RT info:eu-repo/semantics/article
T1 Use of chromatin stability assay, mitochondrial stain JC-1, and fluorometric assessment of plasma membrane to evaluate frozen-thawed ram semen
A1 Martínez Pastor, Felipe
A1 Johannisson, Anders
A1 Gil, Jorge
A1 Kaabi, Mohammed
A1 Anel Rodríguez, Luis
A1 Paz Cabello, Paulino de
A1 Rodríguez Martínez, H.
A2 Biologia Celular
K1 Veterinaria
K1 Semen
K1 Fluorometer
K1 Plasma membrane integrity
K1 Mitochondria
K1 JC-1
K1 Chromatin stability
K1 Sheep-male reproduction
AB 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.
PB Elsesvier
YR 2019
FD 2019-04-17
LK http://hdl.handle.net/10612/10309
UL http://hdl.handle.net/10612/10309
NO Animal Reproduction Science, 2004, vol. 84, n. 1-2
NO P. 121-133
DS BULERIA. Repositorio Institucional de la Universidad de León
RD 27-abr-2024