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

TY - JOUR

T1 - Recombinant human zona pellucida glycoprotein 3 induces calcium influx and acrosome reaction in human spermatozoa

AU - Brewis, I A

AU - Clayton, R

AU - Barratt, C L

AU - Hornby, D P

AU - Moore, H D

PY - 1996/8/1

Y1 - 1996/8/1

N2 - 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.

AB - 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.

KW - Acrosome/drug effects

KW - Animals

KW - CHO Cells

KW - Calcium/metabolism

KW - Cricetinae

KW - Culture Media, Conditioned

KW - Egg Proteins/biosynthesis

KW - Humans

KW - In Vitro Techniques

KW - Intracellular Fluid/metabolism

KW - Ion Transport/drug effects

KW - Male

KW - Membrane Glycoproteins/biosynthesis

KW - Receptors, Cell Surface

KW - Recombinant Proteins/biosynthesis

KW - Signal Transduction

KW - Sperm Capacitation

KW - Spermatozoa/drug effects

KW - Zona Pellucida Glycoproteins

U2 - 10.1093/molehr/2.8.583

DO - 10.1093/molehr/2.8.583

M3 - Article

C2 - 9239670

SN - 1360-9947

VL - 2

SP - 583

EP - 589

JO - Molecular Human Reproduction

JF - Molecular Human Reproduction

IS - 8

ER -