Identification of quantitative trait loci in alcoholism [Elektronische Ressource] / submitted by Eva Drews

Eva Drews

Sujets

• Biologie

Informations

• Publié par: rheinische_friedrich-wilhelms-universitat_bonn
• Publié le: 01 janvier 2009
• Nombre de lectures: 5
• Langue: English
• Poids de l'ouvrage: 2 Mo

Extrait

Identification of Quantitative Trait
Loci in Alcoholism



Thesis
Submitted for a Doctoral Degree in Natural Sciences
(Dr. rer. nat.)
Faculty of Mathematics and Natural Sciences

Rheinische Friedrich Wilhelms University Bonn




Submitted by
Eva Drews





Bonn 2009
Prepared with the consent of the Faculty of Mathematics and Natural Sciences
Rheinische Friedrich Wilhelms University, Bonn.

This thesis is available online on Hochschulschriftenserver of ULB Bonn
http://hss.ulb.uni-bonn.de/diss_online.

Publication year: 2009























1. Reviewer: Prof. Dr. Andreas Zimmer


2. Reviewer: Prof. Dr. Jörg Höhfeld

Date of submission: 12/03/2009
Date of examination: 01/07/2009





























Ich dachte darüber nach, in wie hohem Maße die Naturwissenschaft es sich zum Plan
gemacht hat, absolut alles erklären zu wollen. Worin sich natürlich die Gefahr versteckt,
allem gegenüber, das sich nicht erklären lässt, vollständig blind zu werden.

Maya
oder Das Wunder des Lebens
J. Gaarder
Abbreviations
5-HT 5-hydroxytryptamine-3, serotonin
A Adenine / Adenosine
ADH Alcohol dehydrogenase
ALDH2 Acetaldehyde dehydrogenase 2
AMPA α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid
ANOVA Analysis of variance
bp Base pair
C Cytosine / Cytidine
cAMP Cyclic adenosine monophosphate
Chr Chromosome
cM Centimorgan
cm Centimeter
CPP Conditioned place preference
dB Decibel
DNA Desoxyribonucleic acid
dNTP Desoxyribonucleoside triphosphate
EDTA Ethylenediamine tetraacetic acid
EtOH Ethanol
F Degree of freedom
F1 First filial generation
F2 Second filial generation
FAEE Fatty acid ethyl ester
FAM 6-carboxy-fluorescine
g Gram
G Guanine / Guanosin
GABA γ-aminobutyric acid
h Hour
HEK cells Human embryonic kidney cells
HEX Hexachloro-6-carboxy-fluorescine
HPLC High performance liquid chromatography
i.p. Intraperitoneal
IMP Multiple imputation
kb Kilobase pair
kDa Kilodalton
kg Kilogram
LOD Logarithm of the odds LOG Common logarithm
m Meter
M.musculus Mus musculus (house mouse)
mA Milliampere
MEOS Microsomal ethanol oxidation system
MgCl Magnesium chloride 2
min Minutes
mRNA Messenger RNA
msec Milliseconds
n Sample size
n.s. Not significant
nAChR Nicotinic acetylcholine receptor
NAD (P) Nicotinamide adenine dinucleotide (phosphate)
ng Nanogram
NMDA N-methyl-D-aspartic acid
OD Optical density
P450 Cytochrome P450IIE1-CYP2E1
PA Polyacrylamide
PCR Polymerase chain reaction
pd Postnatal day
Pde6b Phosphodiesterase 6B
PK Protein kinase
QTL Quantitative trait locus / loci
RNA Ribonucleic acid
ROX 6-carboxy-X-rhodamine
sec Seconds
SEM Standard error of the mean
SNP Single nucleotide polymorphism
T Thymine / Thymidine
TAE Tris-acetate-EDTA
TBE Tris-Borate-EDTA
TET Tetrachloro-6-carboxy-fluorescine
Tris Tris (hydroxymethyl) aminomethane
U (Enzyme) Unit
VTA Ventral tegmental area
µl Microlitre
Index I
INDEX
1 INTRODUCTION..................................................................... 1
1.1 Metabolism of alcohol...................... 2
1.2 Neurobiological effects of alcohol .................................................................. 4
1.3 Animal models in alcoholism research........................... 9
1.4 Genetics of alcoholism................................................................................... 11
1.4.1 Linkage versus association studies........................................ 12
1.5 Quantitative trait loci analysis....................................... 13
1.6 Aim of the study.............................................................................................. 15
2 MATERIAL............ 16
2.1 Equipment ....................................................................................................... 16
2.2 Software........... 16
2.3 Chemicals and reagents................................................................................. 17
2.4 Enzymes .......................................... 17
2.5 Molecular weight standards........................................................................... 17
2.6 Oligonucleotides............................................................................................. 17
2.7 Solutions......................................... 18
2.8 Databases and programs............................................... 18
2.9 Mouse strains.................................................................................................. 19

