Vimentin binding to phosphorylated Erk sterically hinders enzymatic dephosphorylation of the kinase

Cleavage fragments of de novo synthesized vimentin were recently reported to interact with phosphorylated Erk1 and Erk2 MAP kinases (pErk) in injured sciatic nerve, thus linking pErk to a signaling complex retrogradely transported on importins and dynein. Here we clarify the structural basis for this interaction, which explains how pErk is protected from dephosphorylation while bound to vimentin. Pull-down and ELISA experiments revealed robust calcium-dependent binding of pErk to the second coiled-coil domain of vimentin, with observed affinities of binding increasing from 180 nM at 0.1 microM calcium to 15 nM at 10 microM calcium. In contrast there was little or no binding of non-phosphorylated Erk to vimentin under these conditions. Geometric and electrostatic complementarity docking generated a number of solutions wherein vimentin binding to pErk occludes the lip containing the phosphorylated residues in the kinase. Binding competition experiments with Erk peptides confirmed a solution in which vimentin covers the phosphorylation lip in pErk, interacting with residues above and below the lip. The same peptides inhibited pErk binding to the dynein complex in sciatic nerve axoplasm, and interfered with protection from phosphatases by vimentin. Thus, a soluble intermediate filament fragment interacts with a signaling kinase and protects it from dephosphorylation by calcium-dependent steric hindrance.     

The effects of estrogen and progesterone on blood glutamate levels: evidence from changes of blood glutamate levels during the menstrual cycle in women

The gonadal steroids estrogen and progesterone have been shown to have neuroprotective properties against various neurodegenerative conditions. Excessive concentrations of glutamate have been found to exert neurotoxic properties. We hypothesize that estrogen and progesterone provide neuroprotection by the autoregulation of blood and brain glutamate levels. Venous blood samples (10 ml) were taken from 31 men and 45 women to determine blood glutamate, estrogen, progesterone, glucose, glutamate-pyruvate transaminase (GPT), and glutamate-oxaloacetate transaminase (GOT) levels, collected on Days 1, 7, 12, and 21 of the female participants' menstrual cycle. Blood glutamate concentrations were higher in men than in women at the start of menstruation (P < 0.05). Blood glutamate levels in women decreased significantly on Days 7 (P < 0.01), 12 (P < 0.001), and 21 (P < 0.001) in comparison with blood glutamate levels on Day 1. There was a significant decrease in blood glutamate levels on Days 12 (P < 0.001) and 21 (P < 0.001) in comparison with blood glutamate levels on Day 7. Furthermore, there was an increase in blood glutamate levels on Day 21 compared with Day 12 (P < 0.05). In women, there were elevated levels of estrogen on Days 7 (P < 0.05), 12, and 21 (P < 0.001), and elevated levels of progesterone on Days 12 and 21 (P < 0.001). There were no differences between men and women with respect to blood glucose concentrations. Concentrations of GOT (P < 0.05) and GPT (P < 0.001) were significantly higher in men than in women during the entire cycle. The results of this study demonstrate that blood glutamate levels are inversely correlated to levels of plasma estrogen and progesterone.     

Glutamine synthetase enhances the clearance of extracellular glutamate by the neural retina

Clearance of synaptic glutamate by glial cells is required for the normal function of excitatory synapses and for prevention of neurotoxicity. Although the regulatory role of glial glutamate transporters in glutamate clearance is well established, little is known about the influence of glial glutamate metabolism on this process. This study examines whether glutamine synthetase (GS), a glial-specific enzyme that amidates glutamate to glutamine, affects the uptake of glutamate. Retinal explants were incubated in the presence of [(14)C]glutamate and glutamate uptake was assessed by measurement of the amount of radioactively labeled molecules within the cells and the amount of [(14)C]glutamine released to the medium. An increase in GS expression in Müller glial cells, caused by induction of the endogenous gene, did not affect the amount of glutamate accumulated within the cells, but led to a dramatic increase in the amount of glutamine released. This increase, which was directly correlated with the level of GS expression, was dependent on the presence of external sodium ions, and could be completely abolished by methionine sulfoximine, a specific inhibitor of GS activity. Our results demonstrate that GS activity significantly influences the uptake of glutamate by the neural retina and suggest that this enzyme may represent an important target for neuroprotective strategies.     

Atrophic thyroid follicles and inner ear defects reminiscent of cochlear hypothyroidism in Slc26a4-related deafness

Thyroid hormone is essential for inner ear development and is required for auditory system maturation. Human mutations in SLC26A4 lead to a syndromic form of deafness with enlargement of the thyroid gland (Pendred syndrome) and non-syndromic deafness (DFNB4). We describe mice with an Slc26a4 mutation, Slc26a4 ^loop/loop , which are profoundly deaf but show a normal sized thyroid gland, mimicking non-syndromic clinical signs. Histological analysis of the thyroid gland revealed defective morphology, with a majority of atrophic microfollicles, while measurable thyroid hormone in blood serum was within the normal range. Characterization of the inner ear showed a spectrum of morphological and molecular defects consistent with inner ear pathology, as seen in hypothyroidism or disrupted thyroid hormone action. The pathological inner ear hallmarks included thicker tectorial membrane with reduced β-tectorin protein expression, the absence of BK channel expression of inner hair cells, and reduced inner ear bone calcification. Our study demonstrates that deafness in Slc26a4 ^loop/loop mice correlates with thyroid pathology, postulating that sub-clinical thyroid morphological defects may be present in some DFNB4 individuals with a normal sized thyroid gland. We propose that insufficient availability of thyroid hormone during inner ear development plays an important role in the mechanism underlying deafness as a result of SLC26A4 mutations.     

