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      Spinal cord regeneration by modulating bone marrow with neurotransmitters and Citicholine: Analysis at micromolecular level

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          Abstract

          Background

          Spinal cord injury results in disruption of brain-spinal cord fibre connectivity, leading to progressive tissue damage at the site of injury and resultant paralysis of varying degrees. The current study investigated the role of autologous bone marrow modulated with neurotransmitters and neurotransmitter stimulating agent, Citicholine, in spinal cord of spinal cord injured rats.

          Methods

          Radioreceptor assay using [3H] ligand was carried out to quantify muscarinic receptor. Gene expression studies were done using Real Time PCR analysis.

          Results

          Scatchard analysis of muscarinic M1 receptor showed significantly decreased B max ( p < 0.001) and K d ( p < 0.01) compared to control and significant reversal ( p < 0.001) in both the treatment groups (spinal cord injury treated with 5HT and GABA, and spinal cord injury treated with Citicholine). Muscarinic M1 receptor gene expression in spinal cord injured group showed significant down regulation ( p < 0.001) compared to control, and both the treatment groups significantly reversed ( p < 0.001) these changes to near control when compared to spinal cord injured group. The confocal microscopic study using specific antibody of muscarinic M1 confirmed the gene expression studies.

          Conclusion

          Thus our results suggest that the neurotransmitters combination along with bone marrow or Citicholine with bone marrow can reverse the muscarinic receptor alterations in the spinal cord of spinal cord injured rats, which is a promising step towards a better therapeutic intervention for spinal cord injury because of the positive role of cholinergic system in regulation of both locomotor activity and synaptic plasticity.

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          Most cited references27

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          Immunofluorescent labeling of cancer marker Her2 and other cellular targets with semiconductor quantum dots.

          Xingyong Wu, Hongjian Liu, Jianquan Liu … (2003)
          Semiconductor quantum dots (QDs) are among the most promising emerging fluorescent labels for cellular imaging. However, it is unclear whether QDs, which are nanoparticles rather than small molecules, can specifically and effectively label molecular targets at a subcellular level. Here we have used QDs linked to immunoglobulin G (IgG) and streptavidin to label the breast cancer marker Her2 on the surface of fixed and live cancer cells, to stain actin and microtubule fibers in the cytoplasm, and to detect nuclear antigens inside the nucleus. All labeling signals are specific for the intended targets and are brighter and considerably more photostable than comparable organic dyes. Using QDs with different emission spectra conjugated to IgG and streptavidin, we simultaneously detected two cellular targets with one excitation wavelength. The results indicate that QD-based probes can be very effective in cellular imaging and offer substantial advantages over organic dyes in multiplex target detection.
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            Acute spinal cord injury, part I: pathophysiologic mechanisms.

            R Dumont, D O Okonkwo, S. Verma … (2015)
            Spinal cord injury (SCI) is a devastating and common neurologic disorder that has profound influences on modern society from physical, psychosocial, and socioeconomic perspectives. Accordingly, the present decade has been labeled the Decade of the Spine to emphasize the importance of SCI and other spinal disorders. Spinal cord injury may be divided into both primary and secondary mechanisms of injury. The primary injury, in large part, determines a given patient's neurologic grade on admission and thereby is the strongest prognostic indicator. However, secondary mechanisms of injury can exacerbate damage and limit restorative processes, and hence, contribute to overall morbidity and mortality. A burgeoning body of evidence has facilitated our understanding of these secondary mechanisms of injury that are amenable to pharmacological interventions, unlike the primary injury itself. Secondary mechanisms of injury encompass an array of perturbances and include neurogenic shock, vascular insults such as hemorrhage and ischemia-reperfusion, excitotoxicity, calcium-mediated secondary injury and fluid-electrolyte disturbances, immunologic injury, apoptosis, disturbances in mitochondrion function, and other miscellaneous processes. Comprehension of secondary mechanisms of injury serves as a basis for the development and application of targeted pharmacological strategies to confer neuroprotection and restoration while mitigating ongoing neural injury. The first article in this series will comprehensively review the pathophysiology of SCI while emphasizing those mechanisms for which pharmacologic therapy has been developed, and the second article reviews the pharmacologic interventions for SCI.
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              Marrow stromal cells migrate throughout forebrain and cerebellum, and they differentiate into astrocytes after injection into neonatal mouse brains.

              Gene Kopen, L D Prockop, Stephen D. Phinney (1999)
              Stem cells are a valuable resource for treating disease, but limited access to stem cells from tissues such as brain restricts their utility. Here, we injected marrow stromal cells (MSCs) into the lateral ventricle of neonatal mice and asked whether these multipotential mesenchymal progenitors from bone marrow can adopt neural cell fates when exposed to the brain microenvironment. By 12 days postinjection, MSCs migrated throughout the forebrain and cerebellum without disruption to the host brain architecture. Some MSCs within the striatum and the molecular layer of the hippocampus expressed glial fibrillary acidic protein and, therefore, differentiated into mature astrocytes. MSCs also populated neuron rich regions including the Islands of Calleja, the olfactory bulb, and the internal granular layer of the cerebellum. A large number of MSCs also were found within the external granular layer of the cerebellum. In addition, neurofilament positive donor cells were found within the reticular formation of the brain stem, suggesting that MSCs also may have differentiated into neurons. Therefore, MSCs are capable of producing differentiated progeny of a different dermal origin after implantation into neonatal mouse brains. These results suggest that MSCs are potentially useful as vectors for treating a variety of central nervous system disorders.
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                Author and article information

                Contributors
                Cheramadathukudiyil Skaria Paulose
                Journal
                Journal ID (nlm-ta): Biomed J
                Journal ID (iso-abbrev): Biomed J
                Title: Biomedical Journal
                Publisher: Chang Gung University
                ISSN (Print): 2319-4170
                ISSN (Electronic): 2320-2890
                Publication date PMC-release: 04 May 2017
                Publication date (Print): April 2017
                Publication date (Electronic): 04 May 2017
                Volume: 40
                Issue: 2
                Pages: 94-100
                Affiliations
                [a ]Molecular Neurobiology and Cell Biology Unit, Centre for Neuroscience, Department of Biotechnology, Cochin University of Science and Technology, Cochin, Kerala, India
                [b ]Pushpagiri Institute of Medical Sciences and Research, Thiruvalla, Kerala, India
                [c ]Center for Neuroscience, Department of Biotechnology, Cochin University of Science and Technology, Cochin, Kerala, India
                Author notes
                [∗ ] Corresponding author. Molecular Neurobiology and Cell Biology Unit, Department of Biotechnology, Cochin University of Science and Technology, Cochin 682 022, Kerala, India. biomncb@ 123456cusat.ac.in cspaulose@ 123456gmail.com
                Article
                Publisher ID: S2319-4170(17)30094-X
                DOI: 10.1016/j.bj.2016.11.006
                PMC ID: 6138792
                PubMed ID: 28521906
                SO-VID: ce7ca539-7213-4465-821a-541efde4a427
                Copyright © © 2017 Chang Gung University. Publishing services by Elsevier B.V.
                License:

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                Date received : 23 November 2015
                Date accepted : 15 November 2016
                Categories
                Subject: Original Article

                Keywords: spinal cord injury,bone marrow cells,neurotransmitters,muscarinic receptor
                Data availability:
                Keywords: spinal cord injury, bone marrow cells, neurotransmitters, muscarinic receptor

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