El propósito primario de cualquier organismo superior con reproducción sexual es pasar sus genes a la siguiente generación vía sus células germinales primordiales (PGCs), células troncales del sistema reproductor. Al transplantar intraespecíficamente las células germinales, éstas colonizan la membrana basal de los túbulos seminíferos del animal receptor en busca del nicho donde se desarrollan las células madre. Es así que la técnica de transplante de PGCs permite que las células madres de origen fetal sean conservadas y divididas en su estado multipotente por largos periodos de tiempo en el animal receptor, preservando el material genético. Así, el presente trabajo de tesis tiene por objetivo evaluar la colonización intraluminal de una suspensión cruda de PGCs de fetos de ratón de la cepa Balb C en los túbulos seminíferos de ratones adultos. Para ello se aislaron las PGCs desde las crestas genitales de fetos machos con dos pasos de digestiones enzimáticas (Tripsina 0.25% y colagenasa tipo IV, 1 mg/ml), transplantándose en los túbulos seminíferos del animal receptor vía rete testis, a los ratones receptores previamente se les disminuyó drásticamente su línea germinal con tratamiento químico con Busulfán (40 mg/kg). Los animales receptores fueron mantenidos por 54 días en un bioterio con 14 horas luz y 10 horas oscuridad. Luego de este periodo los animales fueron sacrificados por dislocación cervical, se les extrajo los testículos y se realizó cortes histológicos seriados de 10 _m de espesor, coloreados con hematoxilina y eosina Y. La evaluación evidenció una colonización intraluminal en 1 de cada 100 túbulos seminíferos evaluados. Lo que demostraría que las PGCs de fetos de 14.5 días post coitum (dpc) si tienen la capacidad de colonizar los túbulos seminíferos de individuos adultos de la misma especie. / --- The aim function of a organism with sexual reproduction is to transmit their genes to the next generation throught their primordial germ cell (PGCs), stem cells of the reproductor system, they colonized basal membrane of seminiferous tubule in the receptor animal, search their nich where they can differentiate to sperms. In the Transplant PGCs Technique, we could preserved and multiplicated cells for long time in their multipotent state in the receptor animal, conserved in their multipotent state. The present Thesis objective is to evaluate the intraluminal colonization of a suspension of PGCs of fetus mice Balb C in semiferous tubule of adult mice. In my research, PGCs were isolated from fetus male mice with two enzimatic digestion steps (Tripsin 0.25% and Colagenase Type IV 1 mg/ml), PGCs were transplanted into the rete testis of the receptor animal that previously were injected with Busulfán (40 mg/kg) to decrease their own spermatogenesis. Mice receptor were maintained in bioterio conditions with 14 hours light and 10 hours dark for fifty four days, they were sacrificed by cervical dislocation, their testis were extracted for and histology procedures of 10 _m, colored with hematoxilin and Y eosin. In this research the intraluminal colonization was identified in 1 of 100 seminiferous tubule. In conclusion, transplantation of PGCs of foetus from 14.5 dpc can colonized the seminiferous tubule.

El propósito primario de cualquier organismo superior con reproducción sexual es pasar sus genes a la siguiente generación vía sus células germinales primordiales (PGCs), células troncales del sistema reproductor. Al transplantar intraespecíficamente las células germinales, éstas colonizan la membrana basal de los túbulos seminíferos del animal receptor en busca del nicho donde se desarrollan las células madre. Es así que la técnica de transplante de PGCs permite que las células madres de origen fetal sean conservadas y divididas en su estado multipotente por largos periodos de tiempo en el animal receptor, preservando el material genético. Así, el presente trabajo de tesis tiene por objetivo evaluar la colonización intraluminal de una suspensión cruda de PGCs de fetos de ratón de la cepa Balb C en los túbulos seminíferos de ratones adultos. Para ello se aislaron las PGCs desde las crestas genitales de fetos machos con dos pasos de digestiones enzimáticas (Tripsina 0.25% y colagenasa tipo IV, 1 mg/ml), transplantándose en los túbulos seminíferos del animal receptor vía rete testis, a los ratones receptores previamente se les disminuyó drásticamente su línea germinal con tratamiento químico con Busulfán (40 mg/kg). Los animales receptores fueron mantenidos por 54 días en un bioterio con 14 horas luz y 10 horas oscuridad. Luego de este periodo los animales fueron sacrificados por dislocación cervical, se les extrajo los testículos y se realizó cortes histológicos seriados de 10 _m de espesor, coloreados con hematoxilina y eosina Y. La evaluación evidenció una