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PubMed2003

Features of the developing brain.

INTRODUCTION: Fetal brain evaluation implies a perfect knowledge of the timing and characteristics of the developing nervous system during gestation. DISCUSSION: The first half of gestation corresponds to the neurulation, differentiation of primary cerebral vesicles, and neuronogenesis. The second half is characterized by the tremendous growth of the cerebral hemispheres and the settlement of gyral formation, while the ventricular system undergoes gradual narrowing. Gyral formation follows an invariable temporospatial schedule and is a good marker of fetal maturation. At the histological level, the fetal cortex displays transient developmental features, a superficial granular layer, and cells of Cajal-Retzius, which disappear at term. Neuroblasts formation has usually ceased by the 16th week of gestation, while neuronal migration continues until the extinction of the germinal layer. At this time, the remnant of the germinal layer constitutes, between the thalami and the caudate nucleus, an eminence of tightly packed cells, called the ganglionic eminence or germinal zone. They disappear by the age of 1 year.

Aging↗

PubMed1996

Cell cycle kinetics in the embryonic mouse corpus striatum.

Cells of the corpus striatum arise from the lateral ganglionic eminence of the telencephalic neuroepithelium. In mice, the length of the cell cycle and that of its component phases were estimated, and the interkinetic nuclear migratory pattern was characterized for lateral ganglionic progenitors on embryonic day 11 and 12 to gain insights into striatal cytogenetic process. An S-phase labeling paradigm using bromodeoxyuridine was employed. To assess regional variation in proliferative patterns, rostral, middle and caudal levels of the ganglionic neuroepithelium were examined separately. The pattern of interkinetic nuclear migration and the duration of G1-, G2-, and M-phases at the rostral and middle levels differed from those at the caudal level. The length of the cell cycle and that of the G1-phase increased during the interval embryonic day 11 to 12, especially at the rostral and middle levels. During the same interval, a sizable secondary proliferative population emerged at all three levels. Two principal conclusions are drawn: Progenitors at the different rostrocaudal levels are heterogeneous with regard to the pattern of cellular proliferation, and ganglionic progenitors are in advance of the cerebral cortical progenitors based on the relative size of the secondary proliferative population and the magnitude of cytokinetic parameters. Thus, proliferative behavior distinguishes progenitor populations at different rostrocaudal levels within the lateral ganglionic neuroepithelium and across the ganglionic and cerebral cortical domains of the telencephalic neuroepithelium.

Animals↗

PubMed2003

Phenotypic expression of monoamines and GABA in the early development of human telencephalon, transient or not transient.

We review the phenotypic expression of molecules involved in monoamine and GABA neurotransmission in the developing human brain. Recent experimental reports have analyzed neurotransmitter signaling before the onset of synaptogenesis, which could act to influence early developmental events such as proliferation, migration, and differentiation of animal brain development. Such signaling may also occur in human development. The expression of molecules involved in neurotransmission in precocious human brain may reflect either the differentiation of a permanent neurotransmitter system of the adult brain or transient expression to serve specific developmental functions different from those in the adult brain. We review the changes observed in the expression of various catecholamine markers such as tyrosine-hydroxylase (TH) immunoreactivity in various neuronal populations of the developing human telencephalon. The specific transporter for serotonin, serotonin transporter (SERT) has been detected in fibers of the internal capsule (IC) during the restricted time period of 12-14 gestational weeks in humans. These serotonin-containing fibers do not correspond to serotoninergic ascending axons from the raphe nuclei. They may be the human counterpart of the thalamo-cortical axons that have been shown to uptake serotonin during the critical period of development of the sensory systems in rodents. GABA phenotypes are expressed in numerous cells of the human ganglionic eminence (GE) and cerebral wall at the end of the embryonic period proper. These results are similar to that described at comparable developmental stages in the mouse and support the hypothesis of an early migration from ganglionic progenitors in humans. But one cannot exclude a transient expression of GABA within the post-mitotic neurons, which could influence early developmental events. In conclusion, data showing the phenotypic expression of molecules in discrete areas of the brain at various points in the protracted human development require careful interpretation.

