title: Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers.
abstract: The objectives of this study were to evaluate the effect of different acrosome reaction (AR) inducers on viability and acrosomal status in llama spermatozoa, by using the FITC-PNA/PI technique and evaluate if there is a positive correlation between the FITC-PNA/PI and the Coomassie blue (CB) staining techniques. After incubating twenty ejaculates in 0.1% collagenase the centrifuged pellets were resuspended in TALP-BSA medium. An aliquot was sonicated to remove the acrosomal content (positive control). The rest of the sample was incubated for 3h at 38 °C with 5% CO2 and 100% humidity.
  
  
  METHODS
  Three aliquots were further incubated 1h with one of the following AR inducers: calcium ionophore, ionomycin or progesterone.
  
  
  METHODS
  One without inducers and the other, incubated with dimethyl sulfoxide (vehicle of the inducing agents). Acrosomes were evaluated at time 0 and after 4h incubation. Calcium ionophore was the most potent agent for inducing the AR (67.2 ± 14.4% live+dead AR sperm) (P < 0.05). These samples showed no motility and viability was very low (0-30%). Both ionomycin and progesterone presented significantly higher (P < 0.05) percentages of total AR sperm than the controls, but had similar percentages of dead reacted sperm to the controls. A positive correlation was observed between the intact acrosome FITC-PNA/PI pattern (live+dead sperm) and the acrosome-present CB pattern (r = 0.64; P = 0.000) in all the evaluated samples.
  
  
  CONCLUSIONS
  the FITC-PNA/PI technique simultaneously evaluates viability and acrosomal status in llama spermatozoa and calcium ionophore could be used as a control of AR.
authors.affiliation: Cátedra de Teriogenología, Instituto de Investigación y Tecnología en Reproducción Animal, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina. Electronic address: ignaciacarretero@fvet.uba.ar.
authors.forename: M I
authors.initials: MI
authors.lastname: Carretero
authors.affiliation: Cátedra de Teriogenología, Instituto de Investigación y Tecnología en Reproducción Animal, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina.
authors.forename: F G
authors.initials: FG
authors.lastname: Fumuso
authors.affiliation: Cátedra de Teriogenología, Instituto de Investigación y Tecnología en Reproducción Animal, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Argentina.
authors.forename: D M
authors.initials: DM
authors.lastname: Neild
authors.affiliation: Cátedra de Física Biológica, Instituto de Investigación y Tecnología en Reproducción Animal, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Argentina.
authors.forename: S M
authors.initials: SM
authors.lastname: Giuliano
authors.affiliation: Cátedra de Química Biológica, Instituto de Investigación y Tecnología en Reproducción Animal, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina.
authors.forename: P
authors.initials: P
authors.lastname: Cetica
authors.affiliation: Cátedra de Teriogenología, Instituto de Investigación y Tecnología en Reproducción Animal, Facultad de Ciencias Veterinarias, Universidad de Buenos Aires, Argentina.
authors.forename: M H
authors.initials: MH
authors.lastname: Miragaya
mesh_terms: D000177:Acrosome / Q000502:physiology*; D020101:Acrosome Reaction / Q000187:drug effects*; D000818:Animals; D061207:Calcium Ionophores / Q000494:pharmacology*; D002161:Camelids, New World / Q000502:physiology*; D015759:Ionomycin / Q000494:pharmacology*; D008297:Male; D011374:Progesterone / Q000494:pharmacology*; D011372:Progestins / Q000494:pharmacology
chemical_list: D061207:Calcium Ionophores; D011372:Progestins; D011374:Progesterone; D015759:Ionomycin
keywords: Acrosome; Calcium ionophore; Ionomycin; Llama; Progesterone; Sperm