GnRH/GnIH e seus receptores no sistema olfato-retinal de zebrafish

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ciclo testicular e espermatogênese de Devario aequipinnatus (cypriniformes, cyprinidae) submetido a diferentes temperaturas

Pós-graduação em Ciência e Tecnologia Animal - FEIS

Ecologia reprodutiva do peixe-donzela, Stegastes fuscus cuvier, 1830 (osteichthyes: pomacentridae) em arrecifes rochosos da praia de Búzios, Rio Grande do Norte, Brasil

The reproductive ecology of fish plays a key role both for rational exploitation methods and for protective measures of natural stocks. The purpose of this study was to analyze the reproductive aspects of the damsel-fish, Stegastes fuscus, during October 2004 to September 2005, in the coastal rocky reefs of Búzios Beach, Nísia Floresta, RN. Fish were captured using hooks and hand nets, during low tide. Reproduction was determined using sexual ratio, mean length of first maturation (L50), absolute fecundity and macroscopic characteristics of gonads. The following parameters were related to gonadosomatic index (GSI): condition factor (CF), hepatosomatic index (HSI), rain fall and temperature. In relation to sex distribution, it was observed that 78% were females and 22% were males. The L50 was 6.2 cm for females and 7.0 for males. Average fecundity was 6832 oocytes. Results showed that S. fuscus had better body condition in the months prior to spawning, particularly during initial and intermediate stages of maturation. Five stages of gonadal maturation were identified through macroscopic analysis: immature, in maturation, mature, spent and resting. The HSI was inversely related to the GSI. This was possibly due to the reproductive cycle of this species which was associated to the dry period of this region. During this period, low rain fall and high temperatures provide an propitious reproductive condition for the study species

Ecologia reprodutiva do peixe-donzela, Stegastes fuscus cuvier, 1830 (osteichthyes: pomacentridae) em arrecifes rochosos da praia de Búzios, Rio Grande do Norte, Brasil

The reproductive ecology of fish plays a key role both for rational exploitation methods and for protective measures of natural stocks. The purpose of this study was to analyze the reproductive aspects of the damsel-fish, Stegastes fuscus, during October 2004 to September 2005, in the coastal rocky reefs of Búzios Beach, Nísia Floresta, RN. Fish were captured using hooks and hand nets, during low tide. Reproduction was determined using sexual ratio, mean length of first maturation (L50), absolute fecundity and macroscopic characteristics of gonads. The following parameters were related to gonadosomatic index (GSI): condition factor (CF), hepatosomatic index (HSI), rain fall and temperature. In relation to sex distribution, it was observed that 78% were females and 22% were males. The L50 was 6.2 cm for females and 7.0 for males. Average fecundity was 6832 oocytes. Results showed that S. fuscus had better body condition in the months prior to spawning, particularly during initial and intermediate stages of maturation. Five stages of gonadal maturation were identified through macroscopic analysis: immature, in maturation, mature, spent and resting. The HSI was inversely related to the GSI. This was possibly due to the reproductive cycle of this species which was associated to the dry period of this region. During this period, low rain fall and high temperatures provide an propitious reproductive condition for the study species

Biologia reprodutiva da Tainha, Mugil curema valenciennes, 1836 (osteichthyes: mugilidade) nas águas costeiras do Rio Grande do Norte

The marine fish white mullet, Mugil curema Valenciennes, 1836 (Osteichthyes: Mugilidae) exhibits a wide geographical distribution, being common in the Brazilian coast and is an important component of the artisanal fisheries. The objective of this study was to investigate the reproductive biology of M. curema in the coastal waters of Rio Grande do Norte. Fish samples were captured on a monthly basis during August, 2008 to July, 2009. The fish specimens were numbered, weighed, measured, dissected and their gonads were removed, weighed, their sex and gonadal development were identified. The length-weight relationship was determined for males and females. The sex ratio, the size at first gonadal maturation and gonadosomatic index (GSI) were calculated and ovarian development was investigated using macroscopic and histological techniques. The fecundity, spawning type and the reproductive period of the species were determined. A total of 366 specimens (186 males and 180 females) were captured. The sex ratio was 1:1 and the females were heavier than males. The estimated values of the angular coefficient for both sexes suggest that the species has isometric growth. The size at which 50% of the population began the process of maturation was 25.9 cm of total length for grouped sex. The macroscopic characteristics of the ovaries showed four stages of development: immature, maturing, mature and spent. However, the microscopic characteristics of the ovaries showed five stages of development: immature, early maturing, late maturing, mature and spent. The development of oocytes indicated five phases: Chromatin-nucleolus (phase I), initial perinucleolar (phase II), final perinuclear (phase II), formation of vitelline vesicle or yolk (phase III), vitellogenic (phase IV) and complete vitellogenesis (phase V). The species has a prolonged spawning period, with two peaks coinciding with the rainy season.

