RiboTag analysis of actively translated mRNAs in Sertoli and Leydig cells in vivo

Elisenda Sanz; Ryan Evanoff; Albert Quintana; Elizabeth Evans; Jeremy A Miller; Chemyong Ko; Paul S Amieux; Michael D Griswold; G Stanley McKnight

doi:10.1371/journal.pone.0066179

Back

RiboTag analysis of actively translated mRNAs in Sertoli and Leydig cells in vivo

Journal article

Open access

RiboTag analysis of actively translated mRNAs in Sertoli and Leydig cells in vivo

Elisenda Sanz, Ryan Evanoff, Albert Quintana, Elizabeth Evans, Jeremy A Miller, Chemyong Ko, Paul S Amieux, Michael D Griswold and G Stanley McKnight

PloS one, Vol.8(6), pp.e66179-e66179

2013

DOI: https://doi.org/10.1371/journal.pone.0066179

Handle:

https://hdl.handle.net/2376/104344

PMCID: PMC3679032

PMID: 23776628

Files and links (1)

url

https://doi.org/10.1371/journal.pone.0066179View

Published (Version of record) Open

Abstract

Sertoli Cells - metabolism

Cell Line

Follicle Stimulating Hormone - metabolism

Animals

Immunoprecipitation

Leydig Cells - metabolism

RNA, Messenger - genetics

Male

Mice

Luteinizing Hormone - metabolism

Reverse Transcriptase Polymerase Chain Reaction

Male spermatogenesis is a complex biological process that is regulated by hormonal signals from the hypothalamus (GnRH), the pituitary gonadotropins (LH and FSH) and the testis (androgens, inhibin). The two key somatic cell types of the testis, Leydig and Sertoli cells, respond to gonadotropins and androgens and regulate the development and maturation of fertilization competent spermatozoa. Although progress has been made in the identification of specific transcripts that are translated in Sertoli and Leydig cells and their response to hormones, efforts to expand these studies have been restricted by technical hurdles. In order to address this problem we have applied an in vivo ribosome tagging strategy (RiboTag) that allows a detailed and physiologically relevant characterization of the "translatome" (polysome-associated mRNAs) of Leydig or Sertoli cells in vivo. Our analysis identified all previously characterized Leydig and Sertoli cell-specific markers and identified in a comprehensive manner novel markers of Leydig and Sertoli cells; the translational response of these two cell types to gonadotropins or testosterone was also investigated. Modulation of a small subset of Sertoli cell genes occurred after FSH and testosterone stimulation. However, Leydig cells responded robustly to gonadotropin deprivation and LH restoration with acute changes in polysome-associated mRNAs. These studies identified the transcription factors that are induced by LH stimulation, uncovered novel potential regulators of LH signaling and steroidogenesis, and demonstrate the effects of LH on the translational machinery in vivo in the Leydig cell.

Metrics

13 Record Views

Details

Title: RiboTag analysis of actively translated mRNAs in Sertoli and Leydig cells in vivo
Creators: Elisenda Sanz - Department of Pharmacology, University of Washington, Seattle, Washington, United States of America
Ryan Evanoff
Albert Quintana
Elizabeth Evans
Jeremy A Miller
Chemyong Ko
Paul S Amieux
Michael D Griswold
G Stanley McKnight
Publication Details: PloS one, Vol.8(6), pp.e66179-e66179
Academic Unit: Molecular Biosciences, School of
Publisher: United States
Grant note: 2 R01 HD010808 / NICHD NIH HHS P50 HD028934 / NICHD NIH HHS U54 HD012629 / NICHD NIH HHS MH086386 / NIMH NIH HHS R01 MH086386 / NIMH NIH HHS U54 HD042454 / NICHD NIH HHS R01 HD010808 / NICHD NIH HHS 1R21HD057790-01 / NICHD NIH HHS U54HD12629 / NICHD NIH HHS
Identifiers: 99900546715401842
Language: English
Resource Type: Journal article