परिधीय तंत्रिका कलम बांधने का काम और मैट्रिक्स मॉडुलन मेल घायल चूहा स्पाइनल कॉर्ड की मरम्मत
Summary
मस्तिष्क और रीढ़ की हड्डी के लिए अभिघातजन्य चोट के साथ संचार बाधित. खो कनेक्टिविटी को पुनर्स्थापित करने के लिए हम एक परिधीय तंत्रिका neurotrophic कारकों और मैट्रिक्स का नियमन करने वाली एंजाइमों के साथ संयोजन में regenerating के फाइबर के लिए बुनियाद निरोधात्मक अणुओं को हटाने के लिए लंबी दूरी की विकास को बढ़ावा देने प्रदान भ्रष्टाचार का उपयोग.
Abstract
Traumatic injury to the spinal cord (SCI) causes death of neurons, disruption of motor and sensory nerve fiber (axon) pathways and disruption of communication with the brain. One of the goals of our research is to promote axon regeneration to restore connectivity across the lesion site. To accomplish this we developed a peripheral nerve (PN) grafting technique where segments of sciatic nerve are either placed directly between the damaged ends of the spinal cord or are used to form a bridge across the lesion. There are several advantages to this approach compared to transplantation of other neural tissues; regenerating axons can be directed towards a specific target area, the number and source of regenerating axons is easily determined by tracing techniques, the graft can be used for electrophysiological experiments to measure functional recovery associated with axons in the graft, and it is possible to use an autologous nerve to reduce the possibility of graft rejection. In our lab we have performed both autologous (donor and recipient are the same animal) and heterologous (donor and recipient are different animals) grafts with comparable results. This approach has been used successfully in both acute and chronic injury situations. Regenerated axons that reach the distal end of the PN graft often fail to extend back into the spinal cord, so we use microinjections of chondroitinase to degrade inhibitory molecules associated with the scar tissue surrounding the area of SCI. At the same time we have found that providing exogenous growth and trophic molecules encourages longer distance axonal regrowth into the spinal cord. Several months after transplantation we perform a variety of anatomical, behavioral and electrophysiological tests to evaluate the recovery of function in our spinal cord injured animals. This experimental approach has been used successfully in several spinal cord injury models, at different levels of injury and in different species (mouse, rat and cat). Importantly, the peripheral nerve grafting approach is effective in promoting regeneration by acute and chronically injured neurons.
Protocol
1) सूक्ष्म सर्जरी के लिए तैयार शल्य चिकित्सा स्टेशन के लिए एक पतला ब्लीच समाधान से पहले बाहर उपकरणों और सहायक सामग्री की स्थापना के साथ स्वच्छ और साफ होने की जरूरत है. उपकरण 1 दिन सर्जरी से पहले autoclaved ज?…
Discussion
- प्रयोग के reproducibility के लिए यह महत्वपूर्ण है कि रीढ़ की हड्डी में चोट के स्तर जानवर से जानवर के अनुरूप हो. इसलिए यह महत्वपूर्ण है कि laminectomy के लिए उपयुक्त कशेरुका प्रक्रिया की पहचान हो. क्योंकि हम hemisection या इस अध्ययन के …
Acknowledgements
इस काम के NIH / NINDS NS26380 अनुदान और NS55976, क्रिस्टोफर और दाना रीव फाउंडेशन और स्पाइनल कॉर्ड अनुसंधान के लिए डैनियल Heumann फंड द्वारा समर्थित किया गया. Drexel विश्वविद्यालय कॉलेज चिकित्सा स्पाइनल कॉर्ड रिसर्च सेंटर के कोर इस काम को पूरा करने के लिए इस्तेमाल किया सुविधाओं के लिए समर्थन प्रदान करता है.
Materials
| Material Name | Type | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| 10-0 silk suture | ArosSurgical | T5A10N10 | ||
| 6-0 silk suture | McKesson | 2693 | ||
| Ampicillin | McKesson | 483549 | ||
| Antibody to cFos | Sigma-Aldrich | F7799 | ||
| Biotinylated dextran Amine | Invitrogen | D7135 | ||
| Buprenorphin (.3mg/ml) | McKesson | 12496075701 | ||
| Chondroitinase ABC | Associates of Cape Cod | 100332-1A | ||
| Euthasol | Webster Veterinary | 07-805-9296 | ||
| Hanks Balanced Salt Solution | Cellgro | 21-021-CV | ||
| Isoflurane | Henry Schein | 209-1966 | ||
| Michel Wound Clips | Fine Science Tools | 12040-02 | ||
| Neurotrace Kit | Invitrogen | N7167 | ||
| True Blue | Sigma-Aldrich | T5891 | ||
| Xenodine | Webster Veterinary | 92201 | ||
| Hot Bead Sterilizer | Fine Science Tools | |||
| Forced Exercise Wheel | Lafayette Instruments | |||
| TreadScan System | Clever System | |||
| Infinite Horizon Impact Device | Precision Systems and Instrumentation | |||
| Magnetic Stimulation Device | MagStim Inc. |
References
- Houle, J. D. Demonstration of the potential for chronically injured neurons to regenerate axons into intraspinal peripheral nerve grafts. Exp. Neurol. 113, 1-9 (1991).
- Ye, J. H., Houle, J. D. Treatment of the chronically injured spinal cord with neurotrophic factors can promote axonal regeneration from supraspinal neurons. Exp. Neurol. 143, 70-81 (1997).
- Houle, J. D., Tom, V., Mayes, D., Wagoner, G., Phillips, N., Silver, J. Combining an autologous peripheral nerve bridge and matrix modification by chondroitinase allows robust functional regeneration across a hemisection lesion of the adult rat spinal cord. J. Neurosci. 26, 7405-7415 (2006).
- Tom, V., Houle, J. D. Intraspinal microinjection of chondroitinase ABC following injury promotes axonal regeneration out of a peripheral nerve graft. Exp. Neurol. 211, 315-319 (2008).
Cite This Article
Houle, J. D., Amin, A., Cote, M., Lemay, M., Miller, K., Sandrow, H., Santi, L., Shumsky, J., Tom, V. Combining Peripheral Nerve Grafting and Matrix Modulation to Repair the Injured Rat Spinal Cord. J. Vis. Exp. (33), e1324, doi:10.3791/1324 (2009).