ABSTRACT
Increased in ovo cortisol content of rainbow trout oocytes from ~3·5 to ~5·0 ng.oocyte(-1) before fertilization enhances the growth of embryos and juveniles and changes the long-term expression pattern of IGF-related genes. This study used embryos reared from oocytes enriched with cortisol and the glucocorticoid receptor (GR) antagonist, RU486, to determine whether the growth-promoting actions of cortisol involve GR protein activation and modulation of gr expression. Whole-mount in situ immunohistofluorescence studies of zygotes showed that enhanced oocyte cortisol increased the immunofluorescent GR signal and activated the relocation of GR from a general distribution throughout the cytoplasm to an accumulation in the peri-nuclear cytoplasm. In ovo cortisol treatment increased the number of embryonic cells within 48-h post-fertilization, and RU486 partially suppressed this cortisol stimulation of cell duplication. In addition, there was complex interplay between the expression of gr and igf system-related genes spatiotemporally in the different treatment groups, suggesting a role for GR in the regulation of the expression of development. Taken together, these findings indicate an essential role for GR in the regulation of epigenomic events in very early embryos that promoted the long-term growth effects of the embryos and juvenile fish. Moreover, the pretreatment of the oocyte with RU486 had a significant suppressive effect on the maternal mRNA transcript number of gr and igf system-related genes in oocytes and very early stage embryos, suggesting an action of antagonist on the stability of the maternal transcriptome.
Subject(s)
Embryo, Nonmammalian/cytology , Hydrocortisone/metabolism , Oncorhynchus mykiss , Oocytes/metabolism , Receptors, Glucocorticoid/agonists , Somatomedins/genetics , Zygote/physiology , Animals , Cell Division/genetics , Cell Division/physiology , Cleavage Stage, Ovum/cytology , Cleavage Stage, Ovum/metabolism , Cleavage Stage, Ovum/physiology , Embryo, Nonmammalian/metabolism , Embryo, Nonmammalian/physiology , Female , Gene Expression Regulation, Developmental , Hydrocortisone/analysis , Oncorhynchus mykiss/embryology , Oncorhynchus mykiss/genetics , Oncorhynchus mykiss/metabolism , Oncorhynchus mykiss/physiology , Oocytes/chemistry , Receptors, Glucocorticoid/metabolism , Receptors, Glucocorticoid/physiology , Receptors, Somatomedin/genetics , Receptors, Somatomedin/metabolism , Signal Transduction/genetics , Somatomedins/metabolism , Time Factors , Up-Regulation/physiology , Zygote/metabolismABSTRACT
The rainbow trout (Oncorhynchus mykiss) is one of the most widely studied of model fish species. Extensive basic biological information has been collected for this species, which because of their large size relative to other model fish species are particularly suitable for studies requiring ample quantities of specific cells and tissue types. Rainbow trout have been widely utilized for research in carcinogenesis, toxicology, comparative immunology, disease ecology, physiology and nutrition. They are distinctive in having evolved from a relatively recent tetraploid event, resulting in a high incidence of duplicated genes. Natural populations are available and have been well characterized for chromosomal, protein, molecular and quantitative genetic variation. Their ease of culture, and experimental and aquacultural significance has led to the development of clonal lines and the widespread application of transgenic technology to this species. Numerous microsatellites have been isolated and two relatively detailed genetic maps have been developed. Extensive sequencing of expressed sequence tags has begun and four BAC libraries have been developed. The development and analysis of additional genomic sequence data will provide distinctive opportunities to address problems in areas such as evolution of the immune system and duplicate genes.
