Endocytosis of androgen-binding protein (ABP) by spermatogenic cells.
To test whether Sertoli cell-secreted ABP could serve as steroid carrier to the germ cell (GC) lineage, radiolabeled ABP and SHBG and gold SHBG were used for binding studies and for internalization studies based on transmission electron microscope analyses and autoradiography of the radiolabeled samples. The data clearly showed that: (1) rat and human germ cells possess a single class of binding sites for rat ABP and human SHBG respectively (Kd of 0.78 and 0.56 nM); (2) 1.7 x 10(10) and 2.7 x 10(10) sites/mg protein was found in the corresponding plasma membrane preparations; (3) the receptor peak was eluted in the same position as dextran blue: 2000 kDa (M(r) = 2 x 106) for labeled rat ABP; (4) in the whole GC lineage, the labeled ligand was internalized through an endocytic pathway involving clathrin coated structures and the distribution was similar throughout the maturation step, however striking differences in the internalization rate were revealed with regard to the maturation step; and (5) this internalization occurred even in ligated seminiferous tubules, via the Sertoli cells cytoplasm. When isolated rat GC were incubated in the presence of ABP, a dose dependent increase in labeled secreted protein was observed for spermatocytes (50-250%) whereas ABP had no effect on spermatids. Addition of steroids and ABP caused a 200 and 50% increase in labeled secreted proteins for spermatocytes and spermatids respectively. 2-D SDS-PAGE analysis revealed that ABP alone increased the secretion of specific spermatocyte proteins whereas steroids in the presence of ABP resulted in the synthesis of new spermatocyte secreted proteins. Taken together these results strongly suggest that ABP may be required for spermatogenesis either as a steroid transmembrane carrier or on its own.
Gerard
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A
Faculté de Médecine de Nancy I, Laboratoire d'Histologie-Embryologie II, Morphogénèse et Reproduction, Vandoeurve-lès-Nancy, France.
D000727:Androgen-Binding Protein / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D004705:Endocytosis; D005089:Exocytosis; D006801:Humans; D008297:Male; D014176:Protein Biosynthesis; D051381:Rats; D012708:Sertoli Cells / Q000378:metabolism*; D012738:Sex Hormone-Binding Globulin / Q000378:metabolism*; D013087:Spermatids / Q000378:metabolism*; D013090:Spermatocytes / Q000378:metabolism*
D000727:Androgen-Binding Protein; D012738:Sex Hormone-Binding Globulin