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PubMed1997

Concurrent cellular output from two proliferative populations in the early embryonic mouse corpus striatum.

In mice, the striatal compartment of the forebrain is established by embryonic day 11 (E11, E0 = day of conception) when a lateral ganglionic eminence emerges surrounding the lateral and ventral margins of the forebrain ventricles. The inception of the striatal compartment is evidence of altered cell cycle kinetics, especially a rapid production of postmitotic cells, within a discrete portion of the telencephalic neuroepithelium. As a step toward understanding the mechanisms which contribute to the development of a cytokinetically distinct striatal compartment, we characterized the rate and pattern of cellular output in the lateral ganglionic neuroepithelium of mice on E11. The data show that the striatal compartment is distinguished by concurrent and equivalent levels of cell output from two proliferative populations: a dominant secondary proliferative population and a smaller, pseudostratified ventricular epithelium. In addition, although the ganglionic neuroepithelium is expanding on E11, 30-35% of the daughter cells produced leave the cell cycle and become postmitotic. These cytogenetic events, occurring in the lateral ganglionic progenitor population, may contribute to the development of a distinct striatal compartment within the telencephalon.

Animals↗

PubMed2001

Evidence for target-specific nerve fiber outgrowth from subpopulations of grafted dopaminergic neurons: a retrograde tracing study using in oculo and intracranial grafting.

Efforts have been made to counteract the symptoms of Parkinson's disease by substituting the loss of dopaminergic neurons with fetal ventral mesencephalic grafts. One of the postulated limiting factors in this treatment is the relatively poor cell survival and limited graft-derived fiber outgrowth. Recent results documenting enhanced survival of grafted dopaminergic neurons showed no positive correlation to enhanced innervation of the striatal target. Therefore this study was undertaken to investigate whether all surviving grafted dopaminergic neurons projected to the striatal target. Hence, fetal ventral mesencephalic tissue was implanted adjacent to mature versus immature striatal tissue using in oculo and intraventricular grafting techniques. In in oculo grafting, fetal ventral mesencephalon was implanted simultaneously with fetal lateral ganglionic eminence (immature striatal target) or to already matured striatal in oculo grafts (mature striatal target). Furthermore, fetal ventral mesencephalon was implanted into the lateral ventricle adjacent to mature dopamine-depleted striatum. The retrograde tracer fluorogold was injected into the striatal portion of the in oculo cografts and into reinnervated areas of the adult brain. Immunohistochemistry revealed that a significantly larger proportion of tyrosine hydroxylase-positive neurons in the ventral mesencephalic graft was innervating in oculo immature striatal tissue, and hence was fluorogold-positive, in comparison with the number of tyrosine hydroxylase-positive neurons innervating mature striatal tissue. Moreover, intracranial transplantations showed that tyrosine hydroxylase-positive neurons were distributed within the grafts in dense clusters of cells. In most clusters tyrosine hydroxylase-positive cells were fluorogold-negative but calbindin-positive. In a few tyrosine hydroxylase-positive cell clusters, neurons were coexpressing fluorogold but were calbindin-negative. In conclusion, significantly more dopamine neurons projected to immature than to mature striatal tissue and thus, a subpopulation of grafted dopaminergic neurons was not projecting into adult striatum. Thus, the results from this study show that further attempts to enhance survival of grafted dopamine neurons in purpose to enhance graft-derived fiber outgrowth and efficacy should also consider different subtypes of dopamine neurons.

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PubMed1996

Chondroitin sulfate proteoglycans in the developing central nervous system. I. cellular sites of synthesis of neurocan and phosphacan.

