C Cheng1, S Gao, J Zhao, S Niu, M Chen, X Li, J Qin, S Shi, Z Guo, A Shen. 1. The Jiangsu Province Key Laboratory of Neuroregeneration, Department of Microbiology and Immunology, Nantong University (Former Nantong Medical College), Nantong, China.
Abstract
AIMS: Postsynaptic density (PSD)-95 is a scaffolding protein linking the N-methyl-D-aspartate receptor with neuronal nitric oxide synthase (nNOS), which contributes to many physiological and pathological actions. We here investigated whether PSD-95 was involved in the secondary response following spinal cord injury (SCI). METHODS: Spinal cord contusion (SCC) and spinal cord transection (SCT) models at thoracic (T) segment 9 (T(9)) were established in adults rats. Real-time polymerase chain reaction, Western blot, immunohistochemistry and immunofluorescence were used to detect the temporal profile and spatial distribution of PSD-95 after SCI. The association between PSD-95 and nNOS in the injured cords was also assessed by coimmmunoprecipation and double immunofluorescent staining. RESULTS: The mRNA and protein for PSD-95 expression were significantly increased at 2 h or 8 h, and then gradually declined to the baseline level, ultimately up-regulated again from 5 days to 7 days for its mRNA level and at 7 days or 14 days for its protein level after either SCC or SCT. PSD-95 immunoreactivity was found in neurones, oligodendrocytes and synaptic puncta of spinal cord tissues within 5 mm from the lesion site. Importantly, injury-induced expression of PSD-95 was colabelled by active caspase-3 (apoptotic marker), Tau-1 (the marker for pathological oligodendrocytes) and nNOS. CONCLUSIONS: Accompanied by the spatio-temporal changes for PSD-95 expression, the association between PSD-95 and nNOS undergoes substantial alteration after SCI. These two molecules are likely to form a complex on apoptotic neurones and pathological oligodendrocytes, which may in turn be involved in the secondary response after SCI.
AIMS: Postsynaptic density (PSD)-95 is a scaffolding protein linking the N-methyl-D-aspartate receptor with neuronal nitric oxide synthase (nNOS), which contributes to many physiological and pathological actions. We here investigated whether PSD-95 was involved in the secondary response following spinal cord injury (SCI). METHODS: Spinal cord contusion (SCC) and spinal cord transection (SCT) models at thoracic (T) segment 9 (T(9)) were established in adults rats. Real-time polymerase chain reaction, Western blot, immunohistochemistry and immunofluorescence were used to detect the temporal profile and spatial distribution of PSD-95 after SCI. The association between PSD-95 and nNOS in the injured cords was also assessed by coimmmunoprecipation and double immunofluorescent staining. RESULTS: The mRNA and protein for PSD-95 expression were significantly increased at 2 h or 8 h, and then gradually declined to the baseline level, ultimately up-regulated again from 5 days to 7 days for its mRNA level and at 7 days or 14 days for its protein level after either SCC or SCT. PSD-95 immunoreactivity was found in neurones, oligodendrocytes and synaptic puncta of spinal cord tissues within 5 mm from the lesion site. Importantly, injury-induced expression of PSD-95 was colabelled by active caspase-3 (apoptotic marker), Tau-1 (the marker for pathological oligodendrocytes) and nNOS. CONCLUSIONS: Accompanied by the spatio-temporal changes for PSD-95 expression, the association between PSD-95 and nNOS undergoes substantial alteration after SCI. These two molecules are likely to form a complex on apoptotic neurones and pathological oligodendrocytes, which may in turn be involved in the secondary response after SCI.
Authors: Jie Liu; Weijie Wu; Jie Hao; Mingchen Yu; Jin Liu; Xinlei Chen; Rong Qian; Feng Zhang Journal: Neurochem Res Date: 2016-11-10 Impact factor: 3.996