Role du calcium dans les processus de couplage stimulus-secretion dans la cellule corticosurrenalienne chez les amphibiens

SIGLECNRS T 59332 / INIST-CNRS - Institut de l'Information Scientifique et TechniqueFRFranc

Urotensine II et somatostatine : les retrouvailles de deux vieilles cousines

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New insight into the molecular evolution of the somatostatin family

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Cloning, sequence analysis and tissue distribution of the mouse and rat urotensin II precursors

International audienceUrotensin II (UII) is a cyclic neuropeptide initially isolated from the caudal neurosecretory system of teleost fish. The recent cloning of the UII precursor in frog and human has demonstrated that the peptide is not restricted to the fish urophysis but that it is also expressed in the central nervous system of tetrapods. Here, we describe the characterization of the cDNAs encoding prepro-UII in mouse and rat. A comparison of the primary structures of mouse and rat UII with those of other vertebrate UII reveals that the sequence of the cyclic region of the molecule (CFWKYC) has been fully conserved. In contrast, the N-terminal flanking domain of prepro-UII has markedly diverged with only 48% sequence identity between the mouse or rat and the human precursors. In situ hybridization histochemistry showed that the prepro-UII gene is predominantly expressed in motoneurons of the brainstem and spinal cord, suggesting that UII may play a role in the control of neuromuscular functions

Selenoprotein T is a key player in ER proteostasis, endocrine homeostasis and neuroprotection

International audienceSelenoprotein T (SELENOT, SELT) is a thioredoxin-like enzyme anchored at the endoplasmic reticulum (ER) membrane, whose primary structure is highly conserved during evolution. SELENOT is abundant in embryonic tissues and its activity is essential during development since its gene knockout in mice is lethal early during embryogenesis. Although its expression is repressed in most adult tissues, SELENOT remains particularly abundant in endocrine organs such as the pituitary, pancreas, thyroid and testis, suggesting an important role of this selenoprotein in hormone production. Our recent studies showed indeed that SELENOT plays a key function in insulin and corticotropin biosynthesis and release by regulating ER proteostasis. Although SELENOT expression is low or undetectable in most cerebral structures, its gene conditional knockout in brain provokes anatomical alterations that impact mice behavior. This suggests that SELENOT also plays an important role in brain development and function. In addition, SELENOT is induced after injury in brain or liver and exerts a cytoprotective effect. Thus, the data gathered during the last ten years of intense investigation of this newly discovered thioredoxin-like enzyme point to an essential function during development and in adult endocrine organs or lesioned brain, most likely by regulating ER redox circuits that control homeostasis and survival of cells with intense metabolic activity

Chromosomal localization of three somatostatin genes in zebrafish. Evidence that the [Pro2]-somatostatin-14 isoform and cortistatin are encoded by orthologous genes

International audienceThere is now evidence for the existence of two somatostatin genes in most vertebrate species, and even three somatostatin genes in teleosts. To help clarify the evolutionary relationships between the different somatostatin isoforms currently known, we characterized the somatostatin loci in a teleost species, the zebrafish Danio rerio, and compared them with the corresponding regions in the human and pufferfish genomes. The occurrence of three somatostatin genes, termed SS1, SS2 and SSII, has been previously demonstrated in the zebrafish. Radiation hybrid mapping assigned these three genes to linkage groups 15, 23 and 2, respectively. Conserved synteny of the zebrafish SS2 gene and the human cortistatin gene was revealed by comparative genomic analysis, indicating that mammalian cortistatin is orthologous to the SS2 variant of non-mammalian species. In contrast, using a similar approach, it was not possible to identify the evolutionary relationships between the atypical SSII gene of zebrafish and the other teleost SSII genes

Impact of gene/genome duplications on the evolution of the urotensin II and somatostatin families

International audienceThe present review describes the molecular evolution of two phylogenetically related families of neuropeptides, the urotensin II (UII) and somatatostatin (SS) families. The UII family consists of four paralogous genes called UII, URP, URP1 and URP2 and the SS family is composed of six paralogous genes named SS1, SS2, SS3, SS4, SS5 and SS6. All these paralogs are present in teleosts, while only four of them, UII, URP, SS1 and SS2 are detected in tetrapods. Comparative genomics showed that most of these genes, namely UII, URP, URP1 and URP2 on the one hand and SS1, SS2 and SS5 on the other hand arose through the 2R. In contrast, the teleost-specific 3R had a much more moderate impact since it only concerned the UII and SS1 genes, which once duplicated, generated a second UII copy and SS4, respectively. The two remaining genes, SS3 and SS6, arose through tandem duplications of the SS1 and SS2 genes respectively, probably in the stem lineage of actinopterygians, before the emergence of teleosts. The history of the UII and SS families has also been marked by massive gene lost, both in tetrapods and in teleosts, but only after the 3R in this latter lineage. Finally, ancestral UII and SS genes are thought to have arisen through tandem duplication of a single ancestral gene, largely before the 1R. An important challenge for the future will be to understand the physiological significance of the molecular diversity of these two families

Molecular cloning of the cDNAs and distribution of the mRNAs encoding two somatostatin precursors in the African lungfishProtopterus annectens

