Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers

Autores
Carretero, Maria Ignacia; Fumuso, Fernanda Gabriela; Neild, Debora Margarita; Giuliano, Susana María; Cetica, Pablo Daniel; Miragaya, Marcelo
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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. TREATMENTS: Three aliquots were further incubated 1h with one of the following AR inducers: calcium ionophore, ionomycin or progesterone. CONTROLS: 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.
Fil: Carretero, Maria Ignacia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina
Fil: Fumuso, Fernanda Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina
Fil: Neild, Debora Margarita. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Giuliano, Susana María. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina
Fil: Cetica, Pablo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina
Fil: Miragaya, Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina
Materia
Llama
Sperm
Acrosome
Calcium Ionophore
Ionomycin
Progesterone
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/44394
Acceder
id CONICETDig_30c1b42a3814f4217f98a3a5fc722dce
oai_identifier_str oai:ri.conicet.gov.ar:11336/44394
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducersCarretero, Maria IgnaciaFumuso, Fernanda GabrielaNeild, Debora MargaritaGiuliano, Susana MaríaCetica, Pablo DanielMiragaya, MarceloLlamaSpermAcrosomeCalcium IonophoreIonomycinProgesteronehttps://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4The 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. TREATMENTS: Three aliquots were further incubated 1h with one of the following AR inducers: calcium ionophore, ionomycin or progesterone. CONTROLS: 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.Fil: Carretero, Maria Ignacia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Fumuso, Fernanda Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Neild, Debora Margarita. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Giuliano, Susana María. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Cetica, Pablo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaFil: Miragaya, Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; ArgentinaElsevier Science2015-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/44394Carretero, Maria Ignacia; Fumuso, Fernanda Gabriela; Neild, Debora Margarita; Giuliano, Susana María; Cetica, Pablo Daniel; et al.; Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers; Elsevier Science; Animal Reproduction Science; 160; 9-2015; 1-110378-43201873-2232CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.anireprosci.2015.06.014info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0378432015001451info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:58:58Zoai:ri.conicet.gov.ar:11336/44394instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:58:58.497CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers
title Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers
spellingShingle Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers
Carretero, Maria Ignacia
Llama
Sperm
Acrosome
Calcium Ionophore
Ionomycin
Progesterone
title_short Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers
title_full Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers
title_fullStr Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers
title_full_unstemmed Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers
title_sort Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers
dc.creator.none.fl_str_mv Carretero, Maria Ignacia
Fumuso, Fernanda Gabriela
Neild, Debora Margarita
Giuliano, Susana María
Cetica, Pablo Daniel
Miragaya, Marcelo
author Carretero, Maria Ignacia
author_facet Carretero, Maria Ignacia
Fumuso, Fernanda Gabriela
Neild, Debora Margarita
Giuliano, Susana María
Cetica, Pablo Daniel
Miragaya, Marcelo
author_role author
author2 Fumuso, Fernanda Gabriela
Neild, Debora Margarita
Giuliano, Susana María
Cetica, Pablo Daniel
Miragaya, Marcelo
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Llama
Sperm
Acrosome
Calcium Ionophore
Ionomycin
Progesterone
topic Llama
Sperm
Acrosome
Calcium Ionophore
Ionomycin
Progesterone
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv 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. TREATMENTS: Three aliquots were further incubated 1h with one of the following AR inducers: calcium ionophore, ionomycin or progesterone. CONTROLS: 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.
Fil: Carretero, Maria Ignacia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina
Fil: Fumuso, Fernanda Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina
Fil: Neild, Debora Margarita. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Giuliano, Susana María. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina
Fil: Cetica, Pablo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina
Fil: Miragaya, Marcelo. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Instituto de Investigacion y Tecnología en Reproducción Animal; Argentina
description 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. TREATMENTS: Three aliquots were further incubated 1h with one of the following AR inducers: calcium ionophore, ionomycin or progesterone. CONTROLS: 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.
publishDate 2015
dc.date.none.fl_str_mv 2015-09
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/44394
Carretero, Maria Ignacia; Fumuso, Fernanda Gabriela; Neild, Debora Margarita; Giuliano, Susana María; Cetica, Pablo Daniel; et al.; Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers; Elsevier Science; Animal Reproduction Science; 160; 9-2015; 1-11
0378-4320
1873-2232
CONICET Digital
CONICET
url http://hdl.handle.net/11336/44394
identifier_str_mv Carretero, Maria Ignacia; Fumuso, Fernanda Gabriela; Neild, Debora Margarita; Giuliano, Susana María; Cetica, Pablo Daniel; et al.; Evaluation of the acrosomal status in Lama glama sperm incubated with acrosome reaction inducers; Elsevier Science; Animal Reproduction Science; 160; 9-2015; 1-11
0378-4320
1873-2232
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.anireprosci.2015.06.014
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0378432015001451
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1842269552927309824
score 13.13397

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