PgmNr Z591: in toto imaging of osteoblast cell cycle dynamics in regenerating zebrafish scales.

Authors:
B. Cox 1 ; V. Tornini 1 ; A. Puliafito 2 ; S. Di Talia 1 ; K. Poss 1


Institutes
1) Duke University, Durham, NC; 2) Candiolo Cancer Institute - FPO, IRCCS, Candiolo, Torino, Italy.


Abstract:

The adult zebrafish can regenerate many tissues, but not all are amenable to live imaging due to their size, location, or time required to regenerate. Zebrafish scales, like fins, contain osteoblasts and epidermis, but they regenerate much more quickly after being plucked or lost during the natural course of the fish’s life, recapitulating their gross structure within three to four days of loss of the original tissue. Thus, they would make ideal models for live imaging of regeneration. We have developed a method for long-term (up to twenty-four hours) live imaging of adult fish using low concentrations of anesthetic, giving us previously unprecedented access to spatiotemporal information about individual and collective cell behavior during regeneration. Through live imaging of transgenic fish expressing the FUCCI cell cycle reporter in osteoblasts (osx:Venus-hGeminin/osx:mCherry-zCdt1), we have identified a transition during regeneration from early uniform proliferation to later growth, which is spatially restricted to the borders of regenerating scales. We have developed computational methods to segment and track hundreds to thousands of osteoblasts, analyze their cell cycle and division patterns over long periods, and infer how individual cell cycle changes underlie tissue-wide growth patterns.



ZFIN Genetics Index
1. sp7