Avaliação ultrassonográfica do aparelho reprodutor em serpentes vivíparas da família Boidae / Ultrasonographic evaluation of reproductive tract in viviparous snakes Boidae family

A reprodução faz parte do ciclo de vida dos animais permitindo a perpetuação e a conservação das espécies. Em serpentes, existe uma escassez de informações técnicas a respeito do ciclo reprodutivo. Este estudo teve o objetivo de avaliar o aparelho reprodutivo por meio da ultrassonografia em serpentes vivíparas cativas da família Boidae, permitindo diagnosticar as diferentes fases reprodutivas. Foram avaliadas ultrassonograficamente onze serpentes adultas de quatro espécies da família Boidae: Eunectes murinus, Boa constrictor constrictor, Corallus hortulanus e Epicrates cenchria pertencentes ao acervo do Museu Biológico do Instituto Butantan, São Paulo Brasil. Para a avaliação ultrassonográfica, as serpentes foram contidas fisicamente com gancho herpetológico e depois manualmente por aproximadamente 15 minutos. A avaliação foi feita aplicando-se gel acústico sobre a pele e posicionando o transdutor na linha lateral-ventral direita e esquerda, em região medial do corpo em sentido crânio-caudal. O exame ultrassonográfico permitiu avaliar todo o ciclo reprodutivo nas serpentes. Nas avaliações ultrassonográficas das fêmeas pode-se definir as fases de desenvolvimento ovariano e ovidutal. Os folículos ovarianos durante a fase pré-vitelogênica foram visualizados como homogêneos e anecogênicos, em forma de "cacho de uva". Já na fase vitelogênica, os folículos estavam maiores e mais ecogênicos seguidos uns dos outros, como um "colar de pérolas". Quando não houve cópula, os folículos foram reabsorvidos dentro do ovário retornando a fase pré-vitelogênica. Na fase pós ovulatória foram visualizados três estágios bem definidos de desenvolvimento fetal dentro do oviduto: 1) logo após a ovulação (e fecundação), somente o vitelo foi visualizado; 2) o vitelo ocupava 60% e o feto 40% do ovo e 3) o feto estava formado e não havia vitelo. Nos machos, os testículos foram visualizados como uma imagem homogênea e hipoecogênica quando se encontravam em estágio...

The reproduction is part of the animal life cycle allowing the perpetuation and conservation of the species. In snakes, there is a shortage of technical information about the reproductive cycle. The objective of this study was to evaluate the reproductive tract by ultrasonography in captive viviparous snakes of the Boidae family, allowing diagnose of the different reproductive stages. Eleven adult snakes of four species of the Boidae family were sonographically evaluated, Eunectes murinus, Boa constrictor constrictor, Corallus hortulanus and Epicrates cenchria belonging to the Biological Museum's collection of the Instituto Butantan, Sao Paulo Brazil. For the sonographic evaluation, snakes were contained physically with herpetologic hook and then manually for about 15 minutes. The evaluation was done by applying acoustic gel on the skin and positioning the transducer on the right and left side-ventral line, in the medial region of the body in the skull tail sense. Ultrasonography allowed the evaluation of the whole reproductive cycle in snakes. In sonographic evaluations of females were defined the stages of ovarian and ovidutal development. The ovarian follicles during the pre-vitellogenic phase were visualized as homogeneous and anecogenic, in a "bunch of grapes" distribution. In the vitellogenic stage follicles were larger and more echogenic, following each other as a "string of pearls". When there was no copula, the follicles were reabsorbed in the ovary returning to pre-vitellogenic phase. In the post ovulatory phase were seen three well-defined stages of fetal development within the oviduct: 1) just after ovulation (and fertilization), only the vitellus was visualized; 2) occupied 60% of the vitellus fetus and 40% egg and 3) the fetus was formed and no vitellus. In males, the testicles were seen as a homogeneous and hypoechoic image during the reproductive stage...

Growth differentiation factor 9 and bone morphogenetic protein 15 expression in previtellogenic oocytes and during early embryonic development of Yellow-tail Kingfish Seriola lalandi

BACKGROUND: During fish oocyte maturation, specific molecules are expressed and accumulated within oocyte until fertilization and embryo development. Special attention have been paid in members of the transforming growth factor (TGF-ß) superfamily; growth differentiation factor 9 (GDF9/gdf9) and bone morphogenetic protein 15 (BMP15/bmp15), which exert regulatory functions during oocyte maturation and follicle development. However, little attention has been paid to the involvement of these molecules during embryogenesis considering its importance for the formation of a good quality egg and subsequent embryo survival. The purpose of this study was to analyze the expression of gdf9 andbmp15 in previtellogenic oocytes and during early embryonic development in Seriola lalandi, a pelagic fish with increasing prospect for its aquaculture development, which however, show high mortality at embryo and larval stages. RESULTS: Through RT-qPCR it was found that gdf9 expression was higher in previtellogenic oocytes decreasing after ovulation. This expression profile agrees with its participation in early stages of the follicular development. The transcripts for bmp15 also showed the highest levels in previtellogenic oocytes, however this expression was lower than obtained with gdf9. Conversely, in recently spawned oocytes mRNA bmp15 levels were highest than observed to gdf9. This, is consequent with the main role proposed for this growth factor at the final fish oocyte maturation: avoid the ovulation of an immature oocyte. During embryo development, low levels of mRNA were detected to gdf9, with an increase in 48 H post-fertilization embryos. The bmp15 expression did not change throughout development and was higher than gdf9 at 16 cells, blastula and appearance embryos stages. CONCLUSIONS: Both (gdf9 and bmp15) expression profiles in previtellogenic oocytes and newly spawned eggs are consistent with the described functions for these growth factors in vertebrate ovarian physiology in early and late stages of the follicular development. So, these genes could be considered as quality biomarkers at these stages. However, further studies of these proteins throughout folliculogenesis, are necessaries to fully understand their functions during the oocyte formation. In addition, the persistent expression of these growth factors during development, allows us to speculate possible roles in embryonic processes, which must also be addressed.

