Osmotic stress stimulates phosphorylation and cellular expression of heat shock proteins in rhesus macaque sperm

Julie A. Cole, Stuart A Meyers

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The cryosurvival of sperm requires cell signaling mechanisms to adapt to anisotonic conditions during the freezing and thawing process. Chaperone proteins heat shock protein 70 (HSP 70) and heat shock protein 90 (HSP 90; recently renamed HSPA and HSPC, respectively) facilitate some of these cell signaling events in somatic cells. Sperm were evaluated for their cellular expression and levels of phosphorylation of both HSP 70 and HSP 90 under anisotonic conditions as a potential model for cell signaling during the cryopreservation of macaque spermatozoa. In order to monitor the level of stress, the motility and viability parameters were evaluated at various time points. Cells were then either prepared for phosphoprotein enrichment or indirect immunocytochemistry. As controls, the phosphoserine, phosphothreonine, and phosphotyrosine levels were measured under capacitation and cryopreservation conditions and were compared with the phosphoprotein levels expressed under osmotic conditions. As expected, there was an increase in the level of tyrosine phosphorylation under capacitation and cryopreservation conditions. There was also a significant increase in the level of all phosphoproteins under hyperosmotic conditions. There was no change in the level of expression of HSP 70 or 90 under osmotic stress conditions as measured by Western blot. The enrichment of phosphoproteins followed by Western immunoblotting revealed an increase in the phosphorylation of HSP 70 but not HSP 90 under osmotic stress conditions. Indirect immunofluorescence localized HSP 70 to the postacrosomal region of sperm, and the level of membrane expression of HSP 70 was significantly affected by anisotonic conditions, as measured by flow cytometry. Taken together, these results suggest a differential role for HSP 70 and HSP 90 during osmotic stress conditions in rhesus macaque sperm.

Original languageEnglish (US)
Pages (from-to)402-410
Number of pages9
JournalJournal of Andrology
Volume32
Issue number4
DOIs
StatePublished - Jul 2011

Fingerprint

HSP70 Heat-Shock Proteins
Osmotic Pressure
Heat-Shock Proteins
Macaca mulatta
Spermatozoa
Phosphorylation
Phosphoproteins
Cryopreservation
HSP90 Heat-Shock Proteins
Western Blotting
Phosphothreonine
Phosphoserine
Phosphotyrosine
Macaca
Indirect Fluorescent Antibody Technique
Freezing
Tyrosine
Flow Cytometry
Immunohistochemistry
Membranes

Keywords

  • Cryopreservation
  • Spermatozoa

ASJC Scopus subject areas

  • Urology
  • Reproductive Medicine
  • Endocrinology, Diabetes and Metabolism
  • Endocrinology

Cite this

Osmotic stress stimulates phosphorylation and cellular expression of heat shock proteins in rhesus macaque sperm. / Cole, Julie A.; Meyers, Stuart A.

In: Journal of Andrology, Vol. 32, No. 4, 07.2011, p. 402-410.

Research output: Contribution to journalArticle

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