Differential Glycosylation Expression in Injured Rat Spinal Cord Treated with Immunosuppressive Drug Cyclosporin-A

Abstract

Glycosylation is ubiquitous throughout the central nervous system and altered following spinal cord injury (SCI). The glial scar that forms following SCI is composed of several chondroitin sulfate proteoglycans, which inhibit axonal regrowth. Cyclosporin-A (CsA), an immunosuppressive therapeutic, has been proposed as a potential treatment after SCI. We investigated CsA treatment in the spinal cord of healthy, contusion injured, and injured CsA-treated rats. Lectin histochemistry using fluorescently labeled lectins, SBA, MAA, SNA-I, and WFA, was performed to identify the terminal carbohydrate residues of glycoconjugates within the spinal cord. SBA staining decreased in gray and white matter following spinal cord injury, whereas staining was increased at the lesion site in CsA-treated animals, indicating an increase in galactose and N -acetylgalactosamine terminal structures. No significant changes in MAA were observed. WFA staining was abundant in gray matter and observed to increase at the lesion site, in agreement with increased expression of chondroitin sulfate proteoglycans. SNA-I-stained blood vessels in all spinal cord regions and dual staining identified a subpopulation of astrocytes in the lesion site, which expressed α-(2,6)-sialic acid. Glycosylation were altered in injured spinal cord treated with CsA, indicating that glycosylation and alteration of particular carbohydrate structures are important factors to consider in the examination of the environment of the spinal cord after injury.

An optimized protocol for combined fluorescent lectin/immunohistochemistry to characterize tissue-specific glycan distribution in human or rodent tissues

“…Tissue-dependent modifications are outlined in the relevant steps. Note: This protocol is a combination and optimization of other methods previously published by ( Brooks and Hall, 2012 ; Mohd Isa et al., 2018 ; Kilcoyne et al., 2019 ; Contessotto et al., 2020 ). CRITICAL: Once the staining procedure starts (after the first wash and throughout the whole procedure), it is crucial to always keep the tissue wet and not allow it to dry.…”

“…Note: This protocol is a combination and optimization of other methods previously published by ( Brooks and Hall, 2012 ; Mohd Isa et al., 2018 ; Kilcoyne et al., 2019 ; Contessotto et al., 2020 ).…”

Mechanism of mesenchymal stem cells in spinal cord injury repair through macrophage polarization

“…Many drugs and treatments have proven effective in experimental studies, but their actual clinical effects are unknown. Beyond macrophages, T cell reduction can improve the recovery of the spinal cord structure and limb function after SCI, but the related mechanisms remain unclear [ 133 – 135 ]. The beneficial and detrimental effects of B cells before and after SCI damage to the BBB remains controversial [ 136 ].…”

Additive‐free Aqueous Dispersions of Two‐Dimensional Materials with Glial Cell Compatibility and Enzymatic Degradability

“…17‐Dec‐05). Primary culture of mixed glial cells (MG) was prepared using three‐day‐old post‐natal rats as described previously [27b] . MG cultures (MGCs) were prepared from spinal cords which were isolated by the “ejection technique” with a minor modification in the spinal cord removal protocol [33] .…”