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samenvatting H2: methods in developmental biology van prof callaerts
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samenvatting van het vak concepts of developmental biology. H2: methods in developmental biology van prof callaerts
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H2 : METHODS IN DEVELOPMENTALBIOLOGY
P. CALLAERTS
Inhoudsopgave
2.1 A LOOK THROUGHOUT THE ANIMAL KINGDOM.....................................................2
2.1.1 LOOKING AT DIFFERENT LEVELS................................................................................................. 3
2.2 LOOKING AT EMBRYOS: FATE MAPPING (=ORGAANKAART).................................................................4
2.2 MANIPULATING EMBRYOS..................................................................................5
2.2.1 LIGATION.............................................................................................................................. 5
2.2.2 TRANSPLANTATION.................................................................................................................. 6
2.2.3 EXPLANTS............................................................................................................................. 6
2.2.4 ABLATION............................................................................................................................. 6
2.2.5 MICROINJECTION..................................................................................................................... 6
2.2.6 CELL MIGRATION.................................................................................................................... 7
2.3 MOLECULES AND EMBRYOS................................................................................7
2.3.1 IN SITU HYBRIDIZATION: LOOKING AT TRANSCRIPTS........................................................................8
2.3.2 IMMUNOHISTOCHEMISTRY: LOOKING AT PROTEINS..........................................................................8
2.4 GENES, GENE NETWORKS AND EMBRYOS............................................................8
2.4.1 GENE REGULATION.................................................................................................................. 8
2.4.1.1 reporter genes............................................................................................................ 9
2.4.1.2 protein-DNA interaction............................................................................................10
2.4.1.3 targeted mutagenesis...............................................................................................10
2.4.2 GENE NETWORKS.................................................................................................................. 12
2.4.2.1 single cell approaches............................................................................................... 12
2.4.2.2 systems biology approaches.....................................................................................12
2.4.3 DISCOVERING/DECIPHERING GENE FUNCTION..............................................................................12
2.4.3.1 loss-of-function approaches......................................................................................12
2.4.3.2 transgenesis............................................................................................................. 15
2.4.3.3 gain-of-function approaches.....................................................................................16
2.4.3.4 genome editing......................................................................................................... 16
2.5 DEVELOPMENT AND EVOLUTION: COMPARATIVE METHODS................................16
2.5.1 MOLECULAR PHYLOGENY........................................................................................................ 16
2.5.2 GENE FAMILIES (E.G. HOX GENES)........................................................................................... 16
2.5.3 CROSS-SPECIES APPROACHES.................................................................................................. 16
1
, 2.1 A LOOK THROUGHOUT THE ANIMAL KINGDOM
8 examples:
1) Coral
Early development of a fertilized egg. (Sperm and
eggs in sea water -> get fertilized -> coral start
growing).
From the egg to the planula = Planktonic phase
when the embryo will settle on a substrate +
undergo metamorphosis it enters the Benthic
phase
2) Snail eggs
- in clusters
- many eggs
- translucent: easy to observate the
development
3) Octopus
- used for research of development of CZS
4) Drosophila melanogaster
- first genetically tractable model organism
5) C. Elegans
- transparent for studying cell number and
programmed cell death
6) Chicken
- study variety of things on developing
embryo
7) Xenopus Laevis
- large eggs easy to observe + manipulate
2
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