Lmx1b controls peptide phenotypes in serotonergic and dopaminergic neurons.

Serotonin (5-HT) neurons synthesize a variety of peptides. How these peptides are controlled during development remains unclear. It has been reported that the co-localization of peptides and 5-HT varies by species. In contrast to the situations in the rostral 5-HT neurons of human and rat brains, several peptides do not coexist with 5-HT in the rostral 5-HT neurons of mouse brain. In this study, we found that the peptide substance P and peptide genes, including those encoding peptides thyrotropin-releasing hormone, enkephalin, and calcitonin gene-related peptide, were expressed in the caudal 5-HT neurons of mouse brain; these findings are in line with observations in rat and monkey 5-HT neurons. We also revealed that these peptides/peptide genes partially overlapped with the transcription factor Lmx1b that specifies the 5-HT cell fate. Furthermore, we found that the peptide cholecystokinin was expressed in developing dopaminergic neurons and greatly overlapped with Lmx1b that specifies the dopaminergic cell fate. By examining the phenotype of Lmx1b deletion mice, we found that Lmx1b was required for the expression of above peptides expressed in 5-HT or dopaminergic neurons. Together, our results indicate that Lmx1b, a key transcription factor for the specification of 5-HT and dopaminergic transmitter phenotypes during embryogenesis, determines some peptide phenotypes in these neurons as well.

Mesencephalic neuronal populations: new insights on the ventral differentiation programs.

The midbrain is a complex structure where different functions are located. This formation is mainly involved in the visual and auditory information process (tectum) and visual movements and motor coordination (tegmentum). Here we display a complete description of midbrain anatomy based on the prosomeric model and of the developmental events that take place to generate this structure. We also summarize the new data about the differentiation and specification of the basal populations of the midbrain. The neural tube suffers the influence of several secondary organizers. These signaling centers confer exact positional information to the neuroblasts. In the midbrain these centers are the Isthmic organizer for the antero-posterior axis and the floor and roof plates for the dorso-ventral axis. This segment of the brain contains, in the dorsal part, structures such as the collicula (superior and inferior), tectal grey and the preisthmic segment, and in the basal plate, neuronal populations such as the oculomotor complex, the dopaminergic substantia nigra and the ventral tegmental area, the reticular formation and the periacueductal grey. Knowledge of the genetic cascades involved in the differentiation programs of the diverse populations will be extremely important to understand not only how the midbrain develops, but how degenerative pathologies, such as Parkinson's disease, occurs. These cascades are triggered by signaling molecules such as Shh, Fgf8 or Wnt1 and are integrated by receptor complexes and transcription factors. These are directly responsible for the induction or repression of the differentiation programs that will produce a specific neuronal phenotype.

Duplicated zebrafish relaxin-3 gene shows a different expression pattern from that of the co-orthologue gene.

Relaxin-3 (Rln3) is thought to function as a neurotransmitter mainly produced in the mammalian nucleus incertus and is involved in different neural processes; among them, the stress response and food intake. Here, we report the expression pattern of the duplicated zebrafish rln3b gene and compare it to the previously analyszd spatial expression pattern of the rln3a gene. Both genes, during the embryogenesis and in the adult fish, are active and show relevant differences in their expression patterns. rln3b is diffusely expressed in the brain until the pharyngula period, when, at 48 h postfertilization (hpf), the expression becomes restricted to the periaqueductal gray, where it persists also at later developmental stages. No expression was observed in the nucleus incertus cells that express the rln3a gene from 72 hpf. In the adult, both genes are expressed in brain, but only rln3b transcript is revealed in testis at the similar expression level, whereas in the other analyzed tissues the transcript levels are lower or absent. Both the putative mature protein sequences are highly conserved, this feature and their differential expression patterns might indicate a sub-functionalization during evolution with the consequent retention of the two paralogues genes.

Patch/matrix patterns of gray matter differentiation in the telencephalon of chicken and mouse.