Morfologia do cromossomo Y, função gonadal e extragonadal em touros de raças sintéticas com alterações na qualidade do sêmen

O motivo deste estudo foi o percentual elevado de descarte de touros por alterações reprodutivas, precisamente qualidade de sêmen, que se apresenta em determinados sistemas produtivos nos quais estão inseridas as raças sintéticas. Através de cinco experimentos foi permitido observar o comportamento da espermatogênese de touros de raças sintéticas do nível cromossômico ao funcional do epitélio seminífero. O experimento I, avaliou por seis meses as características seminais de 12 touros, seis de uma raça pura e seis de uma sintética, contendo em cada grupo touros aptos e inaptos a reprodução classificados por qualidade de sêmen. Os animais do grupo sintético não apresentaram recuperação das características seminais durante o período de avaliação, como os puros, indicando um quadro degenerativo permanente neste conjunto de indivíduos. O experimento II buscou identificar uma relação entre a morfologia do cromossomo Y e tipos de cruzamentos, com a qualidade seminal de touros de duas raças sintéticas, Braford e Brangus-Ibagé. A relação da morfologia do Y com a condição reprodutiva não foi comprovada, no entanto considerando os tipos de cruzamentos para obtenção de touros 3/8, os cruzamentos que utilizam fêmea ¼ com macho ½ sangue filho de Nelore proporcionam um maior percentual de animais considerados inaptos ao exame de sêmen. Este experimento sugere medidas práticas para evitar o cruzamento que traz maiores prejuízos econômicos. O experimento III foi proposto para a avaliar a intensidade de redução de células espermáticas anômalas ao longo do epidídimo em touros de uma raça sintética. A redução da freqüência de gota citoplasmática proximal foi distinta entre os grupos de touros classificados quanto a morfologia espermática e entre as regiões do epidídimo, sendo portanto um indicador sensível da qualidade espermática e classificação da fertilidade potencial de animais de raças híbridas. O experimento IV avaliou a freqüência dos túbulos seminíferos nos diferentes estádios do ciclo espermatogênico em touros de duas raças sintéticas. Nas fases iniciais do ciclo espermatogênico, todos os touros apresentam gametogênese semelhante. Nas fases em que ocorrem as divisões meióticas, os touros inaptos apresentaram maior freqüência e a na fase de maturação das espermátides os touros aptos apresentaram maior freqüência. Estes resultados indicam que nos touros inaptos ocorre um bloqueio na fase das meioses, diminuindo a freqüência dos estádios de maturação das espermátides. O experimento V identificou a expressão diferencial de um fator de crescimento (transforming growth factor alpha- TGFα) no epitélio seminífero de touros Braford e Brangus-Ibagé, aptos e inaptos à reprodução, e também avaliou a freqüência de células de Sertoli nestes animais, como um estudo inicial do controle parácrino da espermatogênese. A maior freqüência da expressão de TGFα foi observada nos estádios I e III, e também na raça Brangus-Ibagé. O número médio das células de Sertoli foi semelhante entre estádios e raças dos touros. Em todos experimentos em que as raças Braford e Brangus-Ibagé foram confrontadas, os resultados obtidos foram diferentes. As maiores freqüências de alterações na qualidade seminal não podem ser extrapoladas para todas as raças sintéticas, cabendo a prerrogativa para cada raça de um estudo específico e ajustado às suas condições de criação.

