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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
    Martínez Pastor, FelipeAutoridad Buleria ORCID
    Johannisson, Anders
    Gil, Jorge
    Kaabi, Mohammed
    Anel Rodríguez, LuisAutoridad Buleria ORCID
    Paz Cabello, Paulino deAutoridad Buleria ORCID
    Rodríguez Martínez, H.
    Facultad/Centro
    Facultad de Ciencias Biologicas y Ambientales
    Área de conocimiento
    Biologia Celular
    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
    Veterinaria
    Palabras clave
    Semen
    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
    http://hdl.handle.net/10612/10309
    Versión del editor
    https://www.sciencedirect.com/science/article/pii/S0378432004000041#!
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    • Artículos de revista (post-print) [2096]
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