为持续给药Neuroactive代理Microinfusion系统的建设和植入。
Summary
随着神经科学的查询变得更加复杂，调查的大脑结构和电路需要提高精度和更高的分辨率的水平。我们已经开发出一种利用与尖端直径为50微米的高硼硅microcannula了大脑内的慢性输液系统的准备和植入的方法。
Abstract
慢性滴注neuroactive代理使用的大脑区域内的实验协议，通常利用一个微型渗透泵系统。代理通常是通过一个直径为0.30毫米或更大的不锈钢套管交付。利用这一口径套管的系统，可以并处创伤导致建筑损坏的利益方面，从而影响结构的完整性和正常运作。随着神经科学的查询变得更加复杂，调查的大脑结构和电路需要提高精度和更高的分辨率的水平。我们已经开发出一种利用与尖端直径为50微米的高硼硅microcannula了大脑内的慢性输液系统的准备和植入的方法。这种技术减少了对当地环境的破坏和减少输液部位的反应性胶质化。 microinfusion系统的配置也能够以符合动物的头骨表面，解除对大脑神经底座的需要，从而促进头皮切口封闭和减少感染的风险。我们表现出持续提供可靠的一个有代表性的分子量使用体外模型，并在大鼠体内研究的染料。
Protocol
简介 neuroactive代理商直接输注允许特定的大脑区域进行研究，同时绕过血脑屏障。这种方法在神经科学中的应用是多种多样的，包括改变离散分区域的大脑活动水平（Berretta等，2004;。Gliddon等，2005;。Kim等，2000），调查的精神科药物的行动（克林顿等人，2006年迪尼迪托等，2004），提供脑部炎症的控制模型（Hauss – Wegrzyniak等，1998; Marchalant等，2007;。罗西等人，2004年），研究的机…
Discussion
各种研究表明，减少交付脑内注射，组织创伤和侮辱血脑屏障套管的大小减少（佩里等人，1993年），炎症减少，免疫反应减弱（Finsen等人减少，1991），反应性胶质化（Nikkhah等，1994）。我们在座的一种neuroactive代理大脑内的慢性交付通过一个直径6倍，比常规使用的交付插管，减少microcannula方法。我们已经证明，该系统可靠地提供了一个有代表性的解决方案随着时间的推移和局部创伤和反应性胶质化水平显着降低…
Materials
| Material Name | Tipo | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| Mini-osmotic pumps (MOPs) | Tool | ALZET | model #1002 | |
| Electrode puller | Tool | Stoelting Co. | ||
| Borosilicate tubing | Surgery | World Precision Instruments | 1B100F-6 | |
| Microforceps | Surgery | World Precision Instruments | ||
| Polyethylene tubing | Surgery | Plastics One | #C312 VT | |
| LumaBond | Reagent | myNeuroLab, Inc |
Referências
- Berretta, S., Lange, N., Bhattacharyya, S., Sebro, R., Garces, J., Benes, F. M. Long-term effectsof amygdala GABA receptor blockade on specific subpopulations of hippocampalinterneurons. Hippocampus. 14, 876-894 (2004).
- Clinton, S. M., Sucharski, I. L., Finlay, J. M. Desipramine attenuates working memoryimpairments induced by partial loss of catecholamines in the rat medial prefrontal cortex. Psychopharmacology. 183, 404-412 (2006).
- Cooley, R., et al. . Stereotaxic surgery in the rat. , (1990).
- Di Benedetto, M., Feliciani, D., D’Addario, C., Izenwasser, S., Candeletti, S. Romualdi P.Effects of the selective norepinephrine uptake inhibitor nisoxetine on prodynorphin geneexpression in rat CNS. Brain Res Mol Brain Res. 127, 115-120 (2004).
- Ding, M., Haglid, K. G., Hamberger, A. Quantitative immunochemistry on neuronal loss,reactive gliosis and BBB damage in cortex/striatum and hippocampus/amygdala aftersystemic kainic acid administration. Neurochemistry international. 36, 313-318 (2000).
- Finsen, B. R., Sorensen, T., Castellano, B., Pedersen, E. B., Zimmer, J. Leukocyte infiltrationand glial reactions in xenografts of mouse brain tissue undergoing rejection in the adultrat brain. A light and electron microscopical immunocytochemical study. JNeuroimmunol. 32, 159-183 (1991).
- Gliddon, C. M., Darlington, C. L., Smith, P. F. Effects of chronic infusion of a GABAA receptoragonist or antagonist into the vestibular nuclear complex on vestibular compensation inthe guinea pig. J Pharmacol Exp Ther. 313, 1126-1135 (2005).
