Development Of Central Neurons Research Articles

Dscam1 Is Required for Normal Dendrite Growth and Branching But Not for Dendritic Spacing inDrosophilaMotoneurons

Down syndrome cell adhesion molecule, Dscam, serves diverse neurodevelopmental functions, including axon guidance and synaptic adhesion, as well as self-recognition and self-avoidance, depending on the neuron type, brain region, or species under investigation. In Drosophila, the extensive molecular diversity that results from alternative splicing of Dscam1 into >38,000 isoforms provides neurons with a unique molecular code for self-recognition in the nervous system. Each neuron produces only a small subset of Dscam1 isoforms, and distinct Dscam1 isoforms mediate homophilic interactions, which in turn, result in repulsion and even spacing of self-processes, while allowing contact with neighboring cells. While these mechanisms have been shown to underlie mushroom body development and spacing of mechanosensory neuron dendrites, here we report that Dscam1 plays no role in adult Drosophila motoneuron dendrite spacing, but is required for motoneuron dendritic growth. Targeted expression of Dscam-RNAi in an identified flight motoneuron did not impact dendrite spacing, but instead produced overgrowth. Increasing the knockdown strength severely reduced dendritic growth and branching. Similarly, Dscam mutant motoneurons in an otherwise control background (MARCM) were completely devoid of mature dendrites. These data suggest that Dscam1 is required cell autonomously for normal adult motoneuron dendrite growth in Drosophila. This demonstrates a previously unreported role of Drosophila Dscam1 in central neuron development, and expands the current understanding that Dscam1 operates as a cell adhesion molecule that mediates homophilic repulsion.

The chemokine SDF-1 differentially regulates axonal elongation and branching in hippocampal neurons

Recent data have shown that the chemokine SDF-1 plays a critical role in several aspects of brain development such as cell migration and axon pathfinding. However, its potential function in the generation of axons and dendrites is poorly characterized. In order to better understand the role of SDF-1 in the development of central neurons, we studied the cellular distribution of the SDF-1 receptor CXCR4 by immunocytochemistry of developing hippocampal neurons and tested the effect of SDF-1 in process patterning at the early stages of neuronal development. We found that CXCR4 immunoreactivity undergoes a striking redistribution during development. At the early stages, from day 2 to day 4 in culture, CXCR4 is particularly concentrated at the leading edge of growing neurites. As the cells mature, staining declines at the tip of the processes and becomes more broadly distributed along axons and, to a lesser extent, dendrites. SDF-1 stimulation of neurons at day 1-2 in culture triggers several effects on neuronal morphogenesis. SDF-1 reduces growth cone number and axonal outgrowth but stimulates axonal branching. These latter two effects are not observed in other neurites. This study unravels a new role for SDF-1/CXCR4 in specifying hippocampal neuron morphology by regulating axonal patterning at an early stage of neuronal development.

Depolarizing Influences Increase Low-Affinity NGF Receptor Gene Expression in Cultured Purkinje Neurons

Multiple cellular and molecular interactions are required for the differentiation and development of central neurons. For example, neural activity may modulate trophic function. In the developing cerebellum, establishment of functional excitatory synaptic connections coincides with the expression of NGF and its receptors. We have previously shown that excitatory signals and NGF act in concert to regulate the survival and morphological differentiation of cerebellar Purkinje cells in culture. To begin investigating the molecular mechanisms by which trophic interactions and neural activity modulate cerebellar development, we have now studied the role of excitatory signals on the expression of both NGF and the p75 glycoprotein (the low-affinity component of the NGF receptor) by cerebellar cells in culture. We used p75 as a model of potential responsiveness, since it is well characterized and conveniently monitored. Expression of the NGF and p75 mRNA's was studied in either mixed, neuron-enriched, or pure glial cultures. Expression of the NGF gene was localized to proliferating glial cells, while expression of p75 was restricted to developing Purkinje cells. To evaluate whether presynaptic activation may potentially modulate trophic factor receptor expression, the expression of the p75 gene was studied in cultures exposed to excitatory signals. Depolarization of cultures with high potassium, veratridine, or exposure to the excitatory neurotransmitter aspartate, resulted in a two- to threefold increase in the expression of both the p75 protein and messenger RNA. These increases did not require the presence of glia, indicating a direct effect of the excitatory signals on the neuronal population. Moreover, message and receptor increased per neuron. Our study suggests that local glia provide trophic support for Purkinje cell development, and that impulse activity modulates Purkinje cell responsiveness by regulating expression of trophic receptor subunits.

Differential regulation of beta III and other tubulin genes during peripheral and central neuron development.

Mammalian peripheral and central neurons differ considerably in the composition and properties of their axonal cytoskeletons. Recent reports of the selective expression of a high molecular weight (HMW) tau protein in neurons with peripherally projecting axons have furthered the idea that the microtubules in central and peripheral neurons are disparate. In the present study, we examined the possibility that the various tubulin genes are differentially expressed in central versus peripheral neurons. To examine this, we compared the expression of four of the beta-tubulin mRNAs (classes beta I, beta II, beta III, beta IV) and the alpha 1-tubulin mRNA in rat dorsal root ganglion (DRG) neurons with their expression in cerebral cortex during postnatal development (P5-90), using northern blots and in situ hybridization. We document both similarities and differences in tubulin gene expression in these two regions of the neuraxis during postnatal development. In both DRG and cortex, the expression of the class beta I- and beta II-tubulin mRNAs and the alpha 1-tubulin mRNA was higher at earlier stages of postnatal development than in the adult. However, class beta IV-tubulin mRNA levels increased during cortical development but decreased during DRG postnatal development. The opposite pattern was found for the neuron-specific class beta III-tubulin gene, the mRNA levels of which were high in cortex, at birth and then decreased with increasing postnatal development. In DRG, the beta III-tubulin mRNA levels generally increased during postnatal development. Beta III-tubulin protein levels were examined qualitatively at different developmental stages (5-90 days) by immunoblotting and immunocytochemistry.(ABSTRACT TRUNCATED AT 250 WORDS)

Neurotrophic effect of naturally occurring long-chain fatty alcohols on cultured CNS neurons

A long-chain fatty alcochol, n-hexacosanol, that we have isolated from the Far-Eastern traditional medicinal plant, Hygrophila erecta, Hochr., is shown to promote the maturation of central neurons. Added at 500 nM to fetal rat brain neurons in culture, it increased both neurite outgrowth by a factor of 4–6 and the number of collaterals, especially in multipolar neurons. The biochemical differentiation of cultured neurons was also strikingly enhanced by this compound: it increased the protein content and almost doubled the activities of two neuron-specific enzymes, phosphate-activated glutaminase and neuron-specific enolase, by 92 and 78%, respectively. Extensive studies with several synthetic long-chain fatty alcohols showed that the neurotrophic activity was maximal for n-hexacosanol. It is suggested that some long-chain fatty alcohols with an appropriate length of hydrocarbon chain might play an important role in central neuron development.