Organotypic Culture of Physiologically Functional Adult Mammalian Retinas
Organotypic Culture of Physiologically Functional Adult Mammalian Retinas
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DOI：
10.1371/journal.pone.0000221
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发表时间：
2007-02-21
期刊：
影响因子：
3.7
通讯作者：
Jakobs, Tatjana C.
中科院分区：
文献类型：
--
作者：
Koizumi, Amane;Zeck, Guenther;Jakobs, Tatjana C.
背景成年哺乳动物视网膜是研究中枢神经系统的重要模型。许多实验需要遗传操作、成像和电生理记录的组合使用，这使得期望使用体外制备。遗憾的是，成年哺乳动物视网膜的组织培养是困难的，主要是因为光感受器的高能量消耗。方法和发现。我们描述了一种用于成年哺乳动物视网膜的间期培养系统，该系统允许通过颗粒介导的转移将基因递送至视网膜神经元来表达。视网膜的形态和功能可保留长达六天，这段时间足以表达许多感兴趣的基因，因此可以通过贴片记录或多电极阵列记录来测量对光和感受野反应的影响。我们发现，各种基因编码的前和突触后标记蛋白正确定位在神经节和无长突细胞。结论.在该系统中，可以在成年哺乳动物视网膜的完整神经回路内研究一个或几个引入的外源基因对神经元功能的影响。该系统足够灵活，可以兼容遗传操作、成像、细胞转染、药理学测定和电生理记录。
Background. The adult mammalian retina is an important model in research on the central nervous system. Many experiments require the combined use of genetic manipulation, imaging, and electrophysiological recording, which make it desirable to use an in vitro preparation. Unfortunately, the tissue culture of the adult mammalian retina is difficult, mainly because of the high energy consumption of photoreceptors. Methods and Findings. We describe an interphase culture system for adult mammalian retina that allows for the expression of genes delivered to retinal neurons by particle-mediated transfer. The retinas retain their morphology and function for up to six days-long enough for the expression of many genes of interest-so that effects upon responses to light and receptive fields could be measured by patch recording or multielectrode array recording. We show that a variety of genes encoding pre- and post-synaptic marker proteins are localized correctly in ganglion and amacrine cells. Conclusions. In this system the effects on neuronal function of one or several introduced exogenous genes can be studied within intact neural circuitry of adult mammalian retina. This system is flexible enough to be compatible with genetic manipulation, imaging, cell transfection, pharmacological assay, and electrophysiological recordings.