Development of high-content assays for kidney progenitor cell expansion in transgenic zebrafish

SUBRAMANIAM, SANKER; CIRIO, MARIA CECILIA; VOLLMER, LAURA L.; GOLDBERG, NATASHA; MCDERMOTT, LEE A; HUKRIEDE, NEIL A.; VOGT, ANDREAS

JOURNAL OF BIOMOLECULAR SCREENING

SAGE PUBLICATIONS INC

Lugar: Thousand Oaks, California.; Año: 2013

1087-0571

Reactivation of genes normally expressed during organogenesis is a characteristic of kidneyregeneration. Enhancing this reactivation could potentially be a therapeutic target to augmentkidney regeneration. The inductive events that drive kidney organogenesis in zebrafish aresimilar to the initial steps in mammalian kidney organogenesis. Therefore, quantifyingembryonic signals that drive zebrafish kidney development is an attractive strategy for thediscovery of potential novel therapeutic modalities that accelerate kidney regeneration. TheLim1 homeobox protein, Lhx1, is a marker of kidney development that is also expressed in theregenerating kidneys after injury. Utilizing a fluorescent Lhx1a-EGFP transgene whosephenotype faithfully recapitulates that of the endogenous protein we developed a high-contentassay for Lhx1a-EGFP expression in transgenic zebrafish embryos employing an artificialintelligence-based image analysis method termed Cognition Network Technology (CNT).Implementation of the CNT assay on high-content readers enabled automated real-time in vivotime-course, dose-response, and variability studies in the developing embryo. The Lhx1a assaywas complemented with a kidney-specific secondary CNT assay that enables directmeasurements of the embryonic renal tubule cell population. The integration of fluorescenttransgenic zebrafish embryos with automated imaging and artificial intelligence-based imageanalysis provides an in vivo analysis system for structure-activity relationship studies and denovo discovery of novel agents that augment innate regenerative processes.