Abstract

BackgroundSeveral studies have been published where sperm plasma membrane integrity correlated to fertility. In this study we describe a simple fluorometer-based assay where we monitored the fluorescence intensity of artificially membrane-ruptured spermatozoa with a fixed time staining with fluorescent DNA dyes.MethodsMembrane-impermeant fluorescent dyes Hoechst 33258 (H258) and propidium iodide (PI) were used to measure the fluorescence of the nucleus in artificially membrane ruptured spermatozoa and membrane-permeant dye Hoechst 33342 (H342) was used to measure fluorescence of intact spermatozoa. The concentration of spermatozoa in insemination doses varied from 31.2 × 106/ml to 50 × 106/ml and the average value was 35 × 106/ml. Each boar was represented by three consecutive ejaculates, collected at weekly intervals. Nonreturn rate within 60 days of first insemination (NR %) and litter size (total number of piglets born) of multiparous farrowings were used as fertility measures.ResultsSperm fluorescence intensity of H258 and H342, but not the fluorescence intensity of PI-stained spermatozoa correlated significantly with the litter size of multiparous farrowings, values being r = - 0.68 (P < 0.01) for H258, r = - 0.69 (P < 0.01) for H342 and r = - 0.38, (P = 0.11) for PI.ConclusionsThe increase in fluorescence values of membrane-ruptured H258 and unruptured H342-stained spermatozoa in boar AI doses can be associated with smaller litter size after AI. This finding indicates that the fluorescence properties of the sperm nucleus could be used to select for AI doses with greater fertilizing potential.

Highlights

  • Several studies have been published where sperm plasma membrane integrity correlated to fertility

  • Acta Veterinaria Scandinavica 2009, 51:53 http://www.actavetscand.com/content/51/1/53 tor testicular function of boars and bulls as spermatozoa are not analyzed in terms of their total integrity [2,3,4]

  • Nonreturn rate within 60 days of first insemination (NR %) and litter size of multiparous farrowings were used as fertility measures

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Summary

Introduction

Several studies have been published where sperm plasma membrane integrity correlated to fertility. Assessing fertilizing potential of an ejaculate generally includes tests of sperm function, as well as evaluation of sperm morphology, motility profiles, concentration, viability, ability to acrosome-react and to penetrate oocytes [1]. Sperm concentration and sperm motility are the three major components of routine sperm quality assessment. Microscopic assessment of sperm morphology, concentration and motility is inexpensive, subjective and of low predictive power to moni-. Assessment of sperm plasma membrane integrity is one of the key parameters in evaluation of spermatozoal quality in relation to fertility in a particular male [5]. One of the major features discriminating dead from live cells is loss in physical integrity of their plasma membranes and loss of motility [7]

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