A long non-coding RNA interacts with Gfra1 and maintains survival of mouse spermatogonial stem cells.

L Li,X Wu,M Wang,Y Xue,Y Jia,L Geng,X Wei

doi:10.1038/cddis.2016.24

Abstract

Spermatogonial stem cells (SSCs) are unique male germline stem cells that support spermatogenesis and male fertility. Long non-coding RNAs (lncRNA) have been identified as key regulators of stem cell fate; however, their role in SSCs has not been explored. Here, we report that a novel spermatogonia-specific lncRNA (lncRNA033862) is essential for the survival of murine SSCs. LncRNA033862 is expressed in early spermatogonia including SSC and was among 805 lncRNAs identified by global expression profiling as responsive to glial cell-derived neurotrophic factor (GDNF), a growth factor required for SSC self-renewal and survival. LncRNA033862 is an antisense transcript of the GDNF receptor alpha1 (Gfra1) that lacks protein coding potential and regulates Gfra1 expression levels by interacting with Gfra1 chromatin. Importantly, lncRNA033862 knockdown severely impairs SSC survival and their capacity to repopulate recipient testes in a transplantation assay. Collectively, our data provide the first evidence that long non-coding RNAs (lncRNAs) regulate SSC fate.

Highlights

With the development of protocols for the long-term propagation of mouse spermatogonial stem cells (SSCs) as clump-forming colonies in vitro,[3,4] multiple exogenous growth factors and chemicals have been identified that support the self-renewal and proliferation of mouse SSCs, including leukemia inhibitory factor,[3] fibroblast growth factor 2 (FGF2),[3,4] epidermal growth factor,[4] insulin growth factor 1 and colony-stimulating factor 1,5 Wnt glycoproteins[6] and low-dose H2O27, among others
In vitro proliferation of SSCs was dependent on glial cell line-derived neurotrophic factor (GDNF): removal of GDNF from the culture medium for a period of 7 days resulted in a significant reduction in cell number and changes in morphology with disappearance of grape shape clumps
Of 55 924 lncRNA transcripts, we identified 805 lncRNA transcripts in SSCs subject to transcript level changes in response to GDNF, which is an essential growth factor required for SSC self-renewal

Summary

Introduction

With the development of protocols for the long-term propagation of mouse SSCs as clump-forming colonies in vitro,[3,4] multiple exogenous growth factors and chemicals have been identified that support the self-renewal and proliferation of mouse SSCs, including leukemia inhibitory factor,[3] fibroblast growth factor 2 (FGF2),[3,4] epidermal growth factor,[4] insulin growth factor 1 and colony-stimulating factor 1,5 Wnt glycoproteins[6] and low-dose H2O27, among others. We have identified a novel spermatogonia-specific lncRNA (lncRNA033862) that controls SSC self-renewal and survival by regulation of the expression of Gfrα[1], the glycosylphosphatidylinositol-linked cell surface receptor gene of GDNF

Methods

Results

Conclusion