Gundela Meyer

Different origins and developmental histories of transient neurons in the marginal zone of the fetal and neonatal rat cortex.

Two major classes of early‐born neurons are distinguished during early corticogenesis in the rat. The first class is formed by the cortical pioneer neurons, which are born in the ventricular neuroepithelium all over the cortical primordium. They appear at embryonic day (E) 11.5 in the lateral aspect of the telencephalic vesicle and cover its whole surface on E12. These cells, which show intense immunoreactivity for calbindin and calretinin, are characterized by their large size and axonal projection. They remain in the marginal zone after the formation of the cortical plate; they project first into the ventricular zone, and then into the subplate and the internal capsule. Therefore, these cells are the origin of the earliest efferent pathway of the developing cortex. Pioneer neurons are only present in prenatal brains. The second class is formed by subpial granule neurons, which form the subpial granular layer (SGL), previously considered to be found exclusively in the human cortex. SGL neurons are smaller than pioneer neurons. They are generated in a transient compartment of the retrobulbar ventricle between E12 and E14, and we propose the hypothesis that they invade the marginal zone, through tangential subpial migration, at different moments of fetal life. SGL neurons contain calbindin, calretinin, and gamma‐aminobutyric acid (GABA), but the GABA‐immunoreactive group becomes inconspicuous before birth. The extracellular matrix‐like glycoprotein reelin, a molecule crucial for cortical lamination, is prenatally expressed by SGL neurons; postnatally, it is present in both Cajal‐Retzius cells and subpial pyriform cells, both derivatives of SGL cells. In the rat, Cajal‐Retzius cells are horizontal neurons that remain only until the end of the first postnatal week. They are located in layer I at a critical distance of approximately 20 μm from the pial surface and express reelin and, only occasionally, calretinin. Subpial pyriform cells coexpress reelin and calretinin and remain in layer I longer than Cajal‐Retzius cells. Both pioneer neurons and subpial granule neurons are specific to the cortex. They mark the limit between the rudimentary cerebral cortex and olfactory bulb in the rat during early corticogenesis. J. Comp. Neurol. 397:493–518, 1998.

Comparative aspects of cerebral cortical development.

This review aims to provide examples of how both comparative and genetic analyses contribute to our understanding of the rules for cortical development and evolution. Genetic studies have helped us to realize the evolutionary rules of telencephalic organization in vertebrates. The control of the establishment of conserved telencephalic subdivisions and the formation of boundaries between these subdivisions has been examined and the very specific alterations at the striatocortical junction have been revealed. Comparative studies and genetic analyses both demonstrate the differential origin and migratory pattern of the two basic neuron types of the cerebral cortex. GABAergic interneurons are mostly generated in the subpallium and a common mechanism governs their migration to the dorsal cortex in both mammals and sauropsids. The pyramidal neurons are generated within the cortical germinal zone and migrate radially, the earliest generated cell layers comprising preplate cells. Reelin‐positive Cajal–Retzius cells are a general feature of all vertebrates studied so far; however, there is a considerable amplification of the Reelin signalling with cortical complexity, which might have contributed to the establishment of the basic mammalian pattern of cortical development. Based on numerous recent observations we shall present the argument that specialization of the mitotic compartments may constitute a major drive behind the evolution of the mammalian cortex. Comparative developmental studies have revealed distinct features in the early compartments of the developing macaque brain, drawing our attention to the limitations of some of the current model systems for understanding human developmental abnormalities of the cortex. Comparative and genetic aspects of cortical development both reveal the workings of evolution.

Prenatal development of reelin-immunoreactive neurons in the human neocortex.

Reelin, the protein defective in reeler mutant mice, is a secreted glycoprotein involved in the architectonic development of the central nervous system, more particularly in the development of neocortical lamination. In mice, reelin mRNA and protein expression are most robust in horizontal neurons of the embryonic marginal zone (MZ). By using monoclonal anti‐reelin antibodies (de Bergeyck et al. [1998] J. Neurosci. Methods), the morphology and evolution of reelin‐expressing neurons were studied in the MZ of the prenatal human neocortex. At 11 gestational weeks (GW), the MZ contained a single layer of reelin‐positive mono‐ or bipolar horizontal Cajal–Retzius (CR) cells. From 14 GW onward, the subpial granular layer (SGL) invaded the MZ, forming a transient layer of undifferentiated, initially reelin‐negative granule cells. In parallel to the emergence of the SGL and the morphological differentiation of the CR cells, a second population of reelin‐positive cells appeared within the SGL. These cells, termed CR‐like cells, were intermediate in size and shape between the CR cells and SGL granule cells. Between 16 GW and 24 GW, the packing density of the reelin‐producing cells remained remarkably stable, despite the continuous growth of the cortical surface. During this period, CR cells settled progressively deeper within the MZ, although they remained in contact with the pial surface through radially ascending processes. Most CR cells disappeared at around 27 GW, in parallel with the dissolution of the SGL. During the last weeks of gestation, reelin was expressed by a few medium‐sized, often horizontal neurons. These observations show that different neuronal populations in the human MZ express reelin and suggest that a possible function of the SGL is to supply reelin‐producing cells through a gradual transformation of reelin‐negative precursor cells into reelin‐immunoreactive CR‐like cells, thus coping with the protracted neurogenesis and dramatic surface expansion of the human neocortex. J. Comp. Neurol. 397:29–40, 1998.

