Expression, regulation and possible function of heat shock genes (hsp47, hsp70, hsp90(alpha) and hsp90(beta)) during normal development and under stress-conditions in zebrafish (cell differentiation, notochords)

Lele, Z.
Ph.D. Thesis : (Thesis)
Registered Authors
Lele, Zsolt
MeSH Terms
The objectives of my thesis work to examine the similarities and differences that exist among the control and stress induced expression patter of the above mentioned heat shock genes during zebrafish embryogenesis. Northern blot analysis revealed stress- and gene-specific differences in the heat and ethanol-induced activation of these genes. Furthermore, spatial differences in the mRNA distribution of hsp genes following heat shock and ethanol-treatment were identified using whole-mount in situ hybridization. The second aspect of my thesis deals with constitutive expression of hsp47 during the embryonic development of zebrafish. Based on biochemical evidence, Hsp47 is regarded as a collagen-specific chaperone. I show that strong expression of hsp47 mRNA is co-incident predominantly with expression of the type II collagen gene (col2al ) in a number of chondrogenic and non-chondrogenic tissues including the notochord, otic vesicle and developing fins. Notochordal expression of both genes is disrupted in floating head (flh) and no tail (ntl) embryos, which lack properly differentiated notochords. Surprisingly, no hsp47 mRNA is detectable in the strongly col2a1-expressing cells of the floor plate and hypochord, indicating that the two genes are not strictly co-regulated. In the third part of my thesis I examined the possible roles Hsp90 might play during zebrafish myogenesis as it was suggested by our previous studies that revealed coordinate expression of hsp90? and myoD, one of the major myogenic regulatory factors (J. Sass, PhD thesis). Utilizing the Hsp9O-binding agent, geldanamycin (GA), I show that GA-treatment during zebrafish gastrulation and early somitogenesis specifically disrupts the development of a group of muscle cells known as the muscle pioneers. However, development of the notochord, which provides many of the signals required for proper somite patterning, is normal in GA-treated embryos. Furthermore, the PKA-dependent mechanism by which responding cells of the paraxial mesoderm are thought to be specified appears to be intact in treated embryos and the myoD-expressing adaxial cell progenitors of muscle pioneers develop. The data indicate that Hsp90 is required during the conversion of adaxial cells into muscle pioneers and that many of the signaling and differentiation events occurring in the pathway prior to this point are unaffected.
Errata / Notes
Ph.D. Thesis, The University of Saskatchewan (Canada)
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes