Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Ram spermatozoa migrating through artificial mucus in vitro have reduced mitochondrial membrane potential but retain their viability

Carmen Martínez-Rodríguez A B , Mercedes Alvarez A C , Elena López-Urueña A C , Susana Gomes-Alves A C , Luis Anel-López A D , Cesar A. Chamorro A E , Luis Anel A C and Paulino de Paz A B F

A ITRA-ULE, INDEGSAL, University of León, 24071, León, Spain.

B Molecular Biology, University of León, 24071, León, Spain.

C Animal Reproduction and Obstetrics, University of León, 24071, León, Spain.

D SaBio IREC (CSIC-UCLM-JCCM), Campus Universitario s. n. 02071 Albacete, Spain.

E Veterinary Anatomy, University of León, 24071, León, Spain.

F Corresponding author. Email: ppazc@unileon.es

Reproduction, Fertility and Development 27(5) 852-864 http://dx.doi.org/10.1071/RD13377
Submitted: 7 November 2013  Accepted: 10 February 2014   Published: 31 March 2014

Abstract

Sperm motility in vitro is one of the most common predictors of fertility in male screening. We propose that a mucus-penetration assay can isolate a cellular subpopulation critical to reproductive success. To this end, a device was designed with three modules (sample, test and collection) and its conditions of use evaluated (length of mucus, incubation time, mucus medium, sperm concentration and position in relation to the horizontal). The number of spermatozoa migrating and the viability and acrosomal status of the spermatozoa not migrating were calculated. The second objective was to evaluate the qualitative parameters of the spermatozoa migrating in 1.6% polyacrylamide for 30 min. The number of spermatozoa migrating and the sperm motility, viability and the acrosomal and mitochondrial status of three sperm populations (fresh, not migrating and migrating) were determined. A higher number of migrating spermatozoa were observed after 60 min of incubation, but this situation adversely affected sperm quality. The methylcellulose-based test showed a significantly lower number of migrating spermatozoa than the polyacrylamide test. The position at an angle of 45° resulted in a higher number of migrating spermatozoa in the polyacrylamide-based test. The sperm counts for three consecutive assays indicated an acceptable repeatability of the method. The viability and acrosomal status of the migrating spermatozoa showed no significant changes with regard to the control when the device was placed at 45°, whereas these parameters showed lower values at 0°. The percentage of high mitochondrial membrane potential spermatozoa was significantly reduced in the population of migrating spermatozoa.

Additional keywords: acrosome status, motility, penetration test, synthetic mucus.


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