Miguel Valdeolmillos

Chemokine Signaling Controls Intracortical Migration and Final Distribution of GABAergic Interneurons

Functioning of the cerebral cortex requires the coordinated assembly of circuits involving glutamatergic projection neurons and GABAergic interneurons. Although much is known about the migration of interneurons from the subpallium to the cortex, our understanding of the mechanisms controlling their precise integration within the cortex is still limited. Here, we have investigated in detail the behavior of GABAergic interneurons as they first enter the developing cortex by using time-lapse videomicroscopy, slice culture, and in utero experimental manipulations and analysis of mouse mutants. We found that interneurons actively avoid the cortical plate for a period of ∼48 h after reaching the pallium; during this time, interneurons disperse tangentially through the marginal and subventricular zones. Perturbation of CXCL12/CXCR4 signaling causes premature cortical plate invasion by cortical interneurons and, in the long term, disrupts their laminar and regional distribution. These results suggest that regulation of cortical plate invasion by GABAergic interneurons is a key event in cortical development, because it directly influences the coordinated formation of appropriate glutamatergic and GABAergic neuronal assemblies.

Biased selection of leading process branches mediates chemotaxis during tangential neuronal migration

Current models of chemotaxis during neuronal migration and axon guidance propose that directional sensing relies on growth cone dynamics. According to this view, migrating neurons and growing axons are guided to their correct targets by steering the growth cone in response to attractive and repulsive cues. Here, we have performed a detailed analysis of the dynamic behavior of individual neurons migrating tangentially in telencephalic slices using high-resolution time-lapse videomicroscopy. We found that cortical interneurons consistently display branched leading processes as part of their migratory cycle, a feature that seems to be common to many other populations of GABAergic neurons in the brain and spinal cord. Analysis of the migratory behavior of individual cells suggests that interneurons respond to chemoattractant signals by generating new leading process branches that are better aligned with the source of the gradient, and not by reorienting previously existing branches. Moreover, experimental evidence revealed that guidance cues influence the angle at which new branches emerge. This model is further supported by pharmacological experiments in which inhibition of branching blocked chemotaxis, suggesting that this process is an essential component of the mechanism controlling directional guidance. These results reveal a novel guidance mechanism during neuronal migration that might be extensively used in brain development.

Actomyosin contraction at the cell rear drives nuclear translocation in migrating cortical interneurons.

Neuronal migration is a complex process requiring the coordinated interaction of cytoskeletal components and regulated by calcium signaling among other factors. Migratory neurons are polarized cells in which the largest intracellular organelle, the nucleus, has to move repeatedly. Current views support a central role for pulling forces that drive nuclear movement. The participation of actomyosin driven forces acting at the nucleus rear has been suggested, however its precise contribution has not been directly addressed. By analyzing interneurons migrating in cortical slices of mouse brains, we have found that nucleokinesis is associated with a precise pattern of actin dynamics characterized by the initial formation of a cup-like actin structure at the rear nuclear pole. Time-lapse experiments show that progressive actomyosin contraction drives the nucleus forward. Nucleokinesis concludes with the complete contraction of the cup-like structure, resulting in an actin spot at the base of the retracting trailing process. Our results demonstrate that this actin remodeling requires a threshold calcium level provided by low-frequency spontaneous fast intracellular calcium transients. Microtubule stabilization with taxol treatment prevents actin remodeling and nucleokinesis, whereas cells with a collapsed microtubule cytoskeleton induced by nocodazole treatment, display nearly normal actin dynamics and nucleokinesis. In summary, the results presented here demonstrate that actomyosin forces acting at the rear side of the nucleus drives nucleokinesis in tangentially migrating interneurons in a process that requires calcium and a dynamic cytoskeleton of microtubules.

Neurons in motion: same principles for different shapes?

The special conformation of the developing nervous system, in which progenitor zones are largely confined to the lumen of the neural tube, places neuronal migration as one of the most fundamental processes in brain development. Previous studies have shown that different neuronal types adopt distinct morphological modes of migration in the developing brain, indicating that neuronal migration might be a diverse process. Here, we review recent data on the molecular mechanisms underlying neuronal migration that suggest that similar signaling principles are responsible for the frequently variable morphology of different types of migrating neuron. According to this idea, the same basic molecular mechanisms found in other cell types, such as fibroblasts, might have been adapted to the special morphological needs of migrating neurons in different regions of the developing brain.

Contact Repulsion Controls the Dispersion and Final Distribution of Cajal-Retzius Cells

Cajal-Retzius (CR) cells play a fundamental role in the development of the mammalian cerebral cortex. They control the formation of cortical layers by regulating the migration of pyramidal cells through the release of Reelin. The function of CR cells critically depends on their regular distribution throughout the surface of the cortex, but little is known about the events controlling this phenomenon. Using time-lapse video microscopy in vivo and in vitro, we found that movement of CR cells is regulated by repulsive interactions, which leads to their random dispersion throughout the cortical surface. Mathematical modeling reveals that contact repulsion is both necessary and sufficient for this process, which demonstrates that complex neuronal assemblies may emerge during development through stochastic events. At the molecular level, we found that contact repulsion is mediated by Eph/ephrin interactions. Our observations reveal a mechanism that controls the even distribution of neurons in the developing brain.

