Inclusão de colesterol na membrana plasmática de espermatozoides caprinos
Autor(a) principal: | |
---|---|
Data de Publicação: | 2013 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/5132 |
Resumo: | The aim of this study was to evaluate the fertilizing capacity, acrosomal integrity, and qualify and quantify by chromatographic techniques the incorporation of cholesterol to the sperm membrane by cyclodextrin in different diluents on cryopreservation of goat sperm. Four males, Saanen (2) and Parda Alpine (2) breeds were used. It was performed a completely randomized design, with each semen sample divided into the following treatments: TG - Tris-glycerol diluent; TGCCC - Cyclodextrin-cholesterol complex (CCC) + Tris-glycerol diluent; TG15CCC - CCC diluted in isosmotic solution (saline) to semen with 15 minutes of incubation before addition of the Tris-glycerol diluent; EE - egg yolk + ethylene glycol diluent; EECCC - CCC + egg yolk + ethylene glycol diluent; EE15CCC - CCC diluted in isosmotic solution (saline) to semen with 15 minutes of incubation before addition of the egg yolk + ethylene glycol diluent. Fresh semen, after its examination, was submitted to the cryopreservation process and stored in a cryogenic cylinder for 10 days. After thawing, the following analysis were performed: acrosomal integrity, sperm fertilizing capacity through perivitelline membrane of hen egg yolk binding test (MPEY) and analysis of sperm motility and vigor. Besides, techniques of gas and thin layer chromatography were conducted to evaluate the incorporation of cholesterol to the sperm membrane. Data were submitted to Lilliefors and Cochran and Bartlett tests to verify normality and homogeneity of variances, respectively. The characteristics that met the assumptions of these tests were submitted to ANOVA and means were compared by Duncan s test at 5% of probability. When data did not meet the assumptions of normality and homogeneity of variances, the means were compared by Kruskal-Wallis test. Pearson s correlation coefficient was calculated in all features.Addition of cyclodextrin-cholesterol complex did not increase binding of sperm to MPEY (P > 0.05). The EE15CCC treatment was superior to EECCC treatment in maintaining the acrosomal integrity but did not differ from control (EE; P > 0.05). In Tris-glycerol diluent, the values observed in the control treatment were higher (P < 0.05) than values of the other treatments (TGCCC and TG15CCC) in maintaining the acrosomal integrity. There was a negative correlation between the binding assay and acrosomal integrity (r = -0.25) and positive correlation between the binding assay and sperm motility (r = 0.20). The sperm motility and acrosomal integrity were negatively correlated (r = -0.26). In quantitative and qualitative evaluation of cholesterol by chromatographic techniques (gas and thin layer) there was no difference between the samples of semen in different treatments (P > 0.05). Both techniques showed no incorporation of cholesterol to spermatozoa. It was concluding that addition of cholesterol-cyclodextrin complex to the medium did not improve the physical aspects of goat semen, or the sperm binding capacity. Pre-incubation of semen with CCC for 15 minutes before addition of ethylene + egg yolk diluent (EE15CCC) did not enhance the integrity of the acrosome in relation to control (EE). The gas and thin layer chromatography showed, respectively, an efficient method for quantitatively and qualitatively determining the cholesterol present in the cryopreserved goat sperm. The concentration of 1 mg of cholesterol-cyclodextrin complex added to the goat semen was not effective for increasing the concentration of cholesterol in sperm. |
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Silveira, Camila Oliveirahttp://lattes.cnpq.br/2921617720071753Costa, Eduardo Paulino dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787237D6Queiroz, Maria Eliana Lopes Ribeiro dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781671U3Guimarães, José Domingoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U6Torres, Ciro Alexandre Alveshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787213D4Pinho, Rogério Oliveirahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4734857T62015-03-26T13:47:12Z2013-06-252015-03-26T13:47:12Z2013-02-22SILVEIRA, Camila Oliveira. Inclusion of cholesterol to goat sperm membrane. 2013. 