Link to the University of Pittsburgh Homepage
Link to the University Library System Homepage Link to the Contact Us Form

Experimental and computational studies of calcium-triggered transmitter release

Cho, Soyoun (2007) Experimental and computational studies of calcium-triggered transmitter release. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

Primary Text

Download (15MB) | Preview


Calcium influx through presynaptic calcium channels triggers transmitter release, but many of the details that underlie calcium-triggered secretion are not well understood. In an attempt to increase our understanding of this process, synaptic transmission at the frog neuromuscular junction has been investigated using physiological experiments and computational modeling. Pharmacological manipulations ((R)-roscovitine and DAP) were used as tools to modulate presynaptic calcium influx and study effects on transmitter release. I showed that (R)-roscovitine predominately slowed deactivation kinetics of calcium current (by 427%), and as a result, increased the integral of calcium channel current evoked by a physiological action potential waveform (by 44%). (R)-roscovitine also increased the quantal content of acetylcholine released from the motor nerve terminals (by 149%) without changing paired-pulse facilitation under low calcium conditions. In contrast, exposure to 3,4-diaminopyridine (which affects transmitter release evoked by partially blocking potassium channels, altering the amplitude of the presynaptic action potential, and indirectly increasing calcium entry) increased paired-pulse facilitation (by 23%). In normal calcium conditions, both pharmacological treatments showed relatively similar effects on paired-pulse facilitation. I used a computational model, constrained by previous reports in the literature and my physiological measurements, to simulate my experimental data. This model faithfully reproduced calcium current with a single action potential, the average number of released synaptic vesicles, and the effects of (R)-roscovitine and DAP on calcium influx and vesicle release. Using this model, I made several predictions about the mechanisms underlying transmitter release. First, calcium ions originating from one or two voltage-gated calcium channels most often contributed to cause the fusion of each vesicle. Second, the calcium channel closest to a vesicle that fuses, provides 77% of calcium ions. My simulation of paired-pulse facilitation using the present model needed more adjustments, and in the process of adjusting the model parameters, various hypotheses that might explain observed short-term synaptic plasticity, including the effects of changes in buffer conditions, the effects of uneven calcium channel distribution, reducing terminal volume by adding vesicles to a storage pool, changes in the second action potential waveform, and possible persistent changes in vesicle release machinery were explored.


Social Networking:
Share |


Item Type: University of Pittsburgh ETD
Status: Unpublished
CreatorsEmailPitt UsernameORCID
Cho, Soyounsoc1@pitt.eduSOC1
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairUrban, Nathaniel
Committee MemberBarrionuevo, GermanGerman@bns.pitt.eduGERMAN
Committee MemberBi, Guo-Qianggqbi@pitt.eduGQBI
Committee MemberAtwood, Harold
Committee MemberStiles, Joel
Committee MemberKandler, Karlkkarl@pitt.eduKKARL
Committee MemberMeriney, Stephen D.meriney@bns.pitt.eduMERINEY
Date: 20 September 2007
Date Type: Completion
Defense Date: 2 August 2007
Approval Date: 20 September 2007
Submission Date: 9 August 2007
Access Restriction: 5 year -- Restrict access to University of Pittsburgh for a period of 5 years.
Institution: University of Pittsburgh
Schools and Programs: Dietrich School of Arts and Sciences > Neuroscience
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: Calcium channel; Monte Carlo simulation; neuromuscular junction; vesicle fusion
Other ID:, etd-08092007-204525
Date Deposited: 10 Nov 2011 19:58
Last Modified: 15 Nov 2016 13:48


Monthly Views for the past 3 years

Plum Analytics

Actions (login required)

View Item View Item