研究生: |
葉雅欣 Yeh, Ya-Hsin |
---|---|
論文名稱: |
CaSR與PMCA1a在斑馬魚側線毛細胞上扮演之功能 Functional study of CaSR and PMCA1a in zebrafish lateral line hair cells |
指導教授: |
林豊益
Lin, Li-Yih |
學位類別: |
碩士 Master |
系所名稱: |
生命科學系 Department of Life Science |
論文出版年: | 2015 |
畢業學年度: | 103 |
語文別: | 中文 |
論文頁數: | 49 |
中文關鍵詞: | 毛細胞 、機械性傳導通道 、鈣離子平衡 、鈣離子感應接受器 、鈣離子通道 |
英文關鍵詞: | Hair cell, MET channel, Calcium homeostasis, CaSR, PMCA1a |
論文種類: | 學術論文 |
相關次數: | 點閱:163 下載:0 |
分享至: |
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報 |
哺乳動物的內耳毛細胞扮演偵測聲音的功能,當音波的震動傳入內耳,會使纖毛束彎曲並進而開啟機械性通道產生聽覺。毛細胞的機械性通道位於纖毛的頂端是一種對鈣離子具有高通透性的陽離子通道。近年來,斑馬魚的側線系統被廣泛研究,且側線毛細胞被做為活體研究的新指標。本篇研究利用掃描式離子選擇電極技術(SIET),以活體的狀況下測量斑馬魚仔魚側線毛細胞機械性通道的功能,進一步觀察PMCA與CaSR在毛細胞的表現以及對機械性通道功能的影響。在本篇實驗中證實PMCA1a與CaSR的蛋白質表現在受精後第96小時斑馬魚仔魚毛細胞靜纖毛的位置。以Morpholino knock down、PMCA抑制劑 (o-vanadate, Eosin-Y)、CaSR促進劑(R568)處理,發現機械性通道的鈣離子流受到抑制,表明了PMCA1a與CaSR對於機械性通道功能的重要性。此外,給予仔魚短期高鈣的環境也會抑制機械性通道鈣離子的流入,顯示環境鈣離子變化時機械性通道的功能會受到調節,而調節機械性通道的功能可能與PMCA1a以及CaSR參與有關。
The sensory hair cells in mammalian inner ear are responsible for sound transduction. Sound waves deflect hair bundles and open the mechanotransducer (MET) channels, which initiate mechanotransduction. The MET channels expressed in stereocilia are cation channels with high Ca2+ permeability. Zebrafish lateral line is recently reported to be a useful in vivo model for studying hair cells. We have demonstrated the scanning ion-electrode technique (SIET) to be a sensitive approach for functionally assaying MET channel in zebrafish. In this study, expression and function of plasma membrane Ca2+ ATPase (PMCA) and calcium-sensing receptor (CaSR) were investigated. The protein expressions of PMCA1a and CaSR were showed in stereocilia of lateral line hair cell of 96 hpf zebrafish larvae. The MET channel mediated Ca2+ influx detected by SIET was suppressed by treatment of PMCA inhibitor (o-vanadate, Eosin Y) and CaSR activator (R568). These results indicated that PMCA1a and CaSR was critical for maintaining the function of MET channel. Furthermore, function of MET channel was decreased in larvae incubated in high Ca2+ water. These results revealed that function of MET channel is altered in different environmental Ca2+ levels. This functional regulation of MET channel may be required the participation of PMCA1a and CaSR.
Alfadda, T.I., Saleh, A.M., Houillier, P. & Geibel, J.P. (2014) Calcium-sensing receptor 20 years later. American journal of physiology. Cell physiology, 307, C221-231.
Amarjargal, N., Mazurek, B., Haupt, H., Andreeva, N., Fuchs, J. & Gross, J. (2008) Effects of SERCA and PMCA inhibitors on the survival of rat cochlear hair cells during ischemia in vitro. Physiological research / Academia Scientiarum Bohemoslovaca, 57, 631-638.
Beurg, M., Nam, J.H., Chen, Q. & Fettiplace, R. (2010) Calcium balance and mechanotransduction in rat cochlear hair cells. Journal of neurophysiology, 104, 18-34.
