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Tenascin glycoproteins in developing neural tissues: only decoration?

Tenascin glycoproteins constitute a growing family of extracellular matrix molecules that are transiently expressed by astrocytes during the development of the central nervous system (CNS). In some areas, tenascin distribution is discrete and tenascin boundaries delineate emerging functional processing units, for example, in the somatosensory cortex. The intact adult CNS displays low levels of expression and up-regulation of tenascin after lesion or in glial tumors. In vitro studies using the purified glycoprotein, bacterially expressed tenascin fusion proteins, monoclonal antibodies, and defined cell culture models have established that tenascin glycoproteins possess cell-binding sites for neural cells, support neuronal migration and the formation of neurites. On the other hand, tenascin also embodies repulsive, inhibitory properties that could serve to conceal neuronal assemblies and to segregate emerging fiber tracts. These functional properties are encoded by distinct domains of the molecule and suggest that tenascin glycoproteins are involved in neural pattern formation and regeneration. This interpretation is discussed with reference to a recent report that the elimination of the tenascin gene does not cause obvious abnormalities of neural tissues.

Animals↗

An evolutionary conserved COUP-TF binding element in a neural-specific gene and COUP-TF expression patterns support a major role for COUP-TF in neural development.

The COUP transcription factors (COUP-TF and ARP-1) are the most highly conserved members of the nuclear receptor superfamily throughout evolution. Previous studies indicated that COUP orphan receptors may be involved in early neurogenesis in Drosophila and zebrafish. Here we identified a neural-specific gene, arrestin, whose transcription can be regulated by endogenous COUPs through a DR-7 element (direct repeat with a 7-base pair spacer) located upstream of the transcription start site. Importantly, the COUP binding site of the arrestin gene promoter is conserved among mouse, bovine, and human. However, the mouse element is also capable of responding to retinoic acid while the element in the human gene does not. Expression of COUP-TF correlates with the known expression sites of the arrestin gene in vivo, notably during the differentiation of the retina. We also show that COUP-TF is expressed in a spatio-temporally defined pattern in the murine central nervous system during embryogenesis. It appears that the expression pattern of COUP-TF is unique in certain regions of the developing brain, which would indicate a novel role for COUP-TF and/or ARP-1, distinct from their role in restricting other hormonal signaling pathways. Together our data suggest that COUPs play a crucial role in controlling a subset of neural-specific programs during development.

Animals↗

Differential expression of SNAP-25 protein isoforms during divergent vesicle fusion events of neural development.

The presynaptic plasma membrane protein SNAP-25 (synaptosome-associated protein of 25 kDa) has been implicated as one of several neural-specific components that direct constitutive fusion mechanisms to the regulated vesicle trafficking and exocytosis of neurotransmitter release. There exist two alternatively spliced isoforms of SNAP-25, a and b, which differ in a putative membrane-interacting domain. We show that these two isoforms have distinct quantitative and anatomical patterns of expression during brain development, in neurons, and in neuroendocrine cells and that the proteins localize differently in neurites of transfected PC12 pheochromocytoma cells. These findings indicate that alternative isoforms of SNAP-25 may play distinct roles in vesicular fusion events required for membrane addition during axonal outgrowth and for release of neuromodulatory peptides and neurotransmitters.

Adrenal Glands↗

Molecular events associated with neural development.

Analyses of the chick embryo using two-dimensional gel electrophoresis or immunohistochemistry with monoclonal antibodies (MAbs) revealed several molecules which were highly concentrated in the nervous tissue. The properties and developmental changes of these substances were investigated. Three acidic proteins (molecular weights 95, 100 and 110 kDa, respectively) appeared characteristically during the embryonic development. One of them predominated alternatively at each developmental stage. They showed a high degree of homology in the peptide map and complete cross-reactivity with any specific antibodies. Therefore these proteins were considered to make a protein family and were collectively named drebrins. MAbs were produced against the membrane fraction of the neural tubes and somites of three-day chick embryos. Five MAbs stained the migrating neural crest cells and their progeny in histochemistry. In addition, the otic vesicle, including the differentiated labyrinth, and subtypes of human lymphocytes were recognized by these MAbs.

