Genetic dissection of the predisposition to feather pecking of laying hens [Elektronische Ressource] : behavioural studies, identification of candidate genes and gene expression analyses / Michal Wysocki

Michal Wysocki

Informations

• Publié par: technische_universitat_munchen
• Publié le: 01 janvier 2007
• Nombre de lectures: 19
• Langue: Deutsch
• Poids de l'ouvrage: 8 Mo

Extrait

Technische Universität München
Lehrstuhl für Tierzucht
Genetic dissection of the predisposition to feather pecking of laying
hens: behavioural studies, identification of candidate genes and
gene expression analyses
Michal Wysocki
Vollständiger Abdruck der von der Fakultät Wissenschaftszentrum Weihenstephan für
Ernährung, Landnutzung und Umwelt der Technischen Universität München zu Erlangung
des akademischen Grades eines
Doktors der Naturwissenschaften
(Dr. rer. nat.)
genehmigten Dissertation.
Vorsitzende: Univ.-Prof. Angelika Schnieke, Ph.D. (Univ. of Edinburgh/UK)
Prüfer der Dissertation:
1. Univ.-Prof. Dr. sc. techn. ETH, Dr. agr. habil. Hans-Rudolf Fries
2. Univ.-Prof. Dr. rer. nat., Dr. agr. habil. Heinrich H. D. Meyer
3. apl. Prof. Dr. rer. nat., Dr. hum. biol. habil. Jerzy Adamski
Die Dissertation wurde am 18.10.2006 bei der Technischen Universität München
eingereicht und durch die Fakultät Wissenschaftszentrum Weihenstephan für Ernährung,
Landnutzung und Umwelt am 21.12.2006 angenommen.The search for truth is in one way hard and
in another way easy. For it is evident that
no one can master it fully nor miss it
wholly. But each adds a little to our
knowledge of Nature, and from all the facts
assembled there arises a certain grandeur.
AristotleContents 5
Contents ...........................................................................................................................5
List of tables and figures .................................................................................................8
List of tables.......................................................................................................8
List of figures.....................................................................................................9
Abbreviations.................................................................................................................10
1 Introduction and goals...........................................................................................12
1.1 Introduction......................................................................................................12
1.2 The goals of this dissertation were....................................................................13
2 Literature review ...................................................................................................14
2.1 Description and definition of feather pecking behaviour ...................................14
2.2 Environmental and endogenous factors involved in feather pecking .................14
2.2.1 Size of the flock..............................................................................................................................14
2.2.2 Social transmission.........................................................................................................................15
2.2.3 Foraging behaviour ........................................................................................................................15
2.2.4 Housing systems.............................................................................................................................16
2.2.5 Light intensity.................................................................................................................................16
2.2.6 Genetic factors and the effect of breed and sex ............................................................................16
2.2.7 Stress and frustration......................................................................................................................18
2.2.8 Nutritional effects and feather eating ............................................................................................18
2.2.9 Neurotransmitters and hormones correlated with feather pecking...............................................18
2.3 Exploratory behaviour......................................................................................19
2.4 Analyses of candidate genes .............................................................................19
2.4.1 Polymorphism analysis ..................................................................................................................20
2.4.2 Gene expression analysis ...............................................................................................................20
2.5 Microarray technique .......................................................................................21
2.5.1 Principles of a microarray experiment ..........................................................................................21
2.5.2 Advantages of the microarray technique.......................................................................................22
2.5.3 Designing and processing the microarray experiment..................................................................22
2.5.4 Statistical analysis of the microarray experiment .........................................................................24
2.6 Realtime quantitative RT-PCR .........................................................................25
2.6.1 Detection methods and chemistries ...............................................................................................25
2.6.2 Quantification of realtime RT-PCR...............................................................................................26
3 Materials and Methods ..........................................................................................27
3.1 Behavioural experiments ..................................................................................27
3.1.1 Animals...........................................................................................................................................27
3.1.2 Hatching conditions........................................................................................................................27
3.1.3 Environmental conditions during the behavioural experiments...................................................28
3.1.4 Experiment #1 and #2 ....................................................................................................................28
3.1.5 Experiments #3-#6 .........................................................................................................................29
3.1.6 Scoring exploratory episodes.........................................................................................................29
3.1.7 Data transformation........................................................................................................................30
3.1.8 Statistical analysis ..........................................................................................................................30
3.2 Identification of candidate genes through literature search................................31
3.3 Acquiring and processing sequences of candidate genes ...................................32
3.4 Isolation and evaluation of genomic DNA ........................................................33
3.4.1 Isolation of genomic DNA from blood .........................................................................................33
3.4.2 Evaluation of genomic DNA .........................................................................................................34
3.5 Panel used for polymorphism analysis..............................................................35Contents 6
3.6 Polymerase chain reaction (PCR) .....................................................................35
3.6.1 Standard PCR .................................................................................................................................35
3.6.2 PCR products purification..............................................................................................................36
3.6.3 Primer design for standard PCR ....................................................................................................36
3.6.4 PCR optimisation ...........................................................................................................................36
3.7 BAC cloning procedure....................................................................................36
3.7.1 Dot-Blot as positive control...........................................................................................................37
3.7.2 Labelling.........................................................................................................................................37
3.7.3 Hybridisation ..................................................................................................................................37
3.7.4 BAC DNA isolation .......................................................................................................................38
3.7.5 Direct sequencing of BAC DNA:..................................................................................................39
3.8 Sequencing reaction .........................................................................................39
3.8.1 Sequencing reaction using the Sanger method (Sanger et al., 1977) ...........................................39
3.8.2 Cleanup of sequencing reaction.....................................................................................................39
3.8.3 Polyacrylamide sequencing gel preparation.................................................