Desenvolvimento gonadal de fêmeas de matrinxã, Brycon amazonicus, submetidas a restrição alimentar

O presente estudo avaliou o efeito de ciclos de restrição alimentar e realimentação (2/3 dias), aplicados durante seis meses antes da desova, no desenvolvimento gonadal de matrinxã. Na ocasião da desova, fêmeas alimentadas diariamente e submetidas ao regime alimentar experimental, selecionadas para a indução hormonal, foram sacrificadas para retirada das gônadas e do fígado, com os quais se calculou o IGS (índice gonadossomático) e o IHS (índice hepatossomático), sendo os ovários processados para análise histológica. Não houve alteração no peso relativo dos ovários e fígado, e o desenvolvimento gonadal não foi afetado pelo esquema alimentar. Os valores de IGS foram de 5,09±4,98% e 9,79±4,17% e os de IHS foram de 0,84±0,07% e 0,91±0,11%, para as fêmeas controle e experimentais, respectivamente, sem diferenças significativas entre os grupos. Os ovários de peixes dos dois grupos apresentaram as mesmas características do estádio maduro, com predominância de ovócitos na fase final de maturação, repletos de vitelo. O estudo indica que a restrição alimentar não afetou a preparação das fêmeas para a reprodução e que ciclos adequados de restrição e realimentação poderão ser aplicados na criação do matrinxã, assegurando menores custos de produção.

Gonadal function arrest in fillies after birth, as reflected by steroid serum concentrations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito de diferentes indutores hormonais sobre o processo de maturação final e ovulação em Leporinus macrocephalus

Pós-graduação em Aquicultura - FCAV

Avaliação da atividade gonadal de machos e fêmeas de pescada branca, Plagioscion squamosissimus (Heckel, 1840), (Pisces, Teleostei, Sciaenidae) no rio Pará (PA)

Fêmeas e machos adultos de P. squamosissimus (Pisces, Teleostei, Sciaenidae) foram coletados mensalmente no Rio Pará, que banha a ilha do Capim (PA) (S 010 34. 971`; W 0480 52.932`), durante o período de fevereiro de 2004 a fevereiro de 2005, correspondendo ao total de 234 espécimes. As gônadas foram coletadas, fixadas e processadas de acordo com os métodos usuais utilizados para processamento em parafina e análise em microscopia de luz. A espécie em estudo é uma das principais fontes de proteína animal para a população local, sendo capturada de forma intensa e ininterrupta ao longo do ano. Com base em informações dos pescadores locais, essa captura parece estar determinando uma aparente diminuição quantitativa e qualitativa em seus estoques locais. A espécie apresenta desova parcelada e o desenvolvimento gonadal foi caracterizado nos estádios de repouso, maturação, maduro e esvaziado ou semi-esvaziado. Gônadas maduras foram encontradas nos meses de dezembro, janeiro e julho. A análise de correlação entre o estádio gonadal maduro e a variação temporal da relação gonadossomática (ΔRGS) indica também a ocorrência de desova nos respectivos meses. Não obstante, as informações obtidas da correlação entre a ΔRGS e a média de oócitos maduros e percentual de espermatozóides por túbulo seminífero, respectivamente, também indicaram haver desova nos respectivos meses, sendo que aparentemente a espécie apresenta uma desova mais intensa ou desova principal entre os meses de dezembro e janeiro (inverno), e outra desova menos intensa ou secundária no mês de julho (verão). Com base no método da morfologia tubular, foram determinados oito estádios do ciclo do epitélio seminífero (CES), sendo que no estádio 1 os túbulos seminíferos são compostos por espermatogônias primárias e cistos de spermatogônias secundárias; o estádio 2 é composto por espermatogônias primárias e secundárias e cistos de espermatócitos; estádio 3 é caracterizado por espermatogônias primárias, secundárias, cistos de espermatócitos e de espermátides jovens ou recém-formadas; estádio 4 com túbulos seminíferos caracterizados pela presença de espermatogônias primárias e secundárias, espermatócitos e por cistos de espermátides jovens e tardias; estádio 5 apresenta todas as células anteriores e é marcado pelo surgimento de espermatozóides no lúmen tubular; estádio 6 tem como características a diminuição dos cistos de células germinativas e considerável aumento do número de espermatozóides no lúmen tubular; os túbulos seminíferos no estádio 7 contêm poucos cistos de células germinativas e se inicia o esvaziamento da massa de espermatozóides do lúmen tubular; estádio 8 é o último do CES e é caracterizado pela aparente desorganização dos cistos remanescentes de células germinativas no túbulo seminífero.

