Regenerating cortical connections in a dish: the entorhino-hippocampal organotypic slice co-culture as tool for pharmacological screening of molecules promoting axon regeneration.

Nature Protocols
José Antonio del Río, Eduardo Soriano

Abstract

We present a method for using long-term organotypic slice co-cultures of the entorhino-hippocampal formation to analyze the axon-regenerative properties of a determined compound. The culture method is based on the membrane interphase method, which is easy to perform and is generally reproducible. The degree of axonal regeneration after treatment in lesioned cultures can be seen directly using green fluorescent protein (GFP) transgenic mice or by axon tracing and histological methods. Possible changes in cell morphology after pharmacological treatment can be determined easily by focal in vitro electroporation. The well-preserved cytoarchitectonics in the co-culture facilitate the analysis of identified cells or regenerating axons. The protocol takes up to a month.

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Citations

Apr 27, 2012·PloS One·Joseph D Steinmeyer, Mehmet Fatih Yanik
Apr 9, 2010·Journal of Neurochemistry·Oscar SeiraJosé Antonio del Río
Aug 28, 2015·Molecular Neurobiology·Sareh PandamoozLeila Dargahi
Sep 11, 2012·Neuron·Sung Ok YoonKun Huang
May 28, 2013·International Journal of Molecular Sciences·Cristina RiggioJosé Antonio Del Río
Sep 26, 2014·The Journal of Biological Chemistry·JinBin TianSung Ok Yoon
Jul 17, 2019·British Journal of Social Work·Cindy A SousaMona El-Zuhairi
Aug 4, 2018·Frontiers in Veterinary Science·Laura Lossi, Adalberto Merighi
Feb 28, 2019·Frontiers in Cellular Neuroscience·Cristina Roselló-BusquetsEduardo Soriano
Apr 23, 2020·Frontiers in Molecular Neuroscience·Cristina Roselló-BusquetsRamon Martínez-Mármol
Mar 12, 2021·Frontiers in Cell and Developmental Biology·Jasmin WeningerEckart Förster

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