We have used in situ hybridization histochemistry to examine the cellular sites of synthesis of two major nervous tissue proteoglycans, neurocan and phosphacan, in embryonic and postnatal rat brain and spinal cord. Both proteoglycans were detected only in nervous tissue. Neurocan mRNA was evident in neurons, including cerebellar granule cells and Purkinje cells, and in neurons of the hippocampal formation and cerebellar nuclei. In contrast, phosphacan message was detected only in astroglia, such as the Golgi epithelial cells of the cerebellum. At embryonic day 13-16, phosphacan mRNA is largely confined to areas of active cell proliferation (e.g., the ventricular zone of the ganglionic eminence and septal area of the brain and the ependymal layer surrounding the central canal of the spinal cord) as well as being present in the roof plate. The distribution of neurocan message is more widespread, extending to the cortex, hippocampal formation, caudate putamen, and basal telencephalic neuroepithelium, and neurocan mRNA is present in both the ependymal and mantle layers of the spinal cord but not in the roof plate. The presence of neurocan mRNA in areas where the proteoglycan is not expressed suggests that the short open reading frame in the 5'-leader of neurocan may function as a cis-acting regulatory signal for the modulation of neurocan expression in the developing central nervous system.

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PubMed1996

The neostriatal mosaic: basis for the changing distribution of neurokinin-1 receptor immunoreactivity during development.

The pattern of neurokinin-1 receptor-like immunoreactivity (NK-1Rir) was mapped in perinatal and adult mouse striatum by using a new polyclonal antiserum. NK-1Rir was detected in the differentiating regions of the ganglionic eminences on embryonic day 12.5 (E12.5). NK-1Rir structures were enriched in the striatal patch compartment between E16.5 and approximately postnatal day 3 (P3); distributed more uniformly, within portions of both the patch and matrix compartments on P7; and enriched in the matrix compartment in the adult. Analysis of the phenotype of NK-1Rir cells on P2, P7, and in the adult suggested that cholinergic cells accounted for the majority of NK-1Rir cells early postnatally, with increasing contributions from somatostatinergic cells later postnatally. In the adult, approximately half of NK-1Rir cells were cholinergic and half were somatostatinergic. The transient enrichment of NK-1R-bearing cells and processes in the patch compartment which contains cells that express substance P (SP), a putative ligand for the NK-1R, may be a consequence of compartment formation or may be functionally important for compartment development.

Amino Acid Sequence↗

PubMed1997

Restricted expression of cadherin-8 in segmental and functional subdivisions of the embryonic mouse brain.

We have cloned full-length cDNA of a novel mouse cadherin ("mCad8"). The deduced amino acid sequence of the mature form of mCad8 shows 98.2% identity with the sequence of human cadherin-8. The expression of mCad8 was studied by in situ hybridization in mouse embryos of 9.5-14 days gestation (E9.5-E14). Results show that mCad8 expression is restricted to particular subdivisions of the early central nervous system (CNS) and to the thymus. In the CNS, mCad8 expression was observed from E11.5. In the telencephalon, mCad8 is expressed by the ventricular layer of the ganglionic eminence, by cortical areas, and by cells at the caudato-pallial angle. In the diencephalon, the margins of one mCad8-positive area correspond to the borders of the ventral thalamic neuromere, as confirmed by mapping the expression of gene regulatory proteins (Dlx-2, Pax-6, and Gbx-2). In the rhombencephalon, two large groups of mCad8-expressing cells were seen in the pons and in an area of the lateral basal plate of the myelencephalon. These groups of cells extend from the intermediate zone to the mantle zone at E12.5 and later form the anlage of the pontine and the facial nuclei. In conclusion, the expression of mCad8 reflects, in part, the neuromeric organization of the early embryonic CNS. In the mantle layer, mCad8 is expressed by developing gray matter structures, such as brain nuclei, suggesting a role for mCad8 in brain morphogenesis.

Amino Acid Sequence↗

PubMed1998

Dopaminergic growth patterns induced by striatal and cortical grafts show differences in sensitivity to increased striatal trophic activity induced by haloperidol.

