Abstract
Purpose: To assess the involvement of cholinergic transmission in the etiology of spinal cord injury (SCI) in a rat model.
 Methods: Male adult rats (Wistar) with body weight ranging from 200 to 250 g were equally allocated into 2 groups: test (SCI) and control (non-SCI). Clipping method was used to induce SCI. Thereafter, motor function was measured using rotarod. Each rat was sacrificed by decapitation, and the cortex was excised for use in the study of the involvement of cholinergic transmission in SCI using real time quantitative polymerase chain reaction (RT-PCR) and western blot analysis (WBA).
 Results: Significant upregulation in acetylcholine esterase (AChE) was observed in the cortex of SCI rats, relative to non-SCI rats (p < 0.005). Results from cholinergic receptor binding studies revealed significantly decreased maximum binding (Bmax) and dissociation constant (kd) values for muscarinic receptors in SCI rats, when compared to non-SCI rats. Moreover, the reduction in intensity of cholinergic receptors was significantly greater in the cerebral cortex of SCI group compared to non-SCI group.
 Conclusion: The results of this study suggested that the reduction in cortical cholinergic transmission impairs motor functions in SCI, and plays a major role in motor deficits in SCI.
Highlights
Spinal cord injury (SCI) is one of the leading causes of severe incapacitation associated with high health care expenses [1,2]
Bmax and kd values for muscarinic receptor type 1 (MRT1), Muscarinic receptor type 3 (MRT3) and total MR were significantly lower in SCI group as compared to non-SCI group
Results from real time quantitative polymerase chain reaction (RT-PCR) assay revealed that the expressions of acetylcholine esterase (AChE) and choline AChE were significantly greater in SCI group compared to non-SCI group
Summary
Spinal cord injury (SCI) is one of the leading causes of severe incapacitation associated with high health care expenses [1,2]. Studies have reported the participation of several cortical areas in the control of motor function in individuals with stroke during the recovery period [1,2,3,4]. Muscarinic acetylcholine receptor plays a vital role in the functioning of sensory and motor structures [3]. Each section of cerebral cortex was treated with phosphate buffer at pH 7.4 for half an hour, and incubated with muscarinic or nicotinic acetylcholine receptor antibody. Comparison of retention times and the expressions of muscarinic and nicotinic receptors in cortical region between both groups (SCI and non-SCI) were analyzed by t-test. Data related to muscarinic receptor binding analysis in cortex for both groups were analyzed using non-parametric test (Mann Whitney test). Pixel intensities in the cortex between both groups were analyzed by non-parametric test
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