Abstract
The organization of the mouse spinal dorsal horn has been delineated in 2D for the six Rexed laminae in our publication Atlas of the Spinal Cord: Mouse, Rat, Rhesus, Marmoset, and Human. In the present study, the tissue clearing technique CLARITY was used to observe the cyto- and chemoarchitecture of the mouse spinal cord in 3D, using a variety of immunohistochemical markers. We confirm prior observations regarding the location of glycine and serotonin immunoreactivities. Novel observations include the demonstration of numerous calcitonin gene-related peptide (CGRP) perikarya, as well as CGRP fibers and terminals in all laminae of the dorsal horn. We also observed sparse choline acetyltransferase (ChAT) immunoreactivity in small perikarya and fibers and terminals in all dorsal horn laminae, while gamma aminobutyric acid (GABA) and glutamate decarboxylase-67 (GAD67) immunoreactivities were found only in small perikarya and fibers. Finally, numerous serotonergic fibers were observed in all laminae of the dorsal horn. In conclusion, CLARITY confirmed the 2D immunohistochemical properties of the spinal cord. Furthermore, we observed novel anatomical characteristics of the spinal cord and demonstrated that CLARITY can be used on spinal cord tissue to examine many proteins of interest.
Highlights
The major characteristics of the neurons of each of the spinal dorsal horn laminae have been studied using standard histological, histochemical, and immunohistochemical staining methods
We aimed to show the cytoand chemoarchitecture of the mouse spinal cord dorsal horn with many protein markers, i.e., calretinin, glycine, nitric oxide, calcitonin gene-related peptide (CGRP), choline acetyltransferase (ChAT), glutamate decarboxylase-67 (GAD67), and serotonin 3D using CLARITY for the first time
Our observations of the cyto- and chemoarchitecture of the dorsal horn were similar to earlier reports that used conventional light and fluorescent microscopy techniques (Figures 1 and 2, Online resources 1-6), confirming that CLARITY can be used reliably to examine spinal cord tissue
Summary
The major characteristics of the neurons of each of the spinal dorsal horn laminae have been studied using standard histological, histochemical, and immunohistochemical staining methods. The ten laminae in the spinal cord gray matter based on cellular shape, size, and density were first described by Rexed in the cat [3, 4], followed by studies in the mouse [2, 5, 6], rat [2, 7, 8], marmoset, rhesus monkey, and human [2]. Six of these laminae (laminae 1-6) are in the dorsal horn. These neurons receive a variety of neurotransmitter inputs and release a variety of neurotransmitters that show different patterns of distribution that is often similar across different species
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