Human disorders of cortical development: from past to present

Epilepsy and mental retardation, originally of unknown cause, are now known to result from many defects including cortical malformations, neuronal circuitry disorders and perturbations of neuronal communication and synapse function. Genetic approaches in combination with MRI and related imaging techniques continually allow a re‐evaluation and better classification of these disorders. Here we review our current understanding of some of the primary defects involved, with insight from recent molecular biology advances, the study of mouse models and the results of neuropathology analyses. Through these studies the molecular determinants involved in the control of neuron number, neuronal migration, generation of cortical laminations and convolutions, integrity of the basement membrane at the pial surface, and the establishment of neuronal circuitry are being elucidated. We have attempted to integrate these results with the available data concerning, in particular, human brain development, and to emphasize the limitations in some cases of extrapolating from rodent models. Taking such species differences into account is clearly critical for understanding the pathophysiological mechanisms associated with these disorders.

Developmental roles of p73 in Cajal-Retzius cells and cortical patterning.

To examine the role of the p53 homolog p73 in brain development, we studied p73-/-, p73+/-, E2F1-/-, and reeler mutant mice. p73 in developing brain is expressed in Cajal-Retzius (CR) cells, the cortical hem, and the choroid plexus. p73-expressing CR cells are lost in p73-/- embryos, although Reelin is faintly expressed in the marginal zone. Ectopic neurons in the p73-/- preplate and cortical hem at embryonic day 12 implicate p73 in the early developmental program of the cortex; however, preplate partition and early cortical plate formation are not disturbed. Postnatal p73-/- mice show a mild hypoplasia of the rostral cortex and a severely disrupted architecture of the posterior telencephalon. In the developing p73-/- hippocampus, the most striking abnormality is the absence of the hippocampal fissure, suggesting a role of p73 in cortical folding. p73+/- mice have a less severe cortical phenotype; they display a dorsal shift of the entorhinal cortex and a reduced size of occipital and posterior temporal areas, which acquire entorhinal-like features such as Reelin-positive cells in layer II. CR cells appear unaffected by heterozygosity. We relate the malformations of the posterior pole in p73 mutant mice to alterations of p73 expression in the cortical hem and suggest that p73 forms part of an early signaling network that controls neocortical and archicortical regionalization. In mice deficient for the transcription factor E2F1, a main activator of the TAp73 (transactivating p73) isoform, we find a defect of the caudal cortical architecture resembling the p73+/- phenotype along with reduced TAp73 protein levels and propose that an E2F1-TAp73 dependent pathway is involved in cortical patterning.

Distribution patterns of estrogen receptor α and β in the human cortex and hippocampus during development and adulthood

The expression of estrogen receptors (ERs) in the developing and adult human brain has not been clearly established, although estrogens are crucial for neuronal differentiation, synapse formation, and cognitive functions. By using immunohistochemistry, we have studied the distribution of ERα and ERβ in human cerebral cortex and hippocampus from early prenatal stages to adult life. ERα was detected in the cortex at 9 gestational weeks (GW), with a high expression in proliferating zones and the cortical plate. The staining intensity decreased gradually during prenatal development but increased again from birth to adulthood. In contrast, ERβ was first detected at 15 GW in proliferating zones, and at 16/17 GW, numerous ERβ immunopositive cells were also observed in the cortical plate. ERβ expression persisted in the adult cortex, being widely distributed throughout cortical layers II–VI. In addition, from around 15 GW to adulthood, ERα and ERβ were expressed in human hippocampus mainly in pyramidal cells of Ammons horn and in the dentate gyrus. Western blotting and immunohistochemistry in the adult cerebral cortex and hippocampus revealed lower protein expression of ERα compared with ERβ. Double immunostaining showed that during fetal life both ERs are expressed in neurons as well as in radial glia, although only ERα is expressed in the Cajal‐Retzius neurons of the marginal zone. These observations demonstrate that the expression of ERα and ERβ displays different spatial‐temporal patterns during human cortical and hippocampal development and suggest that both ERs may play distinct roles in several processes related to prenatal brain development. J. Comp. Neurol. J. Comp. Neurol. 503:790–802, 2007.

Morphology of neurons in the white matter of the adult human neocortex.

