Damage assessment of the equine sperm membranes by fluorimetric technique
To validate a practical technique of simultaneous evaluation of the plasma, acrosomal and mitochondrial membranes in equine spermatozoa three fluorescent probes (PI, FITC-PSA and MITO) were associated. Four ejaculates from three stallions (n=12) were diluted in TALP medium and split into 2 aliquots,...
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| Format: | Article |
| Language: | English |
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Instituto de Tecnologia do Paraná (Tecpar)
2010-12-01
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| Series: | Brazilian Archives of Biology and Technology |
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| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132010000600004 |
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| author | Eneiva Carla Carvalho Celeghini André Furugen Cesar de Andrade Cláudia Fernandes Raphael Juliana Nascimento Janahi Souza Ticianelli Rubens Paes de Arruda |
| author_facet | Eneiva Carla Carvalho Celeghini André Furugen Cesar de Andrade Cláudia Fernandes Raphael Juliana Nascimento Janahi Souza Ticianelli Rubens Paes de Arruda |
| author_sort | Eneiva Carla Carvalho Celeghini |
| collection | DOAJ |
| description | To validate a practical technique of simultaneous evaluation of the plasma, acrosomal and mitochondrial membranes in equine spermatozoa three fluorescent probes (PI, FITC-PSA and MITO) were associated. Four ejaculates from three stallions (n=12) were diluted in TALP medium and split into 2 aliquots, 1 aliquot was flash frozen in liquid nitrogen to induce damage in cellular membranes. Three treatments were prepared with the following fixed ratios of fresh semen: flash frozen semen: 100:0 (T100), 50:50 (T50), and 0:100 (T0). A 150-µL aliquot of diluted semen of each treatment was added of 2 µL of PI, 2 µL of MITO and 80 µL of FITC-PSA; incubated at 38.5ºC/8 min, and sperm cells were evaluated by epifluorescent microscopy. Based in regression analysis, this could be an efficient and practical technique to assess damage in equine spermatozoa, as it was able to determine the sperm percentage more representative of the potential to fertilize the oocyte.<br>Para validar uma técnica prática de avaliação simultânea das membranas plasmática, acrossomal e mitocondrial em espermatozóides eqüinos três sondas fluorescentes (PI, FITC-PSA e MITO) foram associadas. Quatro ejaculados de três garanhões (n=12) foram diluídos em meio TALP e divididos em duas alíquotas, uma alíquota foi submetida a flash frozen em nitrogênio líquido para induzir danos nas membranas celulares. Três tratamentos foram preparados com as seguintes proporções de sêmen fresco: sêmen flash frozen: 100:0 (T100), 50:50 (T50), e 0:100 (T0). Uma amostra de 150 µL de sêmen diluído de cada tratamento foi adicionada de 2 µL de PI, 2 µL de MITO e 80 µL de FITC-PSA; incubadas à 38,5ºC/8 min, e as células espermáticas foram avaliadas por microscopia de epifluorescência. Baseados na análise de regressão esta é uma técnica eficiente e prática para determinar danos em espermatozóides eqüinos, capaz de determinar a porcentagem de espermatozóides mais representativa do potencial fertilizante do ovócito. |
| first_indexed | 2024-12-23T13:49:45Z |
| format | Article |
| id | doaj.art-2568e3fa90f040ccaac19180190183ec |
| institution | Directory Open Access Journal |
| issn | 1516-8913 1678-4324 |
| language | English |
| last_indexed | 2024-12-23T13:49:45Z |
| publishDate | 2010-12-01 |
| publisher | Instituto de Tecnologia do Paraná (Tecpar) |
| record_format | Article |
| series | Brazilian Archives of Biology and Technology |
| spelling | doaj.art-2568e3fa90f040ccaac19180190183ec2022-12-21T17:44:37ZengInstituto de Tecnologia do Paraná (Tecpar)Brazilian Archives of Biology and Technology1516-89131678-43242010-12-015361285129210.1590/S1516-89132010000600004Damage assessment of the equine sperm membranes by fluorimetric techniqueEneiva Carla Carvalho CeleghiniAndré Furugen Cesar de AndradeCláudia Fernandes RaphaelJuliana NascimentoJanahi Souza TicianelliRubens Paes de ArrudaTo validate a practical technique of simultaneous evaluation of the plasma, acrosomal and mitochondrial membranes in equine spermatozoa three fluorescent probes (PI, FITC-PSA and MITO) were associated. Four ejaculates from three stallions (n=12) were diluted in TALP medium and split into 2 aliquots, 1 aliquot was flash frozen in liquid nitrogen to induce damage in cellular membranes. Three treatments were prepared with the following fixed ratios of fresh semen: flash frozen semen: 100:0 (T100), 50:50 (T50), and 0:100 (T0). A 150-µL aliquot of diluted semen of each treatment was added of 2 µL of PI, 2 µL of MITO and 80 µL of FITC-PSA; incubated at 38.5ºC/8 min, and sperm cells were evaluated by epifluorescent microscopy. Based in regression analysis, this could be an efficient and practical technique to assess damage in equine spermatozoa, as it was able to determine the sperm percentage more representative of the potential to fertilize the oocyte.<br>Para validar uma técnica prática de avaliação simultânea das membranas plasmática, acrossomal e mitocondrial em espermatozóides eqüinos três sondas fluorescentes (PI, FITC-PSA e MITO) foram associadas. Quatro ejaculados de três garanhões (n=12) foram diluídos em meio TALP e divididos em duas alíquotas, uma alíquota foi submetida a flash frozen em nitrogênio líquido para induzir danos nas membranas celulares. Três tratamentos foram preparados com as seguintes proporções de sêmen fresco: sêmen flash frozen: 100:0 (T100), 50:50 (T50), e 0:100 (T0). Uma amostra de 150 µL de sêmen diluído de cada tratamento foi adicionada de 2 µL de PI, 2 µL de MITO e 80 µL de FITC-PSA; incubadas à 38,5ºC/8 min, e as células espermáticas foram avaliadas por microscopia de epifluorescência. Baseados na análise de regressão esta é uma técnica eficiente e prática para determinar danos em espermatozóides eqüinos, capaz de determinar a porcentagem de espermatozóides mais representativa do potencial fertilizante do ovócito.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132010000600004Stallionacrosomeplasma membranemitochondriafluorescent probes |
| spellingShingle | Eneiva Carla Carvalho Celeghini André Furugen Cesar de Andrade Cláudia Fernandes Raphael Juliana Nascimento Janahi Souza Ticianelli Rubens Paes de Arruda Damage assessment of the equine sperm membranes by fluorimetric technique Brazilian Archives of Biology and Technology Stallion acrosome plasma membrane mitochondria fluorescent probes |
| title | Damage assessment of the equine sperm membranes by fluorimetric technique |
| title_full | Damage assessment of the equine sperm membranes by fluorimetric technique |
| title_fullStr | Damage assessment of the equine sperm membranes by fluorimetric technique |
| title_full_unstemmed | Damage assessment of the equine sperm membranes by fluorimetric technique |
| title_short | Damage assessment of the equine sperm membranes by fluorimetric technique |
| title_sort | damage assessment of the equine sperm membranes by fluorimetric technique |
| topic | Stallion acrosome plasma membrane mitochondria fluorescent probes |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132010000600004 |
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