Comportamento espermatogonial em ratos após irradiação e submetidos à supressão hormonal

Detalhes bibliográficos
Autor(a) principal: Amanda Vasconcelos de Albuquerque
Data de Publicação: 2013
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFMG
Texto Completo: http://hdl.handle.net/1843/BUBD-9C2GSR
Resumo: Ionizing radiation has been shown to arrest spermatogenesis despite the presence of surviving stem spermatogonia, by blocking their differentiation. This block is a result of damage to the somatic environment and is reversed when gonadotropins and testosterone are suppressed, but the mechanisms are still unknown. We examined spermatogonial differentiation and Sertoli cell factors that regulate spermatogonia after irradiation, during hormone suppression, and after hormone suppression combined with Leydig cell elimination with EDS. The results showed that the numbers and cytoplasmic structure of Sertoli cells are unaffected by irradiation; that only a few Aund spermatogonia and even fewer A1 spermatogonia remained and that immunohistochemical analysis showed that Sertoli cells still produced KITG and GNDF. Some of these cells expressed KIT-receptor, demonstrating that the failure of differentiation was not a result of the absence of the KIT system. Hormone suppression resulted in an increase in Aund spermatogonia within 3 days, a gradual increase in KIT-positive spermatogonia, and differentiation mainly to A3 spermatogonia after 2-weeks. KITLG protein expression did not change after hormone suppression indicating that it is not a factor in the stimulation. However, GDNF increased steadily after hormone suppression, which was unexpected since GDNF is supposed to promote stem spermatogonial self-renewal and not differentiation We conclude that the primary cause of block in spermatogonial development is not due to Sertoli cell factors such (KITLG\GDNF) or the KIT receptor. Since elimination of Leydig cells in addition to hormone suppression resulted in differentiation to the A3 stage within 1 week, Leydig cell factors were not necessary for spermatogonial differentiation.
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spelling Comportamento espermatogonial em ratos após irradiação e submetidos à supressão hormonalBiologia CelularLeydig, Células deEspermatogôniasBiologia celularEspermatogeneseIonizing radiation has been shown to arrest spermatogenesis despite the presence of surviving stem spermatogonia, by blocking their differentiation. This block is a result of damage to the somatic environment and is reversed when gonadotropins and testosterone are suppressed, but the mechanisms are still unknown. We examined spermatogonial differentiation and Sertoli cell factors that regulate spermatogonia after irradiation, during hormone suppression, and after hormone suppression combined with Leydig cell elimination with EDS. The results showed that the numbers and cytoplasmic structure of Sertoli cells are unaffected by irradiation; that only a few Aund spermatogonia and even fewer A1 spermatogonia remained and that immunohistochemical analysis showed that Sertoli cells still produced KITG and GNDF. Some of these cells expressed KIT-receptor, demonstrating that the failure of differentiation was not a result of the absence of the KIT system. Hormone suppression resulted in an increase in Aund spermatogonia within 3 days, a gradual increase in KIT-positive spermatogonia, and differentiation mainly to A3 spermatogonia after 2-weeks. KITLG protein expression did not change after hormone suppression indicating that it is not a factor in the stimulation. However, GDNF increased steadily after hormone suppression, which was unexpected since GDNF is supposed to promote stem spermatogonial self-renewal and not differentiation We conclude that the primary cause of block in spermatogonial development is not due to Sertoli cell factors such (KITLG\GDNF) or the KIT receptor. Since elimination of Leydig cells in addition to hormone suppression resulted in differentiation to the A3 stage within 1 week, Leydig cell factors were not necessary for spermatogonial differentiation.Utilizando-se ratos como modelo experimental sabe-se que, após a irradiação, as espermatogônias remanescentes ficam bloqueadas, não se diferenciam e que esse bloqueio pode ser revertido mediante a supressão da testosterona. No entanto, não se sabia ao certo como ocorre a cinética espermatogonial no momento do bloqueio pós-irradiação e ao longo da supressão hormonal, a participação das células de Sertoli e de Leydig nesse processo e principalmente quais eventos moleculares estão diretamente envolvidos durante o desbloqueio. Foram utilizados ratos LBNF1 irradiados com uma dose suficiente para destruir a espermatogênese e em seguida tratados com uma mistura de análogo de GnRH (comercialmente chamado Acyline) + flutamida, associada ou não ao EDS (sulfonato dimetano etano), droga que elimina especificamente as células de Leydig. O presente trabalho demonstrou, utilizando diferentes técnicas de microscopia e diferentes imunoensaios, que as espermatogônias indiferenciadas e remanescentes no epitélio seminífero após a irradiação são aparentemente saudáveis e ocasionalmente podem vencer o bloqueio e se diferenciar em A1. Esse dado foi confirmado pela presença de células KIT positivas após irradiação em animais que não receberam supressão hormonal. Os níveis de GDNF (fator neurotrófico derivado de célula glial), proteína sintetizada pela célula de Sertoli, são alterados após 4 semanas