54 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2013.http://locus.ufv.br/handle/123456789/5132The aim of this study was to evaluate the fertilizing capacity, acrosomal integrity, and qualify and quantify by chromatographic techniques the incorporation of cholesterol to the sperm membrane by cyclodextrin in different diluents on cryopreservation of goat sperm. Four males, Saanen (2) and Parda Alpine (2) breeds were used. It was performed a completely randomized design, with each semen sample divided into the following treatments: TG - Tris-glycerol diluent; TGCCC - Cyclodextrin-cholesterol complex (CCC) + Tris-glycerol diluent; TG15CCC - CCC diluted in isosmotic solution (saline) to semen with 15 minutes of incubation before addition of the Tris-glycerol diluent; EE - egg yolk + ethylene glycol diluent; EECCC - CCC + egg yolk + ethylene glycol diluent; EE15CCC - CCC diluted in isosmotic solution (saline) to semen with 15 minutes of incubation before addition of the egg yolk + ethylene glycol diluent. Fresh semen, after its examination, was submitted to the cryopreservation process and stored in a cryogenic cylinder for 10 days. After thawing, the following analysis were performed: acrosomal integrity, sperm fertilizing capacity through perivitelline membrane of hen egg yolk binding test (MPEY) and analysis of sperm motility and vigor. Besides, techniques of gas and thin layer chromatography were conducted to evaluate the incorporation of cholesterol to the sperm membrane. Data were submitted to Lilliefors and Cochran and Bartlett tests to verify normality and homogeneity of variances, respectively. The characteristics that met the assumptions of these tests were submitted to ANOVA and means were compared by Duncan s test at 5% of probability. When data did not meet the assumptions of normality and homogeneity of variances, the means were compared by Kruskal-Wallis test. Pearson s correlation coefficient was calculated in all features.Addition of cyclodextrin-cholesterol complex did not increase binding of sperm to MPEY (P > 0.05). The EE15CCC treatment was superior to EECCC treatment in maintaining the acrosomal integrity but did not differ from control (EE; P > 0.05). In Tris-glycerol diluent, the values observed in the control treatment were higher (P < 0.05) than values of the other treatments (TGCCC and TG15CCC) in maintaining the acrosomal integrity. There was a negative correlation between the binding assay and acrosomal integrity (r = -0.25) and positive correlation between the binding assay and sperm motility (r = 0.20). The sperm motility and acrosomal integrity were negatively correlated (r = -0.26). In quantitative and qualitative evaluation of cholesterol by chromatographic techniques (gas and thin layer) there was no difference between the samples of semen in different treatments (P > 0.05). Both techniques showed no incorporation of cholesterol to spermatozoa. It was concluding that addition of cholesterol-cyclodextrin complex to the medium did not improve the physical aspects of goat semen, or the sperm binding capacity. Pre-incubation of semen with CCC for 15 minutes before addition of ethylene + egg yolk diluent (EE15CCC) did not enhance the integrity of the acrosome in relation to control (EE). The gas and thin layer chromatography showed, respectively, an efficient method for quantitatively and qualitatively determining the cholesterol present in the cryopreserved goat sperm. The concentration of 1 mg of cholesterol-cyclodextrin complex added to the goat semen was not effective for increasing the concentration of cholesterol in sperm.O objetivo do presente estudo foi avaliar a capacidade fecundante, integridade acrossomal, além de qualificar e quantificar por técnicas cromatográficas a incorporação do colesterol à membrana plasmática do espermatozoide pela ciclodextrina em diferentes diluentes na criopreservação de espermatozoides caprinos. Foram utilizados quatro machos caprinos das raças Saanen (2) e Parda Alpina (2) seguindo um delineamento inteiramente casualizado, dividido nos seguintes tratamentos: TGcontrole negativo para o diluente a base de Tris-glicerol; TGCCC- Complexo ciclodextrina-colesterol (CCC) + diluente Tris glicerol; TG15CCC- CCC diluído em solução isosmótica ao sêmen (soro fisiológico) com 15 minutos de incubação antes da adição do diluente Tris glicerol; EE- controle negativo para o diluente a base de Gema de ovo + etilenoglicol; EECCC- CCC + diluente Gema de ovo + etilenoglicol; EE15CCC- CCC diluído em solução isosmótica ao sêmen (soro fisiológico) com 15 minutos de incubação antes da adição do diluente Gema de ovo + etilenoglicol. O sêmen fresco, após realização de sua análise física foi submetido ao processo de criopreservação e estocado em botijão de nitrogênio por 10 dias. Após o descongelamento realizou-se análise da integridade acrossomal, capacidade fecundante do espermatozoide por meio do teste de ligação à membrana perivitelina da gema do ovo de galinha (MPGV) e as análises de motilidade progressiva e vigor espermático. Além destes testes foi realizada a avaliação da incorporação do colesterol à membrana plasmática dos espermatozoides pelas técnicas de cromatografia gasosa e de camada delgada. Para verificação da normalidade e homogeneidade dos dados foi empregado, respectivamente, o teste de Lilliefors e Cochran e Bartlett. As características que atenderam as premissas destes testes foram submetidas à ANOVA e as médias foram comparadas pelo teste de Duncan com 5% de probabilidade de erro. Quando as distribuições não atenderam as premissas de normalidade e homogeneidade, as médias foram comparadas pelo teste de Kruskal-Wallis. Realizou-se a correlação simples de Pearson entre todas as características. A adição do complexo ciclodextrina-colesterol não aumentou a ligação dos espermatozoides a MPGV (P>0,05). O tratamento