zebraflash transgenic lines for in vivo bioluminescence imaging of stem cells and regeneration in adult zebrafish

Abstract

The zebrafish has become a standard model system for stem cell and tissue regeneration research, based on powerful genetics, high tissue regenerative capacity and low maintenance costs. Yet, these studies can be challenged by current limitations of tissue visualization techniques in adult animals. Here we describe new imaging methodology and present several ubiquitous and tissue-specific luciferase-based transgenic lines, which we have termed zebraflash, that facilitate the assessment of regeneration and engraftment in freely moving adult zebrafish. We show that luciferase-based live imaging reliably estimates muscle quantity in an internal organ, the heart, and can longitudinally follow cardiac regeneration in individual animals after major injury. Furthermore, luciferase-based detection enables visualization and quantification of engraftment in live recipients of transplanted hematopoietic stem cell progeny, with advantages in sensitivity and gross spatial resolution over fluorescence detection. Our findings present a versatile resource for monitoring and dissecting vertebrate stem cell and regeneration biology.

A Functional Bioluminescent Zebrafish Screen for Enhancing Hematopoietic Cell Homing

“…Note that our bathing dose of D-luciferin (0.5 mM) was considerably lower than other reports that used 50–100 mM for a bathing technique (Chen et al., 2013). Therefore, further studies used an intraperitoneal injection of D-luciferin.…”

“…Our system is very similar to that developed by the Poss Lab using the same ubiquitin promoter fragment (Chen et al., 2013), although their study did not use sedation routinely for BLI acquisition and allowed animals to swim freely (donor BLI was assessed 28 days post-HCT). By optimizing sedation and keeping the animals immobile, we were able to measure the fraction of hematopoietic cells that homed to the WKM 24 hr post-HCT, a unique feature of our approach.…”

Regeneration Genetics

“…Incorporation of sensitive reporters (e.g. FUCCI or luciferase (14; 21)) into forward and reverse genetic screens, as a means to quantify proliferating cells in the regenerate, will facilitate identification of both positive and negative regulators of regeneration.…”

Fish to Learn: Insights into Blood Development and Blood Disorders from Zebrafish Hematopoiesis

“…The zebrafish named ''zebraflash'' is a luciferase-based transgenic line that can provide deep tissue bioluminescence information in a freely swimming organism. 51 This model has been used to track HSPC transplantations in zebrafish and can be utilized to track the migration and the behavior of both the stem cell and its progeny over time.…”