Visualization of 'water secretion' by confocal microscopy in rat salivary glands: possible distinction of para- and transcellular pathway.
Visualization of 'water secretion' by confocal microscopy in rat salivary glands: possible distinction of para- and transcellular pathway.
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通过共聚焦显微镜观察大鼠唾液腺“水分泌”的可视化：旁细胞途径和跨细胞途径的可能区别。
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发表时间：
2002
期刊：
影响因子：
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通讯作者：
M. Murakami
中科院分区：
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作者：
A. Segawa;S. Yamashina;M. Murakami
细胞、组织和器官中水分运输的可视化是形态学中一个重要但仍然困难的任务。利用共聚焦显微镜和液相荧光示踪技术，观察了大鼠腮腺和颌下腺腺泡上皮的水分泌情况，并估计了水在腺泡上皮中的运输途径。全腺体动脉灌注Lucifer黄的共聚焦显微镜显示腺泡基底外侧空间有明亮的荧光。腔内无荧光，经异丙肾上腺素预处理、经导管向腔内灌注荧光右旋糖酐或胶原酶解离组织后可见荧光。在这些条件下，用乙醇刺激液体分泌引起腔内荧光强度迅速下降，表明分泌的水冲洗了腺泡腔内的荧光探针。在受刺激的游离腺泡中，荧光消失15秒，但在30-45秒时重新出现，保持较低的平台水平。假设紧密连接被胶原酶消化“麻痹”，并且细胞旁液体运输不能影响路西法黄的稀释，我们估计CCh最初的水分泌是通过跨细胞途径发生的，而在30-45秒后，额外的水通过细胞旁途径渗透。
Visualization of water transport in cells, tissues and organs is an important, yet still difficult, task in morphological science. By using confocal microscopy and the fluid-phase fluorescent tracer technique, we visualized water secretion and estimated the routes of water transport across the acinar epithelia in rat parotid and submandibular glands. Confocal microscopy of whole glands perfused arterially with Lucifer yellow revealed a bright fluorescence at the basolateral space of acini. Luminal space was devoid of fluorescence, but revealed it after isoproterenol pretreatment, ductal infusion of fluorescent dextrans into the lumen, or tissue dissociation by collagenase. Under these conditions, stimulation of fluid secretion with carbachol caused a rapid decline of the luminal fluorescence intensity, indicating that the secreted water washed out the fluorescent probes in the acinar lumen. In the stimulated dissociated acini, the luminal fluorescence disappeared by 15 sec, but reappeared at 30-45 sec to maintain a low plateau level. By assuming that the tight junction was 'paralyzed' by the collagenase digestion and that the paracellular fluid transport could not influence the dilution of Lucifer yellow, we estimated that the initial water secretion by CCh occurs via the transcellular pathway, while later than 30-45 sec the additional water permeates through the paracellular pathway.