During development, dopaminergic neurons innervate the developing striatal target, forming two different growth patterns: a patchy growth and a diffuse growth. Chronic treatment with the dopamine antagonist haloperidol increase the neurotrophic activity in striatum, but it is not known how this trophic activity influences different patterns of dopaminergic growth. In this paper we have studied dopaminergic growth patterns by evaluating tyrosine hydroxylase (TH)-positive growth from mature midbrain dopaminergic neurons innervating grafted fetal lateral ganglionic eminence (LGE) or fetal cortical tissue implanted into the dorsal striatum. Host dopaminergic neurons innervated LGE grafts with a patchy growth pattern, leaving large portions of the striatal graft noninnervated, and cortical grafts with diffuse, nonpatchy growth, evenly distributed over the total volume of the graft. Both types of growth patterns were enhanced over time, albeit the most pronounced change was found in the nonpatchy pattern in the cortical grafts. When fetal LGE or cortical tissues were transplanted into the dorsal striatum of chronically haloperidol-treated animals, the nonpatchy growth into fetal cortical grafts was enhanced while the patchy growth into fetal striatal tissue was not. Taken together, the results suggest that both the patchy and diffuse growth patterns may be induced from adult midbrain dopaminergic neurons depending on the target of innervation, and that increased striatal trophic activity due to blockade of the dopaminergic input stimulates growth from a subpopulation of dopaminergic neurons that produce the nonpatchy growth.

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PubMed1997

New observations on the development of the gonadotropin-releasing hormone system in the mouse.

In ongoing efforts to study the ontogeny of gonadotropin-releasing hormone (GnRH) neurons, we serendipitously observed that increasing times of incubation in antibodies enhanced signal detection. Here, we describe significant differences in the early migration pattern, population dynamics, and growth cone morphology from published reports. The first immunoreactive GnRH cells were detected in the mouse at E10.75 (7.6 +/- 2.8 cells; morning after mating = E0.5), prior to the closure of the olfactory placode. Although half of these cells were in the medial wall of the olfactory pit, the other half had already initiated their migration, and approximately one quarter had reached the telencephalic vesicle. Although the migratory pattern of the GnRH cells after E11.00 was identical to that described previously, these earliest migrating cells traveled singly rather than in cords, with some reaching the presumptive preoptic area (posterior to the ganglionic eminence) by E11.75. The number of GnRH cells increased significantly (p < 0.05) to 777 +/- 183 at E11.75 and peaked at 1949.6 +/- 161.6 (p < 0.05) at E12.75. The adult population was approximately 800 cells distributed between the central nervous system (CNS) and the nasal region. Hence, the population of GnRH neurons during early development is much larger than previously appreciated; mechanisms for its decline are discussed. Neuritic extensions on the earliest GnRH neurons are short (30-50 microm) and blunt and may represent the leading edge of the moving cell. By E12.75, GnRH axons in the CNS had a ribboned or beaded morphology and increasingly more complex growth cones were noted from this time until the day of birth. The most complex growth cones were associated with apparent choice points along the axons' trajectory. By E13.75, GnRH axons were seen at the presumptive median eminence in all animals, and it was at this stage that the axons began to branch profusely. Branching, as well as the presence of growth cones, continued post-natally. These results provide further insights into the pathfinding mechanisms of GnRH cells and axons.

Animals↗

PubMed2001

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.

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PubMed1995

Gsh-1: a novel murine homeobox gene expressed in the central nervous system.

We report the characterization of Gsh-1, a novel murine homeobox gene. Northern blot analysis revealed a transcript of approximately 2 kb in size present at embryonic days 10.5, 11.5, and 12.5 of development. The cDNA sequence encoded a proline rich motif, a polyalanine tract, and a homeodomain with strong homology to those encoded by the clustered Hox genes. The Gsh-1 expression pattern was determined for days E8.5 to E13.5 by whole mount and serial section in situ hybridizations. Gsh-1 transcription was restricted to the central nervous system. Expression is present in the neural tube and hindbrain as two continuous, bilaterally symmetrical stripes within neural epithelial tissue. In the mesencephalon, expression is seen as a band across the most anterior portion. There is also diencephalon expression in the anlagen of the thalamus and the hypothalamus as well as in the optic stalk, optic recess, and the ganglionic eminence. Moreover, through the use of fusion proteins containing the Gsh-1 homeodomain, we have determined the consensus DNA binding site of the Gsh-1 homeoprotein to be GCT/CA/CATTAG/A.

