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用于实时Ca2+成像的鸡味蕾分离

Isolation of chicken taste buds for real-time Ca2+ imaging.

作者信息

Kudo Ken-ichi, Kawabata Fuminori, Nomura Toumi, Aridome Ayumi, Nishimura Shotaro, Tabata Shoji

机构信息

Department of Bioresource Sciences, Graduate School of Bioresource and Bioenvironmental Sciences, Fukuoka, Japan.

出版信息

Anim Sci J. 2014 Oct;85(10):904-9. doi: 10.1111/asj.12222. Epub 2014 May 19.

Abstract

We isolated chicken taste buds and used a real-time Ca2+ imaging technique to investigate the functions of the taste cells. With RT-PCR, we found that isolated chicken taste bud-like cell subsets express chicken gustducin messenger RNA. Immunocytochemical techniques revealed that the cell subsets were also immunopositive for chicken gustducin. These results provided strong evidence that the isolated cell subsets contain chicken taste buds. The isolated cell subsets were spindle-shaped and approximately 61-75 μm wide and 88-98 μm long, and these characteristics are similar to those of sectional chicken taste buds. Using Ca2+ imaging, we observed the buds' response to 2 mmol/L quinine hydrochloride (a bitter substance) and their response to a mixture of 25 mmol/L L-glutamic acid monopotassium salt monohydrate and 1 mmol/L inosine 5'-monophosphate disodium salt, umami substances. The present study is the first morphological demonstration of isolated chicken taste buds, and our results indicate that the isolated taste buds were intact and functional approaches for examining the taste senses of the chicken using Ca2+ imaging can be informative.

摘要

我们分离出鸡的味蕾,并使用实时Ca2+成像技术来研究味觉细胞的功能。通过逆转录聚合酶链反应(RT-PCR),我们发现分离出的类似鸡味蕾的细胞亚群表达鸡味觉传导素信使核糖核酸。免疫细胞化学技术显示,这些细胞亚群对鸡味觉传导素也呈免疫阳性。这些结果提供了强有力的证据,证明分离出的细胞亚群包含鸡的味蕾。分离出的细胞亚群呈纺锤形,宽约61 - 75μm,长88 - 98μm,这些特征与切片鸡味蕾的特征相似。利用Ca2+成像,我们观察了味蕾对2mmol/L盐酸奎宁(一种苦味物质)的反应以及它们对25mmol/L L-谷氨酸单钾盐一水合物和1mmol/L 5'-肌苷二磷酸二钠盐(鲜味物质)混合物的反应。本研究首次对分离出的鸡味蕾进行了形态学证明,我们的结果表明,分离出的味蕾是完整的,并且使用Ca2+成像检查鸡味觉的功能方法可能会提供有用信息。

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