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Título
Use of chromatin stability assay, mitochondrial stain JC-1, and fluorometric assessment of plasma membrane to evaluate frozen-thawed ram semen
Autor
Facultad/Centro
Área de conocimiento
Datos de la obra
Animal Reproduction Science, 2004, vol. 84, n. 1-2
Editor
Elsesvier
Fecha
2004-08
Descripción física
P. 121-133
Abstract
Cryopreservation of semen imposes deleterious effects on spermatozoa, either killing a certain proportion of cells or causing subtle damages on sperm function in the surviving population, changes not easily revealed by conventional assays. We have tested three functional assessment techniques in frozen-thawed ram semen from six adult rams, cryopreserved following eight different protocols (four extenders, and glycerol being added at two temperatures). Semen samples were thawed and the following analyses were carried out: motility (CASA), membrane integrity (Hoescht 33258 and fluorometry), chromatin status (chromatin stability test and fluorescence-assisted cell sorting, FACS) and mitochondrial activity (JC-1 and FACS). Fluorometry outcome did not correlate with the other parameters and showed large variation, albeit discriminating among cryopreservation techniques (P<0.01). Mitochondrial activity correlated, but with low values, with total and progressive motility. However, good sperm motility and high velocity values were associated to high mitochondrial membrane potential. The chromatin stability assay was also successfully carried out, and had a good relationship with male factor (%COMP αt and SD αt parameters). In conclusion, fluorometric assessment of membrane integrity albeit rendering poor results, merits improvement, being a low-cost and handy technique, especially for work in the field. On the other hand, both assessments of chromatin stability and mitochondrial status (JC-1 staining), combined with FACS, are reliable techniques that can be used for the functional assessment of frozen-thawed ram semen.
Materia
Palabras clave
Semen
Fluorometer
Plasma membrane integrity
Mitochondria
JC-1
Chromatin stability
Sheep-male reproduction
Fluorometer
Plasma membrane integrity
Mitochondria
JC-1
Chromatin stability
Sheep-male reproduction
Idioma
eng
Tipo documental
info:eu-repo/semantics/article
Peer review
SI
URI
Versión del editor
https://www.sciencedirect.com/science/article/pii/S0378432004000041#!
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