Gonadal development in the freshwater crab Sylviocarcinus pictus (H. Milne Edwards, 1853) (Brachyura: Trichodactylidae) from the Guamá river, state of Pará, Brazil

Os estádios de desenvolvimento das gônadas de machos e fêmeas de caranguejos Sylviocarcinus pictus (H. Milne Eduards, 1853) foram descritos por meio de observações macroscópicas e microscópicas (técnica histológica). A descrição histológica foi baseada em 40 espécimes (20 de cada sexo). Foram identificados quatro estádios de desenvolvimento para as fêmeas: imaturo, em maturação, maturo e em reabsorção. As seguintes células foram encontradas: ovogônias, ovócitos em vitelogênese inicial, ovócitos em vitelogênese avançada, células foliculares e folículos pós-ovulatórios. Três estádios de desenvolvimento foram encontrados para os machos: imaturo, em maturação e maturo, com indicação de: espermatogônias, espermatócitos, espermátides, espermatozóides e espermatóforos. Tais dados sugerem o padrão descrito na literatura. O tamanho da maturidade sexual foi de 32,3 mm de largura da carapaça para machos e 31,5 mm para fêmeas. Os estádios gonadais observados macroscopicamente por meio do volume e da coloração das gônadas foram validados pela análise histológica, sendo um critério útil e ágil para a identificação da maturidade sexual para a espécie. O presente estudo oferece informações inéditas sobre a biologia reprodutiva de Sylviocarcinus pictus.

Differences in the Behavior of Luteinizing Hormones of Various Species at the Rat Gonadal Cell Receptor Site

The ability of different LH-like hormones, such as hCG, PMSG/equine (e) CG, ovine (o) LH, eLH, and rat (r) LH, to bind to and stimulate steroidogenesis in two types of rat gonadal cells was studied under the same experimental conditions. In both Leydig and granulosa cells, the maximal steroidogenic responses elicited by optimal doses of different LHs present during a 2-h incubation were comparable. However, if the cells were exposed to the different LHs for a brief period and then subjected to interference with hormone action by removing the unbound hormone from the medium by washing or adding specific antisera, differences were observed in the amount of steroid produced during subsequent incubation in hormone-free medium. Thus, in the case of hCG, either of these procedures carried out at 15 or 30 min of incubation had little inhibitory effect on the amount of steroid produced at 2 h, the latter being similar to that produced by cells incubated in the continued presence of hCG for 2 h. With eCG and rLH, the effect was dramatic, in that there was a total inhibition of subsequent steroidogenic response. In cells exposed to eLH and oLH, inhibition of subsequent steroidogenesis due to either removal of the free-hormone or addition of specific antisera at 15 or 30 min was only partial. Although all of the antisera used were equally effective in inhibiting the steroidogenic response to respective gonadotropins when added along with hormones at the beginning of incubation, differences were observed in the degree of inhibition of this response when the same antisera were added at later times of incubation. Thus, when antisera were added 60 min after the hormone, the inhibition of steroidogenesis was total (100%) for eCG, partial (10–40%) for eLH and oLH, and totally lacking in cells treated with hCG. From this, it appears that hCG bound to the receptor probably becomes unavailable for binding to its antibody with time, while in the case of eCG and other LHs used, the antibody can still inhibit the biological activity of the hormone. Studies with 125I-labeled hormones further supported the conclusion that hCG differs from all other LHs in being most tightly bound and, hence, least dissociable, while eCG and rLH dissociate most readily; oLH and eLH can be placed in between these hormones in the extent of their dissociability. (Endocrinology 116: 597–603,1985)