Bortolozzi, M., Brini, M., Parkinson, N., Crispino, G., Scimemi, P., De Siati, R.D., Di Leva, F., Parker, A., Ortolano, S., Arslan, E., Brown, S.D., Carafoli, E. & Mammano, F. (2010) The novel PMCA2 pump mutation Tommy impairs cytosolic calcium clearance in hair cells and links to deafness in mice. The Journal of biological chemistry, 285, 37693-37703.
Chiu, L.L., Cunningham, L.L., Raible, D.W., Rubel, E.W. & Ou, H.C. (2008) Using the zebrafish lateral line to screen for ototoxicity. Journal of the Association for Research in Otolaryngology : JARO, 9, 178-190.
Cruz, S., Shiao, J.C., Liao, B.K., Huang, C.J. & Hwang, P.P. (2009) Plasma membrane calcium ATPase required for semicircular canal formation and otolith growth in the zebrafish inner ear. The Journal of experimental biology, 212, 639-647.
Donini, A. & O'Donnell, M.J. (2005) Analysis of Na+, Cl-, K+, H+ and NH4+ concentration gradients adjacent to the surface of anal papillae of the mosquito Aedes aegypti: application of self-referencing ion-selective microelectrodes. The Journal of experimental biology, 208, 603-610.
Dumont, R.A., Lins, U., Filoteo, A.G., Penniston, J.T., Kachar, B. & Gillespie, P.G. (2001) Plasma membrane Ca2+-ATPase isoform 2a is the PMCA of hair bundles. The Journal of neuroscience : the official journal of the Society for Neuroscience, 21, 5066-5078.
Esterberg, R., Hailey, D.W., Coffin, A.B., Raible, D.W. & Rubel, E.W. (2013) Disruption of intracellular calcium regulation is integral to aminoglycoside-induced hair cell death. The Journal of neuroscience : the official journal of the Society for Neuroscience, 33, 7513-7525.
Feldman, B., Tuchman, M. & Caldovic, L. (2014) A zebrafish model of hyperammonemia. Molecular genetics and metabolism, 113, 142-147.
Fettiplace, R. (2009) Defining features of the hair cell mechanoelectrical transducer channel. Pflugers Archiv : European journal of physiology, 458, 1115-1123.
Ficarella, R., Di Leva, F., Bortolozzi, M., Ortolano, S., Donaudy, F., Petrillo, M., Melchionda, S., Lelli, A., Domi, T., Fedrizzi, L., Lim, D., Shull, G.E., Gasparini, P., Brini, M., Mammano, F. & Carafoli, E. (2007) A functional study of plasma-membrane calcium-pump isoform 2 mutants causing digenic deafness. Proceedings of the National Academy of Sciences of the United States of America, 104, 1516-1521.
Germana, A., Abbate, F., Gonzalez-Martinez, T., del Valle, M.E., de Carlos, F., Germana, G. & Vega, J.A. (2004) S100 protein is a useful and specific marker for hair cells of the lateral line system in postembryonic zebrafish. Neuroscience letters, 365, 186-189.
Ghysen, A. & Dambly-Chaudiere, C. (2007) The lateral line microcosmos. Genes & development, 21, 2118-2130.
Go, W., Bessarab, D. & Korzh, V. (2010) atp2b1a regulates Ca2+ export during differentiation and regeneration of mechanosensory hair cells in zebrafish. Cell calcium, 48, 302-313.
Hwang, P.P. & Chou, M.Y. (2013) Zebrafish as an animal model to study ion homeostasis. Pflugers Archiv : European journal of physiology, 465, 1233-1247.
Jensen, T.P., Buckby, L.E. & Empson, R.M. (2004) Expression of plasma membrane Ca2+ ATPase family members and associated synaptic proteins in acute and cultured organotypic hippocampal slices from rat. Brain research. Developmental brain research, 152, 129-136.
Jensen, T.P., Filoteo, A.G., Knopfel, T. & Empson, R.M. (2007) Presynaptic plasma membrane Ca2+ ATPase isoform 2a regulates excitatory synaptic transmission in rat hippocampal CA3. The Journal of physiology, 579, 85-99.
Kozel, P.J., Davis, R.R., Krieg, E.F., Shull, G.E. & Erway, L.C. (2002) Deficiency in plasma membrane calcium ATPase isoform 2 increases susceptibility to noise-induced hearing loss in mice. Hearing research, 164, 231-239.
