RT Journal Article
T1 Cooling rate modifies the location of aquaporin 3 in spermatozoa of sheep and goat
A1 Pequeño, Belén
A1 Millán de la Blanca, María Gema
A1 Castaño, Cristina
A1 Toledano Díaz, Adolfo
A1 Esteso, Milagros C.
A1 Alba, Esther
A1 Arrebola, Francisco A.
A1 Ungerfeld, Rodolfo
A1 Martínez Madrid, Carmen Belén
A1 Álvarez Rodríguez, Manuel
A1 Rodríguez Martínez, Heriberto
A1 Santiago Moreno, Julián
AB The freeze-thawing process induces osmotic changes that may affect the membrane domain location of aquaporins’ (AQP) in spermatozoa. Recent studies suggest that changes in AQP3 localization allows better sperm osmo-adaptation, improving the cryoresistance. Ultra-rapid freezing is an alternative cryopreservation technique that requires less equipment than conventional freezing, and it is faster, simpler and can be used in the field. This study aimed to determine the influence of freezing-thawing rates (slow (control) vs. ultra-rapid) on AQP3 expression and location in the spermatozoa from small ruminants (sheep and goats) and its relationship with sperm cryo-damage. Spermatozoa were collected from 10 Merino rams and 10 Murciano-Granadina bucks. The presence and distribution of AQP3 were assessed by Western blotting and immunocytochemistry (ICC), employing a commercial rabbit polyclonal antibody. Sperm motility was CASA system-analyzed, and membrane and acrosome integrity assessed by fluorescence (PI/PNA-FITC). Western blotting did not detect a significant effect of freezing-thawing rate on the amount of AQP3 while ICC found freezing-thawing rate affecting AQP3 location (P < 0.05). In both species, the percentages of spermatozoa showing AQP3 in the post-acrosome region, mid-piece, and principal piece of the tail were greater in samples cryopreserved by slow freezing-thawing (control) than ultra-rapid freezing-thawing rates (P < 0.05). Spermatozoa cryopreserved using ultra-rapid freezing-thawing showed decrease motility, plasma membrane, and acrosome integrity (P < 0.05), which might be related, at least in part, to a lower expression of AQP3. In conclusion, the cooling rate modifies the location of AQP3 in spermatozoa of sheep and goat, which might be associated with sperm cryosurvival.
PB Elsevier
SN 0093-691X
YR 2024
FD 2024-04-19
LK https://hdl.handle.net/20.500.14352/103756
UL https://hdl.handle.net/20.500.14352/103756
LA eng
NO Authorship contribution statement: Belen Pequeno: Formal analysis, Methodology, Writing – original draft. María Gemma Millan de la Blanca: Formal analysis, Methodology. Cristina Castano: Formal analysis, Methodology. Adolfo Toledano-Díaz: Formal analysis, Investigation, Methodology. Milagros Cristina Esteso: Conceptualization, Investigation. Esther Alba: Data curation, Methodology. Francisco A. Arrebola: Investigation, Methodology. Rodolfo Ungerfeld: Conceptualization, Investigation, Writing – review & editing. Belen Martínez-Madrid: Conceptualization, Data curation, Methodology, Writing – review & editing. Manuel Alvarez Rodriguez: Conceptualization, Methodology, Writing – review & editing. Heriberto Rodriguez-Martinez: Conceptualization, Investigation, Writing – review & editing. Julian Santiago-Moreno: Conceptualization, Formal analysis, Funding acquisition, Investigation, Supervision, Writing – review & editing.
NO Ministerio de Ciencia, Innovación y Universidades (España)
DS Docta Complutense
RD 17 abr 2025