Abstract

BAEZ ORE, Luis; SEGURA ZEVALLOS, Carlos; LA ROSA AGAPITO, José  and  SANTIANI, Alexei. Frequency of spermatozoa with caspase 3/7 activation obtained from alpaca epididymis analyzed by flow cytometry. Rev. investig. vet. Perú [online]. 2023, vol.34, n.6, e26956.  Epub Dec 31, 2023. ISSN 1609-9117.  http://dx.doi.org/10.15381/rivep.v34i6.26956.

The study aimed to determine the proportion of alpaca (Vicugna pacos) sperm cells with caspases-3/7 in an activated form. The epididymis of 23 pairs of testicles obtained from the Municipal Slaughterhouse of Huancavelica, Peru, were used. In the laboratory, an extender (fat-free milk, egg yolk and fructose) was used to obtain sperm cells from the caudal part of the epididymis. The cells were washed twice with 1 mL of PBS and centrifuged. It was added 100 µL of CellEvent™ Caspase 3/7 Green Detection Reagent to the product and incubated at 37.5 °C for 30 min, while 0.5 µL of Propidium Iodide was added 10 minutes before the end of the incubation, as a marker of cell death. The reading was done by flow cytometry. The mean ± standard deviation of sperm cells with caspase 3/7 activationwas36.21 ± 11.25%, confidence interval from31.34 to 41.08%and a coefficient of variation of 31.07%. The data presented a normal distribution according to the Kolmogorv-Smirnov test. The correlations between spermatozoa with activated caspases 3/7 with the weight and volume of the testis and with sperm motility and concentration were not significant. It is estimated between 31 to 41% of alpaca epididymal spermatozoa are in the process of apoptosis.

Keywords : spermatozoa; alpaca; flow cytometry; apoptosis; caspases.

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