Culex quinquefasciatus vitellogenesis: morphological and biochemical aspects

The vitellogenic process in Culex quinquefasciatus, which is triggered by a blood meal, involves the synthesis, distribution and storage of the nutrients necessary for embryo development. The fat body of an adult female Cx. quinquefasciatus revealed two cell types: large trophocytes and small, eosinophilic, "oenocyte-like" cells, which show no morphological changes throughout the gonotrophic cycle. Trophocytes, which only begin to synthesise vitellogenin (Vg) 12 h post-blood meal (PBM), undergo a series of morphological changes following engorgement. These changes include the expansion of the rough endoplasmic reticulum (RER) and Golgi complex, which are later destroyed by autophagosomes. At 84 h PBM, trophocytes return to their pre-engorgement morphology. The ovarian follicles of non-blood-fed Cx. quinquefasciatus contain a cluster of eight undifferentiated cells surrounded by follicular epithelium. After engorgement, the oocyte membrane facing the perioocytic space increases its absorptive surface by microvilli development; large amounts of Vg and lipids are stored between 24 and 48 h PBM. Along with yolk storage in the oocyte, follicular cells exhibit the development of RER cisternae and electron-dense granules begin to fill the perioocytic space, possibly giving rise to endochorion. Later in the gonotrophic cycle, electron-dense vesicles, which are possible exochorion precursors, fuse at the apical membrane of follicular cells. This fusion is followed by follicular cell degeneration.

Oogenesis and egg development in triatomines: a biochemical approach

Em triatomíneos, assim como em outros insetos, o acúmulo de vitelo é um processo no qual um tecido extraovariano, o corpo gorduroso, produz proteínas que são empacotadas no interior de um ovo. A principal proteína, sintetizada pelo corpo gorduroso, que é acumulada no interior de um ovócito, é a vitelogenina. Este processo é também conhecido por vitelogênese. Existem crescentes evidências em triatomíneos, que além do corpo gorduroso, o ovário também produz proteínas de vitelo. A forma como estas proteínas de vitelo entram nos ovócitos será aqui comentada. O vitelo é um material complexo composto por proteínas, lipídeos, carboidratos e outros compostos minoritários que são empacotados de uma maneira organizada no interior dos ovócitos. A fertilização dispara a embriogênese, um processo que culmina com o desenvolvimento do embrião. Durante a embriogênese o vitelo será utilizado para a construção de um novo indivíduo, a ninfa de primeiro estádio. O desafio para a próxima década é entender onde e como estas proteínas de vitelo são utilizadas junto com os seus componentes não protéicos, em compasso com o programa genético do embrião, que comanda a diferenciação celular (fase inicial da embriogênese) e diferenciação do embrião (fase final da embriogênese) no interior do ovo.

Seasonal changes in the ovary of a freshwater murrel Channa punctatus (Bloch)

Foram descritas as mudanças histomorfológicas sazonais no ovário de Channa punctatus. O ovário é do tipo assincrônico e o ciclo ovariano apresenta 2 estaçöes no ano (uma no começo de abril e outra de agosto a setembro) nas quais há liberaçäo de ovócitos. O ciclo ovariano anual foi dividido em 5 fases : I) Fase de repouso (fim de novembro a dezembro e início de maio); II) Fase preparatória (janeiro a fevereiro e fim de maio a junho); III) Fase de préovulaçäo (março a julho); IV) Fase de ovulaçäo (começo de abril e agosto a setembro); V) Fase pós-ovulaçäo (outubro ao começo de novembro e fim de abril). As novas populaçöes de ovogônias säo produzidas a partir das ovogônias residuais por multiplicaçäo. Durante a vitelogênese, dois tipos de estruturas foram observadas : vesículas de vitelo que apresentam natureza glicoproteinácea (PAS e Hg-BPB-positivas), e glóbulos de vitelo que säo Sudäo negro B e Hg-BPB-positivas, indicando natureza lipoproteinácea. Foram discutidos problemas relativos à origem das novas populaçöes de ovogônias, extrusöes nucleolares, "nucleos vitelínicos", deposiçäo e natureza dos materiais do vitelo

Vitellogenesis in Manduca sexta

Eggs of Manduca sexta contain four well-characterized protein derived from hemolymph: vitellogenin and lipophorin (very high density lipoproteins); microvitellogenin, a 26,000 dalton female-specific protein lacking lipid and carbohydrate, and insecticyanin, a blue biliprotein composed of four identical 22,000 dalton subunits. In addition, eggs contain a large store of triacyl glycerols. It has been shown that vitellogenin and lipophorin are actively taken up by follicles in vitro. The lipid components of these two proteins together account for only 10% of egg lipid. The follicle actively sequesters intact high density lipophorin, which, inside the oocyte, is stripped of much of its neutral lipid and two molecules of apolipophorin III. On the other hand, low density lipophorin donates diacylglycerol to the oocyte without its protein components being sequestered. Most of the egg lipid is transported from the fat body by a shuttle system involving low density lipophorin.