Biology and Molecular Markers of Malignant Gonadal Germ Cell Tumors

Germ cell tumors occur both in the gonads of both sexes and in extra-gonadal sites during adoles-cence and early adulthood. Malignant ovarian germ cell tumors are rare neoplasms accounting for less than 5% of all cases of ovarian malignancy. In contrast, testicular cancer is the most common malignancy among young males. Most of patients survive the disease. Prognostic factors of gonadal germ cell tumors include histology, clinical stage, size of the primary tumor and residua, and levels of tumor markers. Germ cell tumors include heterogeneous histological subgroups. The most common subgroup includes germinomas (ovarian dysgerminoma and testicular seminoma); other subgroups are yolk sac tumors, embryonal carcinomas, immature teratomas and mixed tumors. The origin of germ cell tumors is most likely primordial germ cells. Factors behind germ cell tumor development and differentiation are still poorly known. The purpose of this study was to define novel diagnostic and prognostic factors for malignant gonadal germ cell tumors. In addition, the aim was to shed further light into the molecular mechanisms regulating gonadal germ cell tumorigenesis and differentiation by studying the roles of GATA transcription factors, pluripotent factors Oct-3/4 and AP-2γ, and estrogen receptors. This study revealed the prognostic value of CA-125 in malignant ovarian germ cell tumors. In addition advanced age and residual tumor had more adverse outcome. Several novel markers for histological diagnosis were defined. In the fetal development transcription factor GATA-4 was expressed in early fetal gonocytes and in testicular carcinoma precursor cells. In addition, GATA-4 was expressed in both gonadal germinomas, thus it may play a role in the development and differentiation of the germinoma tumor subtype. Pluripotent factors Oct-3/4 and AP-2γ were expressed in dysgerminomas, thus they could be used in the differential diagnosis of the germ cell tumors. Malignant ovarian germ cell tumors expressed estrogen receptors and their co-regulator SNURF. In addition, estrogen receptor expression was up-regulated by estradiol stimulation. Thus, gonadal steroid hormone burst in puberty may play a role in germ cell tumor development in the ovary. This study shed further light in to the molecular pathology of malignant gonadal germ cell tumors. In addition, some novel diagnostic and prognostic factors were defined. This data may be used in the differential diagnosis of germ cell tumor patients.

Studies on follicular atresia: role of gonadotropins and gonadal steroids in regulating cathepsin-D activity of preovulatory follicles in the rat

Preovulatory follicular atresia was studied using pregnant mare serum gonadotropin (PMSG)-primed rats (15 IU/rat) which were deprived of hormonal support either by allowing the metabolic clearance of the PMSG or by injecting a specific PMSG antiserum (PMSG a/s). Atresia was monitored by an increase in lysosomal cathepsin-D activity and a decrease in the receptor activity of the granulosa cells (GC) isolated from the preovulatory follicles. It was shown that the increase in lysosomal activity and the decrease in receptor activity seen at 96 h after PMSG (or PMSG plus PMSG a/s) could be arrested both by follicle stimulating hormone (FSH) and luteinizing hormone (LH). Injection of cyanoketone or clomiphene citrate together with FSH/LH prevented this 'rescue' suggesting a role for estrogens in the regulation of atresia. Although the administration of estradiol-17 beta (20 micrograms/rat) together with PMSG a/s could show a 'rescue effect' in terms of reduction in cathepsin-D activity the gonadotropin receptor activities of these granulosa cells were not restored. The injection of dihydrotestosterone (DHT) to 48 h PMSG-primed rats induced atresia as noted by an increase in cathepsin-D activity. However, the exogenous administration of FSH along with DHT prevented this atretic effect suggesting that DHT is not having a direct effect on atresia. Determination of androgen: estrogen content of the granulosa cells and an analysis of the individual profile of androgen and estrogen revealed that the increase in cathepsin-D activity could be correlated only with the decrease in GC estrogen content. This along with the observation that GC showed a loss of estrogen synthesis well before the increase in cathepsin-D activity strongly points out that the lack of estrogen rather than an increase in androgen is the principle factor responsible for the atresia of preovulatory follicles in the rat.

Gonadal cell surface receptor for plasma retinol-binding protein A method for its radioassay and studies on its level during spermatogenesis

A specific membrane receptor for plasma retinol-binding protein has been demonstrated in testicular cells. Prealbumin-2 did not show any specific binding to the membrane. The affinity of retinol-binding protein for receptor drastically decreases upon delivery of retinol and the retinol-binding protein does not enter the cell. The mechanism of delivery of retinol to the target cell by plasma retinol-binding protein has been investigated. The process involves two steps: direct binding of retinol-binding protein to the receptor and uptake of retinol by the target cell with a concomitant drastic reduction in the affinity of the retinol-binding protein to the receptor. Probably the second step of the process needs a cytosolic factor, possibly the cellular retinol-binding protein or an enzyme.The binding of retinol-binding protein to the receptor is saturable and reverible. The interaction shows a Kd value of 2.1 · 10−10 M. The specific binding of a retinol-binding protein with great affinity has been employed in the development of a method for radioassay of the receptor. The receptor level of the gonadal cell has been found to vary with the stage of differentiation. The receptor concentrations in 11-week-old birds and adult birds are comparable. Testoterone treatment of 11-week-old birds produced a substantial increase in the receptor concentration over control, while the protein content increased marginally, indicating that, probably, synthesis of the receptor is specifically induced by testoterone during spermatogenesis, and the concentration of receptor is relatively higher before the formation of the acrosome.