The nervous system is formed by a complex network of neuronal connections. During development, neurons elongate their axons through highly stereotyped anatomical pathways to form precise connections. Defects in these mechanisms are related with neurological disorders. Previous studies have reported that inhibition of the P2X7 receptor, an ionotropic purinergic receptor, promotes axonal growth and branching in cultured neurons. However, little is known about the in vivo mechanism of axonal elongation regulated by P2X7. Here, we detailed a step-by-step method to perform in utero cortical electroporation and quantified the electroporated axons employing accessible and open-source image processing software. This effective surgical procedure manipulates in vivo the gene expression in a discrete population of callosal projection neuron. Thus, a better understanding of the involvement of P2X7 in the in vivo establishment of neuronal circuits might help to clarify the basic biology of several neurodevelopmental disorders and axonal regenerative processes.
Keywords: Axonal growth; Corpus callosum; Electroporator; GFP; In utero electroporation; Neurodevelopment; Neurons; P2X7.
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