Kwong, R.W., Auprix, D. & Perry, S.F. (2014) Involvement of the calcium-sensing receptor in calcium homeostasis in larval zebrafish exposed to low environmental calcium. American journal of physiology. Regulatory, integrative and comparative physiology, 306, R211-221.
Liao, B.K., Deng, A.N., Chen, S.C., Chou, M.Y. & Hwang, P.P. (2007) Expression and water calcium dependence of calcium transporter isoforms in zebrafish gill mitochondrion-rich cells. BMC genomics, 8, 354.
Lieschke, G.J. & Currie, P.D. (2007) Animal models of human disease: zebrafish swim into view. Nature reviews. Genetics, 8, 353-367.
Lin, C.H., Su, C.H. & Hwang, P.P. (2014) Calcium-sensing receptor mediates Ca2+ homeostasis by modulating expression of PTH and stanniocalcin. Endocrinology, 155, 56-67.
Lin, C.Y., Huang, C.C., Wang, W.D., Hsiao, C.D., Cheng, C.F., Wu, Y.T., Lu, Y.F. & Hwang, S.P. (2013a) Low temperature mitigates cardia bifida in zebrafish embryos. PloS one, 8, e69788.
Lin, L.Y., Pang, W., Chuang, W.M., Hung, G.Y., Lin, Y.H. & Horng, J.L. (2013b) Extracellular Ca2+ and Mg2+ modulate aminoglycoside blockade of mechanotransducer channel-mediated Ca2+ entry in zebrafish hair cells: an in vivo study with the SIET. American journal of physiology. Cell physiology, 305, C1060-1068.
Lin, Y.H., Hung, G.Y., Wu, L.C., Chen, S.W., Lin, L.Y. & Horng, J.L. (2015) Anion exchanger 1b in stereocilia is required for the functioning of mechanotransducer channels in lateral-line hair cells of zebrafish. PloS one, 10, e0117041.
Loretz, C.A. (2008) Extracellular calcium-sensing receptors in fishes. Comparative biochemistry and physiology. Part A, Molecular & integrative physiology, 149, 225-245.
Penheiter, A.R., Filoteo, A.G., Croy, C.L. & Penniston, J.T. (2001) Characterization of the deafwaddler mutant of the rat plasma membrane calcium-ATPase 2. Hearing research, 162, 19-28.
Riccardi, D. & Kemp, P.J. (2012) The calcium-sensing receptor beyond extracellular calcium homeostasis: conception, development, adult physiology, and disease. Annual review of physiology, 74, 271-297.
Schultz, J.M., Yang, Y., Caride, A.J., Filoteo, A.G., Penheiter, A.R., Lagziel, A., Morell, R.J., Mohiddin, S.A., Fananapazir, L., Madeo, A.C., Penniston, J.T. & Griffith, A.J. (2005) Modification of human hearing loss by plasma-membrane calcium pump PMCA2. The New England journal of medicine, 352, 1557-1564.
Stauffer, T.P., Guerini, D. & Carafoli, E. (1995) Tissue distribution of the four gene products of the plasma membrane Ca2+ pump. A study using specific antibodies. The Journal of biological chemistry, 270, 12184-12190.
Stawicki, T.M., Esterberg, R., Hailey, D.W., Raible, D.W. & Rubel, E.W. (2015) Using the zebrafish lateral line to uncover novel mechanisms of action and prevention in drug-induced hair cell death. Frontiers in cellular neuroscience, 9, 46.
Street, V.A., McKee-Johnson, J.W., Fonseca, R.C., Tempel, B.L. & Noben-Trauth, K. (1998) Mutations in a plasma membrane Ca2+-ATPase gene cause deafness in deafwaddler mice. Nature genetics, 19, 390-394.
Talarico, E.F., Jr., Kennedy, B.G., Marfurt, C.F., Loeffler, K.U. & Mangini, N.J. (2005) Expression and immunolocalization of plasma membrane calcium ATPase isoforms in human corneal epithelium. Molecular vision, 11, 169-178.
Wada, M., Furuya, Y., Sakiyama, J., Kobayashi, N., Miyata, S., Ishii, H. & Nagano, N. (1997) The calcimimetic compound NPS R-568 suppresses parathyroid cell proliferation in rats with renal insufficiency. Control of parathyroid cell growth via a calcium receptor. The Journal of clinical investigation, 100, 2977-2983.