Luiz Eduardo Maia Nery

Biochemical and physiological adaptations in the estuarine crab Neohelice granulata during salinity acclimation.

Neohelice granulata (Chasmagnathus granulatus) is an intertidal crab species living in salt marshes from estuaries and lagoons along the Atlantic coast of South America. It is a key species in these environments because it is responsible for energy transfer from producers to consumers. In order to deal with the extremely marked environmental salinity changes occurring in salt marshes, N. granulata shows important and interesting structural, biochemical, and physiological adaptations at the gills level. These adaptations characterize this crab as a euryhaline species, tolerating environmental salinities ranging from very diluted media to concentrated seawater. These characteristics had led to its use as an animal model to study estuarine adaptations in crustaceans. Therefore, the present review focuses on the influence of environmental salinity on N. granulata responses at the ecological, organismic and molecular levels. Aspects covered include salinity tolerance, osmo- and ionoregulatory patterns, morphological and structural adaptations at the gills, and mechanisms of ion transport and their regulation at the gills level during environmental salinity acclimation. Finally, this review compiles information on the effects of some environmental pollutants on iono- and osmoregulatory adaptations showed by N. granulata.

Antioxidant Defenses and DNA Damage Induced by UV-A and UV-B Radiation in the Crab Chasmagnathus granulata (Decapoda, Brachyura)¶

Abstract The photoprotector role of pigment dispersion in the melanophores of the crab, Chasmagnathus granulata, against DNA and oxidative damages caused by UV-A and UV-B was investigated. Intact and eyestalkless crabs were used. In eyestalkless crabs, the dorsal epidermis of the cephalothorax (dispersed melanophores) and the epidermis of pereiopods (aggregated melanophores) were analyzed. Intact crabs showed only dispersed melanophores in the two epidermis. Antioxidant enzymes activity and lipoperoxidation content were analyzed after UV-A (2.5 J/cm2) or UV-B (8.6 J/cm2) irradiation. DNA damage was analyzed by single cell electrophoresis (comet) assay, after exposure to UV-B (8.6 J/cm2). UV-A radiation increased the glutatione-S-transferase activity in the pereiopods epidermis of eyestalkless crabs (P < 0.05). UV-B radiation induced DNA damage in the dorsal epidermis of eyestalkless crabs (P < 0.05). In pereiopod epidermis of eyestalkless crabs, there was no significant difference between control and UV-B–exposed crabs. In the pereiopods epidermis of eyestalkless, the control group showed higher scores of DNA damage and ∼50% of cellular viability. Because in eyestalkless and irradiated crabs the cellular viability was ∼5%, it was not possible to observe nuclei for determination of DNA damage. The findings show that melanophores can play a role in the defense against harmful effects of a momentary exposure to UV radiation.

Daily variation of melatonin content in the optic lobes of the crab Neohelice granulata

Melatonin is a biogenic amine, known from almost all phyla of living organisms. In vertebrates melatonin is produced rhythmically in the pinealocytes of the pineal gland, relaying information of the environmental light/dark cycle to the organism. With regard to crustaceans only a handful of studies exist that has attempted to identify the presence and possible daily variation of this substance. We set out to investigate whether in the crab Neohelice granulata melatonin was produced in the optic lobes of these animals and underwent rhythmic fluctuations related to the daily light/dark cycle. Our experimental animals were divided into three groups exposed to different photoperiods: normal photoperiod (12L:12D), constant dark (DD), and constant light (LL). The optic lobes were collected every 4 hours over a 24-h period for melatonin quantification by radioimmunoassay (RIA). N. granulata kept under 12 L:12D and DD conditions, showed daily melatonin variations with two peaks of abundance (p<0.05), one during the day and another, more extensive one, at night. Under LL-conditions no significant daily variations were noticeable (p>0.05). These results demonstrate the presence of a daily biphasic fall and rise of melatonin in the eyestalk of N. granulata and suggest that continuous exposure to light inhibits the production of melatonin synthesis.

