High-density lipoprotein remodeling affects the osmotic properties of plasma in goldfish under critical salinity

To investigate the stress response and physiological adaptations of goldfish (Carassius auratus) to critical salinity (CS) waters, we analyzed high-density lipoprotein (HDL) stoichiometry, stress markers (cortisol, glucose), and plasma osmotic properties (Na⁺, osmolality, water content) using ichthyology, biochemistry, and proteomics approaches. After 21 days of exposure to CS, plasma concentrations of cortisol, glucose, and Na⁺ increased, indicating stress. Total plasma osmolality (Osm_total) and osmolality generated by inorganic (Osm_inorg) and organic osmolytes (Osm_org) also increased, the latter by ~2%. We associated the increase of Osm_org with (1) increased metabolite concentration (glucose), (2) dissociation of HDL particles resulting in increased HDL number per unit plasma volume (~1.5–2-fold) and (3) increased HDL osmotic activity. HDL remodeling may be the reason for the redistribution of bound and free water in plasma, which may contribute to water retention in plasma and, at the same time, to hemodynamic disturbances under CS conditions. The study's findings suggest that HDL remodeling is an important mechanism for maintaining osmotic homeostasis in fish, which is consistent with current capillary exchange models in vertebrates.     

Proteases from the Regenerating Gut of the Holothurian Eupentacta fraudatrix

Four proteases with molecular masses of 132, 58, 53, and 47 kDa were detected in the digestive system of the holothurian Eupentacta fraudatrix. These proteases displayed the gelatinase activity and characteristics of zinc metalloproteinases. The 58 kDa protease had similar protease inhibitor sensitivity to that of mammalian matrix metalloproteinases. Zymographic assay revealed different lytic activities of all four proteases during intestine regeneration in the holothurian. The 132 kDa protease showed the highest activity at the first stage. During morphogenesis (stages 2–4 of regeneration), the highest activity was measured for the 53 and 58 kDa proteases. Inhibition of protease activity exerts a marked effect on regeneration, which was dependent on the time when 1,10-phenanthroline injections commenced. When metalloproteinases were inhibited at the second stage of regeneration, the restoration rates were decreased. However, such an effect proved to be reversible, and when inhibition ceased, the previous rate of regeneration was recovered. When protease activity is inhibited at the first stage, regeneration is completely abolished, and the animals die, suggesting that early activation of the proteases is crucial for triggering the regenerative process in holothurians. The role of the detected proteases in the regeneration processes of holothurians is discussed.     

Rearrangement of the actin-spectrin cytoskeleton during oocyte maturation of the starfish Asterias amurensis

Actin and spectrin localization in the oocytes of the starfish Asterias amurensis at hormonal induction of maturation until the destruction of the germinal vesicular membrane has been investigated by immunocytochemical and immunoblotting methods. In immature oocytes, spectrinlike protein and actin are detected to be colocalized in the undermembranous area of the cytoplasm and nuclear membrane. 1-Methyladenine causes redistribution of these proteins into intracellular structures. The actin-spectrin cytoskeleton rearrangement is shown to start at the animal pole of the oocyte and to spread then to its vegetative pole.     

Subcellular shifts of trimeric G-proteins following activation of Baker’s yeast by glucose

Addition of glucose to a resting cell suspension of the yeastSaccharomyces cerevisiae was accompanied by marked shifts of the Gα-protein subunits from the plasma membrane to the cell interior. This process was rapid with half-times between <10 and 20 s. The decrease of the plasma membrane pool of the G_iα/G_oα- and G_qα/G₁₁α-protein subunits correlated with an increase in acid-sensitive forms of these proteins which was recovered in the mitochondrial and/or lysosomal membrane fraction. In contrast to cells from higher organisms glucose-stimulated yeast exhibits an extremely rapid type of the redistribution (internalization). The question remains’open as to the functional significance of the internalized forms of the G-proteins as these remain sequestered from the plasma membrane. well after glucose has been consumed.     

