The long-term objective of the proposed research is to further the understanding of mammalian spermatogenesis at the molecular level. Results from these studies address issues of both human infertility and contraception. The proposed studies utilize an innovative technique to synchronize spermatogenesis and isolate subtypes of spermatogonia undergoing development, expanding in numbers and acquiring the competence to enter meiosis. Isolation of enough of these cells to carry out molecular studies is possible only because of this innovative technique. Specific spermatogonial subtypes from wild-type and STRA8 knockout mice will be isolated before and after a retinoic acid pulse. The key focus of the proposed studies will be to determine retinoic acid responsive transcripts and then relate the genes for these transcripts to chromatin structure, histone acetylation and enhancer formation. The retinoic acid responsive transcripts will be determined by RNAseq and the chromatin structure will utilize ATAC chromatin accessibility techniques. Retinoic acid receptors and H3K27ac modifications will be determined by CHIP-seq. We will determine the presence of super enhancer sites marked by H3K27ac and examine the epigenetic changes and transcription factor binding sites in these sites. These studies will provide evidence for the molecular changes required for the entry into meiosis and lay the ground-work for future studies relating to the molecular causes of infertility.

Public Health Relevance

This project will further our understanding of normal spermatogenesis and will provide information relevant to infertility and contraception. In particular, the results will be important in understanding how cells enter meiosis.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
2R01HD010808-40
Application #
9445076
Study Section
Cellular, Molecular and Integrative Reproduction Study Section (CMIR)
Program Officer
Moss, Stuart B
Project Start
1977-08-01
Project End
2023-04-30
Budget Start
2018-06-19
Budget End
2019-04-30
Support Year
40
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Washington State University
Department
Veterinary Sciences
Type
Schools of Veterinary Medicine
DUNS #
041485301
City
Pullman
State
WA
Country
United States
Zip Code
99164
Jauregui, Estela J; Mitchell, Debra; Topping, Traci et al. (2018) Retinoic acid receptor signaling is necessary in steroidogenic cells for normal spermatogenesis and epididymal function. Development 145:
Griswold, Michael D; Hogarth, Cathryn (2018) Beyond stem cells: Commitment of progenitor cells to meiosis. Stem Cell Res 27:169-171
Agrimson, Kellie S; Oatley, Melissa J; Mitchell, Debra et al. (2017) Retinoic acid deficiency leads to an increase in spermatogonial stem number in the neonatal mouse testis, but excess retinoic acid results in no change. Dev Biol 432:229-236
Griswold, Michael D (2016) Spermatogenesis: The Commitment to Meiosis. Physiol Rev 96:1-17
Agrimson, Kellie S; Onken, Jennifer; Mitchell, Debra et al. (2016) Characterizing the Spermatogonial Response to Retinoic Acid During the Onset of Spermatogenesis and Following Synchronization in the Neonatal Mouse Testis. Biol Reprod 95:81
Chen, Yao; Ma, Li; Hogarth, Cathryn et al. (2016) Retinoid signaling controls spermatogonial differentiation by regulating expression of replication-dependent core histone genes. Development 143:1502-11
Kent, Travis; Arnold, Samuel L; Fasnacht, Rachael et al. (2016) ALDH Enzyme Expression Is Independent of the Spermatogenic Cycle, and Their Inhibition Causes Misregulation of Murine Spermatogenic Processes. Biol Reprod 94:12
Arnold, Samuel L M; Kent, Travis; Hogarth, Cathryn A et al. (2015) Pharmacological inhibition of ALDH1A in mice decreases all-trans retinoic acid concentrations in a tissue specific manner. Biochem Pharmacol 95:177-92
Hogarth, Cathryn A; Evans, Elizabeth; Onken, Jennifer et al. (2015) CYP26 Enzymes Are Necessary Within the Postnatal Seminiferous Epithelium for Normal Murine Spermatogenesis. Biol Reprod 93:19
Arnold, Samuel L; Kent, Travis; Hogarth, Cathryn A et al. (2015) Importance of ALDH1A enzymes in determining human testicular retinoic acid concentrations. J Lipid Res 56:342-57

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