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成年斑马鱼(Danio rerio)内耳感觉上皮的大体解剖和精细解剖。

Gross and fine dissection of inner ear sensory epithelia in adult zebrafish (Danio rerio).

作者信息

Liang Jin, Burgess Shawn M

机构信息

Genome Technology Branch, National Human Genome Research Institute, NIH, USA.

出版信息

J Vis Exp. 2009 May 8(27):1211. doi: 10.3791/1211.

Abstract

Neurosensory epithelia in the inner ear are the crucial structures for hearing and balance functions. Therefore, it is important to understand the cellular and molecular features of the epithelia, which are mainly composed of two types of cells: hair cells (HCs) and supporting cells (SCs). Here we choose to study the inner ear sensory epithelia in adult zebrafish not only because the epithelial structures are highly conserved in all vertebrates studied, but also because the adult zebrafish is able to regenerate HCs, an ability that mammals lose shortly after birth. We use the inner ear of adult zebrafish as a model system to study the mechanisms of inner ear HC regeneration in adult vertebrates that could be helpful for clinical therapy of hearing/balance deficits in human as a result of HC loss. Here we demonstrate how to do gross and fine dissections of inner ear sensory epithelia in adult zebrafish. The gross dissection removes the tissues surrounding the inner ear and is helpful for preparing tissue sections, which allows us to examine the detailed structure of the sensory epithelia. The fine dissection cleans up the non-sensory-epithelial tissues of each individual epithelium and enables us to examine the heterogeneity of the whole epithelium easily in whole-mount epithelial samples.

摘要

内耳中的神经感觉上皮是听觉和平衡功能的关键结构。因此,了解该上皮的细胞和分子特征很重要,其主要由两种类型的细胞组成:毛细胞(HCs)和支持细胞(SCs)。在这里,我们选择研究成年斑马鱼的内耳感觉上皮,这不仅是因为上皮结构在所有已研究的脊椎动物中高度保守,还因为成年斑马鱼能够再生毛细胞,而哺乳动物在出生后不久就失去了这种能力。我们将成年斑马鱼的内耳作为一个模型系统,来研究成年脊椎动物内耳毛细胞再生的机制,这可能有助于临床治疗因毛细胞损失导致的人类听力/平衡缺陷。在这里,我们展示了如何对成年斑马鱼的内耳感觉上皮进行大体解剖和精细解剖。大体解剖去除内耳周围的组织,有助于制备组织切片,使我们能够检查感觉上皮的详细结构。精细解剖清理每个上皮的非感觉上皮组织,使我们能够在整装上皮样本中轻松检查整个上皮的异质性。

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