Animals↗

Retinoid-regulated gene expression in neural development.

The discovery and development of information surrounding the retinoic acid receptors (RAR and RXR) has ushered in a new era in understanding the molecular mechanism of action of vitamin A in embryonic development and cellular differentiation. The mechanisms involved in the regulation of gene expression by the retinoids is at least partially known and involves binding of the RAR and RXR to retinoic acid response elements. Additional factors, including coregulatory proteins, associated regulatory elements, and cell-specific factors, may also be involved in determining the specificity of retinoid-regulation of gene expression during development. During embryogenesis, retinoids are required for the development of the posterior hindbrain and its associated structures, as well as for the survival and differentiation of certain classes of neurons and neural crest cell derivatives. At least some of the effects of retinoid on hindbrain development are related to the regulation of Hox gene expression. Additional retinoid-regulated genes have been implicated in nervous system development, and the manner in which they lead to phenotypic changes during embryogenesis remains to be determined.

Animals↗

Pax-2 regulatory sequences that direct transgene expression in the developing neural plate and external granule cell layer of the cerebellum.

Expression of Pax-2 in the mouse gastrula is the first marker of the midbrain-hindbrain region. To address roles played by transcription factors in the process of neural plate pattern formation and to facilitate gain-of-function approaches in the study of midbrain-hindbrain and cerebellar development, we characterized regulatory sequences at the Pax-2 locus using an in vivo transgenic mouse reporter assay. An 8.5 kb fragment of genomic DNA located upstream of Pax-2 directed lacZ expression prior to neurulation (7.5 days post-coitum, dpc) in a region fated to become midbrain and hindbrain, and subsequently in developing neuroepithelium. While similar to the pattern of Pax-2 expression, reporter gene activity extended beyond the boundaries of Pax-2 expression, most probably reflecting purdurance of beta-galactosidase activity and an absence of DNA sequences that restrict Pax-2 expression to rhombomere 1 by 9. 5 dpc. In the fetal and neonatal brain, Pax-2-lacZ activity was confined largely to Purkinje cells and the external granule cell layer (EGL) of the cerebellum. A 4 kb regulatory element, in contrast, initiated neural expression at 8.25 dpc in the anterior hindbrain, but recapitulated all later aspects of Pax-2-lacZ activity observed with the larger transgene. These results indicate the presence of regulatory sequences upstream of the Pax-2 locus capable of directing gene expression in the developing midbrain, first rhombomere of the hindbrain, and its principal derivative, the cerebellum. Successful misexpression of Sonic hedgehog demonstrates that Pax-2 regulatory sequences should prove generally useful for transgenic gain-of-function approaches in mice.

Animals↗

msh/Msx gene family in neural development.

The involvement of Msx homeobox genes in skull and tooth formation has received a great deal of attention. Recent studies also indicate a role for the msh/Msx gene family in development of the nervous system. In this article, we discuss the functions of these transcription factors in neural-tissue organogenesis. We will deal mainly with the interactions of the Drosophila muscle segment homeobox (msh) gene with other homeobox genes and the repressive cascade that leads to neuroectoderm patterning; the role of Msx genes in neural-crest induction, focusing especially on the differences between lower and higher vertebrates; their implication in patterning of the vertebrate neural tube, particularly in diencephalon midline formation. Finally, we will examine the distinct activities of Msx1, Msx2 and Msx3 genes during neurogenesis, taking into account their relationships with signalling molecules such as BMP.

Animals↗

Arsenic-induced alterations in embryonic transcription factor gene expression: implications for abnormal neural development.

We examined the morphological and molecular consequences of acute in utero exposure to teratogenic concentrations of arsenate. The treatment produced a dose-related increase in neural tube defects, along with a significant alteration in the pattern of gene expression for several transcription factors (creb, Hox 3.1, Pax3, and Emx-1) that were examined using in situ transcription and antisense RNA amplification procedures. On gestational day 9:0, there was a significant delay in the embryos progression through neural tube closure, accompanied by a significant downregulation of Hox 3.1 expression and a significant upregulation of Pax3, Emx-1, and creb. As both Hox 3.1 and Pax3 serve to regulate N-CAM expression, it is possible that abnormalities associated with N-CAM may compromise neural crest cell migration and normal neural tube closure.