Amino Acid Sequence↗

PubMed2002

Ontogeny of gamma-aminobutyric acid-immunoreactive neuronal populations in the forebrain and midbrain of the sea lamprey.

Although brain organization in lampreys is of great interest for understanding evolution in vertebrates, knowledge of early development is very scarce. Here, the development of the forebrain and midbrain gamma-aminobutyric acid (GABA)-ergic systems was studied in embryos, prolarvae, and small larvae of the sea lamprey using an anti-GABA antibody. Ancillary immunochemical markers, such as proliferating cell nuclear antigen (PCNA), calretinin, and serotonin, as well as general staining methods and semithin sections were used to characterize the territories containing GABA-immunoreactive (GABAir) neurons. Differentiation of GABAir neurons in the diencephalon begins in late embryos, whereas differentiation in the telencephalon and midbrain was delayed to posthatching stages. In lamprey prolarvae, the GABAir populations appear either as compact GABAir cell groups or as neurons interspersed among GABA-negative cells. In the telencephalon of prolarvae, a band of cerebrospinal fluid-contacting (CSF-c) GABAir neurons (septum) was separated from the major GABAir telencephalic band, the striatum (ganglionic eminence) primordium. The striatal primordium appears to give rise to most GABAir neurons observed in the olfactory bulb and striatum of early larval stages. GABAir populations in the dorsal telencephalon appear later, in 15-30-mm-long larvae. In the diencephalon, GABAir neurons appear in embryos, and the larval pattern of GABAir populations is recognizable in prolarvae. A small GABAir cluster consisting mainly of CSF-c neurons was observed in the caudal preoptic area, and a wide band of scattered CSF-c GABAir neurons extended from the preoptic region to the caudal infundibular recess. A mammillary GABAir population was also distinguished. Two compact GABAir clusters, one consisting of CSF-c neurons, were observed in the rostral (ventral) thalamus. In the caudal (dorsal) thalamus, a long band extended throughout the ventral tier. The nucleus of the medial longitudinal fascicle contained an early-appearing GABAir population. The paracommissural pretectum of prolarvae and larvae contained a large group of non-CSF-c GABAir neurons, although it was less compact than those of the thalamus, and a further group was found in the dorsal pretectum. In the midbrain of larvae, several groups of GABAir neurons were observed in the dorsal and ventral tegmentum and in the torus semicircularis. The development of GABAergic populations in the lamprey forebrain was similar to that observed in teleosts and in mouse, suggesting that GABA is a very useful marker for understanding evolution of forebrain regions. The possible relation between early GABAergic cell groups and the regions of the prosomeric map of the lamprey forebrain (Pombal and Puelles [ 1999] J. Comp. Neurol. 414:391-422) is discussed in view of these results and information obtained with ancillary markers.

Aging↗

PubMed2002

Differential localization of high- and low-molecular-weight variants of microtubule-associated protein 2 in the developing rat telencephalon.

Microtubule-associated protein 2 (MAP2) occurs in developing mammalian neuronal tissue as both high- and low-molecular-weight forms with temporally regulated expression. We studied the MAP2 expression in the developing rat telencephalon with monoclonal antibodies that recognized both the high- and low-molecular-weight forms of MAP2 variants or that specifically recognized high-molecular-weight forms of MAP2 variants. Differences in the staining patterns of these antibodies reflected differences in the distribution of the high- and low-molecular-weight MAP2s. The immunoreactive sites of high- and low-molecular-weight MAP2 had a more widespread distribution in the embryonic telencephalon than those of high-molecular-weight MAP2. Many bipolar cells in the ganglionic eminence (GE) and in the intermediate zone (IZ) of the neocortex showed low-molecular-weight MAP2 immunoreactivity, but they showed weak or no high-molecular-weight MAP2 immunoreactivity. Expression of mRNA containing exons common to high- and low-molecular-weight MAP2 was detected in the tangentially ellipsoidal cells in the IZ, but expression of mRNA containing an exon specific to high-molecular-weight MAP2 was not detected in these cells by in situ hybridization. We interpreted these observations as indicating that the bipolar cells contained MAP2c preferentially, but contained MAP2a and MAP2b (MAP2a/b) at a very low or negligible level. The cells that expressed MAP2c preferentially among the MAP2 splicing variants composed 50% of the preplate cells, most of the MAP2-positive cells in the hippocampus and the corpus callosum. Double labeling by DiI staining and Dlx2 immunohistochemistry, or by Dlx2 and MAP2 immunohistochemistry, revealed that most of the Dlx2-positive cells in the IZ expressed MAP2c preferentially at embryonic day 16. Another double-labeling study revealed that most GAD-positive cells in the preplate were MAP2a/b positive, whereas most GAD-positive cells in the IZ expressed MAP2c preferentially, with only a negligible level of MAP2a/b immunoreactivity. We conclude that MAP2 immunoreactivity in the IZ was localized in the tangentially migrating neurons. The tangentially migrating neurons seemed to acquire MAP2a/b immunoreactivity as they entered the preplate or cortical plate and developed into mature neurons. Radially migrating neurons in the IZ were MAP2 negative. After entering to the preplate or the cortical plate, they became MAP2a/b positive as they developed into mature neurons.