The mammalian striatum, a subpallial area, consists of two compartments (patches/striosomes and matrix) that differ in their neuronal birth dates, connectivity, neurochemistry, and molecular make-up. For example, members of the cadherin family of adhesion molecules (cadherin-8 and OL-protocadherin) are differentially expressed by the striosomes and the striatal matrix. A patch/matrix type of organization also has recently been found in the ventral hyperstriatum and the neostriatum of the chicken pallium, where cell clusters of similar birthdates ("isochronic" clusters) are surrounded by a matrix of cells that are born at a different time. Immunostaining with antibodies against cadherins reveals a similar arrangement of cell clusters. In the avian neostriatum, cadherin-7-positive cell clusters ("islands") are surrounded by a matrix of cells that express R-cadherin. The islands coincide, at least in part, with the isochronic cell clusters, as shown by pulse-labeling with bromodeoxyuridine. Likewise, isochronic clusters of the hyperstriatum ventrale relate to patchy heterogeneities in the cadherin-7 immunoreactivity pattern. Cadherins are known to mediate the aggregation and sorting of cells during development in many organs. Their differential expression by isochronic cell populations in the mammal subpallium and avian pallium suggests a common morphogenetic mechanism that regulates the formation of the patch/matrix patterns in these regions.

Expression of calbindin D28K in the dopaminergic mesotelencephalic system in embryonic and fetal human brain.

A subset of tyrosine-hydroxylase (TH) neurons of the substantia nigra (A9) containing calbindin D28K (CaBP) appeared to be less vulnerable to cell death induced by Parkinson's disease than the subset containing dopamine (DA) alone. Because grafting procedures of fetal human neurons are increasingly used in the therapy of Parkinson's disease, it is important to study the development of DA neurons coexpressing CaBP. In humans, the genesis of TH immunoreactivity of A9, of the ventral tegmental area (A10), and of the retrorubral area (A8) occurred during a 2-week period from the 4. 5th gestational week (g.w.) in the ventricular zone of the floor plate and the contiguous basal plate of the mesencephalon and diencephalon, i.e., the prosomeres p1-p3. Double-immunolabeled TH-CaBP neurons were detected from 5.5 g.w. on, in the first wave of DA neuron's migration, and were observed in their final residence in the dorsal A9 by 10.5 g.w. Calretinin immunoreactivity was expressed in TH-immunoreactive (IR) neurons from 10.5 g.w. on. Ascending TH-CaBP-IR axons were observed toward the telencephalon from 6-7 g.w. , reaching the anlage of the nucleus accumbens and amygdaloid complex at 10.5 g.w., but were not detected in the ganglionic eminence at this latter stage. Dopaminergic patches were detected at 13 g.w. in the anlage of the putamen, but no TH-CaBP-IR fibers were observed in the matrix at this stage. In conclusion, even if CaBP immunoreactivity was detected in TH-IR cell bodies during the embryonic period, the TH-CaBP-IR axonal terminal was observed earlier in some limbic-related areas than in the matrix compartment of the basal ganglia in humans.

In vivo studies of brain development by magnetic resonance techniques.

Understanding of the morphological development of the human brain has largely come from neuropathological studies obtained postmortem. Magnetic resonance (MR) techniques have recently allowed the provision of detailed structural, metabolic, and functional information in vivo on the human brain. These techniques have been utilized in studies from premature infants to adults and have provided invaluable data on the sequence of normal human brain development. This article will focus on MR techniques including conventional structural MR imaging techniques, quantitative morphometric MR techniques, diffusion weighted MR techniques, and MR spectroscopy. In order to understand the potential applications and limitations of MR techniques, relevant physical and biological principles for each of the MR techniques are first reviewed. This is followed by a review of the understanding of the sequence of normal brain development utilizing these techniques. MRDD Research Reviews 6:59-67, 2000.