Dopamine inhibits luteinizing hormone synthesis and release in the juvenile European eel: a neuroendocrine lock for the onset of puberty

In various adult teleost fishes, LH ovulatory peak is under a dual neurohormonal control that is stimulatory by GnRH and inhibitory by dopamine (DA). We investigated whether DA could also be involved in the inhibitory control of LH at earlier steps of gametogenesis by studying the model of the European eel, Anguilla anguilla, which remains at a prepubertal stage until the oceanic reproductive migration. According to a protocol previously developed in the striped bass, eels received sustained treatments with GnRH agonist (GnRHa), DA-receptor antagonist (pimozide), and testosterone (T) either alone or in combination. Only the triple treatment with T, GnRHa, and pimozide could trigger dramatic increases in LH synthesis and release as well as in plasma vitellogenin levels and a stimulation of ovarian vitellogenesis. Thus, in the prepubertal eel, removal of DA inhibition is required for triggering GnRH-stimulated LH synthesis and release as well as ovarian development. To locate the anatomical support for DA inhibition, the distribution of tyrosine hydroxylase (TH) in the brain and pituitary was studied by immunocytochemistry. Numerous TH-immunoreactive cell bodies were observed in the preoptic anteroventral nucleus, with a dense tract of immunoreactive fibers reaching the pituitary proximal pars distalis, where the gonadotrophs are located. This pathway corresponds to that mediating the inhibition of LH and ovulation in adult teleosts. To our knowledge, this is the first demonstration of a pivotal role for DA in the control of LH and puberty in a juvenile teleost. These data support the view that DA inhibition on LH secretion is an ancient evolutionary component in the neuroendocrine regulation of reproduction that may have been partially maintained throughout vertebrate evolution.     

Interactive effects of GAs,CKs and growth retardants on the germination of celery seeds

The germination of whole seeds of celery (Apium graveolens L.) was inhibited by paclobutrazol, ancymidol and lower concentrations of uniconazole. The growth retardants daminozide, AMO 1618 and chlormequat chloride inhibited the germination of cut seeds only, indicating that the seed coat prevents the penetration of these compounds at the examined concentrations. Application of a mixture of the gibberellins A4 and A7 (GA 4/7) reversed the inhibition of all the examined growth retardants. Cytokinins (artificial or natural) had no effect when applied alone and did not interact with GA4/7 in the light. However, in the dark the cytokinins at some concentrations and GA4/7 had a synergistic effect in reversing the inhibition caused by growth retardants to whole seeds or in promoting the germination of whole seeds. It is therefore suggested that the major effect on seed of exogenous cytokinins when applied together with GA's is to increase the uptake of gibberellins by the seeds.Abbreviations AMO 1618 (2 isopropyl-5-methyl-4-trimethylammonium chloride-phenyl-1-Piperidinium-carboxylate - ancymidol -cyclopropyl-[4-methoxyphenyl]-5-pyrimidinemeth anol] - chlormequat chloride 2-chloroethyltrimethylommonium chloride - daminozide succinin acid 2,2-diamethyl hydrazide - paclobutrazol [2 RS, 3 RS]-1-(4-chlorophenyl)-4,4-dimethyl-2-(1 H-1,2,4-triazol-1-yl) pentan-3-ol - uniconazole (E)-1-(P-chlorophenyl)-4,4-dimethyl-2-(1,2,4 triazol-1-yl)-1 penten-3-ol     

Evolution of fringing reefs: space and time constraints from the Gulf of Aqaba

This study documents the pattern and rate of reef growth during the late Holocene as revealed by unique geological conditions at the subsiding NW Gulf of Aqaba. We discovered that the modern fringing reef near the city Elat grows on top of a fossil submerged mid-Holocene reef platform. Four coral cores from the fossil platform were dated using the radiocarbon and U-Th methods. The fossil corals range from 5.6±0.1 to 2.4±0.03 ka, constraining the initiation of the modern reef to 2,400 years ago at most. We documented the detailed morphology of the reef using aerial photographs and scuba diving. The survey shows that at its northern end, growth of the 2-km-long reef is inhibited by an active alluvial fan, and it is composed of isolated knolls that are just approaching the sea surface. Towards the south, the knolls are progressively larger and closer together, until they form a continuous reef platform. Along this north-to-south trend we follow the evolution of reef morphology, changes in coral distribution, and the development of a lagoon separated from the open sea. Based on these observations, we suggest a four-stage reef growth model: (a) the reef initiates as coral colonies, forms knolls, and begins to grow upward, limited by the sea surface. (b) Upon reaching the surface, the knolls spread laterally, preferentially parallel to the dominant wave direction assuming an elongated morphology. (c) Continued growth results in adjacent knolls eventually coalescing to form a continuous jagged reef. We interpret the spurs-and-grooves morphology that can be traced across the reef at Elat as remnants of the original trends of knolls. (d) While reef expansion continues, the original knoll trends may be obscured as a massive reef front takes shape. Considering reef growth rates and observations from the modern reef at Elat, this evolution scheme predicts an age range of 10³ years for corals on the reef platform. The range and distribution of radiometric ages we obtained from the fossil reef platform underlying the living Elat reef confirm this hypothesis.