Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Pesquisa Veterinária Brasileira (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2016000800767 |
Resumo: | Abstract: The aim of the study was to cryopreserve the semen of six-banded armadillos (Euphractus sexcinctus) in Tris-yolk and glycerol diluent, and to determine the damage caused by the freezing-thawing process, using fluorescent markers and ultrastructural analysis. Semen samples (n=11) collected from 4 adult six-banded armadillos by electroejaculation were cryopreserved in Tris diluent plus 20% egg yolk and 3% glycerol, in a fast freezing curve. Classical analysis of samples was performed after dilution, refrigeration and thawing, followed by fluorescence analysis, using a combination of fluorescent probes to assess membrane integrity (propidium iodide - PI and Hoechst - H342), and mitochondrial activity (CMXRos - Mito Tracker Red®). We also used the ultrastructural analysis to verify possible morphological alterations caused by cryoinjuries. When compared with fresh samples, we verified a significant decline in all the armadillos' semen parameters after thawing, in which only 6.1% motile sperm were found. However, the percentage of sperm which remained with viable (13%) and functional (24.7%) membranes after thawing suggests that some cells could be live but immotile. Analysis using fluorescent markers revealed that the mitochondria of armadillos' sperm is highly sensible to the freezing protocol and the findings through ultrastructure analysis proved this statement. Additionally, the images obtained by transmission electron microscopy revealed that frozen-thawed sperm presented damaged plasma membrane, nuclear modifications as changes in chromatin and acrossomal changes relative to sperm capacitation. In conclusion, this study is the first attempt to cryopreserve the semen of an armadillo species, and to help us to identify critical points on the freezing-thawing procedure in order to improve the protocol. |
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Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservationUltrastructural and functional damagesspermsix-banded armadilloEuphractus sexcinctuscryopreservationXenarthrassemenmembrane integritymitochondrial activityultrastructure.Abstract: The aim of the study was to cryopreserve the semen of six-banded armadillos (Euphractus sexcinctus) in Tris-yolk and glycerol diluent, and to determine the damage caused by the freezing-thawing process, using fluorescent markers and ultrastructural analysis. Semen samples (n=11) collected from 4 adult six-banded armadillos by electroejaculation were cryopreserved in Tris diluent plus 20% egg yolk and 3% glycerol, in a fast freezing curve. Classical analysis of samples was performed after dilution, refrigeration and thawing, followed by fluorescence analysis, using a combination of fluorescent probes to assess membrane integrity (propidium iodide - PI and Hoechst - H342), and mitochondrial activity (CMXRos - Mito Tracker Red®). We also used the ultrastructural analysis to verify possible morphological alterations caused by cryoinjuries. When compared with fresh samples, we verified a significant decline in all the armadillos' semen parameters after thawing, in which only 6.1% motile sperm were found. However, the percentage of sperm which remained with viable (13%) and functional (24.7%) membranes after thawing suggests that some cells could be live but immotile. Analysis using fluorescent markers revealed that the mitochondria of armadillos' sperm is highly sensible to the freezing protocol and the findings through ultrastructure analysis proved this statement. Additionally, the images obtained by transmission electron microscopy revealed that frozen-thawed sperm presented damaged plasma membrane, nuclear modifications as changes in chromatin and acrossomal changes relative to sperm capacitation. In conclusion, this study is the first attempt to cryopreserve the semen of an armadillo species, and to help us to identify critical points on the freezing-thawing procedure in order to improve the protocol.Colégio Brasileiro de Patologia Animal - CBPA2016-08-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2016000800767Pesquisa Veterinária Brasileira v.36 n.8 2016reponame:Pesquisa Veterinária Brasileira (Online)instname:Colégio Brasileiro de Patologia Animal (CBPA)instacron:EMBRAPA10.1590/S0100-736X2016000800015info:eu-repo/semantics/openAccessSousa,Patrícia C.Santos,Erika A.A.Silva,Andréia M.Bezerra,José A.B.Souza,Ana L.P.Lima,Gabriela L.Oliveira,Moacir F.Silva,Alexandre R.eng2016-10-28T00:00:00Zoai:scielo:S0100-736X2016000800767Revistahttp://www.pvb.com.br/https://old.scielo.br/oai/scielo-oai.phpcolegio@cbpa.org.br||pvb@pvb.com.br0100-736X1678-5150opendoar:2016-10-28T00:00Pesquisa Veterinária Brasileira (Online) - Colégio Brasileiro de Patologia Animal (CBPA)false |
