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研究生:洪敬賢
研究生(外文):Ching-Hsien Hung
論文名稱:紀錄大鼠中耳腔黏膜電生理行為的新方法
論文名稱(外文):A New Method in Studying the Electrophysiology of the Rat Middle Ear Mucosa
指導教授:劉天申
指導教授(外文):Tien-Shen Lew
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生理學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:英文
論文頁數:44
中文關鍵詞:電生理中耳腔黏膜活組織
外文關鍵詞:electrophysiologymiddle ear mucosalive tissue
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目前有關中耳腔黏膜電生理行為的研究資料,都是取材於新鮮組織分離培養的上皮細胞、或是病毒轉植的細胞株 (SV40)。但這些培養的細胞與新鮮活組織的行為可能有相當差異,因此本實驗室嘗試建立一套新方法,來研究新鮮中耳黏膜組織的電生理行為。我們將2到6個月大的大白鼠以thiopental深度麻醉後,直接剝取顳骨,將包圍骨骼的軟組織盡量清除乾淨,然後以電鑽將bullae oticae的外側壁和上壁磨穿,同時擴大開口,並小心地磨開下壁的骨骼,露出內側的中耳腔黏膜。這個露出部位的基底外側 (basolateral) 膜再與一段PE管 (直徑0.86 mm) 密接,而與浸泡標本的溶液完全隔絕。在一系列含有不同濃度鈉、鉀、和氯離子的管腔側 (apical) 溶液的環境下,我們以注入方波電流的方式紀錄橫跨上皮兩側的電位差 (VTE),並同時計算電阻 (RTE) 和短路電流 (ISC)。初步結果 (n = 11) 顯示:在膜兩側均為生理性溶液的環境下,VTE為 -1.4 ± 0.1 mV,RTE為84 ± 5 Ω•cm2,ISC為18 ± 2 μA/cm2;此外,VTE會被ouabain去極化,但不受amiloride影響。管腔側膜氯離子通透度為0.75 ± 0.14 mS/cm2,鉀離子通透度為0.62 ± 0.14 mS/cm2,而細胞旁 (paracellular) 通透度約為10.7 mS/cm2。因此與分離培養的上皮細胞及SV40細胞株相比,新鮮中耳腔黏膜顯得非常不緊密,並且上皮細胞頂側膜電位的主要決定離子是鉀離子和氯離子,而非鈉離子。換言之,新鮮中耳腔黏膜為分泌性,而非吸收性上皮組織。
Present information referring to the electrophysiology of the middle ear mucosa came from cultured epithelial cells or the virus-transfected cell line (SV40). However, the characteristics of live tissue may be different from those of the cultured cells. Therefore in this lab, we tried to establish a new method to study the bioelectric properties of the live mucosa of the middle ear. Adult S.D. rats (2 to 6 months old) were deeply anesthetized with thiopental to harvest the temporal bones. Soft tissue outside the bone was removed as possible. The superior and the lateral walls of the bullae oticae were drilled open. The bony plate on the inferior wall was carefully removed to expose the mucosa inside, which was to be made seal by a section of PE tubing (0.86 mm in diameter). The seal isolated that part of basolateral membrane from the bath solution. The transepithelial potential (VTE) was recorded by current clamping in a series of apical bath solutions with different concentrations of Na+, K+, or Cl-, so as to calculate the transepithelial resistance (RTE) and the short circuit current (ISC). The preliminary results (n = 11) showed that the VTE was around -1.4 ± 0.1 mV in physiological bath solution, the RTE was 84 ± 5 Ω•cm2, and the ISC was 18 ± 2 μA/cm2. Besides, the VTE was depolarized by ouabain, but was not altered by amiloride. Also, the apical chloride conductance was 0.75 ± 0.14 mS/cm2, the apical potassium conductance was 0.62 ± 0.13 mS/cm2, and the paracellular conductance was around 10.7 mS/cm2. Therefore at this moment we conclude that live middle ear epithelia behaves relatively leaky in comparison with cultured mucosal cells or SV40. Moreover, the main contributors for the apical membrane potential seem to be K+ and Cl-, instead of Na+. That is to say, the live epithelia is secretory, not absorptive.
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