TSPYL1 regulates steroidogenic gene expression and male factor fertility in mice

Abstract

Objective: To determine the importance of testis-specific, Y-encoded-like 1 (TSPYL1) in survival and male factor fertility in mice. Design: Experimental prospective study. Setting: Research laboratories in a university medical faculty. Animals: We generated Tspyl1 knockout (KO) mouse lines by CRISPR/Cas9. The lines were maintained by pairing heterozygous mice to provide wild-type control and KO males for comparison. Intervention(s): None. Main Outcome Measure(s): Mendelian ratio, body and testis weight, histology, sperm motility, mating tests, pregnancy outcome, transcript levels of genes for testosterone production, and serum testosterone level. Result(s): A variable percentage of Tspyl1 KO mice survived beyond weaning depending on the genetic background. Growth around weaning was retarded in KO mice, but the testes-to-body weight ratio remained normal and complete spermatogenesis was revealed in testis histology. Sperm was collected from the cauda epididymis, and a significantly smaller percentage of sperm was progressively motile (22.3% AE 18.3%, n ¼ 14 samples) compared with wild type (58.9% AE 11.5%, 11 samples). All 11 KO mice tested had defective mounting behavior. From 11 KO males paired with a total of 88 females, only one litter was born, compared with 53 litters sired by 11 age-matched wild-type males. Expression of Star, Cyp11a1, Cyp17a1, Hsd3b6, and Hsd17b3 in the KO testis was significantly reduced, while serum testosterone level was within the normal range. Conclusion(s): TSPYL1 is critical for survival and reproductive success in mice. TSPYL1 enhances the expression of key steroidogenic genes in the mouse testis. (Fertil Steril Sci Ò 2020;1:115-23. Ó2020 by American Society for Reproductive Medicine.

Coding and regulatory transcriptome comparisons between fertile and infertile spermatozoa identify RNA signatures of male infertility

“…Similarly, the loss of heat shock protein 5 (Hsf5) in zebrafish resulted in disrupted meiosis (Saju et al, 2018). Tspyl5 knockout male mice mated with highly reduced litter size, indicating the importance of Tspyl5 in reproductive success in mice (Peng et al, 2020). During neonatal testis development, Dio‐3‐deficient spermatogonia resulted in reduced Sertoli cell number, cell proliferation and reduced testes growth (Martinez et al, 2019).…”

TSPYL1 as a Critical Regulator of TGFβ Signaling through Repression of TGFBR1 and TSPYL2

“…Tspyl1 KO line Δ463 on pure C57BL/6N background was maintained by breeding between Tspyl1 +/− heterozygotes. [13] PBMC isolation: The protocol was approved by the Institutional Review Board (IRB Ref: UW 22-154). PBMC were isolated from buffy coats provided by the Hong Kong Red Cross using Lymphoprep (Thermo Fisher Scientific).…”

“…MEF derived from each embryo were cultured individually and genotyped with primers as previously described. [ 13 ] MEF were subcultured using the standard 3T3 protocol. Briefly, 3 × 10 5 cells were plated on a 5‐cm plate and cultured for 3 days.…”

“…Tspyl1 KO line Δ463 on pure C57BL/6N background was maintained by breeding between Tspyl1 +/− heterozygotes. [ 13 ]…”

“…Tspyl1 knockout (KO) mice also have retarded growth and most die around the weaning stage, but there is no testis dysgenesis. [ 13 ] Despite the vital importance of TSPYL1, little is known about its function.…”