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从豚鼠鼻腺中分离腺泡。

Isolation of acini from nasal glands of the guinea-pig.

作者信息

Sunose H, Zhang W, Ishigaki M, Katori Y, Suzuki M, Ikeda K, Takasaka T, Saito Y, Nishiyama A

机构信息

Department of Otolaryngology, Tohoku University School of Medicine, Sendai, Japan.

出版信息

Acta Physiol Scand. 1994 Jul;151(3):377-84. doi: 10.1111/j.1748-1716.1994.tb09757.x.

Abstract

A procedure for isolating the acinar cells of the serous gland in the mammalian nasal septum has been developed. This technique is characterized by meticulous and selective isolation with minimal contamination by the surface epithelial cells and employs enzymatic treatment with collagenase. The isolated cells were confirmed to be serous gland acini as shown by negative staining with Alcian blue and a high electron density of the granules. The acini were more than 90% viable as judged by trypan blue exclusion. Ultrastructural integrity of the cells was well maintained following the isolation procedure. Application of acetylcholine to the isolated acini induced an inward current in a whole-cell patch clamp and increased intracellular Ca2+ concentration measured by fura-2. These acetylcholine responses were completely blocked by atropine. These physiological findings directly demonstrated that nasal gland acini possess muscarinic-activated receptors as previously suggested. These isolated cells hold promise for the in vitro study of secretory mechanisms in the mammalian nasal gland.

摘要

已开发出一种用于分离哺乳动物鼻中隔浆液性腺泡细胞的方法。该技术的特点是精细且选择性地分离,表面上皮细胞污染最小,并采用胶原酶进行酶处理。经阿利新蓝阴性染色和颗粒的高电子密度显示,分离出的细胞被确认为浆液性腺泡。通过台盼蓝排斥法判断,腺泡的存活率超过90%。分离过程后,细胞的超微结构完整性得到良好维持。在全细胞膜片钳中,向分离出的腺泡施加乙酰胆碱可诱导内向电流,并增加用fura-2测量的细胞内Ca2+浓度。这些乙酰胆碱反应被阿托品完全阻断。这些生理学发现直接证明,鼻腺泡如先前推测的那样具有毒蕈碱激活受体。这些分离出的细胞有望用于哺乳动物鼻腺分泌机制的体外研究。

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