Orientador: Irene Bastos Franceschini Vicentini / Banca: Antonio Marcos Orsi / Banca: Maíra Aparecida Stefanini / Banca: Margarida Maria Barros Ferreira Lima / Banca: Maria Terezinha Siqueira Bombonato / Resumo: O presente trabalho realizou o estudo dos aspectos estruturais do hepatopâncreas, ovários e da caracterização hormonal de fêmeas de Macrobrachium amazonicum, nos diferentes estágios de maturação gonadal. Para tanto utilizaram-se 150 fêmeas adultas subdivididas entre os cinco diferentes estágios de maturação ovariana. Após serem capturados os animais foram pesados e mortos por choque térmico. Ovários e hepatopâncreas foram coletados e pesados individualmente para obtenção dos índices gonadossomático (IGS) e hepatossomático (IHS). Para as análises de microscopia de luz e eletrônica de transmissão, ovários e hepatopâncreas foram fixados em solução de Karnovsky e seguiram os procedimentos de rotina de inclusão. Os ovários, hepatopâncreas e hemolinfa destinados à quantificação hormonal foram armazenados em ependorfes e preservados em nitrogênio líquido a -70ºC, para posterior processamento de radioimunoensaio visando avaliação das concentrações de estradiol, progesterona e testosterona. Os resultados relacionados ao IGS e IHS demonstram uma correlação inversa entre os ovários e o hepatopâncreas nos diferentes estágios de maturação gonadal. Em relação ao desenvolvimento ovariano, observa-se a distribuição de cinco tipos celulares ao longo dos cinco estágios de maturação gonadal. Desta forma encontram-se as ovogônias ou células germinativas mais jovens no estágio I de maturação ovariana. As células pré-vitelogênicas, observadas a partir do estágio II, apresentam vesículas de retículo endoplasmático rugoso. Essas vesículas são importantes nas fases de vitelogênese endógena e exógena. Os ovócitos em vitelogênese inicial, característicos do estágio III, apresentam a deposição de vitelo endógeno. As células em vitelogênese avançada, próprias do estágio IV de maturação gonadal, são o alvo principal da vitelogênese ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This study described the structural aspects of the hepatopancreas and ovaries, and the hormonal features of females of the freshwater prawn Macrobrachium amazonicum, during the reproductive cycle. The specimens were captured and killed by thermic chock. The gonadosomatic (GSI) and hepatosomatic (HSI) indices were calculated as the percentage of the weight of the gonad and hepatopancreas to total body weight, respectively. Ovaries and hepatopancreas were fixed in Karnovsky solution and destined to light and ultrastructural microscopy studies. Small portions of the ovary, hepatopancreas and hemolymph were frozen in liquid nitrogen, for later steroid quantification (estradiol, progesterone and testosterone). The relationships between GSI and HSI demonstrate an inverse correlation between ovary and hepatopancreas during the maturation cycle. The oocytes distribution is variable depending on the ovarian maturation degree. The ovary in stage I consists of oogonia. The ovary in stage II consists mainly by previtellogenic oocytes that possess vesicles of the endoplasmic reticulum. These vesicles are important to endogenous and exogenous vitelogenesis. Initial vitellogenic oocytes are mainly observed in the ovary in stage III and possess vitelline reserve, called as endogenous vitelogenesis. Ovary in stage IV is composed of late vitellogenic oocytes that uptake exogenous vitellogenin. Mature oocytes present a corion formation. The hepatopancreas is composed by tubules that are lined by a pseudostratified epithelium, which consists of five cell types, identified as E (embryonic), F (fibrillar), B (blisterlike), R (resorptive) and M (basal). These cells present relationships with digestion and nutrient storage, which are very important for the reproductive processes ...(Complete abstract, click electronic access below) / Doutor

Purified brush border membrane vesicles (BBMV) were isolated to characterize primary cellular transport mechanisms for white shrimp. The ultimate goal is to determine the effective components of a shrimp’s diet, thereby enhancing growth, as well as nutrient content. Juvenile shrimp are dependent on plant material as a food source. Potassium is a key component of plants, thus it may play a role in nutrient transport. In addition, divalent metals have been shown to act as co-transporters in several other organisms, thus they may serve as a transport mechanism for shrimp. Fresh, live, white or brown shrimp were obtained, and from them 15-30 hepatopancreases were dissected to prepare the BBMV. Methods for preparing BBMV were based on the Mg2+ precipitation technique developed by Kessler et al., (1978) and Biber et al. (1981) for mammalian eipithelia and applied to crustaceans. The results suggest that there is a sodium/potassium-dependent glucose transport system that resembles the SGLT1 system of vertebrates, except the shrimp transporter can accept both sodium and potassium as cofactors, while the vertebrate system is restricted to sodium stimulation. Potassium showed strong stimulation of L-histidine uptake by shrimp BBMV, suggesting that a crustacean isoform of the insect potassium-dependent carrier protein (KAAT1) might be present in shrimp, and contribute to amino acid uptake. Amino acids also appear to form bis-complexes with divalent metals, that are transported by an analog of the dipeptide transporter (PEPT1). The metals appear to be accommodated, with varying affinities. PEPT1 has been described as a very non-specific carrier process because it transports such a wide range of di- and tripeptide combinations.

Glucose is transported in crustacean hepatopancreas and intestine by Na+-dependent co-transport, while Na+-dependent D-fructose influx has only been described for the hepatopancreas. It is still unclear if the two sugars are independently transported by two distinct cotransporter carrier systems. In this study lobster (Homarus americanus) hepatopancreas brush border membrane vesicles (BBMV) were used to characterize, in detail, the cation-dependency of both D-[3H] glucose and D-[3H] fructose influxes, while in vitro perfused intestines were employed to determine the nature of cation-dependent sugar transport in this organ. Over the sodium concentration range of 0-100 mM, both 3H-D-glucose and 3H-D-fructose influxes (0.1 mM; 1 min uptakes) by hepatopancreatic BBMV were hyperbolic functions of [Na+], exhibiting Km values of 2.30 ± 0.59 and 2.58 ± 0.95 mM, respectively. D-[3H] glucose and fructose influxes by hepatopancreatic BBMV over a potassium concentration range of 15-100 mM were hyperbolic functions of [K+], exhibiting Km values of 9.85 ± 0.41 and 12.6 ± 0.80 mM respectively. Both sugars displayed significant (p < 0.01) Na+/K+-dependent and Na+-independent uptake processes. Transepithelial 25 μM D-[3H] glucose and D-[3H] fructose fluxes across lobster intestine over a luminal sodium and potassium concentration range of 0 – 50 mM and 5-100 mM, respectively, were hyperbolic functions of luminal [Na+] and [K+]. As with hepatopancreatic sugar transport, transepithelial intestinal sugar transport exhibited both significant (p < 0.01) Na+/K+-dependent and Na+-independent processes. Results suggest that both D-glucose and D-fructose are transported by a single carrier process in each organ with sodium being the preferred cation for both sugars in the hepatopancreas, and potassium being the preferred cation for both sugars in the intestine.