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KEY ROLE OF TESTIS SPECIFIC PROTEIN Y ENCODED (TSPY) IN BOVINE EARLY EMBRYO DEVELOPMENT

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Title: KEY ROLE OF TESTIS SPECIFIC PROTEIN Y ENCODED (TSPY) IN BOVINE EARLY EMBRYO DEVELOPMENT
Author: Rho, NaYoung
Department: Department of Biomedical Sciences
Program: Biomedical Sciences
Advisor: Favetta, LauraKing, Allan
Abstract: It is well known that male and female embryos show sexual dimorphic gene expression. To further explore this, the expression pattern and developmental effect of an autosomal gene, thyroid hormone receptor (THR) and a multicopy Y-chromosome linked gene, testis specific protein Y-encoded (TSPY), were examined. Bovine embryos of known sex produced by in vitro fertilization (IVF) with sexed semen and cultured to the blastocyst stage were examined for cleavage and blastocyst percentages. mRNA and protein levels were measured by digital droplet PCR (ddPCR) and immunocytochemistry at the 2-,4-.8- cell and blastocyst stages for both THR and TSPY. TSPY RNA was knocked down by microinjection with un-injected and sham injected zygotes as controls. Post-knockdown development was monitored as were mRNA and protein levels for THR and TSPY. Absolute TSPY copy number was determined by ddPCR in sperm used for IVF and in the resulting embryos. TSPY protein levels and localization, as well as TSPY mRNA levels were determined for sperm. The percentage of the fertilized oocyte that cleaved was significantly higher between the male and female groups while the percentage that reached blastocyst significantly higher between male and female groups and different among the male groups from different bulls. THR mRNA and protein levels were higher in male embryos than female ones. TSPY DNA, RNA and proteins were not present in female embryos or in X chromosome-bearing sperm. TSPY copy number, mRNA and protein varied among male embryos and the Y-bearing sperm from different bulls. TSPY knockdown had no effect on female embryos. No significant effects were detected in male control embryos, however, TSPY knockdown male embryos had significantly lower levels of both TSPY and THR’s mRNA and protein. Development of TSPY knockdown males stopped before the blastocyst stage. The present study showed 1) sex dimorphism in the expression of the Y-linked gene TSPY and the autosomal gene THR in early bovine embryo development, and 2) TSPY knockdown had no effect on female embryo groups, but prevented male embryo development, 3) TSPY knockdown lowered THR expression in males. It was concluded that TSPY and THR are critical for early embryo development.
URI: http://hdl.handle.net/10214/17780
Date: 2020-01-20
Rights: Attribution-NoDerivatives 4.0 International
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Attribution-NoDerivatives 4.0 International Except where otherwise noted, this item's license is described as Attribution-NoDerivatives 4.0 International