Recombinant human zona pellucida glycoprotein 3 induces calcium influx and acrosome reaction in human spermatozoa

I A Brewis (Lead / Corresponding author), R Clayton, C L Barratt, D P Hornby, H D Moore

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62 Citations (Scopus)


Recombinant human ZP3 (rhuZP3) generated by Chinese hamster ovary cells transfected with a plasmid containing human ZP3 cDNA was used to study the acrosome reaction (AR) and intracellular calcium fluxes in capacitated human spermatozoa. Conditioned medium containing rhuZP3 significantly induced the AR (P < or = 0.005) in 59.4 +/- 4.7% of spermatozoa (control = 8.5 +/- 3.1%) and caused complete acrosomal loss in a further 17.2 +/- 3.8% of cells (control = 3.7 +/- 0.7%; mean +/- SEM, n = 5). Sperm motility was not affected and acrosomal exocytosis in response to rhuZP3 was also shown to be time-dependent. Basal concentrations of sperm intracellular calcium were measured (82 +/- 7 nM; mean +/- SEM, n = 9). A transient increase in intracellular calcium (typically up to 400-450 nM) occurred within 1 min of rhuZP3 addition and was followed by sustained lower values of calcium (200-400 nM). These responses were dependent on the amount of rhuZP3. This is the first report of zona protein-induced changes in intracellular calcium levels in human spermatozoa. The results support the premise that ZP3 is an agonist of the human sperm AR and that rhuZP3 generated in a eukaryotic cell is effective in this respect.

Original languageEnglish
Pages (from-to)583-9
Number of pages7
JournalMolecular Human Reproduction
Issue number8
Publication statusPublished - 1 Aug 1996


  • Acrosome/drug effects
  • Animals
  • CHO Cells
  • Calcium/metabolism
  • Cricetinae
  • Culture Media, Conditioned
  • Egg Proteins/biosynthesis
  • Humans
  • In Vitro Techniques
  • Intracellular Fluid/metabolism
  • Ion Transport/drug effects
  • Male
  • Membrane Glycoproteins/biosynthesis
  • Receptors, Cell Surface
  • Recombinant Proteins/biosynthesis
  • Signal Transduction
  • Sperm Capacitation
  • Spermatozoa/drug effects
  • Zona Pellucida Glycoproteins


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