Increase in muscarinic stimulation-induced Ca2+ response by adenovirus-mediated Stim1-mKO1 gene transfer to rat submandibular acinar cells in vivo.
Increase in muscarinic stimulation-induced Ca2+ response by adenovirus-mediated Stim1-mKO1 gene transfer to rat submandibular acinar cells in vivo.
复制标题
通过腺病毒介导的 Stim1-mKO1 基因转移到体内大鼠下颌下腺泡细胞，增加毒蕈碱刺激诱导的 Ca2 反应。
DOI：
10.1016/j.bbrc.2013.08.080
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Tanimura A
中科院分区：
文献类型：
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作者：
Morita T;Nezu A;Tojyo Y;Tanimura A
腺病毒已被用于体内基因转移到唾液腺细胞。它们用于研究唾液腺泡细胞的功能受到严重炎症反应和液体分泌腺泡细胞破坏的限制。在本研究中，低剂量的腺病毒通过逆行导管注射到下颌下腺来表达Stim 1-mKO 1。该方法成功地增加了毒蕈碱刺激诱导的腺泡细胞中的Ca 2+反应，而没有炎症或唾液分泌减少。这种增加的Ca 2+反应在弱毒蕈碱刺激时是显著的，并且归因于从内部存储增加的Ca 2+释放和增加的Ca 2+进入。Stim 1-mKO 1表达细胞的基础Ca 2+水平高于mKO 1表达和非表达细胞。暴露透化的下颂腺泡细胞，其中Ca 2+浓度固定在50 nM，肌醇1，4，5-三磷酸（IP 3）产生类似的影响释放钙从存储在Stim 1-mKO 1表达和非表达细胞。本发明所述的温和基因转移方法的低毒性和对腺泡细胞的相对特异性特别适用于研究体内唾液腺泡细胞的分子功能，并且将来可能应用于增加实验动物和人的唾液分泌。
Adenoviruses have been used for gene transfer to salivary gland cellsin vivo. Their use to study the function of salivary acinar cells was limited by a severe inflammatory response and by the destruction of fluid-secreting acinar cells. In the present study, low doses of adenovirus were administered to express Stim1-mKO1 by retrograde ductal injection to submandibular glands. The approach succeeded in increasing muscarinic stimulation-induced Ca2+responses in acinar cells without inflammation or decreased salivary secretions. This increased Ca2+response was notable upon weak muscarinic stimulation and was attributed to increased Ca2+release from internal stores and increased Ca2+entry. The basal Ca2+level was higher in Stim1-mKO1-expressing cells than in mKO1-expressing and non-expressing cells. Exposure of permeabilized submandibular acinar cells, where Ca2+concentration was fixed at 50 nM, to inositol 1,4,5-trisphosphate (IP3) produced similar effects on the release of Ca2+from stores in Stim1-mKO1-expressing and non-expressing cells. The low toxicity and relative specificity to acinar cells of the mild gene transfer method described herein are particularly useful for studying the molecular functions of salivary acinar cellsin vivo, and may be applied to increase salivary secretions in experimental animals and human in future.