Caractérisation de la voie Sonic hedgehog dans le cerveau de rongeur au cours du développement et chez l'adulte

Ce travail suggère l'existence d'une voie Hedgehog (hh) fonctionnelle dans le cerveau adulte. Il décrit l'identification et la localisation des transcrits des gènes codant pour les protéines hh, leur récepteur Ptc/Smo, le facteur de transcription Gli1 et d'autres composantes de la voie dans le cerveau et les tissus périphériques de rat adulte. Des cartographies détaillées de la localisation des cellules exprimant Shh, Ptc et Smo ont été établies dans le cerveau et la moelle épinière de rat adulte ainsi que dans le cervelet postnatal. Leur co-localisation dans un nombre limite d'aires cérébrales suggère que le modèle ternaire Shh/Ptc/Smo classiquement reconnu chez l'embryon n'est pas l'unique modèle de fonctionnement de la voie chez l'adulte, où Shh et Ptc pourraient fonctionner indépendamment de Smo, tandis que Smo lui-même pourrait transmettre l'activité d'un ligand encore non identifié. Dans le cervelet post-natal, mes travaux décrivent l'expression de Shh, Ptc, Smo et Gli1 dans la couche des cellules de Purkinje et dans les zones de prolifération des neuroblastes cérébelleux, suggérant un rôle de la voie Shh dans la prolifération et/ou la différenciation des précurseurs du cervelet ainsi que dans la genèse de médulloblastomes. L'injection intrastriatale de la protéine recombinante myrShhN induit la transcription du récepteur Ptc au niveau de la zone sous- ventriculaire, une aire de neurogenèse persistant chez l'adulte. Ces observations suggèrent un rôle de la voie Shh dans la neurogenèse chez l'adulte. Les gènes codant pour des protéines morphogènes de l'os (BMP), impliquées dans le développement du tube neural durant l'embryogenèse, sont exprimés dans le cerveau adulte suggérant que la voie des BMP pourrait, chez l'adulte, interagir avec la voie Shh, comme précédemment décrit chez l'embryon. La modulation de la transcription du récepteur BMPRII dans un modèle d'ischémie cérébrale supporte l'idée que cette voie est fonctionnelle dans le système nerveux adulte.This work suggests the existence of the functional Hedgehog (hh) signaling pathway in the adult brain. It describes the identification and the localization of the transcripts of genes coding for hh, their receptor Ptc/Smo, the transcription factor Gli1 and other components of the pathway in the brain and peripheral tissues of the adult rat. The detailed mapping of the Shh, Ptc and Smo transcripts was established in the adult brain and spinal cord and in the developing cerebellum. Their colocalization is restricted to a very limited number of adult brain areas, suggesting that a model of the hh signaling in the adult might be different of that previously proposed in the embryo. Ptc could transduce Shh activities in the absence of Smo and Smo itself might bind another not identified yet ligand. The developmental regulation of Shh, Ptc, Smo and Gli1 in the cerebellum suggests a possible role for Shh signaling in the control of various cell populations within the cerebellum, particularly in granule cell precursor proliferation and/or differentiation that might be impaired in proliferative states such as medulloblastomas. The injection of the recombinant protein myrShhN into the adult rat striatum induce a Ptc transcription in the subventricular zone, containing progenitors that can proliferate and differentiate into new neurons and glia in the adult brain. These observations suggest a role for the Shh signaling in the adult neurogenesis. The bone morphogenetic proteins (BMPs) implicated in the neural tube patterning during embryogenesis are expressed also in the adult brain, suggesting that the BMP pathway could interact with the Shh pathway in the adult as in the embryo.The up-regulation of the BMPRII receptor after transient global cerebral ischemia in rat suggests a modulation of the BMP signaling during neuronal plasticity or repair that occur upon brain injury.ORSAY-PARIS 11-BU Sciences (914712101) / SudocSudocFranceF

Regional distribution of Sonic Hedgehog, patched, and smoothened mRNA in the adult rat brain.

