Midbrain dopaminergic and hindbrain serotonergic neurons play an important role in the modulation of behavior and are involved in a series of neuropsychiatric disorders. Despite the importance of ...these cells, little is known about the molecular mechanisms governing their development. During embryogenesis, midbrain dopaminergic neurons are specified rostral to the midbrain-hindbrain organizer (MHO), and hindbrain serotonergic neurons are specified caudal to it. We report that in transgenic mice in which Otx2 and accordingly the MHO are shifted caudally, the midbrain dopaminergic neuronal population expands to the ectopically positioned MHO and is enlarged. Complementary, the extension of the hindbrain serotonergic cell group is decreased. These changes are preserved in adulthood, and the additional, ectopic dopaminergic neurons project to the striatum, which is a proper dopaminergic target area. In addition, in mutants in which Otx2 and the MHO are shifted rostrally, dopaminergic and serotonergic neurons are relocated at the newly positioned MHO. However, in these mice, the size ratio between these two cell populations is changed in favor of the serotonergic cell population. To investigate whether the position of the MHO during embryogenesis is also of functional relevance for adult behavior, we tested mice with a caudally shifted MHO and report that these mutants show a higher locomotor activity. Together, we provide evidence that the position of the MHO determines the location and size of midbrain dopaminergic and hindbrain serotonergic cell populations in vivo. In addition, our data suggest that the position of the MHO during embryogenesis can modulate adult locomotor activity.
Parvalbumin (PV) belongs to the large family of EF‐hand calcium‐binding proteins and is an excellent marker for a subpopulation of GABAergic neocortical interneurons. During cortical development, PV ...first appears on postnatal day (P)8, in the infragranular layers; after P14, it also becomes apparent within the supragranular layers. However, nothing is known about the factors controlling its expression, which could involve functional activity, neuronal connectivity and/or neurotrophic factors. It being difficult to manipulate these parameters in vivo, their role may be more readily assessed in organotypic cultures, which are deprived of their subcortical afferents and efferents, and hence of subcortically derived neurotrophic factors and extrinsic functional activity. We prepared slices of the rat brain on P3, P5, P7 and P9, maintained them in culture for 2–5 weeks, and compared the temporal and spatial distribution pattern of PV‐immunoreactivity within these slices with the in vivo situation. We found, first, that during late postnatal in vivo development and ageing, the number of PV‐immunoreactive neurons in the parietal cortex decreases significantly, and second, that the expression of PV‐immunoreactivity in the parietal cortex was markedly influenced by the phase of postnatal development at which slice cultures were explanted. In those removed on P7 and P9, the number of PV‐immunoreactive cells, as well as the temporal and spatial distribution pattern of PV‐immunoreactivity corresponded to the in vivo situation, but in explants obtained on P3 or P5, PV‐immunoreactivity remained confined to layer V of the cortex, reminiscent of the expression profile manifested at the end of the second postnatal week in vivo. Also, the number of PV‐immunoreactive cells in these cultures was significantly lower than in explants at the later stages. Our results indicate that the onset of PV‐expression in the parietal cortex depends upon extrinsic cortical factors subsisting prior to P7. Once the production of this protein has been initiated, such influences are no longer required.
The anatomical distributions of the mitochondrial enzyme cytochrome oxidase (CO) and of the calcium binding protein parvalbumin (PV) were studied in the striate cortex of adult and neonate New World ...monkeys (Callithrix jacchus). In the adult marmoset, both proteins were found in laminar arrangements similar to those described for the macaque monkey, with prominent bands of PV-like immunoreactive (PV-LI) puncta in layers IV and IIIb, and fairly evenly distributed PV-LI nonpyramidal neurons. Furthermore, the pattern of CO activity in area 17 of the neonate marmoset was almost identical to the CO pattern described in neonate macaque and squirrel monkeys. It came, therefore, as a surprise to find that the adult pattern of PV-like immunoreactivity (PV-LI) in the marmoset striate cortex arises from a neonatal pattern strikingly different from that seen in any developmental stage of the macaque, or in any other mammal studied so far. In the deep layers IV through VI of the neonate marmoset, a large number of PV-LI neurons was stained in bandlike patterns, their number in layers IV and V exceeding the number of PV-LI neurons present in these layers of the adult marmoset area 17. Staining of layers IV and VI was restricted to area 17 and involved nonpyramidal cells and their processes. The stained band of layer V, in contrast, continued throughout most of the neocortex. In area 17, an estimated 10 to 20% of the stained cells in layer V exhibited pyramidal shapes. The findings show that the expression of PV by visual cortical cells occurs before birth and suggest that the comparatively early onset of PV expression is not dependent on the onset of textured vision. The exuberant number of stained cells in some layers, and particularly the staining of pyramidal cells, in the neonate marmoset, suggest that a considerable number of cells possesses the stainability for PV-LI only transiently, i.e., in the marmoset, these cells have a specific demand for parvalbumin during this phase of their development.
Postnatal development of the rat cortex is characterized by the gradual development of many calcium-dependent processes which demand a precise control of the intracellular levels of this cation; when ...the balance is disturbed, neuronal death ultimately ensues. Calretinin (CR), a calcium-binding protein, has been postulated to have neuroprotective capacity by buffering intracellular calcium. This putative relationship between CR and neuroprotection is still, however, a controversial issue. With a view to shedding further light on this subject, we studied the temporal and spatial distribution of CR in five different regions (the frontal- sensorimotor-, parietal-, temporal- and occipital region) of the rat cortex during postnatal development. Qualitative and quantitative immunocytochemistry of newborn, 5-, 10-, 15-, 20- and 30-day-old and adult rats revealed a profound increase in the density of the CR-positive neurons during the first two postnatal weeks in all regions examined. At the end of this period, CR-immunoreactive cells decreased sharply to adult levels. Cell classes exhibiting transient CR-immunoreactivity during the first two postnatal weeks included cells in layer I (amongst which were Cajal-Retzius cells), the subplate and pyramidal-like cells in the upper portion of layer V, most of them in the motor cortices. The above-described dynamics of CR expression were reflected also in the biochemical analysis performed (immunoblotting, ELISA). The temporal and spatial correlation with calcium-dependent events such as synaptogenesis, neurite elongation and remodelling in further support the contention that CR may play a neuroprotective role during postnatal development of the rat cortex.
