TY - JOUR
T1 - A triple stain method in conjunction with an in-depth screening of cryopreservation effects on post-thaw sperm in dogs
AU - Ramos Angrimani, Daniel de Souza
AU - Bicudo, Luana de Cassia
AU - Luceno, Nuria Llamas
AU - Leemans, Bart
AU - Nichi, Marcilio
AU - Vannucchi, Camila Infantosi
AU - Van Soom, Ann
PY - 2022/4
Y1 - 2022/4
N2 - In order to accurately analyze the possible side effects of sperm cryopreservation, an in-depth screening of post-thaw sperm status is necessary. Thus, this study aimed to identify thorough effects of sperm cryopreservation, by evaluating the integrity of all specific structures of the canine spermatozoa. Thirteen (n = 13) mature dogs of different breeds were selected. Six dogs (n = 6) were subjected to sperm cryopreservation, whereas seven dogs (n = 7) were used as semen donors to validate a simultaneous assessment of sperm plasmatic, acrosomal, and mitochondrial membranes (triple stain) by fluorescent probes. Fresh and post-thaw semen samples were evaluated through a computer-assisted analysis of sperm motility, sperm morpho-functional evaluation, triple stain and sperm DNA integrity. Post-thaw semen samples had lower total and progressive motility, as well as higher percentage of minor and major defects. Moreover, post-thaw samples had higher percentage of sperm with plasma membrane and mitochondrial damage but intact acrosome, and also sperm with simultaneous damaged plasma, acrosomal and mitochondrial membranes. Furthermore, post-thaw sperm had higher protamination deficiency and DNA fragmentation. In conclusion, cryopreservation has a broad impact in sperm morphology and function, altering motility patterns, plasma, acrosome and mitochondrial membranes integrity, as well as sperm DNA.
AB - In order to accurately analyze the possible side effects of sperm cryopreservation, an in-depth screening of post-thaw sperm status is necessary. Thus, this study aimed to identify thorough effects of sperm cryopreservation, by evaluating the integrity of all specific structures of the canine spermatozoa. Thirteen (n = 13) mature dogs of different breeds were selected. Six dogs (n = 6) were subjected to sperm cryopreservation, whereas seven dogs (n = 7) were used as semen donors to validate a simultaneous assessment of sperm plasmatic, acrosomal, and mitochondrial membranes (triple stain) by fluorescent probes. Fresh and post-thaw semen samples were evaluated through a computer-assisted analysis of sperm motility, sperm morpho-functional evaluation, triple stain and sperm DNA integrity. Post-thaw semen samples had lower total and progressive motility, as well as higher percentage of minor and major defects. Moreover, post-thaw samples had higher percentage of sperm with plasma membrane and mitochondrial damage but intact acrosome, and also sperm with simultaneous damaged plasma, acrosomal and mitochondrial membranes. Furthermore, post-thaw sperm had higher protamination deficiency and DNA fragmentation. In conclusion, cryopreservation has a broad impact in sperm morphology and function, altering motility patterns, plasma, acrosome and mitochondrial membranes integrity, as well as sperm DNA.
KW - Casa
KW - Canine
KW - DNA fragmentation
KW - DNA protamination
KW - Triple stain
UR - https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=d7dz6a2i7wiom976oc9ff2iqvdhv8k5x&SrcAuth=WosAPI&KeyUT=WOS:000819660200007&DestLinkType=FullRecord&DestApp=WOS_CPL
U2 - 10.1016/j.cryobiol.2021.12.001
DO - 10.1016/j.cryobiol.2021.12.001
M3 - Article
C2 - 34902341
SN - 0011-2240
VL - 105
JO - Cryobiology
JF - Cryobiology
ER -