Animals↗

PubMed2003

Evidence that Sema3A and Sema3F regulate the migration of GABAergic neurons in the developing neocortex.

The ganglionic eminence (GE) supplies neurons containing gamma-aminobutyric acid (GABA) to the pallium of the telencephalon. We investigated the molecular guidance mechanisms of GE cell migration in the neocortex and found neuropilin-1 (Npn-1) or neuropilin-2 (Npn-2) on the GE cells. Ectopic Sema3A or -3F expression by COS1 cell clusters placed on embryo neocortical slices reduced the cell migration but did not block it completely. However, the cell migration was almost completely blocked by COS1 cell clusters expressing both Sema3A and -3F. The direction of cell migration could be reversed by placing Sema3A- and -3F-coexpressing COS1 cell clusters at the distal cut end of the neocortical slices. Further slice experiments revealed that migration of half of the GE cells in the neocortex was regulated by Sema3A and that migration of the other half of the GE cells in the neocortex was regulated by Sema3F. When the cells responding to Sema3A were diverted by ectopic Sema3A expression in vivo, Dlx2-positive cells were found predominantly in the lower intermediate zone (IZ). When the cells responding to Sema3F were diverted by ectopic Sema3F expression in vivo, Dlx2-positive cells were found predominantly in the upper IZ. It was speculated that the semaphorin-neuropilin interactions distribute the GABAergic GE cells evenly in the neocortex as well as guide the GE cells from the GE to the neocortex. The Sema3A expression site under the subplate extended dorsally as the embryo developed. The Sema3A expression seemed to block the Npn-1-positive GE cells in the neocortex from entering the cortical plate (CP) and guide them to the dorsal cortex and the hippocampus. Sema3F expression in the CP continued through the embryonic stages. The expression seemed to block Npn-2-positive GE cells in the neocortex from entering the CP and make them migrate into the lower IZ. Finally, the semaphorin-neuropilin interactions sorted GABAergic inteneurons into the CP and white matter neurons into the IZ.

Animals↗

PubMed2003

Dlx-1 and Dlx-2 expression in the adult mouse brain: relationship to dopaminergic phenotypic regulation.

Expression of the homeodomain-containing transcription factors Dlx-1 and Dlx-2 in the lateral (LGE) and medial (MGE) ganglionic eminences, subpallial embryonic structures, is required for generation of telencephalic interneurons. LGE- and MGE-derived progenitors migrate and populate a number of forebrain structures, including the cortex, hippocampus, and olfactory bulb (OB). Previous reports focusing on embryogenesis of telencephalic neurons in Dlx-1 and Dlx-2 null mice suggested a specific role for these genes in expression of the OB dopamine (DA) phenotype. We have investigated whether these genes also are expressed in adult brain, especially in those pallial derivatives, such as the OB, hippocampus, and possibly cortex, where neurogenesis continues in adults. With a highly sensitive, nonradioactive in situ hybridization technique and both DLX-2 and pan DLX antisera, widespread expression of both genes was found in adult mouse fore- but not mid- or hindbrain. The adult unilateral naris closure paradigm was employed to establish a causative role for Dlx in regulating tyrosine hydroxylase (TH) expression; TH is the first enzyme in DA biosynthesis. TH mRNA, but not Dlx expression, was significantly down-regulated in the OB ipsilateral to closure. These findings suggest that Dlx-1 and -2 do not play a direct role in DA phenotypic differentiation and TH gene regulation in adult OB. The widespread expression of Dlx mRNA and protein in the adult brain suggests that these genes may have additional roles in mature animals.

