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Impacto da idade e do segmento epididim??rio na concentra????o e na distribui????o de tamanho de ves??culas extracelularesModesto, Karina Ribeiro 31 March 2017 (has links)
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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.
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Obten??o e conserva??o de espermatozoides de cutia (Dasyprocta leporina Linneaus, 1753) do semi?rido brasileiro / Obtainment and conservantion of agouti (Dasyprocta leporina) sperm from brazilian semi-aridCastelo, Thib?rio de Souza 26 February 2015 (has links)
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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.
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