The freshwater prawn Macrobrachium rosenbergii is a tropical crustacean with characteristics similar to those of lobsters and crayfish. Adult males develop through three morphological typessmall (SC), yellow (YC), and blue claws (BC)-with each representing a level in the dominance hierarchy of a group, BC males being the most dominant. We are interested in understanding the role played by neuropeptides in the mechanisms underlying aggressive behavior and the establishment of dominance hierarchies in this type of prawn. SIFamides are a family of arthropod peptides recently identified in the central nervous system of insects and crustaceans, where it has been linked to olfaction, sexual behavior, and gut endocrine functions. One of the six SIFamide isoforms, GYRKPPFNGSIFamide (Gly-SIFamide), is highly conserved among decapod crustaceans such as crabs and crayfish. We wanted to determine whether Gly-SIFamide plays a role in modulating aggression and dominant behavior in the prawn. To do this, we performed behavioral experiments in which interactions between BC/YC pairs were recorded and quantified before and after injecting Gly-SIFamide directly into the circulating hemolymph of the living animal. Behavioral data showed that aggression among interacting BC/YC prawns was enhanced by injection of Gly-SIFamide, suggesting that this neuropeptide does have a modulatory role for this type of behavior in the prawn.

Biogenic amines are implicated in several mental disorders, many of which involve social interactions. Simple model systems, such as crustaceans, are often more amenable than vertebrates for studying mechanisms underlying behaviors. Although various cellular responses of biogenic amines have been characterized in crustaceans, the mechanisms linking these molecules to behavior remain largely unknown. Observed effects of serotonin receptor agonists and antagonists in abdomen posture, escape responses, and fighting have led to the suggestion that biogenic amine receptors may play a role in modulating interactive behaviors. As a first step in understanding this potential role of such receptors, we have cloned and fully sequenced two serotonin receptors, 5-HT 1Mac and 5-HT 2Mac , from the CNS of the freshwater prawn Macrobrachium rosenbergii, and have mapped their CNS immunohistochemical distribution. 5-HT 1Mac was found primarily on the membranes of subsets of cells in all CNS ganglia, in fibers that traverse all CNS regions, and in the cytoplasm of a small number of cells in the brain, circum-and subesophageal ganglia (SEG), most of which also appear to contain dopamine. The pattern of 5-HT 2Mac immunoreactivity was found to differ significantly, being found mostly in the central neuropil area of all ganglia, in glomeruli of the brain's olfactory lobes, and in the cytoplasm of a small number of neurons in the SEG, thoracic and some abdominal ganglia. The observed differences in terms of localization, distribution within cells, and intensity of immunoreactive staining throughout the prawn's CNS suggest that these receptors are likely to play different roles.

There is ample evidence linking octopamine (OA) and tyramine (TA) to several neurophysiological functions in arthropods. In our laboratory we use the freshwater prawn Macrobrachium rosenbergii to study the neural basis of aggressive behavior. As a first step towards understanding the possible role of these amines and their receptors in the modulation of interactive behaviors, we have cloned a putative octopamine/tyramine receptor. The predicted sequence of the cloned OA/TAMac receptor consists of 1,579 base pairs (bp), with an open reading frame of 1,350 bp that encodes a 450 amino acid protein. This putative protein displays sequence identities of 70% to an Aedes aegypti mosquito TA receptor, followed by 60% to a Stegomyia aegypti mosquito OA receptor, 59% and 58% to the migratory locust TA-1 and -2 receptors respectively, and 57% with the silkworm OA receptor. We also mapped the OA/TAMac receptor distribution by in-situ hybridization to the receptor’s mRNA, and by immunohistochemistry to its protein. We observed stained cell bodies for the receptor’s mRNA, mainly in the midline region of the thoracic and in the abdominal ganglia, as well as diffuse staining in the brain ganglia. For the receptor’s protein, we observed extensive punctate staining within the neuropil and on the membrane of specific groups of neurons in all ganglia throughout the CNS, including the brain, the midline region and neuropiles of the thoracic ganglia, and ventral part and neuropiles of the abdominal ganglia. The same pattern of stained cells was observed on the thoracic and abdominal ganglia in both in-situ hybridization and immunohistochemistry experiments. Diffuse staining observed with in-situ hybridization also coincides with punctate staining observed in brain, SEG, thoracic, and abdominal ganglia in immunohistochemical preparations. This work provides the first step towards characterizing the neural networks that mediate octopaminergic signaling in prawn.

Urbanization adjacent to rivers has increased in recent years and is considered a source of environmental contamination. The resulting increase in number of urban rivers in highly populated areas, such as the Caribbean island of Puerto Rico, has led to the appearance of synthetic as well as naturally occurring chemicals not previously observed nor regularly monitored in freshwater habitats. Some of these chemicals, such as heavy metals and plasticizers, have been shown to affect endocrine, respiratory, and nervous system function in animals and humans, even at relatively low concentrations. The purpose of this study was to measure concentrations of such emergent contaminants on rivers of urbanized areas on the northeast of Puerto Rico, as one element in the assessment of the impact of urbanism on water quality in these communities. To accomplish this, we used Inductively Coupled Plasma and Gas Chromatography Mass Spectrometry to measure amounts of heavy metals and phthalates, respectively, in superficial water of three rivers of Puerto Rico: Mameyes (non-urban), Río Piedras (urban river without a dam), and La Plata (urban river with a dam). The urban rivers had significantly higher concentrations of heavy metals arsenic, barium, cadmium, manganese, and antimony, when compared with the reference non-urban river. Manganese was the only metal found in concentrations higher than limits established by the EPA for drinking water. Of eight phthalates amenable to measurement with the chosen protocol and instrumentation, only dibutyl phthalate was detected, only in the La Plata river, and at concentrations ranging from 3 to 8 parts-per-billion. These findings suggest that urbanism close to rivers of Puerto Rico is likely having an impact on water quality and thus further study to identify the potential sources, as well as the inclusion of these emergent contaminants on the list of chemicals regularly monitored by government agencies is justified.

The kidney androgen-regulated protein (KAP) gene is exclusively expressed in proximal tubules of mouse kidney and in the uterus of pregnant females before they give birth. It displays an exquisite and differential regulation of expression by steroid and thyroid hormones (THs) in different proximal tubule segments. Whereas the pars recta (PR cells) responds to thyroid and sexual hormones, the pars convoluta (PCT cells) represents a truly androgen-dependent compartment because expression occurs only in the presence of androgens and functional androgen receptors. Nevertheless, different hypothyroidism models have indicated that TH might also contribute to the androgenic response in PCT cells. In the present study, we aimed to determine the molecular mechanisms that ultimately control KAP expression in these cells. Using several genetically deficient mouse models and different pharmacologic and hormonal treatments, we determined that thyroid and GH modulate CCAAT/enhancer binding protein alpha and beta levels that, in turn, control KAP expression in PCT cells in a developmentally dependent manner. We demonstrated that these factors bind to sites in the proximal KAP promoter, thereby collaborating with androgens for full KAP expression. Finally, we propose that TH and GH, acting through CCAAT/enhancer binding protein, may constitute a general regulatory mechanism of androgen-dependent genes in mouse kidney.