NGF and NT-3 have differing effects on the growth of dorsal root axons in developing mammalian spinal cord.

The functions of neurotrophins in relation to axon growth and branching during development of the nervous system are unknown. In order to address this question, we have investigated the influences of systemically administered mouse nerve growth factor (mNGF) and human recombinant neurotrophin-3 (hrNT-3) on dorsal root axon growth in the spinal cord of embryonic rats. As anticipated, mNGF has a marked influence on growth of dorsal root axons. In mNGF-treated animals, dorsal root axons in the developing dorsal funiculi and axon collaterals in developing gray matter are substantially longer than those of age-matched controls. Furthermore, growth cones of some dorsal root axons have more than twice the surface area of controls. These effects of NGF are highly selective. Dorsal root axons that occupy a lateral position in white matter and that normally give off collaterals to superficial dorsal horn are prominently affected. Axons that run medially in dorsal columns and that give off collaterals to laminae III and IV and the ventral horn are not demonstrably influenced by treatment with exogenous mNGF. In contrast to the striking effects of mNGF on dorsal root axon growth, the influences of hrNT-3 were considerably more complex. Administration of hrNT-3 increased the mean soma area of DRG neurons, particularly those at the larger end of the size spectrum, consistent with its hypothesized role as a growth factor for proprioceptive sensory neurons. However, in striking contrast to the actions of mNGF, hrNT-3 consistently inhibited axon collateral growth in gray matter at early developmental stages. At later stages, we could not discern a clear-cut influence of hrNT-3 on dorsal root axon growth and branching. We conclude that the ability of mNGF to stimulate axon growth in both white and gray matter is consistent with the idea that mNGF regulates the developing axonal projections of DRG neurons in vivo. In contrast, systemically administered hrNT-3 inhibits the axon collateralizations of DRG neurons in gray matter at early developmental stages. We hypothesize that this inhibitory effect may be related to disruption of a chemotropic gradient of NT-3, or to the widespread expression of the NT-3 receptor trkC, on non-neuronal cells.

On the development of the pyramidal tract in the rat. II. An anterograde tracer study of the outgrowth of the corticospinal fibers.

An anterograde tracer study has been made of the developing corticospinal tract (CST) in the rat using wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP). Analysis of normal Rager stained material revealed that corticospinal axons reach upper cervical spinal cord levels at the day of birth (PO). Postnatal rats ranging in age from one (P1) to fourteen (P14) days received multiple WGA-HRP injections into the cortex of their left hemisphere and were allowed to survive for 24 h. The first labeled CST fibers caudally extend into the third thoracic spinal cord segment at P1; into the eighth thoracic segment at P3; into the first or second lumbar segment at P7 and into the second to third sacral segment at Pg. Thus the outgrowth of the leading 'pioneer' fibers of the CST is completed at P9 but later developing axons are continuously added even beyond P9. Quantitative analysis of the amount of label along the length of the outgrowing CST revealed a characteristic pattern of labeling varying with age. The most striking features of that pattern are: the formation of two standing peaks at the level of the cervical and lumbar enlargements respectively and the transient presence of a smaller running peak which moves caudally with the front of the outgrowing bundle. The standing peaks are ascribed to the branching of the axon terminals at both intumescences, whereas the running peak probably arises by the accumulation of tracer within the growth cones at the tips of the outgrowing CST axons. Factors such as the number of axons, the varying axon diameters, the branching collaterals, the presence of varicosities, the transport rate of the tracer, the uptake of the tracer at the injection site, which possibly may affect the amount of label present in both the entire bundle and in the individual axons are discussed. Current research is focused upon an analysis of the relation between the site of injection within the cortex and the pattern of labeling of the CST. A delay of two days was found between the arrival of the CST axons at a particular spinal cord level and their outgrowth into the adjacent spinal gray. However, combined HRP and electronmicroscopic experiments are necessary to determine the factors behind the maturation of the CST as well as the maturation of the spinal gray.