …........ 68
3.1 Purification and immunodetenction of N-terminal CGA fragments …………….......... 69
3.2 Comparison CGA sequences fro