A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits

Scientific Reports
Jonathan T TingEd Lein

Abstract

The powerful suite of available genetic tools is driving tremendous progress in understanding mouse brain cell types and circuits. However, the degree of conservation in human remains largely unknown in large part due to the lack of such tools and healthy tissue preparations. To close this gap, we describe a robust and stable adult human neurosurgically-derived ex vivo acute and cultured neocortical brain slice system optimized for rapid molecular-genetic manipulation. Surprisingly, acute human brain slices exhibited exceptional viability, and neuronal intrinsic membrane properties could be assayed for at least three days. Maintaining adult human slices in culture under sterile conditions further enabled the application of viral tools to drive rapid expression of exogenous transgenes. Widespread neuron-specific labeling was achieved as early as two days post infection with HSV-1 vectors, with virally-transduced neurons exhibiting membrane properties largely comparable to uninfected neurons over this short timeframe. Finally, we demonstrate the suitability of this culture paradigm for optical manipulation and monitoring of neuronal activity using genetically encoded probes, opening a path for applying modern molecular-genetic to...Continue Reading

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Citations

Sep 26, 2019·Frontiers in Molecular Neuroscience·Larissa KrausPawel Fidzinski
Jan 30, 2020·Journal of Neurophysiology·Kevin LeeJohanna M Montgomery
Apr 1, 2020·Neuro-oncology Advances·Alastair J KirbyGerald T Finnerty
Mar 3, 2020·Frontiers in Synaptic Neuroscience·Guanxiao QiDirk Feldmeyer
Dec 30, 2020·The American Journal of Bioethics : AJOB·Henry T Greely
Dec 7, 2019·Brain Research·Mohammed A Mostajo-RadjiAlex A Pollen
Feb 13, 2020·Developmental Cell·Ryann M Fame, Maria K Lehtinen
Oct 3, 2020·Cortex; a Journal Devoted to the Study of the Nervous System and Behavior·Joana LourençoAlberto Bacci
Apr 1, 2021·Neuron·Lucas T GraybuckBosiljka Tasic
May 5, 2021·Nature Communications·Homeira Moradi ChamehTaufik A Valiante
Jun 16, 2021·Cellular and Molecular Life Sciences : CMLS·Egor DzyubenkoDirk M Hermann
Sep 6, 2020·Cell Stem Cell·Aparna BhaduriTomasz J Nowakowski
Dec 11, 2021·Frontiers in Neuroscience·Glaucia M AlmeidaAdriano Sebollela

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Methods Mentioned

BETA
dissection
dissecting
Fluorescence imaging

Software Mentioned

Elements
Image J
NIS
Igor Pro

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