Confocal imaging and k-means clustering of GABAB and mGluR mediated modulation of Ca2+ spiking in hippocampal neurons

Swain, Sarpras and Gupta, Rishikesh Kumar and Ratnayake, Kasun and Devi, Priyanka Pantula and Singh, Ranjana and Jana, Soumya and Mitra, Kishalay and Karunarathne, Ajith and Giri, Lopamudra (2018) Confocal imaging and k-means clustering of GABAB and mGluR mediated modulation of Ca2+ spiking in hippocampal neurons. ACS Chemical Neuroscience. ISSN 1948-7193 (In Press)

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Imaging cytosolic calcium in neurons is emerging as a new tool in neurological disease diagnosis, drug screening, and toxicity testing. Ca2+ oscillation signatures show a significant variation depending on GPCR targeting agonists. Quantification of Ca2+ spike trains in ligand induced Ca2+ oscillations remain challenging due to their inherent heterogeneity in primary culture. Moreover, there is no framework available for identification of optimal number of clusters and distance metric to cluster Ca2+ spike trains. Using quantitative confocal imaging and clustering analysis, we show the characterization of Ca2+ spiking in GPCR targeting drug-treated primary culture of hippocampal neurons. A systematic framework for selection of the clustering method instead of an intuition-based method was used to optimize the cluster number and distance metric. The results discern neurons with diverse Ca2+ response patterns, including higher amplitude fast spiking and lower spiking responses, and their relative percentage in a neuron population in absence and presence of GPCR-targeted drugs. This framework was employed to control clustering patterns of Ca2+ spiking using GABAB and mGluR targeting drugs. This approach can be used for unbiased measurement of neural activity and identification of spiking population with varying amplitude and frequencies, providing a platform for high-content drug screening.

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Item Type: Article
Subjects: Electrical Engineering
Divisions: Department of Electrical Engineering
Depositing User: Team Library
Date Deposited: 31 Jul 2018 08:42
Last Modified: 31 Jul 2018 08:42
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