- Hauss-Wegrzyniak, B., Dobrzanski, P., Stoehr, J. D., Wenk, G. L. Chronic neuroinflammation in rats reproduces components of the neurobiology of Alzheimer’s disease. Brain Res. 780, 294-303 (1998).
- Kim, H. S., Choi, H. S., Lee, S. Y., Oh, S. Changes of GABA(A) receptor binding and subunitmRNA level in rat brain by infusion of subtoxic dose of MK-801. Brain Res. 880, 28-37 (2000).
- Kim, S. Y., Chudapongse, N., Lee, S. M., Levin, M. C., Oh, J. T., Park, H. J., Ho, I. K. Proteomicanalysis of phosphotyrosyl proteins in morphine-dependent rat brains. Brain Res MolBrain Res. 133, 58-70 (2005).
- Lockman, P. R., McAfee, G., Geldenhuys, W. J., Schyf, C. J., Abbruscato, T. J., Van Allen, D. D. der Schyf CJ, Abbruscato TJ, Allen DD.Brain uptake kinetics of nicotine and cotinine after chronic nicotine exposure. J Pharmacol Exp Ther. 314, 636-642 (2005).
- MA, K. C., Chang, Z. H., Shih, H., Zhu, J. H., Wu, J. Y. The compensatory ‘rebound’ of reactiveastrogliosis: glial fibrillary acidic protein immunohistochemical analysis of reactiveastrogliosis after a puncture wound to the brain of rats with portocaval anastomosis. Acta neuropathologica. 82, 72-77 (1991).
- Marchalant, Y., Rosi, S., Wenk, G. L. Anti-inflammatory property of the cannabinoid agonistWIN-55212-2 in a rodent model of chronic brain inflammation. Neurociência. 144, 1516-1522 (2007).
- Naert, G., Ixart, G., Tapia-Arancibia, L., Givalois, L. Continuous i.c.v. infusion of brainderivedneurotrophic factor modifies hypothalamic-pituitary-adrenal axis activity,locomotor activity and body temperature rhythms in adult male rats. Neurociência. 139, 779-789 (2006).
- Nikkhah, G., Olsson, M., Eberhard, J., Bentlage, C., Cunningham, M. G., Bjorklund, A. Amicrotransplantation approach for cell suspension grafting in the rat Parkinson model: adetailed account of the methodology. Neurociência. 63, 57-72 (1994).
- Perry, V. H., Andersson, P. B., Gordon, S. Macrophages and inflammation in the centralnervous system. Trends Neurosci. 16, 268-273 (1993).
- Radecki, D. T., Brown, L. M., Martinez, J., Teyler, T. J. BDNF protects against stress-inducedimpairments in spatial learning and memory and LTP. Hippocampus. 15, 246-253 (2005).
- Rosi, S., Ramirez-Amaya, V., Hauss-Wegrzyniak, B., Wenk, G. L. Chronic brain inflammationleads to a decline in hippocampal NMDA-R1 receptors. J Neuroinflammation. 1, 12-12 (2004).
- Takahashi, M., Kakita, A., Futamura, T., Watanabe, Y., Mizuno, M., Sakimura, K., Castren, E., Nabeshima, T., Someya, T., Nawa, H. Sustained brain-derived neurotrophic factor upregulationand sensorimotor gating abnormality induced by postnatal exposure tophencyclidine: comparison with adult treatment. J Neurochem. 99, 770-780 (2006).
- Williams, L. R., Vahlsing, H. L., Lindamood, T., Varon, S., Gage, F. H., Manthorpe, M. A smallgaugecannula device for continuous infusion of exogenous agents into the brain. Experimental neurology. 95, 743-754 (1987).
- Zhang, X., Lee, T. H., Xiong, X., Chen, Q., Davidson, C., Wetsel, W. C. Ellinwood EH.Methamphetamine induces long-term changes in GABAA receptor alpha2 subunit andGAD67 expression. Biochem Biophys Res Commun. 351, 300-305 (2006).
Tags
ConstructionImplantationMicroinfusion SystemSustained DeliveryNeuroactive AgentsPrecisionLocalized TraumaInflammationCustomizeResearch AnimalMicrocannula DiameterAccuracyBrain RegionDelivery CannulaSculptedSurface Of The SkullClosure Of Scalp IncisionRisk Of InfectionComfort To The AnimalAssembled In Air-free EnvironmentGlass MicropipettesHeat SourceSpecial SkillsVideo Instruction
Citar este artigo
Cunningham, M. G., O’Connor, R. P., Wong, S. E. Construction and Implantation of a Microinfusion System for Sustained Delivery of Neuroactive Agents.. J. Vis. Exp. (13), e716, doi:10.3791/716 (2008).