Development of new inverse-type calcium probe for analysis of inhibitory neuronal activities

2015 Fiscal Year Final Research Report

• Project/Area Number: 26560461
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Basic / Social brain science
• Research Institution: Kyushu University
• Principal Investigator: Kuge Sayuri 九州大学, 理学(系)研究科(研究院), 学術研究員 (50260104)
• Co-Investigator(Renkei-kenkyūsha): ISHIHARA Takeshi 九州大学, 大学院理学研究院, 教授 (10249948) ; TERAMOTO Takayuki 九州大学, 大学院理学研究院, 准教授 (90571836) ; NAGAI Takeharu 大阪大学, 産業科学研究所 (20311350) ; MATSUDA Tomoki 大阪大学, 産業科学研究所 (50419206)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: カルシウムプローブ / 神経活動 / 線虫 / インバース

Outline of Final Research Achievements

Fluorescent Ca2+ indicators, which have been improved for the last several decades are ideal only for monitoring the activation of neurons. To understand precise functions of the neuronal network, a fluorescent Ca2+ indicator that is suitable for monitoring the neuronal inhibition would be necessary. Previously reported "inverse-pericam" has unique property; the fluorescence intensity gets 7-fold dimmer upon Ca2+ binding. After creating a large mutant library of "inverse-pericam" by error-prone PCR, a variant, IP2.0, which shows nearly 20-folds dynamic range in vitro, was found. When IP2.0 was expressed in AWC neuron of C. elegans, the increase of fluorescence of IP2.0 during the stimulation of isoamyl alcohol was observed. These results suggest that odor stimuli decreases Ca2+ concentration in AWCON neurons and so IP2.0 will be an indispensable tool for studying neuronal inhibition.

Free Research Field

神経科学