Cajal-Retzius cells in early postnatal mouse cortex selectively express functional metabotropic glutamate receptors.

Glutamate receptors have been linked to the regulation of several developmental events in the CNS. By using cortical slices of early postnatal mice, we show that in layer I cells, glutamate produces intracellular calcium ([Ca2+]i) elevations mediated by ionotropic and metabotropic glutamate receptors (mGluRs). The contribution of mGluRs to these responses was demonstrated by application of tACPD, an agonist to groups I and II mGluRs, which evoked [Ca2+]i increases that could be reversibly blocked by MCPG, an antagonist to groups I and II mGluRs. In the absence of extracellular Ca2+, repetitive applications of tACPD or quisqualate, an agonist to group I mGluRs, elicited decreasing [Ca2+]i responses that were restored by refilling a thapsigargin‐sensitive Ca2+ store. The use of specific group I mGluR agonists CHPG and DHPG indicated that the functional mGluR in layer I was of the mGluR1 subtype. Subtype specific antibodies confirmed the presence of mGlur1α, but not mGluR5, in Cajal‐Retzius (Reelin‐immunoreactive) neurons.

Prenatal thalamic waves regulate cortical area size prior to sensory processing

The cerebral cortex is organized into specialized sensory areas, whose initial territory is determined by intracortical molecular determinants. Yet, sensory cortical area size appears to be fine tuned during development to respond to functional adaptations. Here we demonstrate the existence of a prenatal sub-cortical mechanism that regulates the cortical areas size in mice. This mechanism is mediated by spontaneous thalamic calcium waves that propagate among sensory-modality thalamic nuclei up to the cortex and that provide a means of communication among sensory systems. Wave pattern alterations in one nucleus lead to changes in the pattern of the remaining ones, triggering changes in thalamic gene expression and cortical area size. Thus, silencing calcium waves in the auditory thalamus induces Rorβ upregulation in a neighbouring somatosensory nucleus preluding the enlargement of the barrel-field. These findings reveal that embryonic thalamic calcium waves coordinate cortical sensory area patterning and plasticity prior to sensory information processing.

Regulation by tolbutamide and diazoxide of the electrical activity in mouse pancreatic β‐cells recorded in vivo

The glucose‐dependence of β‐cell electrical activity and the effects of tolbutamide and diazoxide were studied in anaesthetized mice. In untreated animals there was a direct relationship between glycaemia and the burst pattern of electrical activity. Animals with high glucose concentration showed continuous electrical activity. The application of insulin led to a steady decrease in blood glucose concentration and a transition from continuous to oscillatory activity at 7.7±0.1 mM glucose (mean±s.d.) and a subsequent transition from oscillatory to silent at 4.7±0.6 mM glucose. At physiological blood glucose concentrations the electrical activity was oscillatory. The injection of tolbutamide (1800 mg kg−1) transformed this oscillatory pattern into one of continuous electrical activity. The increased electrical activity was associated with a decrease in blood glucose concentration from 7.1±0.9 (control) to 5.5±1.0 mM (10 min after tolbutamide injection). The effects of tolbutamide are consistent with a direct blocking effect on the KATP channel that leads to membrane depolarization. The injection of diazoxide (6000 mg kg−1) hyperpolarized the cells and transformed the oscillatory pattern into a silent one. This is consistent with a direct stimulant effect by diazoxide on the KATP channel. The use of tolbutamide or diazoxide correspondingly led to the lengthening or shortening of the active phase of electrical activity, respectively. This indicates that in vivo, such activity can be modulated by the relative degree of activation or inhibition of the KATP channel. These results indicate that under physiological conditions, tolbutamide and diazoxide have direct and opposite effects on the electrical activity of pancreatic β‐cells, most likely through their action on KATP channels. This is consistent with previous work carried out on in vitro models and explains the drugs hypo‐ and hyperglycaemic effects.

Impact of Thalamocortical Input on Barrel Cortex Development

The development of cortical maps requires the balanced interaction between genetically determined programs and input/activity-dependent signals generated spontaneously or triggered from the environment. The somatosensory pathway of mice provides an excellent scenario to study cortical map development because of its highly organized cytoarchitecture, known as the barrel field. This precise organization makes evident even small alterations in the cortical map layout. In this review, we will specially focus on the thalamic factors that control barrel field development. We will summarize the role of thalamic input integration and identity, neurotransmission and spontaneous activity in cortical map formation and early cross-modal plasticity.

Functional NMDA and GABAA receptors in pioneer neurons of the cortical marginal zone.

Transient pioneer neurons in the neocortical marginal zone generate an early corticofugal axonal projection at E12–E16 ( Meyer et al. 1998 ). We have analysed the functional activity of glutamate and GABA receptors in such cells by measuring changes in intracellular calcium concentrations ([Ca2+]i). The activation of GABAA receptors with muscimol, as well as bath application of glutamate, lead to increases in [Ca2+]i in pioneer neurons. The stimulatory action of glutamate is mostly produced through the NMDA‐type of ionotropic receptors. Metabotropic glutamate receptor activation has no effect on [Ca2+]i. Consistent with such results, immunocytochemical studies showed a prominent expression of GABAA and NMDA receptors in pioneer neurons. The activation of such receptors may be implicated in the remodelling of pioneer neurons during development.