empregando o complexo ciclodextrina-colesterol (CCC) diluído em solução isosmótica ao sêmen com 15 minutos de incubação antes da adição do diluente a base gema de ovo + etilenoglicol (EE15CCC) foi superior aos valores médios do tratamento EECCC na manutenção da integridade acrossomal, porém não diferiu dos valores do tratamento controle para este diluente (EE; P >0,05). No diluente Tris-Glicerol, os valores observados no tratamento controle foi superior (P<0,05) aos valores médios dos demais tratamentos (TGCCC e TG15CCC) na manutenção da integridade acrossomal. Houve correlação negativa entre o teste de ligação e a integridade do acrossoma (r= -0,25) e positiva entre o teste de ligação e a motilidade espermática progressiva (r= 0,20). A motilidade espermática progressiva e a integridade do acrossoma apresentaram correlação negativa (r= -0,26). Na avaliação quantitativa e qualitativa do colesterol pelas técnicas cromatográficas (gasosa e camada delgada) não se verificou diferença entre as amostras do sêmen nos diferentes tratamentos (P>0,05). Ambas as técnicas demonstraram que não houve incorporação do colesterol aos espermatozoides. Conclui-se que o complexo ciclodextrina-colesterol no meio diluidor não melhorou os aspectos físicos do sêmen caprino pós-descongelamento e a capacidade de ligação à membrana perivitelina da gema do ovo de galinha. A pré-incubação do sêmen com o CCC por 15 minutos antes da adição do diluente a base de Etilenoglicol+ gema de ovo (EE15CCC) não proporcionou um aumento na integridade do acrossoma a ponto de diferir dos valores do tratamento controle (EE). A cromatografia gasosa e de camada delgada demonstraram, respectivamente, um eficiente método quantitativo e qualitativo para determinar o colesterol presente no espermatozoide caprino criopreservados. A concentração de 1 mg do complexo ciclodextrina-colesterol adicionada ao sêmen caprino não foi eficaz em aumentar a concentração de colesterol presente no espermatozoide.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de ViçosaMestrado em Medicina VeterináriaUFVBRBiotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. deColesterolCiclodextrinaCromatografia de camada delgadaCholesterolCiclodextrinesThin-layer chromatographyCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMALInclusão de colesterol na membrana plasmática de espermatozoides caprinosInclusion of cholesterol to goat sperm membraneinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf765049https://locus.ufv.br//bitstream/123456789/5132/1/texto%20completo.pdfdcb0cc188aa67726f24e856817503e00MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain94584https://locus.ufv.br//bitstream/123456789/5132/2/texto%20completo.pdf.txtf99b7e3b6a8ef669ae03d96b06848e51MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3594https://locus.ufv.br//bitstream/123456789/5132/3/texto%20completo.pdf.jpg37ecf4dc41f2215473792649b6bb08e5MD53123456789/51322016-04-11 23:10:36.946oai:locus.ufv.br:123456789/5132Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-12T02:10:36LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Inclusão de colesterol na membrana plasmática de espermatozoides caprinos |
dc.title.alternative.eng.fl_str_mv |
Inclusion of cholesterol to goat sperm membrane |
title |
Inclusão de colesterol na membrana plasmática de espermatozoides caprinos |
spellingShingle |
Inclusão de colesterol na membrana plasmática de espermatozoides caprinos Silveira, Camila Oliveira Colesterol Ciclodextrina Cromatografia de camada delgada Cholesterol Ciclodextrines Thin-layer chromatography CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL |
title_short |
Inclusão de colesterol na membrana plasmática de espermatozoides caprinos |
title_full |
Inclusão de colesterol na membrana plasmática de espermatozoides caprinos |
title_fullStr |
Inclusão de colesterol na membrana plasmática de espermatozoides caprinos |
title_full_unstemmed |
Inclusão de colesterol na membrana plasmática de espermatozoides caprinos |
title_sort |
Inclusão de colesterol na membrana plasmática de espermatozoides caprinos |
author |
Silveira, Camila Oliveira |
author_facet |
Silveira, Camila Oliveira |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/2921617720071753 |
dc.contributor.author.fl_str_mv |
Silveira, Camila Oliveira |
dc.contributor.advisor-co1.fl_str_mv |
Costa, Eduardo Paulino da |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787237D6 |
dc.contributor.advisor-co2.fl_str_mv |
Queiroz, Maria Eliana Lopes Ribeiro de |
dc.contributor.advisor-co2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781671U3 |
dc.contributor.advisor1.fl_str_mv |
Guimarães, José Domingos |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U6 |
dc.contributor.referee1.fl_str_mv |
Torres, Ciro Alexandre Alves |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787213D4 |
dc.contributor.referee2.fl_str_mv |
Pinho, Rogério Oliveira |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4734857T6 |
contributor_str_mv |
Costa, Eduardo Paulino da Queiroz, Maria Eliana Lopes Ribeiro de Guimarães, José Domingos Torres, Ciro Alexandre Alves Pinho, Rogério Oliveira |
