Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-09-08T11:57:20Z No. of bitstreams: 1 KarinaRibeiroModestoDissertacao2017.pdf: 6705375 bytes, checksum: 161944cc6fbeb260f63db5fea9e61056 (MD5) / Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-09-08T11:57:33Z (GMT) No. of bitstreams: 1 KarinaRibeiroModestoDissertacao2017.pdf: 6705375 bytes, checksum: 161944cc6fbeb260f63db5fea9e61056 (MD5) / Made available in DSpace on 2017-09-08T11:57:33Z (GMT). No. of bitstreams: 1 KarinaRibeiroModestoDissertacao2017.pdf: 6705375 bytes, checksum: 161944cc6fbeb260f63db5fea9e61056 (MD5) Previous issue date: 2017-03-31 / Recent discoveries points towards seminal plasma playing a major role in directing processes on mammalian spermatozoa as well as influencing offspring development prior to and after birth. Amongst the candidates for such role, recent data has brought the extracellular vesicles (EVs) into the field. Aiming to characterize EVs present on the intraluminal fluid of different segments of the epididymis, such as cauda and caput in male Wistar rats, this research assessed the influence of sexual developmental stages on the concentration and size distribution of epididymal EVs. The individuals were also grouped in terms of sexual developmental stages, clustered as ???prior to puberty, ???puberty???, and ???post-puberty???. TRPS based analysis showed a significant shifts in concentration and diameter of such EVs between clustering groups as well as epididymal segment groups. Caput epididymal EVs were found presenting reduced concentration and reduced diameter when compared to cauda epididymis EVs. Regarding the more sexually developed individuals, those showed higher EVs counts than ???prior to puberty??? ones. Additional findings suggest that post-puberty epididymal EVs are enlarged in comparison to the two other groups. Such data might serve a purpose of better comprehending the processes regarding spermatic maturation and capacitation (fertilization), offering findings that bring up clinically important issues regarding male epigenesis and father-to-offspring effects. / Evid??ncias crescentes t??m apontado para a import??ncia do conte??do presente no plasma seminal de mam??feros sobre as c??lulas esperm??ticas bem como sobre o desenvolvimento da prole durante os per??odos pr?? e p??s-natal. Entre os mediadores aparentemente envolvidos nesse processo, podemos citar as ves??culas extracelulares (VEs). Com o objetivo de caracterizar as VEs presentes no l??quido intraluminal de diferentes compartimentos do epid??dimo, cabe??a e cauda, de ratos machos da linhagem Wistar, o presente trabalho avaliou a influ??ncia da idade sobre a concentra????o e a distribui????o de tamanho dessas ves??culas nos compartimentos do epid??dimo. Para isso, utilizou-se tr??s grupos de ratos em idades distintas, sendo um grupo com indiv??duos em idade pr??-p??bere, um grupo com indiv??duos em idade p??bere e um grupo com indiv??duos em idade p??s-p??bere. A contagem de part??culas por detec????o de pulso resistivo ajust??vel (TRPS) revelou diferen??as importantes na concentra????o e no di??metro dessas VEs, tanto em fun????o do segmento anat??mico epididim??rio, quanto em fun????o da idade dos indiv??duos. A cabe??a do epid??dimo apresentou menor concentra????o de part??culas em compara????o com a cauda, bem como part??culas com di??metro menor. Com rela????o a idade, os ratos mais velhos apresentaram maior concentra????o de VEs quando comparados com os ratos imaturos sexualmente. Al??m disso, foi observado que as VEs presentes no ducto epididim??rio dos ratos p??s-p??beres possui tamanho maior que as VEs dos outros dois grupos experimentais. Os resultados aqui obtidos podem servir para uma melhor compreens??o dos processos que englobam tanto a matura????o e capacita????o esperm??tica em si, oferecendo propriedades clinicamente relevantes, quanto processos da espermatog??nese masculina e poss??veis efeitos epigen??ticos transgeracionais.