Animals↗

Multiple roles of canonical Wnt signaling in cell cycle progression and cell lineage specification in neural development.

Signaling by the canonical Wnt pathway has multiple functions in stem cells. It can either control stem cell expansion or--as we have recently demonstrated with neural crest stem cells-influence cell lineage decisions by promoting specific fates at the expense of others. Thus, the role of canonical Wnt in stem cells is dependent on cell--intrinsic properties that determine how a cell responds to Wnt. The molecular basis for the functional diversity of Wnt in different stem cell types remains to be elucidated.

Cell Cycle↗

The role of class I HLH genes in neural development--have they been overlooked?

Helix-loop-helix (HLH) genes encode for transcription factors affecting a whole variety of developmental programs, including neurogenesis. At least seven functional classes (denoted I to VII) of HLH genes exist, (1) with subclass members exhibiting homo- and heterodimerisation for proper DNA binding and transcriptional regulation of downstream target genes. In the developing nervous system, members of class II, V and VI have been most extensively studied concerning their roles in neural programming. In contrast, the function of class I proteins (such as E12 and E47) is poorly defined and the orthodox view relegates them to general dimerisation duties that are necessary for the activity of the other classes. However, closer scrutiny of the spatiotemporal expression patterns of class I factors, combined with recent biochemical evidence, would suggest that class I proteins possess specific functions during early neural differentiation. This essay supports this possibility, in addition to putting forward the hypothesis that, outside their general dimerisation activity, class I genes have independent roles in regulating neurogenesis.

Animals↗

Expression of Tsga10 sperm tail protein in embryogenesis and neural development: from cilium to cell division.

Tsga10 has been localised in sperm tail as a fibrous sheath protein. In this study, we showed its expression during developmental stages of mouse embryo, in adult mice brain, and in some malignancies. RT-PCR and immunohistochemistry study show that Tsga10 expression starts in 4.5-7.5 dpc mouse embryos and continues throughout embryogenesis. Then we showed that the Tsga10 is expressed in adult brain and in the cells with neural crest origin, olfactory epithelium, and human germ cell tumour. It is expressed with two transcripts in sperm and whole embryos but just with the long transcript in brain embryo as a result of its exon 16 splicing. Our finding of the Tsga10 perinuclear localisation and its expression pattern suggests that it may be involved in active cell division, differentiation, and migrating cells. The results of the experiments in this project hypothesize the presence of Tsga10 protein wherever there is a conserved ciliary structure.

Alternative Splicing↗

Ephrin-B3, a ligand for the receptor EphB3, expressed at the midline of the developing neural tube.

The ephrins are a family of ligands that bind to Eph family receptor tyrosine kinases, and have been implicated in axon guidance and other patterning processes during vertebrate development. We describe here the identification and characterization of murine ephrin-B3. The cDNA encodes a 340 amino acid transmembrane molecule, most closely related to the two other known transmembrane ligands, ephrin-B1 and ephrin-B2. In addition to homology in their extracellular receptor binding domains, these transmembrane ligands share striking homology between their cytoplasmic domains, with 31 of the last 34 amino acids of ephrin-B3 being identical to ephrin-B2, suggesting functional interactions of the cytoplasmic tail. While most Eph family ligands are promiscuous in their interactions with Eph receptors, binding studies with the five receptors known to bind other transmembrane ligands only revealed a high affinity interaction of ephrin-B3 with EphB3, with a dissociation constant of approximately 1 nM. In situ hybridization of mouse embryos showed ephrin-B3 is expressed prominently at the dorsal and ventral midline of the neural tube, particularly in the floor plate, a structure with key functions in patterning the nervous system. The isolation of this ligand may help to elucidate the molecular basis of patterning activities at the neural tube midline.

Animals↗

Patterning cascades in the neural tube. Neural development.

The vertebrate central nervous system comprises an intricate array of neurons generated in a highly organized way. Examination of the genes expressed and required at early stages of neural differentiation reveals that a coordinated signalling cascade transforms progenitor cells into discrete neuronal subsets.

Animals↗

The Wnt-activated Xiro1 gene encodes a repressor that is essential for neural development and downregulates Bmp4.

