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Kinetics and molecular mechanisms of calcium -regulated exocytosis of catecholamines.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Kinetics and molecular mechanisms of calcium -regulated exocytosis of catecholamines./
作者:	Wang, Chih-Tien.
出版者:	Ann Arbor : ProQuest Dissertations & Theses, : 2001,
面頁冊數:	135 p.
附註:	Source: Dissertations Abstracts International, Volume: 63-10, Section: B.
Contained By:	Dissertations Abstracts International63-10B.
標題:	Cellular biology. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3033282
ISBN:	9780493462653

Kinetics and molecular mechanisms of calcium -regulated exocytosis of catecholamines.
Wang, Chih-Tien.

Kinetics and molecular mechanisms of calcium -regulated exocytosis of catecholamines. - Ann Arbor : ProQuest Dissertations & Theses, 2001 - 135 p.

Source: Dissertations Abstracts International, Volume: 63-10, Section: B.

Thesis (Ph.D.)--The University of Wisconsin - Madison, 2001.

This item must not be sold to any third party vendors.

The kinetics and molecular mechanisms underlying Ca2+-regulated exocytosis were studied with real time amperometry. PC12 cells transfected with proteins of interest were used as the model system. Several cellular and molecular manipulations modulated Ca2+-regulated exocytosis. This study focused on synaptotagmin effects on release kinetics and fusion pore dynamics. Transfection of synaptotagmin I did not affect the overall kinetics of exocytosis, while transfection of synaptotagmin IV slowed exocytosis. Fusion dynamics were found to be regulated by synaptotagmin I and IV in different directions. Synaptotagmin I impaired fusion pore dilation, whereas synaptotagmin IV promoted fusion pore expansion. Mutation analysis showed that C2B domains in both synaptotagmin I and IV play an important role in determining the stability of fusion pores. Surprisingly, transfection of synaptotagmin IV also produced long stand-alone feet, which represented transient fusion pores in a kiss-and-run process. The average amplitude of these stand-alone feet was ∼0.3 pA, ten times smaller than that of prespike feet corresponding to open fusion pores preceding full dilation. The number of stand-alone feet was positively correlated to that of amperometric spikes, suggesting that the kiss-and-run mechanism is compatible with complete fusion of vesicles. Substitution of aspartate by serine in position 230 in the C2A domain of synaptotagmin I resulted in long stand-alone feet similar to those seen in synaptotagmin IV transfected cells, suggesting that the C2A domain of synaptotagmin I plays an important role in regulating the duration of transient fusion pores. Additional mutation analysis showed that both C2A and C2B domains of synaptotagmin IV were involved in regulating transient pores. These findings from PC12 cells may provide an understanding for the role of synaptotagmins in synaptic transmission.

ISBN: 9780493462653Subjects--Topical Terms:

3172791
Cellular biology.
Subjects--Index Terms:

Calcium-regulated

Kinetics and molecular mechanisms of calcium -regulated exocytosis of catecholamines.
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520 $a The kinetics and molecular mechanisms underlying Ca2+-regulated exocytosis were studied with real time amperometry. PC12 cells transfected with proteins of interest were used as the model system. Several cellular and molecular manipulations modulated Ca2+-regulated exocytosis. This study focused on synaptotagmin effects on release kinetics and fusion pore dynamics. Transfection of synaptotagmin I did not affect the overall kinetics of exocytosis, while transfection of synaptotagmin IV slowed exocytosis. Fusion dynamics were found to be regulated by synaptotagmin I and IV in different directions. Synaptotagmin I impaired fusion pore dilation, whereas synaptotagmin IV promoted fusion pore expansion. Mutation analysis showed that C2B domains in both synaptotagmin I and IV play an important role in determining the stability of fusion pores. Surprisingly, transfection of synaptotagmin IV also produced long stand-alone feet, which represented transient fusion pores in a kiss-and-run process. The average amplitude of these stand-alone feet was ∼0.3 pA, ten times smaller than that of prespike feet corresponding to open fusion pores preceding full dilation. The number of stand-alone feet was positively correlated to that of amperometric spikes, suggesting that the kiss-and-run mechanism is compatible with complete fusion of vesicles. Substitution of aspartate by serine in position 230 in the C2A domain of synaptotagmin I resulted in long stand-alone feet similar to those seen in synaptotagmin IV transfected cells, suggesting that the C2A domain of synaptotagmin I plays an important role in regulating the duration of transient fusion pores. Additional mutation analysis showed that both C2A and C2B domains of synaptotagmin IV were involved in regulating transient pores. These findings from PC12 cells may provide an understanding for the role of synaptotagmins in synaptic transmission.
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