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Hippocampal slice preparation in rats acutely suppresses immunoreactivity of microtubule‐associated protein (Map2) and glycogen levels without affecting numbers of glia or levels of the glutamate transporter VGlut1

INTRODUCTION: With its preservation of cytoarchitecture and synaptic circuitry, the hippocampal slice preparation has been a critical tool for studying the electrophysiological effects of pharmacological and genetic manipulations. To analyze the maximum number of slices or readouts per dissection, l...

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Autores principales: Stein, Liana R., Zorumski, Charles F., Izumi, Yukitoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5516609/
https://www.ncbi.nlm.nih.gov/pubmed/28729941
http://dx.doi.org/10.1002/brb3.736
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author Stein, Liana R.
Zorumski, Charles F.
Izumi, Yukitoshi
author_facet Stein, Liana R.
Zorumski, Charles F.
Izumi, Yukitoshi
author_sort Stein, Liana R.
collection PubMed
description INTRODUCTION: With its preservation of cytoarchitecture and synaptic circuitry, the hippocampal slice preparation has been a critical tool for studying the electrophysiological effects of pharmacological and genetic manipulations. To analyze the maximum number of slices or readouts per dissection, long incubation times postslice preparation are commonly used. We were interested in how slice integrity is affected by incubation postslice preparation. METHODS: Hippocampal slices were prepared by three different methods: a chopper, a vibratome, and a rotary slicer. To test slice integrity, we compared glycogen levels and immunohistochemistry of selected proteins in rat hippocampal slices immediately after dissection and following 2 and 4 hr of incubation. RESULTS: We found that immunoreactivity of the dendritic marker microtubule‐associated protein 2 (Map2) drastically decreased during this incubation period, whereas immunoreactivity of the glutamate transporter VGlut1 did not significantly change with incubation time. Astrocytic and microglial cell numbers also did not significantly change with incubation time whereas glycogen levels markedly increased during incubation. CONCLUSION: Immunoreactivity of the dendritic marker Map2 quickly decreased after dissection with all the slicing methods. This work highlights a need for caution when using long incubation periods following slice preparation.
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spelling pubmed-55166092017-07-20 Hippocampal slice preparation in rats acutely suppresses immunoreactivity of microtubule‐associated protein (Map2) and glycogen levels without affecting numbers of glia or levels of the glutamate transporter VGlut1 Stein, Liana R. Zorumski, Charles F. Izumi, Yukitoshi Brain Behav Original Research INTRODUCTION: With its preservation of cytoarchitecture and synaptic circuitry, the hippocampal slice preparation has been a critical tool for studying the electrophysiological effects of pharmacological and genetic manipulations. To analyze the maximum number of slices or readouts per dissection, long incubation times postslice preparation are commonly used. We were interested in how slice integrity is affected by incubation postslice preparation. METHODS: Hippocampal slices were prepared by three different methods: a chopper, a vibratome, and a rotary slicer. To test slice integrity, we compared glycogen levels and immunohistochemistry of selected proteins in rat hippocampal slices immediately after dissection and following 2 and 4 hr of incubation. RESULTS: We found that immunoreactivity of the dendritic marker microtubule‐associated protein 2 (Map2) drastically decreased during this incubation period, whereas immunoreactivity of the glutamate transporter VGlut1 did not significantly change with incubation time. Astrocytic and microglial cell numbers also did not significantly change with incubation time whereas glycogen levels markedly increased during incubation. CONCLUSION: Immunoreactivity of the dendritic marker Map2 quickly decreased after dissection with all the slicing methods. This work highlights a need for caution when using long incubation periods following slice preparation. John Wiley and Sons Inc. 2017-05-30 /pmc/articles/PMC5516609/ /pubmed/28729941 http://dx.doi.org/10.1002/brb3.736 Text en © 2017 The Authors. Brain and Behavior published by Wiley Periodicals, Inc. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Research
Stein, Liana R.
Zorumski, Charles F.
Izumi, Yukitoshi
Hippocampal slice preparation in rats acutely suppresses immunoreactivity of microtubule‐associated protein (Map2) and glycogen levels without affecting numbers of glia or levels of the glutamate transporter VGlut1
title Hippocampal slice preparation in rats acutely suppresses immunoreactivity of microtubule‐associated protein (Map2) and glycogen levels without affecting numbers of glia or levels of the glutamate transporter VGlut1
title_full Hippocampal slice preparation in rats acutely suppresses immunoreactivity of microtubule‐associated protein (Map2) and glycogen levels without affecting numbers of glia or levels of the glutamate transporter VGlut1
title_fullStr Hippocampal slice preparation in rats acutely suppresses immunoreactivity of microtubule‐associated protein (Map2) and glycogen levels without affecting numbers of glia or levels of the glutamate transporter VGlut1
title_full_unstemmed Hippocampal slice preparation in rats acutely suppresses immunoreactivity of microtubule‐associated protein (Map2) and glycogen levels without affecting numbers of glia or levels of the glutamate transporter VGlut1
title_short Hippocampal slice preparation in rats acutely suppresses immunoreactivity of microtubule‐associated protein (Map2) and glycogen levels without affecting numbers of glia or levels of the glutamate transporter VGlut1
title_sort hippocampal slice preparation in rats acutely suppresses immunoreactivity of microtubule‐associated protein (map2) and glycogen levels without affecting numbers of glia or levels of the glutamate transporter vglut1
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5516609/
https://www.ncbi.nlm.nih.gov/pubmed/28729941
http://dx.doi.org/10.1002/brb3.736
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