TY - JOUR
T1 - Planar cell polarity (PCP) protein vangl2 regulates ectoplasmic specialization dynamics via its effects on actin microfilaments in the testes of male rats
AU - Chen, Haiqi
AU - Mruk, Dolores D.
AU - Lee, Will M.
AU - Cheng, C. Yan
N1 - Funding Information:
This work was supported by Grant R01 HD056034 (to C.Y.C.) and Grant U54 HD029990, Project 5 (to C.Y.C.) from the National Institutes of Health, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Natural Science Foundation of China/Research Grants Council (NSFC/RGC) Joint Research Scheme (N-HKU 717/12 to W.M.L.), General Research Fund from RGC (771513 to W.M.L.), andCRCGSeed Funding, University ofHongKong (to W.M.L.).
Publisher Copyright:
Copyright © 2016 by the Endocrine Society.
PY - 2016/5
Y1 - 2016/5
N2 - Planar cell polarity (PCP) proteins confer polarization of a field of cells (eg, elongating/elongated spermatids) within the plane of an epithelium such as the seminiferous epithelium of the tubule during spermatogenesis. In adult rat testes, Sertoli and germ cells were found to express PCP core proteins (eg, Van Gogh-like 2 [Vangl2]), effectors, ligands, and signaling proteins. Vangl2 expressed predominantly by Sertoli cells was localized at the testis-specific, actin-rich ectoplasmic specialization (ES) at the Sertoli-spermatid interface in the adluminal compartment and also Sertoli-Sertoli interface at the blood-testis barrier (BTB) and structurally interacted with actin, Ncadherin, and another PCP/polarity protein Scribble. Vangl2 knockdown (KD) by RNA interference in Sertoli cells cultured in vitro with an established tight junction-permeability barrier led to BTB tightening, whereas its overexpression using a full-length cDNA construct perturbed the barrier function. These changes were mediated through an alteration on the organization actin microfilaments at the ES in Sertoli cells, involving actin-regulatory proteins, epidermal growth factor receptor pathway substrate 8, actin-related protein 3, and Scribble, which in turn affected the function of adhesion protein complexes at the ES during the epithelial cycle of spermatogenesis. Using Polyplus in vivo-jetPEI reagent as a transfection medium to silence Vangl2 in the testis in vivo by RNA interference with high efficacy, Vangl2 KD led to changes in F-actin organization at the ES in the epithelium, impeding spermatid and phagosome transport and spermatid polarity, meiosis, and BTB dynamics. For instance, step 19 spermatids remained embedded in the epithelium alongside with step 9 and 10 spermatids in stages IX-X tubules. In summary, the PCP protein Vangl2 is an ES regulator through its effects on actin microfilaments in the testis.
AB - Planar cell polarity (PCP) proteins confer polarization of a field of cells (eg, elongating/elongated spermatids) within the plane of an epithelium such as the seminiferous epithelium of the tubule during spermatogenesis. In adult rat testes, Sertoli and germ cells were found to express PCP core proteins (eg, Van Gogh-like 2 [Vangl2]), effectors, ligands, and signaling proteins. Vangl2 expressed predominantly by Sertoli cells was localized at the testis-specific, actin-rich ectoplasmic specialization (ES) at the Sertoli-spermatid interface in the adluminal compartment and also Sertoli-Sertoli interface at the blood-testis barrier (BTB) and structurally interacted with actin, Ncadherin, and another PCP/polarity protein Scribble. Vangl2 knockdown (KD) by RNA interference in Sertoli cells cultured in vitro with an established tight junction-permeability barrier led to BTB tightening, whereas its overexpression using a full-length cDNA construct perturbed the barrier function. These changes were mediated through an alteration on the organization actin microfilaments at the ES in Sertoli cells, involving actin-regulatory proteins, epidermal growth factor receptor pathway substrate 8, actin-related protein 3, and Scribble, which in turn affected the function of adhesion protein complexes at the ES during the epithelial cycle of spermatogenesis. Using Polyplus in vivo-jetPEI reagent as a transfection medium to silence Vangl2 in the testis in vivo by RNA interference with high efficacy, Vangl2 KD led to changes in F-actin organization at the ES in the epithelium, impeding spermatid and phagosome transport and spermatid polarity, meiosis, and BTB dynamics. For instance, step 19 spermatids remained embedded in the epithelium alongside with step 9 and 10 spermatids in stages IX-X tubules. In summary, the PCP protein Vangl2 is an ES regulator through its effects on actin microfilaments in the testis.
UR - http://www.scopus.com/inward/record.url?scp=84969857954&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84969857954&partnerID=8YFLogxK
U2 - 10.1210/en.2015-1987
DO - 10.1210/en.2015-1987
M3 - Article
C2 - 26990065
AN - SCOPUS:84969857954
SN - 0013-7227
VL - 157
SP - 2140
EP - 2159
JO - Endocrinology
JF - Endocrinology
IS - 5
ER -