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Abstract

Developmental Biology

Clonal Analysis of the Neonatal Mouse Heart using Nearest Neighbor Modeling

Published: August 22nd, 2020

DOI:

10.3791/61656

1Department of Medicine, Duke University, 2Center for Genomic and Computational Biology, Duke University, 3Department of Statistical Science, Duke University, 4Regeneration Next, Duke University

* These authors contributed equally

Abstract

By replacing lost or dysfunctional myocardium, tissue regeneration is a promising approach to treat heart failure. However, the challenge of detecting bona fide heart regeneration limits the validation of potential regenerative factors. One method to detect new cardiomyocytes is multicolor lineage tracing with clonal analysis. Clonal analysis experiments can be difficult to undertake, because labeling conditions that are too sparse lack sensitivity for rare events such as cardiomyocyte proliferation, and diffuse labeling limits the ability to resolve clones. Presented here is a protocol to undertake clonal analysis of the neonatal mouse heart by using statistical modeling of nearest neighbor distributions to resolve cardiomyocyte clones. This approach enables resolution of clones over a range of labeling conditions and provides a robust analytical approach for quantifying cardiomyocyte proliferation and regeneration. This protocol can be adapted to other tissues and can be broadly used to study tissue regeneration.

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Keywords Clonal Analysis

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