Multicolor Labeling and Tracing of Pancreatic Beta-Cell Proliferation in Zebrafish

Methods Mol Biol. 2020:2128:159-179. doi: 10.1007/978-1-0716-0385-7_12.

Abstract

During embryogenesis, beta-cells arise from the dorsal and ventral bud originating in the endoderm germ layer. As the animal develops to adulthood, the beta-cell mass dramatically increases. The expansion of the beta-cell population is driven by cell division among the embryonic beta-cells and supplanted by neogenesis from post-embryonic progenitors. Here, we describe a protocol for multicolor clonal analysis in zebrafish to define the contribution of individual embryonic beta-cells to the increase in cell numbers. This technique provides insights into the proliferative history of individual beta-cells in an islet. This insight helps in defining the replicative heterogeneity among individual beta-cells during development. Additionally, the ability to discriminate individual cells based on unique color signatures helps quantify the volume occupied by beta-cells and define the contribution of cellular size to the beta-cell mass.

Keywords: Brainbow; Heterogeneity; Lineage tracing; Quiescence; Single cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Cell Lineage
  • Cell Proliferation*
  • Cell Tracking / methods*
  • Cloning, Molecular / methods
  • Color
  • Genes, Reporter
  • Image Processing, Computer-Assisted / methods*
  • Insulin-Secreting Cells / chemistry
  • Insulin-Secreting Cells / cytology*
  • Integrases
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Microscopy, Confocal / methods*
  • Models, Animal
  • Staining and Labeling / methods*
  • Zebrafish

Substances

  • Luminescent Proteins
  • Cre recombinase
  • Integrases