Evidence that rodent epididymal sperm contain the Mr ∼94,000 glucocorticoid receptor but lack the Mr ∼90,000 heat shock protein

Scott H. Kaufmann, William W Wright, Sam Okret, Ann Charlotte Wikström, Jan Åke Gustafsson, Nancy L. Shaper, Joel H. Shaper

Research output: Contribution to journalArticle

Abstract

Monoclonal antibodies directed against four different polypeptide epitopes on the Mr ∼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 Mr ∼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 Mr ∼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 Mr ∼94,000 steroid-binding subunit of the GcR and an immunologically related Mr ∼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 Mr ∼94,000 GcR subunit in intact cells.

Original languageEnglish (US)
Pages (from-to)3074-3084
Number of pages11
JournalEndocrinology
Volume130
Issue number5
StatePublished - May 1992

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HSP90 Heat-Shock Proteins
Glucocorticoid Receptors
Spermatozoa
Rodentia
Steroids
Peptides
Spermatids
Spermatogenesis
Glucocorticoids
Lamin Type B
Nuclear Matrix-Associated Proteins
Lamin Type A
Type I DNA Topoisomerase
Messenger RNA
Spermatocytes
Poly(ADP-ribose) Polymerases
Liver
Germ Cells
Northern Blotting
Histones

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Kaufmann, S. H., Wright, W. W., Okret, S., Wikström, A. C., Gustafsson, J. Å., Shaper, N. L., & Shaper, J. H. (1992). Evidence that rodent epididymal sperm contain the Mr ∼94,000 glucocorticoid receptor but lack the Mr ∼90,000 heat shock protein. Endocrinology, 130(5), 3074-3084.

Evidence that rodent epididymal sperm contain the Mr ∼94,000 glucocorticoid receptor but lack the Mr ∼90,000 heat shock protein. / Kaufmann, Scott H.; Wright, William W; Okret, Sam; Wikström, Ann Charlotte; Gustafsson, Jan Åke; Shaper, Nancy L.; Shaper, Joel H.

In: Endocrinology, Vol. 130, No. 5, 05.1992, p. 3074-3084.

Research output: Contribution to journalArticle

Kaufmann, SH, Wright, WW, Okret, S, Wikström, AC, Gustafsson, JÅ, Shaper, NL & Shaper, JH 1992, 'Evidence that rodent epididymal sperm contain the Mr ∼94,000 glucocorticoid receptor but lack the Mr ∼90,000 heat shock protein', Endocrinology, vol. 130, no. 5, pp. 3074-3084.
Kaufmann, Scott H. ; Wright, William W ; Okret, Sam ; Wikström, Ann Charlotte ; Gustafsson, Jan Åke ; Shaper, Nancy L. ; Shaper, Joel H. / Evidence that rodent epididymal sperm contain the Mr ∼94,000 glucocorticoid receptor but lack the Mr ∼90,000 heat shock protein. In: Endocrinology. 1992 ; Vol. 130, No. 5. pp. 3074-3084.
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abstract = "Monoclonal antibodies directed against four different polypeptide epitopes on the Mr ∼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 Mr ∼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 Mr ∼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 Mr ∼94,000 steroid-binding subunit of the GcR and an immunologically related Mr ∼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 Mr ∼94,000 GcR subunit in intact cells.",
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AU - Wright, William W

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AU - Wikström, Ann Charlotte

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AU - Shaper, Nancy L.

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N2 - Monoclonal antibodies directed against four different polypeptide epitopes on the Mr ∼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 Mr ∼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 Mr ∼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 Mr ∼94,000 steroid-binding subunit of the GcR and an immunologically related Mr ∼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 Mr ∼94,000 GcR subunit in intact cells.

AB - Monoclonal antibodies directed against four different polypeptide epitopes on the Mr ∼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 Mr ∼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 Mr ∼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 Mr ∼94,000 steroid-binding subunit of the GcR and an immunologically related Mr ∼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 Mr ∼94,000 GcR subunit in intact cells.

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