A Method to Culture GABAergic Interneurons Derived from the Medial Ganglionic Eminence
Understanding the mechanisms guiding interneuron development is a central aspect of the current research on cortical/hippocampal interneurons, which is highly relevant to brain function and pathology. In this methodological study we have addressed the setup of protocols for the reproducible culture...
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doaj-bb3661304bbd4907b819175d6f0e2b212020-11-24T22:43:20ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022018-01-011110.3389/fncel.2017.00423308210A Method to Culture GABAergic Interneurons Derived from the Medial Ganglionic EminenceSira A. Franchi0Romina Macco1Veronica Astro2Diletta Tonoli3Elisa Savino4Flavia Valtorta5Kristyna Sala6Martina Botta7Ivan de Curtis8Cell Adhesion Unit San Raffaele Scientific Institute and San Raffaele University, Milan, ItalyCell Adhesion Unit San Raffaele Scientific Institute and San Raffaele University, Milan, ItalyCell Adhesion Unit San Raffaele Scientific Institute and San Raffaele University, Milan, ItalyCell Adhesion Unit San Raffaele Scientific Institute and San Raffaele University, Milan, ItalyNeuropsychopharmacology Unit, Division of Neuroscience, San Raffaele Scientific Institute and San Raffaele University, Milan, ItalyNeuropsychopharmacology Unit, Division of Neuroscience, San Raffaele Scientific Institute and San Raffaele University, Milan, ItalyCell Adhesion Unit San Raffaele Scientific Institute and San Raffaele University, Milan, ItalyCell Adhesion Unit San Raffaele Scientific Institute and San Raffaele University, Milan, ItalyCell Adhesion Unit San Raffaele Scientific Institute and San Raffaele University, Milan, ItalyUnderstanding the mechanisms guiding interneuron development is a central aspect of the current research on cortical/hippocampal interneurons, which is highly relevant to brain function and pathology. In this methodological study we have addressed the setup of protocols for the reproducible culture of dissociated cells from murine medial ganglionic eminences (MGEs), to provide a culture system for the analysis of interneurons in vitro. This study includes the detailed protocols for the preparation of the dissociated cells, and for their culture on optimal substrates for cell migration or differentiation. These cultures enriched in interneurons may allow the investigation of the migratory behavior of interneuron precursors and their differentiation in vitro, up to the formation of morphologically identifiable GABAergic synapses. Live imaging of MGE–derived cells plated on proper substrates shows that they are useful to study the migratory behavior of the precursors, as well as the behavior of growth cones during the development of neurites. Most MGE-derived precursors develop into polarized GABAergic interneurons as determined by axonal, dendritic, and GABAergic markers. We present also a comparison of cells from WT and mutant mice as a proof of principle for the use of these cultures for the analysis of the migration and differentiation of GABAergic cells with different genetic backgrounds. The culture enriched in interneurons described here represents a useful experimental system to examine in a relatively easy and fast way the morpho-functional properties of these cells under physiological or pathological conditions, providing a powerful tool to complement the studies in vivo.http://journal.frontiersin.org/article/10.3389/fncel.2017.00423/fullGABAergic interneuronsmedial ganglionic eminencesneuritesgrowth conesRac GTPases |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Sira A. Franchi Romina Macco Veronica Astro Diletta Tonoli Elisa Savino Flavia Valtorta Kristyna Sala Martina Botta Ivan de Curtis |
spellingShingle |
Sira A. Franchi Romina Macco Veronica Astro Diletta Tonoli Elisa Savino Flavia Valtorta Kristyna Sala Martina Botta Ivan de Curtis A Method to Culture GABAergic Interneurons Derived from the Medial Ganglionic Eminence Frontiers in Cellular Neuroscience GABAergic interneurons medial ganglionic eminences neurites growth cones Rac GTPases |
author_facet |
Sira A. Franchi Romina Macco Veronica Astro Diletta Tonoli Elisa Savino Flavia Valtorta Kristyna Sala Martina Botta Ivan de Curtis |
author_sort |
Sira A. Franchi |
title |
A Method to Culture GABAergic Interneurons Derived from the Medial Ganglionic Eminence |
title_short |
A Method to Culture GABAergic Interneurons Derived from the Medial Ganglionic Eminence |
title_full |
A Method to Culture GABAergic Interneurons Derived from the Medial Ganglionic Eminence |
title_fullStr |
A Method to Culture GABAergic Interneurons Derived from the Medial Ganglionic Eminence |
title_full_unstemmed |
A Method to Culture GABAergic Interneurons Derived from the Medial Ganglionic Eminence |
title_sort |
method to culture gabaergic interneurons derived from the medial ganglionic eminence |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Cellular Neuroscience |
issn |
1662-5102 |
publishDate |
2018-01-01 |
description |
Understanding the mechanisms guiding interneuron development is a central aspect of the current research on cortical/hippocampal interneurons, which is highly relevant to brain function and pathology. In this methodological study we have addressed the setup of protocols for the reproducible culture of dissociated cells from murine medial ganglionic eminences (MGEs), to provide a culture system for the analysis of interneurons in vitro. This study includes the detailed protocols for the preparation of the dissociated cells, and for their culture on optimal substrates for cell migration or differentiation. These cultures enriched in interneurons may allow the investigation of the migratory behavior of interneuron precursors and their differentiation in vitro, up to the formation of morphologically identifiable GABAergic synapses. Live imaging of MGE–derived cells plated on proper substrates shows that they are useful to study the migratory behavior of the precursors, as well as the behavior of growth cones during the development of neurites. Most MGE-derived precursors develop into polarized GABAergic interneurons as determined by axonal, dendritic, and GABAergic markers. We present also a comparison of cells from WT and mutant mice as a proof of principle for the use of these cultures for the analysis of the migration and differentiation of GABAergic cells with different genetic backgrounds. The culture enriched in interneurons described here represents a useful experimental system to examine in a relatively easy and fast way the morpho-functional properties of these cells under physiological or pathological conditions, providing a powerful tool to complement the studies in vivo. |
topic |
GABAergic interneurons medial ganglionic eminences neurites growth cones Rac GTPases |
url |
http://journal.frontiersin.org/article/10.3389/fncel.2017.00423/full |
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