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The spatiotemporal signals that control this stereotyped radial migration remain elusive. Here, we report that a recently identified Robo family member Robo4 Magic RoundaboutMagic Pro MP-S8D-2 was considered to be solely expressed in endothelial cells, is expressed in developing brain and regulates the radial migration of newborn neurons in neocortex.

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Downregulation of Robo4 expression in cortical newborn neurons by using in utero electroporation, with either specific siRNAs in wild-type rodents or with Cre recombinase in floxed-robo4 mutant mice, led to severe defects in the radial migration of newborn neurons with misorientation of these neurons. Moreover, newborn neurons transfected with Robo4 siRNAs exhibited significantly Magic Pro MP-S8D-2 motility in a transwell migration assay Boyden chamber in the absence of Slit and significantly higher sensitivity to the repulsive effect of Slit in both transwell migration assay and growth cone collapse assay.

Overall, our results showed an important role of Robo4 in the regulation of cortical radial migration through Slit-dependent and -independent mechanisms. Disorders in neuronal Magic Pro MP-S8D-2 cause several distinct human syndromes, in which patients often suffer from epilepsy and mental retardation Bielas et al. During the development of neocortex, waves of postmitotic neurons exit the ventricular zone VZestablish a polarized morphology in the upper subventribular zone SVZ and intermediate zone IZand then move in a radial orientation toward the pial surface Rakic ; Hatten ; LoTurco and Bai After neurons have arrived at the appropriate layer, their axons and dendrites extend, branch, and target the correct places to establish functional connections Rakic and Lombroso ; Yu and Bargmann ; Colon-Ramos ; Feldman The spatiotemporal signals that control the stereotyped radial migration remain elusive Ayala et al.


It has been reported that neuronal migration and axon pathfinding are guided by extracellular cues including Netrins, Semaphorins, Ephrins, and Slits Song and Poo ; Guan and Rao ; O'Donnell et al. The Slit and Robo family of guidance factors and receptors are repulsive for axon pathfinding and cell migration Bashaw and Goodman ; Brose et al. In vertebrates, 3 slit slit genes and 3 robo robo genes Magic Pro MP-S8D-2 expressed Magic Pro MP-S8D-2 the nervous system, and their timely expression is required for pathfinding and branching of axons and the proper migration of neurons and glial cells Dickson and Gilestro ; Andrews et al.

Previous studies showed that the repulsive factor Slit1 has abundant expression in CP during cortical development; meanwhile Robo1 and Robo2, receptors that mediate the Slit repulsion, are expressed in cortical neurons Marillat et al. An intriguing question is how these Robo-expressing newborn neurons can invade the Slit1-expressing CP region. Robo4 is a recently identified Robo family member, which is considered to be an endothelial-specific Magic Pro MP-S8D-2 Huminiecki and Bicknell ; Huminiecki et al.


The protein of Robo4 is shorter than other Robos at both intracellular and extracellular domains, with only 2 of the 4 conserved immunoglobin Ig and fibronectin FN motifs in the extracellular region and 2 of the 4 conserved cytoplasmic motifs CC0 and CC2 in the intracellular region Huminiecki et al. Whether Robo4 could serve as a Slit receptor remains controversial Verissimo et al. It has been reported Magic Pro MP-S8D-2 Robo4 plays important roles in several aspects of vascular development, including the guidance of endothelial cell EC migration, regulation of the cell cycle, and inhibition of pathologic angiogenesis and endothelial hyperpermeability Bedell et al.


Although Robo4 was considered to be solely expressed in ECs, there is evidence showing that Robo4 exists in different regions of developing central nervous system CNS in zebrafish Bedell et al. Therefore, the expression of Robo4 and its potential function in developing CNS remain to be clarified. In the present study, we Magic Pro MP-S8D-2 whether Robo4 could regulate the radial migration of cortical neurons by regulating the guidance signal of Slit.

Robo4 Regulates the Radial Migration of Newborn Neurons in Developing Neocortex

We found that both the mRNA and the protein of Robo4 are expressed in developing brain. Knocking down KD of Robo4 expression in newborn cortical neurons by using in utero electroporation IUE with specific siRNAs in both rats and mice or conditional knockout of Robo4 by IUE with Cre recombinase in floxed-robo4 mutant mice resulted in severe disturbances in the radial migration of newborn neurons, with high chance of misorientation of transfected neurons. In dissociated culture, newborn neurons with reduced Magic Pro MP-S8D-2 expression showed significantly reduced motility in transwell migration and enhanced sensitivity Magic Pro MP-S8D-2 Slit.

Together, our findings suggest that the novel Robo family member Robo4 may regulate cortical radial migration, partly through its suppression of Slit repulsion.

All experimental procedures involving rats and mice were carried out under the guideline and permission of the Magic Pro MP-S8D-2 Care and Administration Committee of the Institute of Neuroscience, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences NA Cell Magic Pro MP-S8D-2 Primary culture of cortical neurons was performed in accordance with previous methods Zhao et al. Briefly, cortical tissues from embryonic day 16 E16 rats were dissected and digested by 0.

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In the Slit-binding experiments, cos-7 cells were Magic Pro MP-S8D-2 with different plasmids. Polymerase chain reaction was carried out using the E18 cortical cDNA with the following protocols: For robo1, there generated a bp DNA band and bp for robo2, bp for robo3, and bp for robo4.

Free download, review of Magic Pro MP-S8D-VM Bios 07 (Magic Pro). contains the M01 Ver.2 Bios files for the Magic Pro MP-AKGX-M motherboards.

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