Use of “MGE Enhancers” for Labeling and Selection of Embryonic Stem Cell-Derived Medial Ganglionic Eminence (MGE) Progenitors and Neurons

Ying-Jiun J Chen,Len A Pennacchio,Arnold R Kriegstein,Yoshiki Sasai,Daniel Vogt,Teruko Danjo,Arturo Alvarez-Buylla,Yanling Wang,Stewart Anderson,John L R Rubenstein,Scott C Baraban,Shanni N Silberberg,Cory R Nicholas,Joshua L Pollack,Axel Visel,Marcel Daadi

doi:10.1371/journal.pone.0061956

Abstract

The medial ganglionic eminence (MGE) is an embryonic forebrain structure that generates the majority of cortical interneurons. MGE transplantation into specific regions of the postnatal central nervous system modifies circuit function and improves deficits in mouse models of epilepsy, Parkinson's disease, pain, and phencyclidine-induced cognitive deficits. Herein, we describe approaches to generate MGE-like progenitor cells from mouse embryonic stem (ES) cells. Using a modified embryoid body method, we provided gene expression evidence that mouse ES-derived Lhx6+ cells closely resemble immature interneurons generated from authentic MGE-derived Lhx6+ cells. We hypothesized that enhancers that are active in the mouse MGE would be useful tools in detecting when ES cells differentiate into MGE cells. Here we demonstrate the utility of enhancer elements [422 (DlxI12b), Lhx6, 692, 1056, and 1538] as tools to mark MGE-like cells in ES cell differentiation experiments. We found that enhancers DlxI12b, 692, and 1538 are active in Lhx6-GFP+ cells, while enhancer 1056 is active in Olig2+ cells. These data demonstrate unique techniques to follow and purify MGE-like derivatives from ES cells, including GABAergic cortical interneurons and oligodendrocytes, for use in stem cell-based therapeutic assays and treatments.

Highlights

Cortical interneuron dysfunction may contribute to the risk of developing autism, epilepsy, bipolar disorder, schizophrenia, and dementia [1,2,3,4,5]
Cortical interneurons are born in the progenitor zones of the medial ganglionic eminence (MGE), the caudal ganglionic eminence (CGE) and preoptic area (POA), and migrate tangentially into the cortex [6,7,8]
Since previous studies had been successful in expanding neural stem cells in serum-free or serum-containing media with the addition of epidermal growth factor (EGF) and basic fibroblast growth factor [41,42], we tested these different protocols for MGE cells

Summary

Introduction

Cortical interneuron dysfunction may contribute to the risk of developing autism, epilepsy, bipolar disorder, schizophrenia, and dementia [1,2,3,4,5]. Cortical interneurons are born in the progenitor zones of the medial ganglionic eminence (MGE), the caudal ganglionic eminence (CGE) and preoptic area (POA), and migrate tangentially into the cortex [6,7,8] (abbreviations are listed in Table S1 in File S2). Several transcription factors, such as Dlx1&2, Nkx and Lhx, regulate interneuron development. Dlx12/2 mice are viable, but, due to lateonset interneuron loss, develop cortical dysrhythmias and epilepsy [9]. Lhx is required for differentiation of Parvalbumin+ and Somatostatin+ interneurons [18,19]

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Results

Conclusion