Spermatogonial Gene Networks Selectively Couple to Glutathione and Pentose Phosphate Metabolism but Not Cysteine Biosynthesis

David Prokai; Ashutosh Pudasaini; Mohammed Kanchwala; Andrew T Moehlman; Alexandrea E Waits; Karen M Chapman; Jaideep Chaudhary; Jesus Acevedo; Patrick Keller; Xing Chao; Bruce R Carr; F Kent Hamra

doi:10.1016/j.isci.2020.101880

Spermatogonial Gene Networks Selectively Couple to Glutathione and Pentose Phosphate Metabolism but Not Cysteine Biosynthesis

iScience. 2020 Dec 1;24(1):101880. doi: 10.1016/j.isci.2020.101880. eCollection 2021 Jan 22.

Authors

David Prokai¹, Ashutosh Pudasaini^{2

3}, Mohammed Kanchwala⁴, Andrew T Moehlman⁵, Alexandrea E Waits⁵, Karen M Chapman⁶, Jaideep Chaudhary⁷, Jesus Acevedo^{2

8}, Patrick Keller^{2

8}, Xing Chao^{4

9

10}, Bruce R Carr^{1

2}, F Kent Hamra^{2

8}

Affiliations

¹ Division of Reproductive Endocrinology and Infertility, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
² Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
³ GenomeDesigns Laboratory, LLC, 314 Stonebridge Drive, Richardson, TX 75080, USA.
⁴ McDermott Center for Human Growth and Development, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
⁵ Department of Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
⁶ Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
⁷ Advanced Imaging Research Center, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
⁸ Cecil H. & Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
⁹ Department of Bioinformatics, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
¹⁰ Department of Population and Data Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.

Abstract

In adult males, spermatogonia maintain lifelong spermatozoa production for oocyte fertilization. To understand spermatogonial metabolism we compared gene profiles in rat spermatogonia to publicly available mouse, monkey, and human spermatogonial gene profiles. Interestingly, rat spermatogonia expressed metabolic control factors Foxa1, Foxa2, and Foxa3. Germline Foxa2 was enriched in Gfra1^Hi and Gfra1^Low undifferentiated A-single spermatogonia. Foxa2-bound loci in spermatogonial chromatin were overrepresented by conserved stemness genes (Dusp6, Gfra1, Etv5, Rest, Nanos2, Foxp1) that intersect bioinformatically with conserved glutathione/pentose phosphate metabolism genes (Tkt, Gss, Gc l c , Gc l m, Gpx1, Gpx4, Fth), marking elevated spermatogonial GSH:GSSG. Cystine-uptake and intracellular conversion to cysteine typically couple glutathione biosynthesis to pentose phosphate metabolism. Rat spermatogonia, curiously, displayed poor germline stem cell viability in cystine-containing media, and, like primate spermatogonia, exhibited reduced transsulfuration pathway markers. Exogenous cysteine, cysteine-like mercaptans, somatic testis cells, and ferroptosis inhibitors counteracted the cysteine-starvation-induced spermatogonial death and stimulated spermatogonial growth factor activity in vitro.

Keywords: Biological Sciences; Developmental Biology; Systems Biology.

Abstract

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