title: Use of hypometabolic TRIS extenders and high cooling rate refrigeration for cryopreservation of stallion sperm: presence and sensitivity of 5' AMP-activated protein kinase (AMPK).
abstract: This study evaluated the effect of the use of hypometabolic TRIS extenders in the presence or the absence of AMPK activators as well as the utilization of high cooling rates in the refrigeration step on the freezability of stallion sperm. Twelve ejaculates were cryopreserved using Botucrio® as a control extender and a basic TRIS extender (HM-0) separately supplemented with 10 mM metformin, 2mM 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR), 2 mM Adenosine monophosphate (AMP), 40 μM compound C AMPK inhibitor or 2 mM AMP+40 μM compound C. Our results showed that the utilization of a hypometabolic TRIS extender supplemented or not with AMP or metformin significantly improves stallion sperm freezability when compared with a commercial extender. Additionally, high cooling rates do not affect stallion sperm quality after cooling and post-thawing. Finally, stallion spermatozoa present several putative AMPK sperm isoforms that do not seem to respond to classical activators, but do respond to the Compound C inhibitor.
authors.affiliation: Instituto de Ciencia Animal, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Chile.
authors.forename: Alex
authors.initials: A
authors.lastname: Córdova
authors.affiliation: Instituto de Ciencia Animal, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Chile.
authors.forename: Pablo
authors.initials: P
authors.lastname: Strobel
authors.affiliation: Instituto de Ciencia Animal, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Chile.
authors.forename: Andrés
authors.initials: A
authors.lastname: Vallejo
authors.affiliation: Instituto de Ciencia Animal, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Chile.
authors.forename: Pamela
authors.initials: P
authors.lastname: Valenzuela
authors.affiliation: Haras Militar Pupunahue, DGFER-Ejército de Chile, Chile.
authors.forename: Omar
authors.initials: O
authors.lastname: Ulloa
authors.affiliation: Instituto de Farmacología y Morfofisiología, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Chile.
authors.forename: Rafael A
authors.initials: RA
authors.lastname: Burgos
authors.affiliation: Instituto de Ciencias Clínicas, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Chile.
authors.forename: Bruno
authors.initials: B
authors.lastname: Menarim
authors.affiliation: Unitat Reproducció Animal, Universitat Autònoma de Barcelona, Spain.
authors.forename: Joan Enric
authors.initials: JE
authors.lastname: Rodríguez-Gil
authors.affiliation: Ross University School of Veterinary Medicine, Basseterre, West Indies.
authors.forename: Marcelo
authors.initials: M
authors.lastname: Ratto
authors.affiliation: Instituto de Ciencia Animal, Facultad de Ciencias Veterinarias, Universidad Austral de Chile, Chile. Electronic address: alfredoramirez@uach.cl.
authors.forename: Alfredo
authors.initials: A
authors.lastname: Ramírez-Reveco
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000249:Adenosine Monophosphate / Q000378:metabolism; D000620:Aminoimidazole Carboxamide / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000378:metabolism*; D006736:Horses / Q000502:physiology*; D007004:Hypoglycemic Agents / Q000378:metabolism; D008297:Male; D008687:Metformin / Q000378:metabolism; D012265:Ribonucleotides / Q000378:metabolism; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D014325:Tromethamine / Q000378:metabolism*
chemical_list: D003451:Cryoprotective Agents; D007004:Hypoglycemic Agents; D012265:Ribonucleotides; D014325:Tromethamine; D000620:Aminoimidazole Carboxamide; D000249:Adenosine Monophosphate; D008687:Metformin; D055372:AMP-Activated Protein Kinases; C031143:AICA ribonucleotide
keywords: AMPK; Cryopreservation; Equine; Hypometabolism; Spermatozoa