In the early Xenopus embryo, the Xiro homeodomain proteins of the Iroquois (Iro) family control the expression of proneural genes and the size of the neural plate. We report that Xiro1 functions as a repressor that is strictly required for neural differentiation, even when the BMP4 pathway is impaired. We also show that Xiro1 and Bmp4 repress each other. Consistently, Xiro1 and Bmp4 have complementary patterns of expression during gastrulation. The expression of Xiro1 requires Wnt signaling. Thus, Xiro1 is probably a mediator of the known downregulation of Bmp4 by Wnt signaling.

Animals↗

Mouse GLI3 regulates Fgf8 expression and apoptosis in the developing neural tube, face, and limb bud.

The zinc finger transcription factor GLI3 is considered a repressor of vertebrate Hedgehog (Hh) signaling. In humans, the absence of GLI3 function causes Greig cephalopolysyndactyly syndrome, affecting the development of the brain, eye, face, and limb. Because the etiology of these malformations is not well understood, we examined the phenotype of mouse Gli3-/- mutants as a model to investigate this. We observed an up-regulation of Fgf8 in the anterior neural ridge, isthmus, eye, facial primordia, and limb buds of mutant embryos, sites coinciding with the human disease. Intriguingly, endogenous apoptosis was reduced in Fgf8-positive areas in Gli3-/- mutants. Since SHH is thought to be involved in Fgf8 regulation, we compared Fgf8 expression in Shh-/- and Gli3-/-;Shh-/- mutant embryos. Whereas Fgf8 expression was almost absent in Shh-/- mutants, it was up-regulated in Gli3-/-;Shh-/- double mutants, suggesting that SHH is not required for Fgf8 induction, and that GLI3 normally represses Fgf8 independently of SHH. In the limb bud, we provide evidence that ectopic expression of Gremlin in Gli3-/- mutants might contribute to a decrease in apoptosis. Together, our data reveal that GLI3 limits Fgf8-expression domains in multiple tissues, through a mechanism that may include the induction or maintenance of apoptosis.

Animals↗

Expression of KIF3C kinesin during neural development and in vitro neuronal differentiation.

KIF3A, KIF3B and KIF3C are kinesin-related motor subunits of the KIF3 family that associate to form the kinesin-II motor complex in which KIF3C and KIF3B are alternative partners of KIF3A. We have analysed the expression of Kif3 mRNAs during prenatal murine development. Kif3c transcripts are detectable from embryonic day 12.5 and persist throughout development both in the CNS and in some peripheral ganglia. Comparison of the expression patterns of the Kif3 genes revealed that Kif3c and Kif3a mRNAs colocalize in the CNS, while only Kif3a is also present outside the CNS. In contrast, Kif3b is detectable in several non-neural tissues. We have also performed immunocytochemical analyses of the developing rat brain and have found the presence of the KIF3C protein in selected brain regions and in several fibre systems. Using neuroblastoma cells as an in vitro model for neuronal differentiation, we found that retinoic acid stimulated the expression of the three Kif3 and the kinesin-associated protein genes, although with different time courses. The selective expression of Kif3c in the nervous system during embryonic development and its up-regulation during neuroblastoma differentiation suggest a role for this motor during maturation of neuronal cells.

Animals↗

All-trans retinoic acid affects the expression of orphan receptors COUP-TF I and COUP-TF II in the developing neural tube.

The effect of all-trans retinoic acid (RA) on chicken ovalbumin upstream promoter-transcription factor (COUP-TF) I and COUP-TF II expression in the developing cervical spinal cord and telencephalon was examined using embryonic day 11 and 13 mice. All-trans RA treatment results in changes in expression of COUP-TF I and COUP-TF II genes in the spinal cord and telencephalon. COUP-TF I mRNA levels were reduced in the spinal cord ventricular zone (VZ) after treatment, whereas in the telencephalon, mRNA levels were unaffected in the VZ and increased in the intermediate zone (IZ). COUP-TF II mRNA levels were increased in the spinal cord VZ after treatment, while in the telencephalon, IZ mRNA levels were decreased in the E11 embryos and increased in E13 embryos.

Animals↗