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Sectioning and Counting of Motor Neurons in the L3 to L6 Region of the Adult Mouse Spinal Cord

Austin, A; Beresford, L; Price, G; Cunningham, T; Kalmar, B; Yon, M; (2022) Sectioning and Counting of Motor Neurons in the L3 to L6 Region of the Adult Mouse Spinal Cord. Current Protocols , 2 (5) , Article e428. 10.1002/cpz1.428. Green open access

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Abstract

Histology is the study of the microscopic structure of tissues. This protocol permits the generation of frozen transverse sections of lumbar spinal cord regions L3 to L6. It enables counting of murine ventral horn lumbar motor neurons in a reproducible manner. Methods include spinal column dissection, hydraulic extrusion, and histological processing. The preparation for cryo-sectioning includes embedding lumbar spinal cord in optimal cutting temperature (OCT) medium. The correct orientation of the tissue is critical as calculating the amount of tissue to discard saved time overall. Specific details regarding section thickness and mounting are described. These requirements not only allow optimum coverage of specific regions but also ensure that no individual motor neuron was counted twice. The Nissl bodies of the motor neurons were stained using gallocyanin. The sections obtained are all of a comparable area and quality assurance is consistent. The specificity of the staining enables the scientist to identify and reliably quantify lumbar motor neurons. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Euthanasia of mouse and isolation of spinal cord. Basic Protocol 2: Hydraulic extrusion of the spinal cord. Basic Protocol 3: Identification of the lumbar region. Basic Protocol 4: Embedding cord in OCT. Basic Protocol 5: Collection of frozen sections onto slides. Basic Protocol 6: Gallocyanin staining. Basic Protocol 7: Motor neuron counting.

Type: Article
Title: Sectioning and Counting of Motor Neurons in the L3 to L6 Region of the Adult Mouse Spinal Cord
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/cpz1.428
Publisher version: https://doi.org/10.1002/cpz1.428
Language: English
Additional information: © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Keywords: adult mouse spinal cord, gallocyanin staining, mouse lumbar, neuron counting, Animals, Anterior Horn Cells, Lumbosacral Region, Mice, Motor Neurons, Spinal Cord
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Institute of Prion Diseases
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology > Department of Neuromuscular Diseases
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Institute of Prion Diseases > MRC Prion Unit at UCL
URI: https://discovery.ucl.ac.uk/id/eprint/10165324
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