G3BP2, a stress granule assembly factor, is dispensable for spermatogenesis in mice

PeerJ. 2022 Jun 28:10:e13532. doi: 10.7717/peerj.13532. eCollection 2022.

Abstract

Background: Spermatogenesis is a complex process that includes mitosis, meiosis, and spermiogenesis. During spermatogenesis, genetic factors play a vital role inthe formation of properly functioning sperm. GTPase-activating protein (SH3 domain)-binding protein 2 (G3BP2) is known to take part in immune responses, mRNA transport, and stress-granule assembly. However, its role in male fertility is unclear. Here, we generated a G3bp2 conditional knockout (cKO) mouse model to explore the function of G3BP2 in male fertility.

Methods: Polymerase chain reaction (PCR) and western blotting (WB) were used to confirm testis-specific G3bp2 knockout. Hematoxylin-eosin (HE) staining to observe testicular morphology and epididymal structure. Computer-aided sperm analysis (CASA) to detect sperm concentration and motility. Terminal deoxynucleotidyl transferase-dUTP nick-end labeling (TUNEL) assay was used to detect apoptotic cells.

Results: We found that cKO male mice are fertile with the normal morphology of the testis and sperm. Additionally, CASA of the semen from cKO mice showed that they all had a similar sperm concentration and motility. In addition, sperm from these mice exhibited a similar morphology. But the tunnel assay revealed increased apoptosis in their testes relative to the level in the wild type (WT).

Conclusion: Together, our data demonstrate that G3BP2 is dispensable for spermatogenesis and male fertility in mice albeit with the increased germ-cell apoptosis.

Keywords: Apoptosis; CRISPR-Cas9; Conditional knockout; G3bp2; Germ cell; Male fertility; Reproductive health; Sperm; Spermatogenesis; Testis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Male
  • Mice
  • Mice, Knockout
  • Semen*
  • Spermatogenesis / genetics
  • Stress Granules*
  • Testis / metabolism

Substances

  • G3BP2 protein, mouse

Grants and funding

This work was supported by The National Key Research and Development Program of China (No. 2021YFC2700200 to F.S); The Key Research and Development Program of Ningxia Hui Autonomous Region (No. 2021BEG02029 to F.S). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.