Revealing Natural Intracellular Peptides in Gills of Seahorse Hippocampus reidi.

The seahorse is a marine teleost fish member of the Syngnathidae family that displays a complex variety of morphological and reproductive behavior innovations and has been recognized for its medicinal importance. In the Brazilian ichthyofauna, the seahorse Hippocampus reidi is among the three fish species most used by the population in traditional medicine. In this study, a protocol was performed based on fast heat inactivation of proteases plus liquid chromatography coupled to mass spectrometry to identify native peptides in gills of seahorse H. reidi. The MS/MS spectra obtained from gills allowed the identification of 1080 peptides, of which 1013 peptides were present in all samples and 67 peptide sequences were identified in an additional LC-MS/MS run from an alkylated and reduced pool of samples. The majority of peptides were fragments of the internal region of the amino acid sequence of the precursor proteins (67%), and N- and C-terminal represented 18% and 15%, respectively. Many peptide sequences presented ribosomal proteins, histones and hemoglobin as precursor proteins. In addition, peptide fragments from moronecidin-like protein, described with antimicrobial activity, were found in all gill samples of H. reidi. The identified sequences may reveal new bioactive peptides.

Effects of ocean acidification and salinity variations on the physiology of osmoregulating and osmoconforming crustaceans.

Survival, osmoregulatory pattern, oxygen consumption, energy spent on metabolism, ammonia excretion, type of oxidized energy substrate, and hepatosomatic index were evaluated in decapods (an osmoregulating crab, Callinectes danae, and an osmoconforming seabob shrimp, Xiphopenaeus kroyeri) exposed to carbon dioxide-induced water acidification (pH 7.3, control pH 8.0) and different salinities (20, 25, 30, 35, and 40‰) for 3 days. Compared to the animals kept at controlled pH, exposure to reduced pH resulted in the loss of osmoregulatory capacity in C. danae at all salinities, except for some hyporegulation at 40‰, and reduced oxygen consumption and ammonia excretion at 20 and 40‰. Xiphopenaeus kroyeri remained an osmoconformer in all evaluated conditions, except for some hyporegulation at 40‰, and when exposed to the reduced pH, it presented changes in oxygen consumption at all salinities and reductions in ammonia excretion at 20 and 35‰ compared to the control animals. Both species use protein as the main energy substrate and decrease the hepatosomatic index when exposed to reduced pH relative to the control. The observed changes may be associated with changes in the activity of enzymes related to osmoregulation, the use of amino acids as osmotic effectors of cell volume control and recovery, and the Bohr effect, and, because the gills are multifunctional organs related to osmoregulation, the changes may be related to acid-base control, nitrogen excretion, and respiration, with a change in one of these functions bringing about changes in the others.

Prolonged dark adaptation changes connexin expression in the mouse retina.

In the retina, ambient light levels influence the cell coupling provided by gap junction (GJ) channels, to compensate the visual function for various lighting conditions. However, the effects of ambient light levels on expression of connexins (Cx), the proteins that form the GJ channels, are poorly understood. In the present study, we first determined whether gene expression of specific Cx (Cx26, Cx31.1, Cx36, Cx37, Cx40, Cx43, Cx45, Cx50, and Cx57) was affected by prolonged dark adaptation. Cx mRNA relative levels were determined in mouse retinas dark adapted for 3 hr, 1 day, and 7 days by using quantitative real-time PCR. Transcript levels of some Cx were repressed after 3 hr (Cx57), 1 day (Cx45), or 7 days (Cx36 and Cx43) of dark adaptation; others were increased after 1 day (Cx50) or 7 days (Cx31.1 and Cx37); and two of them (Cx26 and Cx40) were not significantly altered. The second aim was to determine whether prolonged dark adaptation affects protein expression of two important Cx in retina: neuronal Cx36 and glial Cx43. We were able to demonstrate that important changes in protein distribution and expression also took place in retina during long-term dark adaptation. Given their localization, the specific alterations in Cx expression may reflect their distinct response to ambient light levels.