International audienceThe occurrence of two somatostatin precursors, PSS1 and PSS2, yielding S-14 (SS1) and the variant [Pro2, Met13]S-14 (SS2), has been recently reported in the frog Rana ridibunda. The evolutionary significance of frog PSS2 is unclear because its sequence exhibits very little similarity with other known vertebrate somatostatin precursors. In the present study, we report on the characterization of two somatostatin precursor cDNAs from the brain of the African lungfish Protopterus annectens. One of the cDNAs encodes a 115-amino-acid protein that contains the SS1 sequence at its C-terminal extremity and thus is clearly homologous to PSS1. Comparison with other vertebrate PSS1 showed that lungfish PSS1 is more closely related to PSS1 from tetrapods than to PSS1 from fish. The other cDNA encodes a 109-amino-acid protein that contains a somatostatin variant [Pro2]S-14 at its C-terminal extremity. Sequence analysis of this second precursor indicated that it is the lungfish counterpart of frog PSS2. Northern blot analysis showed that lungfish PSS1 mRNA is widely distributed in the central nervous system and in peripheral organs, including the pancreas and gastrointestinal tract. In contrast, PSS2 mRNA was primarily found in the central nervous system but not in the pancreas or gut. In situ hybridization studies showed that the two genes are differentially expressed in various regions of the lungfish brain. The present data indicate that the PSS2 gene, initially discovered in frog, appeared early in vertebrate evolution, before the emergence of the tetrapod lineage. The recent isolation of a [Pro2]S-14 variant in the sturgeon, whose sequence is identical to that of lungfish SS2, suggests that the PSS2 gene may actually be present in the genome of all Osteichthyii

Expression and Processing of the [Pro 2 ,Met 13 ]Somatostatin-14 Precursor in the Intermediate Lobe of the Frog Pituitary

International audienceThe biosynthesis of various hypothalamic neuropeptides has been previously reported in anterior pituitary cells but not in intermediate lobe cells. We have recently demonstrated the occurrence of two somatostatin isoforms in the frog brain, namely somatostatin-14 (SS1) and [Pro(2),Met(13)]somatostatin-14 (SS2). In the present study, we demonstrate that the gene encoding the SS2 precursor (PSS2) is actively expressed in the intermediate lobe of the frog pituitary. High concentrations of PSS2 mRNA have been detected by Northern blot analysis and in situ hybridization in the frog pars intermedia but not in the pars distalis or pars nervosa. The distribution of PSS1- and PSS2-derived peptides has been investigated by immunohistochemistry using two antisera directed against SS1 and the sequence 54-66 of PSS2 (PSS2(54-66)), respectively. The SS1 antiserum stained only a network of fibers in the neural lobe and a few nerve processes in the intermediate lobe. In contrast, the PSS2(54-66) antiserum produced intense labeling of melanotrope cells in the pars intermedia. Biochemical characterization of the immunoreactive materials present in pituitary extracts was performed by combining high-performance liquid chromatography analysis and RIA detection. The SS1 RIA revealed the existence of two major immunoreactive peaks that exhibited the same retention times as synthetic SS1 and SS2. The PSS2(54-66) RIA detected a single peak that likely corresponds to the N-flanking peptide of SS2 (PSS2(1-66)). The present study reveals that melanotrope cells of the frog pituitary selectively express the PSS2 gene and fully process PSS2 to generate the mature somatostatin variant SS2. Taken together, these data provide the first evidence that the gene encoding a hypophysiotropic neuropeptide is intensely expressed in the intermediate lobe of the pituitary

Characterization of the cDNA encoding a somatostatin variant in the chicken brain: Comparison of the distribution of the two somatostatin precursor mRNAs

International audienceAlthough the existence of two somatostatin variants (SS1 and SS2) has now been demonstrated in the brain of mammals, amphibians, and fish, only one isoform of somatostatin (SS1) has been characterized to date in the brain of birds. Here we report cloning of the cDNA encoding a 101-amino-acid protein (PSS2) that encompasses the somatostatin variant [Pro(2)]somatostatin-14 (SS2) at its C-terminus. Sequence analysis indicated that chicken PSS2 is more closely related to fish PSS2 than to mammalian cortistatin precursors. Northern blot analysis showed that the chicken PSS1 gene is expressed in the central nervous system (CNS) and in the pancreas, whereas the PSS2 gene is expressed only in the CNS and not in peripheral organs. In situ hybridization histochemistry revealed that, in the chicken brain, PSS1 mRNA is more widely distributed than PSS2 mRNA. In particular, PSS1 mRNA expression was found in the hippocampus, the hyperstriatum, the preoptic area, the ventricular hypothalamic nuclei, the optic tectum, and several nuclei of the mesencephalon and rhombencephalon. In contrast, the distribution of PSS2 mRNA was restricted to a few regions of the brain, including the paraolfactory lobe, the paleostriatum, and some nuclei of the mesencephalon and rhombencephalon. The fact that the PSS1 and PSS2 genes are differently expressed in the brain and in peripheral organs indicates that, in chicken, the two somatostatin variants likely exert distinct functions. In particular, the observation that PSS1 mRNA, but not PSS2 mRNA, occurs in the preoptic area and in the ventral hypothalamic nuclei suggests that, of the two somatostatin isoforms, only SS1 acts as a hypophysiotropic factor