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-02-17T22:37:29Z No. of bitstreams: 1 ThiberioDeSouzaCastelo_TESE.pdf: 2616733 bytes, checksum: 7ec74d06a9aa91a8661ccee8c3a034fe (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-02-19T22:29:56Z (GMT) No. of bitstreams: 1 ThiberioDeSouzaCastelo_TESE.pdf: 2616733 bytes, checksum: 7ec74d06a9aa91a8661ccee8c3a034fe (MD5) / Made available in DSpace on 2016-02-19T22:29:56Z (GMT). No. of bitstreams: 1 ThiberioDeSouzaCastelo_TESE.pdf: 2616733 bytes, checksum: 7ec74d06a9aa91a8661ccee8c3a034fe (MD5) Previous issue date: 2015-02-26 / O objetivo foi estabelecer protocolos para obten??o e conserva??o de espermatozoides de cutias (Dasyprocta leporina) criadas em cativeiros no semi-?rido brasileiro, visando sua explora??o sustent?vel. Para tanto tr?s experimentos foram realizados. No primeiro, foi avaliada a influ?ncia da intera??o entre dois tipos de sonda (ondas senoidais e quadr?ticas) e dois protocolos de estimula??o (cont?nuo e seriado) sobre a efici?ncia da colheita de s?men de cutias por eletroejacula??o. A intera??o mais eficiente na obten??o de ejaculados com espermatozoides foi a que utilizou eletrodos em aneis associado com est?mulos em s?rie (4/7; 57%, P<0,05). No segundo, foi realizada a compara??o da a??o crioprotetora de diferentes subst?ncias (glicerol, etilenoglicol, dimetilsulf?xido, dimetilformamida) na criopreserva??o de espermatozoides epididim?rios. Relativo a isso, os maiores valores de motilidade (39,5?4,6%), vigor (2,9?0,2) e integridade de membrana (30,6?4,5%) foram obtidos nas amostras criopreservadas com glicerol quando comparada as amostras contendo etilenoglicol e dimetilformamida, contudo, sem diferen?a (P>0,05) quando comparada as criopreservadas com dimetilsulfoxido. Finalmente, foi analisado o efeito dos m?todos de obten??o de espermatozoides (eletroejacula??o vs colheita epididim?ria) sobre a qualidade dos espermatoz?ides criopreservados. Como resultado as amostras obtidas por lavagem epididm?ria apresentaram motilidade de 25,0?10,9% e vigor 2,4?0,8 e as obtidas por eletroejacula??o obtiveram 31,2?14,2% de motilidade e 2,2?0,7 de vigor, por?m sem diferen?as significativas (P>0,05), denotando a possibilidade de aplicar diretamente, com sucesso, o protocolo de criopreserva??o de espermatozoides epididimarios de cutias (Dasyprocta leporina) para ejaculados da mesma esp?cie. Na presente tese, foram alcan?ados avan?os significativos na obten??o e processamento de espermatozoides de cutias, possibilitando a forma??o de bancos de germoplasmas a partir de amostras oriundas tanto dos epid?dimos quanto por eletroejacula??o. / The general objective of this thesis was to stablish protocols to obtain and conserve agouti (Dasyprocta leporina) sperm bred in captivity in the Brazilian semi-arid, aiming its sustainable production. The thesis was divided in three experiments. In the first one, we studied the influence of the interaction between two probes (quadratics and sine waves) and two stimulation protocols (continuous and in series) on the agouti sperm collection by electroejaculation efficiency. The most efficient interaction on this obtainment was the one with probes with rings associated with stimuli in series (4/7; 57%, P<0.05). In the second experiment we compared the cryoprotectant effects of different substances (glycerol, ethyleneglycol, dimethylsulfoxide, dimethylformamide) on epididymis sperm cryopreservation. The highest values on motility (39.5?4.6%), vigor (2.9?0.2) and membrane integrity (30.6?4.5%) were observed on the samples cryopreserved using glycerol when compared to those with ethyleneglycol and dimethylformamide, but there was no difference (P>0.05) when compared to the samples cryopreserved with dimethylsulfoxide. At last, we studied the effects of the methods to obtain sperm (electroejaculation vs epididymal collection) on post thawing sperm quality. The samples obtained by epididymal retrograde flushing showed values for motility of 25.0?10.9% and vigor 2.4?0.8, and those obtained by electroejaculation had 31.2?14.2% of motility and vigor of 2.2?0.7, however, without statistical difference (P>0.05), which shows the possibility to successfully use the epididymal sperm cryopreservation protocol on agouti ejaculated sperm. In conclusion, significant advances on obtainment and processing of agouti sperm were made, allowing the establishment of germplasm banks from sperm samples obtained from the epididymis or by electroejaculation.