近红外（NIR）光增加线粒体功能的标志物的表达，在鼠标前庭上皮细胞
Summary
线粒体功能障碍是细胞衰老的标志。本文使用的非侵入性的近红外（NIR）处理，以改善在老化小鼠前庭感觉上皮线粒体功能。
Abstract
策略随着年龄的增加衰减下降平衡功能主要是专注于物理疗法，包括平衡工作和锻炼。然而，这些方法没有解决平衡下降的根本原因。使用小鼠，近红外光（NIR）的细胞上，在前庭感觉上皮的代谢的影响进行了评估。收集的数据表明，这种简单和安全的干预可能保护这些脆弱的细胞从自然老化的有害影响。的mRNA从分离的前庭外周感觉上皮（壶腹嵴和椭圆囊斑）萃取，随后转录成cDNA文库。这个库然后探查无处不抗氧化剂（SOD-1）的表达。抗氧化剂的基因表达，然后用于量化细胞代谢。用年轻的经颅交付NIR（4周）及以上（8 – 9个月）小鼠，并简要治疗方案（90秒/天5天AYS），这项工作表明NIR单独可足以改善线粒体功能的前庭感觉上皮。由于目前没有可用的，负担得起的，非侵入性的治疗方法，提高前庭毛细胞功能，外部NIR辐射的应用程序提供，以抵消对细胞在矿井前庭感觉上皮代谢老化的影响，潜在的战略。
Introduction
余额递减性能和后续的瀑布是常见的，而自然老化1的不幸往往定义功能。这种下降的影响，既可以是自然和社会，和显著降低生活质量的老年人。作为回应，物理治疗和康复一直是研究的重点为瀑布，但尚未有一致的减少重复跌倒的发生率有关。与此同时，工作调查在周边或中央前庭系统（负责保持平衡的系统）是稀少，并且针对这些系统的潜在的治疗策略的变化和不平衡有限的根本原因。
在年龄相关的神经变性病症，包括年龄相关性黄斑变性2-4，阿耳茨海默氏病模型5-8，和帕金森氏病9-12最近的工作已经表明SI的神经保护作用mple非侵入性的应用的近红外（NIR）光。另外，在前庭系统，NIR已被用于提高在体外 13前庭初级传入神经 元的活性。而没有很好理解NIR光的机构，利用NIR大多数的研究表明，NIR刺激线粒体复合物IV（细胞色素C氧化酶）14-17，以促进细胞代谢。在前庭感觉上皮I型毛细胞的表皮下板是致密的线粒体18，因此可以代表用于治疗NIR治疗作用的部位。
这里，经颅的简要的，非侵入性的治疗方案应用于NIR可用于测量细胞代谢（和暗示线粒体功能）在小鼠前庭感觉上皮进行说明。还讨论了为制备的前庭感觉上皮和它表明NIR增加一个ubiquito的表达我们的抗氧化剂（超氧化物歧化酶1）中的感觉上皮-先前证明是对耳蜗毛细胞存活19重要。
Protocol
伦理学声明：下面列出的所有程序批准了悉尼动物伦理委员会的大学。 1.动物注：1和8 – 9个月大的小鼠（C57 / BL6）是从动物资源中心（珀斯，澳大利亚）中获得。老鼠被安置在博世灭鼠设施，在悉尼大学。 家鼠在上一个12/12小时亮/暗循环以获得食物和水随意标准鼠标笼中。 鸿沟小鼠各年龄组为近红外（NIR）治疗，或假处理（对照?…
Representative Results
比较近红外治疗的年轻年长的影响（4周）和（8 – 9个月）小鼠，我们测得的抗氧化剂超氧化物歧化酶1（SOD-1）的表达年轻的（N = 16）和老年人（N = 20）小鼠是NIR治疗，假治疗，或NIR封; 图2示出了相比于年轻假处理的动物一个显著增加β肌动蛋白标准化的SOD-1的表达超过2倍于年轻NIR处理的动物（P <0.01），年轻的NIR阻断动物（P <0.01）。较旧的NIR处理的动物也显示出超过一个时与旧NI…
Discussion
这里所描述的代表性的结果表明，当与假治疗的小鼠相比较简短颅递送NIR光的（90秒/天5天）足以提高抗氧化剂的表达在老年小鼠的水平。而发射的热可以代表线粒体和/或神经元激活的一个来源，据报道为大鼠前庭传入神经24 -我们的热量由NIR发射测量LED器件是<0.2℃，90秒，因此不太可能导致这里描述的变化。此外，与报告所强调以上，这里使用的NIR治疗不直接向感觉上皮或前庭传?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
笔者要感谢保罗·维悌希博士和吉纳维芙方女士的mRNA的提取和PCR援助，以及加内特凋谢和罗德尼·威廉斯纪念基金会的支持。
Materials
| Name of Material/ Equipment | Company | Catalog Number | Comments/Description |
| Quantum WARP 10 | Quantum Devices | 2070N030-A | |
| Screw top microtubules | Quality Scientific Plastics | 520-GRD-Q | |
| Ketamine | Parnell, Alexandria Australia | ||
| Standard Pattern Scissors | FST | 14001-12 | |
| Carbon steel Surgical Blades #22 | Livingstone | SBLDCL 22 | |
| Friedman-Pearson Rongeurs | FST | 16221-14 | |
| Stereo microscope | Leica Microsystems | A60S | |
| Dumont #5 SF Forceps | FST | 11252-00 | |
| Isolate II RNA Micro Kit | Bioline | BIO-52075 |
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Cite This Article
Zhang, L., Tung, V. W. K., Mathews, M., Camp, A. J. Near Infrared (NIr) Light Increases Expression of a Marker of Mitochondrial Function in the Mouse Vestibular Sensory Epithelium. J. Vis. Exp. (97), e52265, doi:10.3791/52265 (2015).