Here we describe a detailed analysis of the expression of neurochondrin ( ncdn) in the developing and adult mouse brain. Ncdn is first expressed in the hindbrain and spinal cord at embryonic day 10.5 ...(E10.5) followed by expression in the midbrain at E11.5. By E18 ncdn is also expressed in the diencephalon and telencephalon. However, strongest expression is still observed in the hindbrain. In adults, the expression in the forebrain is as strong as in the hindbrain. Ncdn is highly expressed in the hippocampus, piriform cortex, septum, amygdaloid complex, medial geniculate nucleus, inferior colliculus, cerebellar nuclei and the nuclei of the Vth, VIIth, and XIIth cranial nerves.
The occurrence of Cajal-Retzius (CjRe) cells was studied in four cortical areas of five normal adult humans using antibodies against calretinin. Calretinin immunofluorescence and autofluorescence of ...lipofusin granules in CjRe cells were visualized by dual channel confocal laser scanning microscopy. Three types of CjRe cells existed in the adult human cortex: horizontal, triangular and multipolar, and their number did not decrease with ageing. Horizontal CjRe cells were found in all cortical areas; they contained no lipofusin or less than other cells. Triangular CjRe cells with descending dendrites were less numerous. Multipolar CjRe cells were rare and contained more lipofuscin. We conclude that calretinin-immunoreactivity can be used to study CjRe cell morphology in normal and diseased adult human brain.
In pheochromocytoma (PC12) cells nerve growth factor (NGF) and epidermal growth factor (EGF) activate similar receptor tyrosine kinase signaling pathways but evoke strikingly different biological ...outcomes: NGF induces differentiation and EGF acts as a mitogen. A novel approach was developed for identifying transcription factor activities associated with NGF-activated, but not EGF-activated, signaling, using random oligonucleotide clones from a DNA recognition library to isolate specific DNA binding proteins from PC12 nuclear extracts. A protein complex from NGF-treated, but not EGF-treated, cells was identified that exhibits increased mobility and DNA binding activity in gel mobility shift assays. The binding complex was identified in supershift assays as Fra-2/JunD. The clones used as probes contain either AP-1 or cAMP response element binding (CREB) recognition elements. Time course experiments revealed further differences in NGF and EGF signaling in PC12 cells. NGF elicits a more delayed and sustained ERK phosphorylation than EGF, consistent with previous reports. Both growth factors transiently induce c-fos, but NGF evokes a greater response than EGF. NGF specifically increases Fra-1 and Fra-2 levels at 4 and 24 hr. The latter is represented in Western blots by bands in the 40-46 kDa range. NGF, but not EGF, enhances the upper bands, corresponding to phosphorylated Fra-2. These findings suggest that prolonged alterations in Fra-2 and subsequent increases in Fra-2/JunD binding to AP-1 and CREB response elements common among many gene promoters could serve to trigger broadly an NGF-specific program of gene expression.
Neurons of area 17, the primary visual cortex, project to various anatomically and physiologically different extrastriate areas and subcortical regions. In the present investigation, we addressed the ...question of whether the efferent neurons in area 17 can contribute to functional diversity between these regions. We approached this question by analyzing the dendritic morphology of neurons in area 17 projecting to area MT, area 19DM, and the superior colliculus in the new world simian primate Callithrix jacchus, because dendritic morphology is an important factor in determining physiological properties of nerve cells. Retrograde transport of fluorochromes injected into the target regions, and intracellular injections of Lucifer yellow in the prelabelled neurons, revealed the following. 1) Morphologically identical large pyramidal cells in layer VI of area 17 project to all three targets. Some of them possess axon collaterals to two or all three targets, suggesting that they provide common information to all three areas. 2) Pyramidal cells in layer IIIc projecting to area MT form a morphologically homogeneous population. 3) Three small to medium-sized pyramidal cell types in layers IIIa-c, spiny stellate cells in layer IIIc, and another large pyramidal cell type in layer VI project to area 19DM. 4) Pyramidal cells in the lower two-thirds of layer V in area 17 project to the superior colliculus. In conclusion, we have shown that in Callithrix one efferent pathway may originate from several cell types. However, with the exception of the large cells in layer VI, efferent cells projecting to area MT, area 19DM, and the superior colliculus were morphologically distinct. This suggests that functional differences between brain regions could arise in part from morphological heterogeneity between and within the efferent cell populations.
Parvalbumin-immunoreactive interneurons are surrounded by perineuronal nets, containing molecules of the extracellular matrix (e.g. tenascin-R). Furthermore, they seem to have a special cytoskeleton ...composed of, among others, ankyrin
R and
β
Rspectrin. In the present developmental study we showed that the intracellular markers parvalbumin, ankyrin
R and
β
Rspectrin Villosa agglutinin, an extracellular marker for perineuronal nets, appeared in the second postnatal week. In the third postnatal week, ankyrin
R and
β
Rspectrin were present in the parvalbumin-positive interneurons. Tenascin-R appeared in a similar topographic distribution as the intracellular markers. The adult pattern was established upon the end of the fourth postnatal week. Our results indicate that cytoskeletal maturity maybe a prerequisite for the organization of perineuronal nets of extracellular matrix.
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