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出生后小鼠听泡的解剖:中耳骨的分离及组织学分析

Dissection of the Auditory Bulla in Postnatal Mice: Isolation of the Middle Ear Bones and Histological Analysis.

作者信息

Sakamoto Ayako, Kuroda Yukiko, Kanzaki Sho, Matsuo Koichi

机构信息

Laboratory of Cell and Tissue Biology, Keio University School of Medicine.

Department of Otolaryngology Head and Neck Surgery, Keio University School of Medicine.

出版信息

J Vis Exp. 2017 Jan 4(119):55054. doi: 10.3791/55054.

Abstract

In most mammals, auditory ossicles in the middle ear, including the malleus, incus and stapes, are the smallest bones. In mice, a bony structure called the auditory bulla houses the ossicles, whereas the auditory capsule encloses the inner ear, namely the cochlea and semicircular canals. Murine ossicles are essential for hearing and thus of great interest to researchers in the field of otolaryngology, but their metabolism, development, and evolution are highly relevant to other fields. Altered bone metabolism can affect hearing function in adult mice, and various gene-deficient mice show changes in morphogenesis of auditory ossicles in utero. Although murine auditory ossicles are tiny, their manipulation is feasible if one understands their anatomical orientation and 3D structure. Here, we describe how to dissect the auditory bulla and capsule of postnatal mice and then isolate individual ossicles by removing part of the bulla. We also discuss how to embed the bulla and capsule in different orientations to generate paraffin or frozen sections suitable for preparation of longitudinal, horizontal, or frontal sections of the malleus. Finally, we enumerate anatomical differences between mouse and human auditory ossicles. These methods would be useful in analyzing pathological, developmental and evolutionary aspects of auditory ossicles and the middle ear in mice.

摘要

在大多数哺乳动物中,中耳的听小骨,包括锤骨、砧骨和镫骨,是最小的骨骼。在小鼠中,一个名为听泡的骨质结构容纳着听小骨,而听囊则包裹着内耳,即耳蜗和半规管。小鼠的听小骨对听力至关重要,因此在耳鼻喉科学领域备受研究人员关注,但其代谢、发育和进化与其他领域也高度相关。骨代谢改变会影响成年小鼠的听力功能,各种基因缺陷小鼠在子宫内听小骨的形态发生会出现变化。尽管小鼠的听小骨很小,但如果了解其解剖方位和三维结构,对其进行操作是可行的。在此,我们描述如何解剖出生后小鼠的听泡和听囊,然后通过去除部分听泡来分离单个听小骨。我们还讨论了如何以不同方位包埋听泡和听囊,以生成适合制备锤骨纵切面、水平面或额状面切片的石蜡或冰冻切片。最后,我们列举了小鼠和人类听小骨之间的解剖差异。这些方法将有助于分析小鼠听小骨和中耳的病理、发育及进化方面。

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