Hormonal Control of Meiosis in Starfishes

Development of starfish oocytes is blocked at the prophase stage of the first meiotic division. The resumption of meiotic divisions occurs under the effect of the maturation hormone 1-methyladenine (1-MeA), which binds to a specific receptor of the oocyte cell surface. New data in the literature on endocellular signal mechanisms taking part in conduction of the regulatory signal modulated by 1-MeA are adduced in the review. Data on the properties of the 1-MeA receptor are presented and mechanisms of biosynthesis of 1-MeA are considered. The main focus is on processes occurring in the oocyte during the first minutes after the impact of the hormone, before the destruction of the germinal vesicle. A hypothetical pattern of transduction of the hormonal signal is proposed.     

Identification and Isolation of GTP-Binding Regulatory Protein from Starfish (Asterias amurensis) Oocyte Plasma Membranes

A method for isolating a GTP-binding regulatory protein from starfish oocytes is described. The protein consists of three subunits with molecular weights of 40, 37, and about 8 kDa. It is shown that the 40-kDa subunit has a high GTPase activity and is susceptible to ADP-ribosylation by pertussis toxin. The latter property of this subunit proved to decrease upon its incubation with nonhydrolyzable GTP analogues. These data provide evidence that the plasma membrane of starfish oocytes contains a 40-kDa GTP-binding protein with properties characteristic of the alpha subunit of the inhibitory G _i protein. The role of this protein in the transmembrane signal transmission from the 1-methyladenine receptor to intracellular effectors is discussed.     

Muscle regeneration in the holothurian Stichopus japonicus

The regeneration of longitudinal muscle bands (LMBs) in the sea cucumber Stichopus japonicus was studied using light and electron microscopic and immunocytochemical methods. Previous investigations of holothurian organs showed the presence of some cytoskeletal proteins which were specific for LMBs only. One of them, the 98 KDa protein, was isolated by means of SDS-electrophoresis and used as an antigen to obtain polyclonal antibodies. When tested on paraffin sections of sea cucumber organs, the antibodies were shown to interact only with coelomic epithelial cells covering the LMBs. The antibodies were used to study LMB regeneration after transverse cutting. During regeneration no signs of myocyte dedifferentiation or mitotic division were observed. In the wound region, damaged myocytes degenerated and muscle bundles desintegrated. However, the coelomic epithelial cells dedifferentiated and began to invade the LMB. Just beneath the surface these cells formed clusters (muscle bundle rudiments). The number and size of the clusters gradually increased, the cells lengthened and developed contractile filaments. These observations suggest that new muscle bundles arise from coelomic epithelial cells covering the LMBs. The migration of coelomic epithelial cells into the damaged LMBs and their myogenic transformation are the basic mechanism of holothurian muscle regeneration.     

The distribution of the Wnt5 protein in the tissues of the holothurian Eupentacta fraudatrix (Djakonov et Baranova, 1958) (Holothuroidea: Dendrochirotida) in the norm and during regeneration

The Wnt5 protein localization in holothurian Eupentacta fraudatrix tissues was examined in the norm and during regeneration. In healthy E. fraudatrix, Wnt5 was found in solitary cells of the hypodermis and in the radial nerve cords. During regeneration, the number of Wnt5 positive cells increased. They were observed in the connective tissue of the body wall and the pharyngeal bulb, in nervous system tissues, the coelomic epithelium, and amoebocytes. The Wnt5 protein may participate in regulating the regeneration in the holothurian E. fraudatrix; it probably modulates cell migration, extracellular matrix reorganization, and neurogenesis.     

Effect of 1-Methyladenine on Localization of Gαi Protein in Starfish Oocytes

Localization and quantitative dynamics of _i subunit of G protein was studied by electron immunocytochemistry and immunoenzyme assay after hormonal induction of oocyte maturation in starfish Asterias amurensis. G_i protein was chiefly localized in the plasma membrane of immature oocytes; 1-methyladenine induced redistribution of the _i protein from the plasma membrane to intracellular structure up to the breakdown of the germinal vesicle.