Abstract
Neuronal development is regulated by a complex combination of environmental and genetic factors. Assessing the relative contribution of each component is a complicated task, which is particularly difficult in regards to the development of γ-aminobutyric acid (GABA)ergic cortical interneurons (CIs). CIs are the main inhibitory neurons in the cerebral cortex, and they play key roles in neuronal networks, by regulating both the activity of individual pyramidal neurons, as well as the oscillatory behavior of neuronal ensembles. They are generated in transient embryonic structures (medial and caudal ganglionic eminences -MGE and CGE) that are very difficult to efficiently target using in utero electroporation approaches. Interneuron progenitors migrate long distances during normal embryonic development, before they integrate in the cortical circuit. This remarkable ability to disperse and integrate into a developing network can be hijacked by transplanting embryonic interneuron precursors into early post-natal host cortices. Here, we present a protocol that allows genetic modification of embryonic interneuron progenitors using focal ex vivo electroporation. These engineered interneuron precursors are then transplanted into early post-natal host cortices, where they will mature into easily identifiable CIs. This protocol allows the use of multiple genetically encoded tools, or the ability to regulate the expression of specific genes in interneuron progenitors, in order to investigate the impact of either genetic or environmental variables on the maturation and integration of CIs.
Highlights
The function of neuronal networks relies in the existence of a balanced complement of excitatory projection neurons and inhibitory interneurons
Using the procedure presented here, we tested whether the survival of cortical interneurons during early postnatal stages is regulated by activity in a cell autonomous manner
We describe a widely accessible methodology to genetically modify the activity of cortical interneurons (CIs) precursors to study the impact of intrinsic activity on CI maturation, and/or the effect of activity modulated CIs on the assembly/function of the integrated cortical circuits
Summary
The function of neuronal networks relies in the existence of a balanced complement of excitatory projection neurons and inhibitory interneurons. One of the great advantages of these methods is the potential to label and genetically modify the isolated progenitors and follow their differentiation in detail, to detect cell-autonomous changes. These methods are unable to offer information regarding the interactions between developing interneurons and an active network. Interneuron progenitors isolated from embryonic ganglionic eminences are able to survive, disperse and integrate into the host network upon transplantation into the cortex[7,8] This method has been used to reduce the severity of epileptic seizures in genetic mouse models, and has been proposed as a possible new therapy for different neurodevelopmental disorders[9,10].
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