Thesis
Investigating the role of retinoic acid (RA) in the non-human primate testis
Washington State University
Master of Science (MS), Washington State University
12/2017
DOI:
https://doi.org/10.7273/000004195
Handle:
https://hdl.handle.net/2376/125243
Abstract
Retinoic Acid (RA) is essential for germ cell development in the rodent testis. In the absence of RA, germ cell development is halted. RA triggers undifferentiated A spermatogonia to transition into differentiating A1 spermatogonia, which is commonly referred to as the A to A1 transition. This transition is marked by the co-expression of STRA8 and KIT (both RA responsive markers) and by the expression of retinoic acid receptor gamma (RARG) directly prior to this transition, but loss of expression of this receptor shortly after the transition occurs. Whether or not an equivalent of this transition exists in the primate testis and/or whether RA is involved is unknown. We attempted to stain non-human primate testis with the mouse STRA8 antibody, but no staining occurred. Next, we produced a rabbit polyclonal antibody against the full-length recombinant primate STRA8 (pSTRA8) protein. Using immunohistochemistry, we stained non-human primate testis with pSTRA8. Our preliminary data suggests that a subpopulation of primate spermatogonia and primary spermatocytes may be RA responsive as they express pSTRA8 in specific stages along the length of the seminiferous tubules. Specifically, Stage VII-IX contains a ring of pSTRA8 immuno-positive primary spermatocytes along the outside of the tubule, which are the stages where entry into meiosis occurs. Additionally, we performed an organotypic culture of fresh non-human primate explant tissue and treated with DMSO (vehicle control), BDAD (inhibitor of RA synthesis) only, and either BDAD + all-trans RA or BDAD + 13cis RA. The inhibition of RA synthesis or the addition of RA in culture with non-human primate testis tissue altered the RA degrading enzyme, Cyp26a1, expression suggesting the cells in the non-human primate testis are responding appropriately to the treatment. However, more work needs to be done to measure Stra8 expression. The expression of pSTRA8 in a subpopulation of primate spermatogonia and in primary spermatocytes implies that RA may play a role in primate spermatogonial differentiation, as in the rodent testis involving a similar A to A1 transition, as well as entry into meiosis. This data aids in our investigation of the role of RA in spermatogonial differentiation in the non-human primate testis.
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Details
- Title
- Investigating the role of retinoic acid (RA) in the non-human primate testis
- Creators
- Angel Brooke Thalhofer
- Contributors
- Michael D Griswold (Advisor) - Washington State University, Molecular Biosciences, School of
- Awarding Institution
- Washington State University
- Academic Unit
- Molecular Biosciences, School of
- Theses and Dissertations
- Master of Science (MS), Washington State University
- Publisher
- Washington State University
- Identifiers
- 99900896438501842
- Language
- English
- Resource Type
- Thesis