title: A complex containing alpha6beta1-integrin and phosphorylated focal adhesion kinase between Sertoli cells and elongated spermatids during spermatid release from the seminiferous epithelium.
abstract: Spermiation is the final step of spermatogenesis and culminates in the disengagement (release) of elongated spermatids from Sertoli cells into the seminiferous tubule lumen. Spermiation failure, wherein spermatids are retained by Sertoli cells instead of releasing, occurs after hormone suppression. The mechanisms involved in spermatid disengagement and retention are not well understood. We previously showed that beta(1)-integrin is associated with spermatids until the point of disengagement, but the ectoplasmic specialisation junction (ES) is not. The aims of this paper are to further characterise the complex that is present immediately prior to spermatid disengagement by identifying the alpha-integrin form dimerised with beta(1)-integrin, localising focal adhesion kinase (FAK) and determining if microtubules are involved. Adult Sprague-Dawley rats received testosterone and oestradiol implants and an FSH antibody for 7 days to suppress testicular testosterone and FSH and induce spermiation failure. Control rats were treated with saline. Immunohistochemical analysis showed that alpha(6)-integrin and a phosphorylated form of FAK (FAK-Tyr(397)) are present between late spermatids and Sertoli cells after ES removal, until the point of disengagement, and both proteins remain associated with retained spermatids after spermiation failure induced by hormone suppression. Using dual-label immunofluorescence, tubulins (and thus microtubules) were observed to co-localise with ES, but were neither associated with elongated spermatids just prior to release nor with retained spermatids following hormone suppression. These results suggest that microtubules are not involved in the final release of spermatids from Sertoli cells. We conclude that spermatid release during spermiation is mediated by a 'disengagement complex' containing alpha(6)beta(1)-integrin and phospho-FAK, the function of which can be affected by gonadotrophin suppression.
authors.affiliation: Prince Henry's Institute of Medical Research, Monash Medical Centre, Clayton, Victoria 3168, Australia.
authors.forename: Amanda
authors.initials: A
authors.lastname: Beardsley
authors.forename: David M
authors.initials: DM
authors.lastname: Robertson
authors.forename: Liza
authors.initials: L
authors.lastname: O'Donnell
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D015153:Blotting, Western / Q000379:methods; D004958:Estradiol / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000032:analysis* / Q000378:metabolism; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D039121:Integrin alpha6beta1 / Q000032:analysis* / Q000378:metabolism; D008297:Male; D008870:Microtubules / Q000737:chemistry; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D012670:Seminiferous Epithelium / Q000737:chemistry*; D012708:Sertoli Cells / Q000737:chemistry*; D013087:Spermatids / Q000737:chemistry* / Q000187:drug effects; D013091:Spermatogenesis / Q000187:drug effects / Q000502:physiology*; D013739:Testosterone / Q000037:antagonists & inhibitors / Q000494:pharmacology; D014404:Tubulin / Q000032:analysis
chemical_list: D015415:Biomarkers; D039121:Integrin alpha6beta1; D014404:Tubulin; D013739:Testosterone; D004958:Estradiol; D005640:Follicle Stimulating Hormone; D051416:Focal Adhesion Protein-Tyrosine Kinases