Stimulation of a single presynaptic terminal using confocal spot uncaging

2014 Fiscal Year Final Research Report

• Project/Area Number: 24650218
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Fusional basic brain science
• Research Institution: Doshisha University
• Principal Investigator: SAKABA Takeshi 同志社大学, 脳科学研究科, 教授 (80609511)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 神経科学

Outline of Final Research Achievements

We have used the confocal spot uncaging technique to stimulate a single presynaptic terminal. Specifically, caged Ca compound was introduced to the presynaptic cell via a patch pipette, and the compound was uncaged with spot illumination so that Ca was elevated only at a single presynaptic terminal. We have applied this to the synapses between cerebellar interneurons, and were able to elicit transmitter release and calculate the number of releasable synaptic vesicles at a single bouton (Trigo et al., 2012, PNAS). In addition, we have performed direct patch clamp recordings from the cerebellar Purkinje cell terminal, and applied Ca ungaging, capacitance measurements and postsynaptic recordings. Postsynaptic receptors were saturated and the postsynaptic currents seemed not to be a good indicator for transmitter release. Rather, we suggest that capacitance measurements provide a reasonable readout of the amounts of transmitter release.

Free Research Field

神経生理学