de supressão da testosterona, coincidindo com o elevado número de espermatogônias indiferenciadas que se dividiram nesse mesmo período, possivelmente para repor o estoque perdido pós-irradiação. Entretanto, outra proteína também sintetizada pela célula de Sertoli, KITLG, não apresentou qualquer alteração durante a supressão hormonal. A morte das células de Leydig, no entanto, interferiu diretamente na taxa de diferenciação das espermatogônias, pois a adição de EDS antecipou em uma semana o aparecimento das espermatogônias que foram obtidas após supressão hormonal por duas semanas e sem o EDS. Assim, nós concluímos que as células de Leydig, muito mais do que as células de Sertoli, são influenciadoras diretas sobre o desbloqueio da diferenciação espermatogonial pós-irradiação.Universidade Federal de Minas GeraisUFMGHelio Chiarini GarciaMarvin L. MeistrichAugusto Barbosa ReisElizete RizzoGuilherme Mattos Jardim CostaHugo Pereira GodinhoAmanda Vasconcelos de Albuquerque2019-08-10T17:42:11Z2019-08-10T17:42:11Z2013-07-30info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/1843/BUBD-9C2GSRinfo:eu-repo/semantics/openAccessporreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMG2019-11-14T11:33:28Zoai:repositorio.ufmg.br:1843/BUBD-9C2GSRRepositório InstitucionalPUBhttps://repositorio.ufmg.br/oairepositorio@ufmg.bropendoar:2019-11-14T11:33:28Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
dc.title.none.fl_str_mv Comportamento espermatogonial em ratos após irradiação e submetidos à supressão hormonal
title Comportamento espermatogonial em ratos após irradiação e submetidos à supressão hormonal
spellingShingle Comportamento espermatogonial em ratos após irradiação e submetidos à supressão hormonal
Amanda Vasconcelos de Albuquerque
Biologia Celular
Leydig, Células de
Espermatogônias
Biologia celular
Espermatogenese
title_short Comportamento espermatogonial em ratos após irradiação e submetidos à supressão hormonal
title_full Comportamento espermatogonial em ratos após irradiação e submetidos à supressão hormonal
title_fullStr Comportamento espermatogonial em ratos após irradiação e submetidos à supressão hormonal
title_full_unstemmed Comportamento espermatogonial em ratos após irradiação e submetidos à supressão hormonal
title_sort Comportamento espermatogonial em ratos após irradiação e submetidos à supressão hormonal
author Amanda Vasconcelos de Albuquerque
author_facet Amanda Vasconcelos de Albuquerque
author_role author
dc.contributor.none.fl_str_mv Helio Chiarini Garcia
Marvin L. Meistrich
Augusto Barbosa Reis
Elizete Rizzo
Guilherme Mattos Jardim Costa
Hugo Pereira Godinho
dc.contributor.author.fl_str_mv Amanda Vasconcelos de Albuquerque
dc.subject.por.fl_str_mv Biologia Celular
Leydig, Células de
Espermatogônias
Biologia celular
Espermatogenese
topic Biologia Celular
Leydig, Células de
Espermatogônias
Biologia celular
Espermatogenese
description Ionizing radiation has been shown to arrest spermatogenesis despite the presence of surviving stem spermatogonia, by blocking their differentiation. This block is a result of damage to the somatic environment and is reversed when gonadotropins and testosterone are suppressed, but the mechanisms are still unknown. We examined spermatogonial differentiation and Sertoli cell factors that regulate spermatogonia after irradiation, during hormone suppression, and after hormone suppression combined with Leydig cell elimination with EDS. The results showed that the numbers and cytoplasmic structure of Sertoli cells are unaffected by irradiation; that only a few Aund spermatogonia and even fewer A1 spermatogonia remained and that immunohistochemical analysis showed that Sertoli cells still produced KITG and GNDF. Some of these cells expressed KIT-receptor, demonstrating that the failure of differentiation was not a result of the absence of the KIT system. Hormone suppression resulted in an increase in Aund spermatogonia within 3 days, a gradual increase in KIT-positive spermatogonia, and differentiation mainly to A3 spermatogonia after 2-weeks. KITLG protein expression did not change after hormone suppression indicating that it is not a factor in the stimulation. However, GDNF increased steadily after hormone suppression, which was unexpected since GDNF is supposed to promote stem spermatogonial self-renewal and not differentiation We conclude that the primary cause of block in spermatogonial development is not due to Sertoli cell factors such (KITLG\GDNF) or the KIT receptor. Since elimination of Leydig cells in addition to hormone suppression resulted in differentiation to the A3 stage within 1 week, Leydig cell factors were not necessary for spermatogonial differentiation.
publishDate 2013
dc.date.none.fl_str_mv 2013-07-30
2019-08-10T17:42:11Z
2019-08-10T17:42:11Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/1843/BUBD-9C2GSR
url http://hdl.handle.net/1843/BUBD-9C2GSR
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Minas Gerais
UFMG
publisher.none.fl_str_mv Universidade Federal de Minas Gerais
UFMG
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFMG
instname:Universidade Federal de Minas Gerais (UFMG)
instacron:UFMG
instname_str Universidade Federal de Minas Gerais (UFMG)
instacron_str UFMG
institution UFMG
reponame_str Repositório Institucional da UFMG
collection Repositório Institucional da UFMG
repository.name.fl_str_mv Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)
repository.mail.fl_str_mv repositorio@ufmg.br
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