dc.title.none.fl_str_mv |
Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation |
title |
Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation |
spellingShingle |
Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation Sousa,Patrícia C. Ultrastructural and functional damages sperm six-banded armadillo Euphractus sexcinctus cryopreservation Xenarthras semen membrane integrity mitochondrial activity ultrastructure. |
title_short |
Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation |
title_full |
Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation |
title_fullStr |
Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation |
title_full_unstemmed |
Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation |
title_sort |
Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation |
author |
Sousa,Patrícia C. |
author_facet |
Sousa,Patrícia C. Santos,Erika A.A. Silva,Andréia M. Bezerra,José A.B. Souza,Ana L.P. Lima,Gabriela L. Oliveira,Moacir F. Silva,Alexandre R. |
author_role |
author |
author2 |
Santos,Erika A.A. Silva,Andréia M. Bezerra,José A.B. Souza,Ana L.P. Lima,Gabriela L. Oliveira,Moacir F. Silva,Alexandre R. |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Sousa,Patrícia C. Santos,Erika A.A. Silva,Andréia M. Bezerra,José A.B. Souza,Ana L.P. Lima,Gabriela L. Oliveira,Moacir F. Silva,Alexandre R. |
dc.subject.por.fl_str_mv |
Ultrastructural and functional damages sperm six-banded armadillo Euphractus sexcinctus cryopreservation Xenarthras semen membrane integrity mitochondrial activity ultrastructure. |
topic |
Ultrastructural and functional damages sperm six-banded armadillo Euphractus sexcinctus cryopreservation Xenarthras semen membrane integrity mitochondrial activity ultrastructure. |
description |
Abstract: The aim of the study was to cryopreserve the semen of six-banded armadillos (Euphractus sexcinctus) in Tris-yolk and glycerol diluent, and to determine the damage caused by the freezing-thawing process, using fluorescent markers and ultrastructural analysis. Semen samples (n=11) collected from 4 adult six-banded armadillos by electroejaculation were cryopreserved in Tris diluent plus 20% egg yolk and 3% glycerol, in a fast freezing curve. Classical analysis of samples was performed after dilution, refrigeration and thawing, followed by fluorescence analysis, using a combination of fluorescent probes to assess membrane integrity (propidium iodide - PI and Hoechst - H342), and mitochondrial activity (CMXRos - Mito Tracker Red®). We also used the ultrastructural analysis to verify possible morphological alterations caused by cryoinjuries. When compared with fresh samples, we verified a significant decline in all the armadillos' semen parameters after thawing, in which only 6.1% motile sperm were found. However, the percentage of sperm which remained with viable (13%) and functional (24.7%) membranes after thawing suggests that some cells could be live but immotile. Analysis using fluorescent markers revealed that the mitochondria of armadillos' sperm is highly sensible to the freezing protocol and the findings through ultrastructure analysis proved this statement. Additionally, the images obtained by transmission electron microscopy revealed that frozen-thawed sperm presented damaged plasma membrane, nuclear modifications as changes in chromatin and acrossomal changes relative to sperm capacitation. In conclusion, this study is the first attempt to cryopreserve the semen of an armadillo species, and to help us to identify critical points on the freezing-thawing procedure in order to improve the protocol. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-08-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2016000800767 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2016000800767 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0100-736X2016000800015 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Colégio Brasileiro de Patologia Animal - CBPA |
publisher.none.fl_str_mv |
Colégio Brasileiro de Patologia Animal - CBPA |
dc.source.none.fl_str_mv |
Pesquisa Veterinária Brasileira v.36 n.8 2016 reponame:Pesquisa Veterinária Brasileira (Online) instname:Colégio Brasileiro de Patologia Animal (CBPA) instacron:EMBRAPA |
instname_str |
Colégio Brasileiro de Patologia Animal (CBPA) |
instacron_str |
EMBRAPA |
institution |
EMBRAPA |
reponame_str |
Pesquisa Veterinária Brasileira (Online) |
collection |
Pesquisa Veterinária Brasileira (Online) |
repository.name.fl_str_mv |
Pesquisa Veterinária Brasileira (Online) - Colégio Brasileiro de Patologia Animal (CBPA) |
repository.mail.fl_str_mv |
colegio@cbpa.org.br||pvb@pvb.com.br |
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1754122236117123072 |