dc.subject.por.fl_str_mv |
Colesterol Ciclodextrina Cromatografia de camada delgada |
topic |
Colesterol Ciclodextrina Cromatografia de camada delgada Cholesterol Ciclodextrines Thin-layer chromatography CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL |
dc.subject.eng.fl_str_mv |
Cholesterol Ciclodextrines Thin-layer chromatography |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::REPRODUCAO ANIMAL |
description |
The aim of this study was to evaluate the fertilizing capacity, acrosomal integrity, and qualify and quantify by chromatographic techniques the incorporation of cholesterol to the sperm membrane by cyclodextrin in different diluents on cryopreservation of goat sperm. Four males, Saanen (2) and Parda Alpine (2) breeds were used. It was performed a completely randomized design, with each semen sample divided into the following treatments: TG - Tris-glycerol diluent; TGCCC - Cyclodextrin-cholesterol complex (CCC) + Tris-glycerol diluent; TG15CCC - CCC diluted in isosmotic solution (saline) to semen with 15 minutes of incubation before addition of the Tris-glycerol diluent; EE - egg yolk + ethylene glycol diluent; EECCC - CCC + egg yolk + ethylene glycol diluent; EE15CCC - CCC diluted in isosmotic solution (saline) to semen with 15 minutes of incubation before addition of the egg yolk + ethylene glycol diluent. Fresh semen, after its examination, was submitted to the cryopreservation process and stored in a cryogenic cylinder for 10 days. After thawing, the following analysis were performed: acrosomal integrity, sperm fertilizing capacity through perivitelline membrane of hen egg yolk binding test (MPEY) and analysis of sperm motility and vigor. Besides, techniques of gas and thin layer chromatography were conducted to evaluate the incorporation of cholesterol to the sperm membrane. Data were submitted to Lilliefors and Cochran and Bartlett tests to verify normality and homogeneity of variances, respectively. The characteristics that met the assumptions of these tests were submitted to ANOVA and means were compared by Duncan s test at 5% of probability. When data did not meet the assumptions of normality and homogeneity of variances, the means were compared by Kruskal-Wallis test. Pearson s correlation coefficient was calculated in all features.Addition of cyclodextrin-cholesterol complex did not increase binding of sperm to MPEY (P > 0.05). The EE15CCC treatment was superior to EECCC treatment in maintaining the acrosomal integrity but did not differ from control (EE; P > 0.05). In Tris-glycerol diluent, the values observed in the control treatment were higher (P < 0.05) than values of the other treatments (TGCCC and TG15CCC) in maintaining the acrosomal integrity. There was a negative correlation between the binding assay and acrosomal integrity (r = -0.25) and positive correlation between the binding assay and sperm motility (r = 0.20). The sperm motility and acrosomal integrity were negatively correlated (r = -0.26). In quantitative and qualitative evaluation of cholesterol by chromatographic techniques (gas and thin layer) there was no difference between the samples of semen in different treatments (P > 0.05). Both techniques showed no incorporation of cholesterol to spermatozoa. It was concluding that addition of cholesterol-cyclodextrin complex to the medium did not improve the physical aspects of goat semen, or the sperm binding capacity. Pre-incubation of semen with CCC for 15 minutes before addition of ethylene + egg yolk diluent (EE15CCC) did not enhance the integrity of the acrosome in relation to control (EE). The gas and thin layer chromatography showed, respectively, an efficient method for quantitatively and qualitatively determining the cholesterol present in the cryopreserved goat sperm. The concentration of 1 mg of cholesterol-cyclodextrin complex added to the goat semen was not effective for increasing the concentration of cholesterol in sperm. |
publishDate |
2013 |
dc.date.available.fl_str_mv |
2013-06-25 2015-03-26T13:47:12Z |
dc.date.issued.fl_str_mv |
2013-02-22 |
dc.date.accessioned.fl_str_mv |
2015-03-26T13:47:12Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
SILVEIRA, Camila Oliveira. Inclusion of cholesterol to goat sperm membrane. 2013. 54 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2013. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/5132 |
identifier_str_mv |
SILVEIRA, Camila Oliveira. Inclusion of cholesterol to goat sperm membrane. 2013. 54 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2013. |
url |
http://locus.ufv.br/handle/123456789/5132 |
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Universidade Federal de Viçosa |
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Mestrado em Medicina Veterinária |
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UFV |
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BR |
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Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de |
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Universidade Federal de Viçosa |
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