Melatonin as a Signaling Molecule for Metabolism Regulation in Response to Hypoxia in the Crab Neohelice granulata

Melatonin has been identified in a variety of crustacean species, but its function is not as well understood as in vertebrates. The present study investigates whether melatonin has an effect on crustacean hyperglycemic hormone (CHH) gene expression, oxygen consumption (VO2) and circulating glucose and lactate levels, in response to different dissolved-oxygen concentrations, in the crab Neohelice granulata, as well as whether these possible effects are eyestalk- or receptor-dependent. Melatonin decreased CHH expression in crabs exposed for 45 min to 6 (2, 200 or 20,000 pmol·crab−1) or 2 mgO2·L−1 (200 pmol·crab−1). Since luzindole (200 nmol·crab−1) did not significantly (p > 0.05) alter the melatonin effect, its action does not seem to be mediated by vertebrate-typical MT1 and MT2 receptors. Melatonin (200 pmol·crab−1) increased the levels of glucose and lactate in crabs exposed to 6 mgO2·L−1, and luzindole (200 nmol·crab−1) decreased this effect, indicating that melatonin receptors are involved in hyperglycemia and lactemia. Melatonin showed no effect on VO2. Interestingly, in vitro incubation of eyestalk ganglia for 45 min at 0.7 mgO2·L−1 significantly (p < 0.05) increased melatonin production in this organ. In addition, injections of melatonin significantly increased the levels of circulating melatonin in crabs exposed for 45 min to 6 (200 or 20,000 pmol·crab−1), 2 (200 and 20,000 pmol·crab−1) and 0.7 (200 or 20,000 pmol·crab−1) mgO2·L−1. Therefore, melatonin seems to have an effect on the metabolism of N. granulata. This molecule inhibited the gene expression of CHH and caused an eyestalk- and receptor-dependent hyperglycemia, which suggests that melatonin may have a signaling role in metabolic regulation in this crab.

Infrared Radiation Influence on Molt and Regeneration of Neohelice granulata Dana, 1851 (Grapsidae, Sesarminae)

This paper analyzes the influence of infrared radiation (IR) on regeneration, after autotomy of limb buds of Neohelice granulata and consequently the time molt. Eyestalks were ablated to synchronize the start of molt. Afterward, animals were autotomized of five pereopods and divided into control and irradiated groups. The irradiated group was treated for 30u2003min daily until molt. Limb buds from five animals of days 4, 16 and 20 were collected and histological sections were made from them. These sections were photographed and chitin and epithelium content measured. Another group was made, and after 15u2003days limb buds were extracted to analyze mitochondrial enzymatic activity from complex I and II. The irradiated group showed a significant reduction in molt time (19.38u2003±u20031.22u2003days) compared with the control group (32.69u2003±u20031.57u2003days) and also a significant increase in mitochondrial complex I (388.9u2003±u200327.94%) and II (175.63u2003±u20037.66%) in the irradiated group when compared with the control group (100u2003±u200317.90; 100u2003±u20037.82, respectively). However, these effects were not acompanied by histological alterations in relation to chitin and epithelium. This way, it was possible to demonstrate that IR increases complex I and II activity, reduces the time molt and consequently increases the appendage regeneration rate.

Crustacean Chromatophore: Endocrine Regulation and Intracellular Signalling Systems

Crustacean body coloration plays a major role in communication, cryptic and thermoregulatory behavior (Thurman 1990). The pigments ommochromes, melanins, carotenoids, purines, and pterines are synthesized and/or stored within specialized epidermal cells, or epidermal appendices, such as hair or cuticle. The so-called pigment cells or chromatophores possess multiple branching processes, and may also be present in the epidermal lining of internal organs (nerve cord, intestine, and gonads). Chromatophores may be grouped according to their pigment color and internal structure (Rao 1985). Melanophores possess black or brown granules, erythrophores have red color, xanthophores are yellow, leucophores possess white granules, and iridophores are iridescent due to light reflection. Although color pattern is genetically determined, many crustaceans have the ability to change body coloration in response to exogenous or endogenous stimuli. Most species display a slowly changeable color pattern associated with ontogenetic, dietary, or seasonal determinants. These slow and long-lasting changes result from alterations in chromatophore number and/or pigment amount within the cells, and will not be discussed here. The Malocostraca, particularly some species of amphipods, euphausiids, mysids, stomatopods, isopods, and decapods, can also display rapid color changes as an immediate response to environmental changes, such as background color, light intensity, or social context.