Animals↗

PubMed2004

Identification and morphogenesis of the eminentia thalami in the zebrafish.

This study documents early zebrafish brain expression patterns (2-5 days postfertilization) of proliferating neural (PCNA) as well as early-determined (Pax6, Zash-1a, Zash-1b, neurogenin1, neuroD) and differentiating (Hu-proteins) neuronal cells. These patterns are used to outline the spatiotemporal local dynamics of secondary neurogenesis as well as neuronal migration and differentiation in the region of the eminentia thalami. The analysis presented not only allows identification for the first time of the eminentia thalami in the zebrafish model system (because it forms a neurogenin1/neuroD-guided locus of neurogenesis in contrast to adjacent preoptic region and ventral thalamus) but furthermore shows that the entopeduncular complex is a derivative of the embryonic zebrafish eminentia thalami, which has never been reported for a teleost before. An analysis of the relevant literature shows that the mammalian entopeduncular nucleus/avian paleostriatum primitivum/reptilian globus pallidus clearly are part of the basal ganglia (i.e., the pallidum). In amniote embryos, an anterior entopeduncular area is recognized at the base of the medial ganglionic eminence (i.e., the future pallidum; part of alar plate of prosomere 5), separate from the more posterior eminentia thalami (alar prosomere 4). There is a comparable periventricular eminentia thalami in (young and adult) amphibians and teleosts. However, the migrated anterior entopeduncular nucleus of anuran amphibians likely is homologous to part of the pallidum of other vertebrates and has no developmental relationship to the eminentia thalami. In contrast, the migrated teleostean entopeduncular complex does not correspond to a pallidal division but is indeed the adult derivative of the early-recognized eminentia thalami as shown in this study.

Animals↗

PubMed2004

Expression of Cux-1 and Cux-2 in the subventricular zone and upper layers II-IV of the cerebral cortex.

Little is known about how neurons in the different layers of the mammalian cerebral cortex are specified at the molecular level. Expression of two homologues of the Drosophila homeobox Cut gene, Cux-1 and Cux-2, is strikingly specific to the pyramidal neurons of the upper layers (II-IV) of the murine cortex, suggesting that they may define the molecular identity of these neurons. An antibody against Cux-1 labels the nucleus of most of the postmitotic upper layer neurons but does not label parvoalbumin-positive cortical interneurons that derive from the medial ganglionic eminence. Cux-1 and Cux-2 represent early markers of neuronal differentiation; both genes are expressed in postmitotic cortical neurons from embryonic stages to adulthood and in the proliferative regions of the developing cortex. In precursors cells, Cux-1 immunoreactivity is weak and diffuse in the cytoplasm and nucleus of ventricular zone (VZ) cells, whereas it is nuclear in the majority of bromodeoxyuridine (BrdU)-positive subventricular zone (SVZ) dividing cells, suggesting that Cux-1 function is first activated in SVZ cells. Cux-2 mRNA expression is also found in the embryonic SVZ, overlapping with BrdU-positive dividing precursors, but it is not expressed in the VZ. A null mutation in Pax-6 disrupts Cux-2 expression in the SVZ and Cux-1 and Cux-2 expression in the postmigratory cortical neurons. Thus, these data support the existence of an intermediate neuronal precursor in the SVZ dedicated to the generation of upper layer neurons, marked specifically by Cux-2. The patterns of expression of Cux genes suggest potential roles as determinants of the neuronal fate of the upper cortical layer neurons.

Animals↗