In vertebrates, Sonic Hedgehog (Shh), Desert Hedgehog (Dhh), and Indian Hedgehog (Ihh) genes encode a family of morphogen proteins that are implicated in a wide range of signaling activities, particularly during embryonic development. These secreted proteins are proposed to mediate their effects on target cells by interacting with their putative receptor, Patched (Ptc), and with a seven-pass transmembrane protein, Smoothened (Smo). However, the roles that these signaling molecules may play in adult tissues, particularly in brain, are not yet clearly defined. Therefore, we investigated the expression of these genes in adult rat tissues. Northern blot analysis revealed expression of Shh, Dhh, and Ihh genes in peripheral tissues, whereas Shh transcript was also identified in brain. It is interesting that northern blot analysis with probes derived from the mouse Ptc and Smo genes revealed the expression of a 7.9-kb and a 3.7-kb transcript, respectively, in all brain tissues examined. In situ hybridization experiments using specific digoxigenin-labeled riboprobes showed expression of Ptc and Smo transcripts in discrete brain areas. Shh-positive cells were observed in restricted regions of the brain. Within the cerebellum, Shh, Ptc, and Smo transcripts were colocalized in the Purkinje cell layer. These data suggest that, besides its roles in determining cell fate and patterning during embryogenesis, the hedgehog signaling pathway may have also important roles in the adult brain

Distribution of bone morphogenetic protein and bone morphogenetic protein receptor transcripts in the rodent nervous system and up-regulation of bone morphogenetic protein receptor type II in hippocampal dentate gyrus in a rat model of global cerebral ischemia.

Bone morphogenetic proteins belong to the transforming growth factor-beta superfamily and act through serine/threonine kinase type I and type II receptors such as bone morphogenetic protein receptor type I and type II. In order to further understand the roles that these factors exert in the nervous system, we have examined the expression pattern of seven bone morphogenetic proteins and bone morphogenetic protein receptor type I and II transcripts in the brain and spinal cord of rodent. Whereas bone morphogenetic protein receptor type I expression was low in rat brain, in situ hybridization studies performed with specific digoxigenin-labelled riboprobes revealed the presence of bone morphogenetic protein receptor type II-positive cells throughout the brain, with a notable localization in dopaminergic cells of the substantia nigra. Bone morphogenetic protein receptor type II transcripts were also expressed by large motoneuron-like cells located in the ventral horn of the spinal cord and by sensory neurons of dorsal root ganglia. In addition, we observed a significant up-regulation of bone morphogenetic protein receptor type II in the granule cells of the dentate gyrus 48 h after transient global cerebral ischemia in rat suggesting that modulation of this receptor intervenes during neuronal plasticity or repair that occur upon brain injury. Among the potential ligands for this receptor, bone morphogenetic protein-6 and bone morphogenetic protein-7 were expressed in meninges and the choroid plexus, while bone morphogenetic protein-4-expressing cells were spatially and temporally regulated in myelinated structures during development and in the adult suggesting its expression in oligodendrocytes.These data clearly indicate that besides their roles in bone and embryonic tissues, bone morphogenetic proteins and their receptors may have also important functions in adult neural tissues

Intrastriatal sonic hedgehog injection increases Patched transcript levels in the adult rat subventricular zone.

The morphogen sonic hedgehog (Shh) is implicated in neural tissue patterning and the growth of brain structures during embryogenesis and postnatal development and is also present in the adult brain. Shh signals through interaction with the tumour suppressor Patched (Ptc). This receptor for Shh is associated with Smoothened (Smo), a protein with high homology to the G-protein coupled receptors. However, little is known about the transduction mechanisms implicated in Shh signalling in the adult brain. The study described here shows that injection of aminoterminal myristoylated Shh (myrShhN) into the adult rat striatum robustly increases the levels of Ptc transcripts in selective brain areas including the subventricular zone (SVZ). The adult SVZ contains cell progenitors, which can proliferate and differentiate into new neurons and glia. In the myrShhN injected animals, proliferation and differentiation of these SVZ precursor cells were not affected as demonstrated by BrdU incorporation and immunohistochemistry performed with specific antibodies for nestin (uncommitted neural progenitors), PSA-NCAM (migrating neuroblasts) or GFAP (astrocytes). Together with the presence of Smo expressing cells and amino-terminal Shh (ShhN) protein in SVZ area of untreated animals, the data presented here supports the hypothesis that the Shh pathway may be activated in the adult brain, and that a niche for Shh signalling exists within the adult SVZ