Abstract

Progesterone released from the cumulus cells of the oocyte causes a number of physiological responses in human sperm cells including hyperactivation, acrosome reaction and chemotaxis. We employed a validated sperm mobility assay, which involves measuring the ability of sperm to penetrate an inert cell separation solution over time, to assess the ability of progesterone to enhance the mobility of boar spermatozoa. Cells maximally penetrate the solution over 50 minutes. 100nM progesterone significantly (P = 0.01) increased the mobility of non-capacitated sperm cells causing a doubling in the rate at which the cells penetrated through the cell separation solution (control half maximal penetration rate (Km) = 18.0±2.2; +100nM progesterone Km = 8.8±0.8min). Similarly, capacitated cells penetrated at a rate (Km = 19.2±3.0 min) not significantly different from non-capacitated cells and 100nM progesterone also significantly increased the rate of penetration of capacitated cells (Km = 9.5±1.0 min, P<0.05). The T-type voltage gated calcium channel blocker mibefradil (30mM) significantly inhibited both the control and progesterone enhanced mobility in non-capacitated and capacitated sperm. Only capacitated cells showed a significant increase in the acrosome reaction in response to 100nM progesterone (control non-reacted = 75±4%, +100nM progesterone non- reacted = 47±10%). Western blot analysis confirmed that there was an increase in the total protein tyrosine phosphorylation levels in capacitated cells. In conclusion, we have demonstrated that 100nM progesterone accelerates the mobility of boar sperm cells through an inert cell separation solution in an extracellular calcium dependent manner.

Highlights

  • Progesterone is released from cumulus cells surrounding the oocyte and has been demonstrated to act as a chemoattractant for human and rabbit sperm [1,2,3,4]

  • There were no significant differences between the gradients for 100nM progesterone treated and control sperm for either capacitated or non-capacitated sperm indicating that there was no interaction between hormone treatment and time and that the relationship between absorbance and time could be modelled by a simple additive regression model without variable interaction

  • Progesterone has been demonstrated to induce a number of calcium dependent mechanisms in sperm including hyperactivation [7, 30], protein tyrosine phosphorylation [31, 32], acrosome reaction [33] and chemotaxis [34]

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Summary

Introduction

Progesterone is released from cumulus cells surrounding the oocyte and has been demonstrated to act as a chemoattractant for human and rabbit sperm [1,2,3,4]. This chemoattractant property is thought to be necessary in order to guide the sperm to the oocyte, appears to only occur in capacitated cells [5, 6]. Progesterone has been demonstrated to induce a rapid rise in intracellular calcium in sperm in other species [13,14,15]. Acrosome reacted sperm are unable to fertilise this may serve to select against inferior sperm [20]

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