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

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.

Acetylated oligopeptide and N‐acetylcysteine protect against iron overload‐induced dentate gyrus hippocampal degeneration through upregulation of Nestin and Nrf2/HO‐1 and downregulation of MMP‐9/TIMP‐1 and GFAP

“…Histopathological examinations of the hippocampus of the iron overload rats by light microscope showed shrinkage in DG with distortion in the granular cell layer and an increased number of degenerated cells and nuclei, indicating that iron overload can lead to disturbance in the normal structure of the hippocampus. These results have been illustrated by both H & E and gallocyanin stains, with the latter specifically binding to the granular structures of nucleic acid 61 . In the same way, the iron degenerating effects were also shown by structural and ultrastructural observations.…”

“…These findings may be explained by the fact that both NAC and AOP were able to ameliorate the oxidative stress with the latter specifically binding to the granular structures of nucleic acid. 61 In the same way, the iron degenerating effects were also shown by structural and ultrastructural observations. The immunohistochemical examination of anti-GFAP showed strong expression in the DG region of the iron-treated group.…”