EVIDENCE THAT RODENT EPIDIDYMAL SPERM CONTAIN THE M(R)SIMILAR-TO-94,000 GLUCOCORTICOID RECEPTOR BUT LACK THE M(R)SIMILAR-TO-90,000 HEAT-SHOCK PROTEIN

被引:26
作者
KAUFMANN, SH
WRIGHT, WW
OKRET, S
WIKSTROM, AC
GUSTAFSSON, JA
SHAPER, NL
SHAPER, JH
机构
[1] JOHNS HOPKINS UNIV, SCH HYG & PUBL HLTH, DEPT POPULAT DYNAM, DIV REPROD BIOL, BALTIMORE, MD 21205 USA
[2] KAROLINSKA INST, HUDDINGE UNIV HOSP, DEPT MED NUTR, S-14186 HUDDINGE, SWEDEN
[3] NOVUM, S-14186 HUDDINGE, SWEDEN
[4] JOHNS HOPKINS UNIV, SCH MED, DEPT PHARMACOL & MOLEC SCI, BALTIMORE, MD 21205 USA
关键词
D O I
10.1210/en.130.5.3074
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Monoclonal antibodies directed against four different polypeptide epitopes on the M(r) approximately 94,000 steroid-binding subunit of the rat liver cytosolic glucocorticoid receptor (GcR) were used to probe Western blots of epididymal spermatozoa from rats and mice. Two sperm polypeptides with apparent molecular weights of 94,000 (indistinguishable in size from the liver GcR subunit) and 150,000 reacted with these antibodies. Other polypeptides that are present in a wide variety of somatic cells [lamin-A, -B, and -C; topoisomerase-I; poly(ADP-ribose) polymerase; the 62-kilodalton internal nuclear matrix protein; the nucleolar protein B23; and histone H1] could not be detected in these preparations of spermatozoa, thus appearing to rule out contamination by somatic cells. Rat and mouse pachytene spermatocytes and round spermatids contained much lower amounts of the M(r) approximately 94,000 and 150,000 polypeptides. These results suggested that the steroid-binding subunit of the GcR might be accumulated late in spermatogenesis. Consistent with this view, a 6-kilobase mRNA (identical in size to a mRNA detected in mouse somatic cell lines) was detected when Northern blots of mouse round spermatid RNA were probed with a cDNA to the steroid-binding GcR subunit. Although the results described above suggest the presence of GcR in rodent sperm, high affinity binding of glucocorticoids to epididymal sperm could not be detected in a whole cell binding assay. Further analysis revealed that the M(r) approximately 90,000 heat shock protein (hsp90), a component reportedly required for high affinity ligand binding to the GcR, was present in early germ cells, but absent from rodent epididymal sperm. These results suggest that the M(r) approximately 94,000 steroid-binding subunit of the GcR and an immunologically related M(r) approximately 150,000 polypeptide are specifically accumulated during the later stages of rodent spermatogenesis, but are not assembled into receptor complexes capable of binding steroid. In addition, these results support the view that hsp90 is required for high affinity binding of glucocorticoids to the M(r) approximately 94,000 GcR subunit in intact cells.
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页码:3074 / 3084
页数:11
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