mTORC1/rpS6 regulates blood-testis barrier dynamics and spermatogenetic function in the testis in vivo

Stephen Y.T. Li, Ming Yan, Haiqi Chen, Tito Jesus, Will M. Lee, Xiang Xiao, C. Yan Cheng

Research output: Contribution to journalArticlepeer-review

29 Scopus citations

Abstract

The blood-testis barrier (BTB), conferred by Sertoli cells in the mammalian testis, is an important ultrastructure that supports spermatogenesis. Studies using animal models have shown that a disruption of the BTB leads to meiotic arrest, causing defects in spermatogenesis and male infertility. To better understand the regulation of BTB dynamics, we report findings herein to understand the role of ribosomal protein S6 (rpS6), a downstream signaling protein of mammalian target of rapamycin complex 1 (mTORC1), in promoting BTB disruption in the testis in vivo, making the barrier “leaky.” Overexpression of wild-type rpS6 (rpS6-WT, the full-length cDNA cloned into the mammalian expression vector pCI-neo) and a constitutively active quadruple phosphomimetic mutant cloned into pCI-neo (p-rpS6-MT) vs. control (empty pCI-neo vector) was achieved by transfecting adult rat testes with the corresponding plasmid DNA using a Polyplus in vivo-jetPEI transfection reagent. On the basis of an in vivo functional BTB integrity assay, p-rpS6-MT was found to induce BTB disruption better than rpS6-WT did (and no effects in empty vector control), leading to defects in spermatogenesis, including loss of spermatid polarity and failure in the transport of cells (e.g., spermatids) and organelles (e.g., phagosomes), to be followed by germ exfoliation. More important, rpS6-WT and p-rpS6-MT exert their disruptive effects through changes in the organization of actin- and microtubule (MT)-based cytoskeletons, which are mediated by changes in the spatiotemporal expression of actin- and MT-based binding and regulatory proteins. In short, mTORC1/rpS6 signaling complex is a regulator of spermatogenesis and BTB by modulating the organization of the actin- and MT-based cytoskeletons.

Original languageEnglish (US)
Pages (from-to)E174-E190
JournalAmerican Journal of Physiology - Endocrinology and Metabolism
Volume314
Issue number2
DOIs
StatePublished - Feb 2018
Externally publishedYes

Keywords

  • Blood-testis barrier
  • Ectoplasmic specialization
  • Germ cell
  • Mammalian target of rapamycin complex 1
  • Ribosomal protein S6
  • Sertoli cell
  • Spermatogenesis
  • Testis

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Physiology
  • Physiology (medical)

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