SummaryNeurons in the human cerebral cortical white matter below motor, visual, auditory and prefrontal orbital areas have been studied with the Golgi method, immunohistochemistry and diaphorase histochemistry. The majority of white matter neurons are pyramidal cells displaying the typical polarized, spiny dendritic system. The morphological variety includes stellate forms as well as bipolar pyramidal cells, and the expression of a certain morphological phenotype seems to depend on the position of the neuron. Spineless nonpyramidal neurons with multipolar to bitufted dendritic fields constitute less than 10% of the nuerons stained for microtubule associated protein (MAP-2). Only 3% of the MAP-2 immunoreactive neurons display nicotine adenine dinucleotide-diaphorase activity. The white matter pyramidal neurons are arranged in radial rows continuous with the columns of layer VI neurons. Neuron density is highest below layer VI, and decreases with increasing distance from the gray matter. White matter neurons are especially abundant below the primary motor cortex, and are least frequent below the visual cortex area 17. In contrast to other mammalian species, the white matter neurons in man are not only present during development, but persist throughout life.

DeltaNp73 regulates neuronal survival in vivo

Apoptosis occurs widely during brain development, and p73 transcription factors are thought to play essential roles in this process. The p73 transcription factors are present in two forms, the full length TAp73 and the N-terminally truncated DeltaNp73. In cultured sympathetic neurons, overexpression of DeltaNp73 inhibits apoptosis induced by nerve growth factor withdrawal or p53 overexpression. To probe the function of DeltaNp73 in vivo, we generated a null allele and inserted sequences encoding the recombinase Cre and green fluorescent protein (EGFP). We show that DeltaNp73 is heavily expressed in the thalamic eminence (TE) that contributes neurons to ventral forebrain, in vomeronasal neurons, Cajal-Retzius cells (CRc), and choroid plexuses. In DeltaNp73−/− mice, cells in preoptic areas, vomeronasal neurons, GnRH-positive cells, and CRc were severely reduced in number, and choroid plexuses were atrophic. This phenotype was enhanced when DeltaNp73–positive cells were ablated by diphtheria toxin expression. However, ablation of cells that express DeltaNp73 and Wnt3a did neither remove all CRc, nor did they abolish Reelin secretion or generate a reeler-like cortical phenotype. Our data emphasize the role of DeltaNp73 in neuronal survival in vivo and in choroid plexus development, the importance of the TE as a source of neurons in ventral forebrain, and the multiple origins of CRc, with redundant production of Reelin.

THE PALEOCORTICAL VENTRICLE IS THE ORIGIN OF REELIN-EXPRESSING NEURONS IN THE MARGINAL ZONE OF THE FOETAL HUMAN NEOCORTEX

The subpial granular layer (SGL) is a transient cell layer in the cortical marginal zone during the period of neuronal migration into the cortical plate. The origin of the SGL has been studied by immunocytochemistry for calretinin (CR) and reelin in human foetuses from 11 to 40 gestational weeks (GW). At 11 GW, the paleocortical ventricle, a rostral dilatation of the lateral ventricle, gives rise to two fountainheads: a medial fountainhead provides neurons for the marginal zone (MZ) of the rostral cortex and rostral hippocampal rudiment, while multiple cell streams migrate from a lateral fountainhead into the MZ of the paleocortex and insula. The latero‐medial gradient of neuronal packing density in the neocortical MZ indicates that migration extends farther into the neocortex. Neurons express CR already in the retrobulbar ventricular zone; they express reelin only as they approach the MZ of the paleocortex and rostral archicortex. At 16/17 GW, large numbers of CR‐immunoreactive granule cells originate from the same fountainheads, and then direct medially, toward the surface of the anterior perforated substance, and laterally, into the paleocortical MZ, from where they continue into the neocortical SGL following a ventrolateral to dorsomedial gradient. From 13 to 18 GW, reelin is expressed by a subpopulation of granule cells and by Cajal–Retzius‐like neurons. By 22 GW, the paleocortical ventricle undergoes regression and no longer supplies the SGL. Our results show that the paleocortical ventricle gives rise to a stream of neurons which extends over the cortical MZ as the subpial granular layer. The fact that SGL derivatives express reelin suggests that this transient cell layer may play a significant role in the establishment of the complex cytoarchitecture of the cerebral cortex.

VDAC and ERα interaction in caveolae from human cortex is altered in Alzheimer's disease

Voltage-dependent anion channel (VDAC) is a mitochondrial porin also found in the neuronal membrane (pl-VDAC), where its function may be related to redox homeostasis and apoptosis. Murine models have evidenced pl-VDAC into caveolae in a complex with estrogen receptor alpha (mERalpha), which participates in neuroprotection against amyloid beta (Abeta), and whose integration into this hydrophobic domain remains unclear. Here, we have demonstrated in caveolae of human cortex and hippocampus the presence of pl-VDAC and mERalpha, in a complex with scaffolding caveolin-1 which likely provides mERalpha stability at the plasma membrane. In Alzheimers disease (AD) brains, VDAC was accumulated in caveolae, and it was observed in dystrophic neurites of senile plaques, whereas ERalpha was expressed in astrocytes surrounding the plaques. Together with previous data in murine neurons demonstrating the participation of pl-VDAC in Abeta-induced neurotoxicity, these data suggest that the channel may be involved in membrane dysfunctioning observed in AD neuropathology.