Index II
3 METHODS............................................................................. 21
3.1 Behavioural experiments............................................... 21
3.1.1 Animals................................................. 21
3.1.2 Experimental design.............................. 21
3.1.3 Alcohol-induced hypothermia................................................. 22
3.1.4 Chronic alcohol consumption and alcohol withdrawal ............................................ 22
3.1.5 Alcohol preference................................................................. 24
3.1.6 Stress-induced alcohol drinking............. 25
3.1.7 Statistical data analysis of behavioural data.......................... 25
3.2 Biomolecular experiments............................................................................. 26
3.2.1 Mouse tissue preparation...................... 26
3.2.2 Isolation of genomic DNA 26
3.2.3 Normalization of genomic DNA.............................................................................. 26
3.2.4 Selection of the microsatellite marker set............................................................... 27
3.2.5 Genotyping and fragment length analysis.............................. 27
3.2.5.1 Polymerase chain reaction ..................................................................28
3.2.5.2 Fragment lengths analysis...................................29
3.2.6 Detection and mapping of quantitative traits.......................... 30
4 RESULTS.............................................................................. 33
4.1 Behavioural analysis of the parental mouse strains ................................... 33
4.1.1 Acute ethanol-induced hypothermia and tolerance................ 33
4.1.2 Ethanol dependence and withdrawal ..................................... 34
4.1.3 Ethanol preference and consumption.................................... 36
4.1.4 Stress-induced ethanol drinking............. 37
4.2 Behavioural analysis of the first filial generation ........................................ 39
4.2.1 Acute ethanol effects and tolerance....................................... 39
4.2.2 Ethanol dependence and withdrawal..... 40
4.2.3 Ethanol preference and consumption.................................... 41
4.2.4 Stress-induced ethanol drinking............................................. 42
4.3 Analysis of the second filial generation ....................................................... 43
4.3.1 Data quality control................................................................ 43
4.3.2 Mapping of quantitative traits................................................. 44 Index III
4.3.2.1 Acute ethanol-induced hypothermia and tolerance ...........................................................45
4.3.2.2 Ethanol withdrawal ...............................................................................50
4.3.2.3 Ethanol preference and consumption.................................................53
4.3.2.4 Stress-induced ethanol drinking..........................................................54
4.3.2.5 Summary of the two-dimensional-QTL analysis ................................................................57
5 DISCUSSION ........................................................................ 59
5.1 Alcohol effects on body temperature regulation......... 59
5.2 Alcohol and anxiety........................................................ 62
5.2.1 Chronic alcohol consumption and withdrawal ................................ 63
5.3 Alcohol preference and consumption........................... 65
5.4 Alcohol and stress.......................................................................................... 67
5.5 Somatic alcohol dependence ........................................................................ 68
6 SUMMARY AND OUTLOOK................ 70
7 REFERENCES...................................................................... 71
8 SUPPLEMENT...... 80
8.1 Supplemental material.................................................................................... 80
8.2 Publ