Isolation and Characterization of the Olfactory Epithelial Cells of the Catfish
Isolation and Characterization of the Olfactory Epithelial Cells of the Catfish
复制标题
鲶鱼嗅觉上皮细胞的分离和表征
DOI：
10.1093/chemse/3.4.381
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发表时间：
1978
期刊：
影响因子：
3.5
通讯作者：
P. Cancalon
中科院分区：
文献类型：
--
作者：
P. Cancalon
从鲶鱼（Ictalurus punctatus）的嗅觉粘膜中获得了一种富含嗅觉受体细胞的制剂。参照Cohen等人(1)的方法，在Ca+ +、Mg+ +游离培养基中依次用胰蛋白酶、DNase、胰蛋白酶抑制剂、EDTA处理组织。在机械破坏分离的嗅觉薄片后，在Ficoll梯度上离心分离细胞。通过扫描电镜（SEM）对比原位和体外制备，对各类型细胞进行形态学鉴定。在10%的Ficoll上收集小的圆形细胞。这些细胞的性质尚不清楚，但其中一部分肯定是基底细胞，它们已被证明是不断再生的嗅觉神经元的前体。呼吸细胞主要落在20%的Ficoll上。在33%的Ficoll上收集含有60%支撑细胞的部分。嗅觉细胞以轴突、树突和几根纤毛为特征，在37%的Ficoll上发现。这个部分也含有高达40%的支撑细胞。嗅觉细胞分离的产率为20%。活体染色和合成RNA的能力表明最终制备的存活率为70%至80%。通过测量一系列氨基酸与嗅觉细胞中富集的制剂的结合活性，进一步鉴定了这些细胞。在结合程度和这些氨基酸在体内记录的电生理活动之间确定了良好的相关性。虽然最终的嗅细胞悬浮液不纯，但它构成了嗅觉受体位点研究的第一步。
A preparation enriched in olfactory receptor cells has been obtained from the olfactory mucosa of the catfish (Ictalurus punctatus). The tissue was treated successively with trypsin, DNase, trypsin inhibitor, EDTA in Ca+ +, Mg+ +free medium according to a method derived from that of Cohen,et al.(1). After mechanical disruption of the isolated olfactory lamellae, the cells were isolated by centrifugation on a Ficoll gradient. Each type of cell was morphologically identified by comparingin situandin vitropreparations by SEM. Small round cells were collected on 10% Ficoll. The nature of these cells is not known but part of them are certainly basal cells which have been shown(2)to be the precursors of the constantly regenerating olfactory neurons. Respiratory cells settled mainly on 20% Ficoll. A fraction containing 60% sustentacular cells was collected on 33% Ficoll. Olfactory cells characterized by an axon, a dendrite and several cilia, were found on 37% Ficoll. This fraction also contains up to 40% sustentacular cells. A yield of 20% was measured for olfactory cell isolation. Vital staining and ability to synthesize RNA indicate a viability of the final preparation of 70% to 80%. Further identification of the cells was performed by measuring the binding activity of a series of amino acids to a preparation enriched in olfactory cells. A good correlation was determined between the extent of the binding and the reported electrophysiological activities of these amino acids recordedin vivo. Although the final olfactory cell suspension is not pure, it constitutes the first step in the study of the olfactory receptor sites.