Expression of Glial Cell Line-derived Neurotrophic Factor in the Human Intervertebral Disc.

Junichi Yamada, Koji Akeda, Tomohiko Sano, Tatsuya Iwasaki, Norihiko Takegami, Akihiro Sudo

Spine 2020 Jul 01

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Biochemical and immunohistochemical analyses by the human intervertebral disc (IVD) cells and tissues. To examine the expression of glial cell line-derived neurotrophic factor (GDNF) and its receptors, GDNF family receptor (GFR) α1 and rearranged during transfection (RET) in the human IVD cells and the tissues with the early and advanced stages of degeneration. The neurotrophin family, including nerve growth factor, has been reported to be expressed in the IVDs and plays a role in hyperalgesia and neuronal sensitization. Despite having properties similar to the nerve growth factor, the expression of GDNF in the IVD remains unknown. Human IVD cells were cultured in monolayer. Immunohistochemical analyses and western blotting were performed to examine the protein levels of GDNF and its receptors. To examine the effect of proinflammatory cytokines, cells were cultured in the presence of interleukin-1β (IL-1β). The immunohistochemical expression of these proteins was also evaluated using human IVD tissues with different stages of degeneration. Immunofluorescent reactivity against anti-GDNF, GFRα1, and RET antibodies was identified in human IVD cells. In protein extracts from IVD cells, those protein expressions were also identified by Western blot. IL-1β significantly stimulated the mRNA expression of GDNF compared with that of the control group. There was no significant effect of IL-1β on the mRNA expression of GFRα1 and RET. The percentage of GDNF-immunopositive cells in advanced degenerated discs was significantly higher than that in early degenerated discs, whereas those of GFRα1 and RET showed no significant differences. GDNF and its receptors were constitutively expressed in the human IVD cells. GDNF expression was significantly enhanced by proinflammatory stimuli, and in the microenvironment with advanced tissue degeneration. N/A.

Citation

Junichi Yamada, Koji Akeda, Tomohiko Sano, Tatsuya Iwasaki, Norihiko Takegami, Akihiro Sudo. Expression of Glial Cell Line-derived Neurotrophic Factor in the Human Intervertebral Disc. Spine. 2020 Jul 01;45(13):E768-E775