Abstract
The main scope of this manuscript is to analyse the dynamics of mitochondrial activity in boar sperm subjected to 'in vitro' capacitation (IVC) and subsequent progesterone-induced 'in vitro' acrosome reaction (IVAR). This was determined after analysis of the rhythm of O(2) consumption and concomitant changes in the mitochondria activity-specific JC-1 staining. Results showed that IVC, and especially IVAR, was concomitant with a peak in O(2) consumption (from 1.61 ± 0.08 nmol O(2)/min/10(7) viable sperm at 0 h of incubation to 2.62 ± 0.12 nmol O(2) /min/10(7) viable sperm after 5 min of IVAR induction). These results were accompanied by parallel changes in the mean intensity of JC-1 staining. Based on JC-1, mitochondrial activation followed a nucleated pattern, with specific, activation starting points at the midpiece from which mitochondrial activation was spread. Moreover, four separate sperm subpopulations were detected following the JC-1 orange-red/green ratio, and the observed changes in the mean JC-1 staining during IVC and IVAR were related to concomitant changes in both the orange-red/green JC-1 ratio and the percentage of sperm included in each subpopulation. All of these results indicate that IVC and the first minutes of IVAR are accompanied by a progressive increase in mitochondrial activity, which reached a peak coincidental with the achievement of IVAR. Moreover, results suggest the presence of separate sperm subpopulations, which show a different mitochondrial sensitivity to IVC and IVAR. Finally, mitochondrial activation, at least under JC-1 staining, seems to originate in concrete nucleation points at the midpiece, thus suggesting thus a well-coordinated pattern in boar-sperm mitochondrial activity modulation.
Highlights
The main aim of this work is to describe the changes in mitochondrial activity which are concomitant to IVC and subsequent ‘in vitro’, progesterone-induced acrosome reaction (IVAR) in boar spermatozoa
Evaluation of capacitation status during IVC and IVAR The induction of IVC induced a progressive decline in the percentage of viability, which was concomitant to an increase in the percentage of altered acrosomes (Table 1)
The results shown here highlighted a new perspective in the dynamics of mitochondrial function during the attainment of IVC and subsequent IVAR of boar sperm
Summary
The main aim of this work is to describe the changes in mitochondrial activity which are concomitant to IVC and subsequent ‘in vitro’, progesterone-induced acrosome reaction (IVAR) in boar spermatozoa. For this purpose, IVC- and IVAR-linked changes in mitochondrial activity through two separate techniques were analysed. The first one was the quantification of changes in the rhythm of oxygen consumption. The second one was the study of mitochondrial activity through the JC-1 staining procedure, analysing this through quantification of JC-1 intensity in digitalized, confocal laser microscope images
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