colonización intraluminal en 1 de cada 100 túbulos seminíferos evaluados. Lo que demostraría que las PGCs de fetos de 14.5 días post coitum (dpc) si tienen la capacidad de colonizar los túbulos seminíferos de individuos adultos de la misma especie. / The aim function of a organism with sexual reproduction is to transmit their genes to the next generation throught their primordial germ cell (PGCs), stem cells of the reproductor system, they colonized basal membrane of seminiferous tubule in the receptor animal, search their nich where they can differentiate to sperms. In the Transplant PGCs Technique, we could preserved and multiplicated cells for long time in their multipotent state in the receptor animal, conserved in their multipotent state. The present Thesis objective is to evaluate the intraluminal colonization of a suspension of PGCs of fetus mice Balb C in semiferous tubule of adult mice. In my research, PGCs were isolated from fetus male mice with two enzimatic digestion steps (Tripsin 0.25% and Colagenase Type IV 1 mg/ml), PGCs were transplanted into the rete testis of the receptor animal that previously were injected with Busulfán (40 mg/kg) to decrease their own spermatogenesis. Mice receptor were maintained in bioterio conditions with 14 hours light and 10 hours dark for fifty four days, they were sacrificed by cervical dislocation, their testis were extracted for and histology procedures of 10 _m, colored with hematoxilin and Y eosin. In this research the intraluminal colonization was identified in 1 of 100 seminiferous tubule. In conclusion, transplantation of PGCs of foetus from 14.5 dpc can colonized the seminiferous tubule.

Orientador: Selma Maria de Almeida-Santos / Resumo: O conjunto de características manifestadas por uma espécie que levam ao sucesso reprodutivo pode ser chamado de estratégias reprodutivas. Isto inclui, por exemplo, o período reprodutivo, o modo reprodutivo, a sincronia reprodutiva entre machos e fêmeas, o tempo de fertilização e a frequência reprodutiva. O gênero Lachesis é composto de quatro espécies ovíparas restritas às florestas úmidas da América Central e América do Sul. Popularmente conhecidas como surucucu, são um grupo de importância médica, tal como Bothrops e Crotalus. Lachesis muta é a espécie com a maior distribuição, e ocorre na América no Sul, na Mata Atlântica e na Floresta Amazônica. Ao contrário de Bothrops e Crotalus, estudos sobre biologia reprodutiva para o gênero Lachesis são, em sua maioria, desenvolvidos em cativeiro e com foco no comportamento reprodutivo. Propomos, então, descrever em dois capítulos aspectos da biologia reprodutiva da espécie Lachesis muta investigados a partir da anatomia e morfologia de seus órgãos reprodutivos ao longo das estações do ano. No Capítulo 1, investigamos a biologia reprodutiva de L. muta da Floresta Amazônica e da Mata Atlântica brasileira a partir de dados de biometria e histologia de animais preservados em coleções biológicas e provenientes da natureza. No Capítulo 2, descrevemos o oviduto feminino em diferentes estágios do ciclo reprodutivo. Comparando tanto as duas populações quanto a espécie com outros crotalíneos neotropicais, nosso objetivo foi investigar possív... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The set of characteristics manifested by a species leading to reproductive success can be called reproductive strategies. These include, for example, the reproductive period, the reproductive mode, the reproductive synchrony between males and females, the time of fertilization, and the reproductive frequency. The genus Lachesis comprises four oviparous species restricted to Central and South American rainforests and popularly known as bushmasters, and is a group of medical importance, as are Bothrops and Crotalus. Lachesis muta is the most widely distributed and occurs in South America, at the Atlantic Forest and the Amazon Rainforest.Unlike Bothrops and Crotalus, reproductive biology studies for the genus Lachesis weremostly developed in captivity, focusing mainly on reproductive behavior. We propose to describe in two chapters aspects of the reproductive biology of the Lachesis muta species investigated from the anatomy and morphology of its reproductive organs over the seasons of the year. In Chapter 1, we investigate the reproductive biology of L. muta by biometric and histology data from biological collections, focusing on individuals collected in the wild. In Chapter 2, we describe the female oviduct at different stages of the reproductive cycle. Comparing both populations and species with the other neotropical crotalines, we aim to investigate possibles patterns for the Viperidae group, as well as the hypothesis of the influence of abiotic factors on the snake reproduc... (Complete abstract click electronic access below) / Mestre