Influence of seminal plasma during cooling of boar spermatozoa on dna integrity, sperm subpopulations and calcium influx under in vitro capacitation.

Abstract

Artificial insemination (AI) allows for rapid genetic improvement, being widely used throughout the world in pig farms. Higher reproductive rates achieved with the use of cooled semen in relation to cryopreserved semen make this the most used form of storage in the majority of AIs performed. However, capacitation-like changes, DNA damage and changes on motility patterns are consequences commonly reported as a result of the handling and storage of spermatozoa. The effect of seminal plasma on these changes have been the goal of several studies, in which, however, a duality action was verified. The current study was designed to evaluate the effect of seminal plasma during cooling upon DNA integrity, sperm motile subpopulations and intracellular calcium movements under in vitro capacitating conditions. DNA integrity will be assessed by Sperm Chromatin Dispersion Assay (SCSA). The evaluation of changes on motility parameters and proportions of each sperm subpopulation will be assessed by CASA analysis, based on previous determinations in the raw ejaculate. Furthermore, intracellular calcium levels in the head and the tail will be analysed using two calcium markers with different physic-chemical properties, Fluo-3 and Rhod-5N. All these tests will shed new light into the role of seminal plasma during sperm capacitation.