Isolation of GABAergic Cortical Neurons and Implications for Cell Transplantation Strategies in the Nervous System

The correct layer placement of interneurons and pyramidal neurons during corticogenesis is required for precise neuronal activity and subsequent functions of the neocortex. Interneurons are generated in the medial ganglionic eminence (MGE) and migrate from the ventral to dorsal telencephalon to reside within the developing cortical plate. Transplantation strategies are valuable in understanding the cellular and molecular basis of interneuron development, and more recently in the potential for cell-based therapies of neurological disorders. With the advancement of transgenic technologies, interneurons can be fluorescently labelled to aid separation from other cell types in brain tissue and visualization in host tissues post-transplantation. This chapter presents a method to generate a dissociated cell preparation from embryonic MGE and utilizes fluorescence-activated cell sorting (FACS) to isolate migrating interneurons from progenitors and non-interneuron cell types. The FACS protocol is optimized to increase the purity and quantity of sorted interneurons that can be recovered and used for further analysis. This chapter also details how to perform in utero transplantation of dissociated MGE cells into the lateral ventricle of an embryonic brain and methods for analysis of the host brain tissue post-transplantation. These experimental approaches can be applied to understanding interneuron circuit formation and cell-based therapies to rectify GABAergic dysfunction in the neocor...
Source: Springer protocols feed by Neuroscience - Category: Neuroscience Source Type: news