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Hypotonic resistance of boar spermatozoa: sperm subpopulations and relationship with epididymal maturation and fertility

¹ Union Nationale des Coopératives d’Elevage et d’Insémination Animale, 13 rue Jouet, 94704 Cedex Maisons Alfort, France² UMR 6175 INRA, CNRS-Université de Tours-Haras Nationaux, Station de Physiologie de la Reproduction et des Comportements, Institut National de la Recherche Agronomique, 37380 Nouzilly, France³ Unité de recherche Mathématiques et Informatique Appliquées, INRA Domaine de Vilvert, 78352 Cedex Jouy-en-Josas, France

Correspondence should be addressed to X Druart; Email: xavier.druart{at}tours.inra.fr

Hypotonic resistance of boar spermatozoa was investigated by measuring the ratio of live/dead spermatozoa (SYBR-14/propidium iodide) by flow cytometry after hypotonic stress. The survival rate of ejaculated spermatozoa incubated in hypotonic solutions ranging from 3 to 330 mmol/kg followed a sigmoid curve that fitted a simple logistic model. The critical osmolality value (Osm_crit) at which 50% of spermatozoa died was determined with this model. Hypotonic resistance of spermatozoa increased with temperature between 15 and 39 °C and decreased after hydrogen superoxide treatment, but was not modified during 8 days of preservation in Beltsville thawing solution. Hypotonic resistance markedly decreased during epididymal maturation and after ejaculation as Osm_crit at 15 °C was 54.7±3.2, 68.5±10.6, 116.7±2.1 and 194.3±3.7 mmol/kg for the caput, corpus, cauda and ejaculated spermatozoa respectively. Hypo-osmotic stress of 100 mmol/kg revealed a sperm subpopulation exhibiting increased hypotonic resistance compared with the whole ejaculate (Osm_crit=67.8±2.1 mmol/kg). Consistent differences were observed between lean and standard breeds (Pietrain versus Large White) and between boars within the same breed. According to data collected by artificial insemination centers during a large-scale field trial, hypotonic resistance of ejaculates was found to be positively correlated with in vivo fertility.