Gene Function, Expression and Regulation in Zebrafish

Eunice Kennedy Shriver National Institute Of Child Health & Human Development

Investigators

Linked publications, trials & patents

Abstract

From 9/1/2021 through 8/25/2022 Feldman supported zebrafish research of ten labs/other customers, conducted independent research and engaged in institutional service. Support of Zebrafish Research Led by Labs & Other Customers, Principal Projects: Dever Lab (NICHD): Translation of Distinct RNA Populations by Eif1 and Eif5. Feldman advised Dever and performed several microinjection experiments to explore differences in phenotypes resulting from ectopic expression of either Eif1 or Eif5 in zebrafish embryos and the possibility that restoration of a balanced Eif1/Eif5 ratio can ameliorate these phenotypes. Sackett Lab (NICHD): Assessing Expression and Function of Zebrafish Alpha and Beta Tubulin Isotypes. The degree to which specific tubulin isotypes and/or their post-translational modification are essential for specific aspects of development in any organism remains a surprisingly open question. Feldman assisted Sackett in finding and recruiting two postbac students with undergraduate training in reputable zebrafish labs. Since the arrival of these students in the summer of 2022, he has been training and mentoring them through an ambitious project to systematically knock-out each zebrafish alpha and beta tubulin isotype in F0 embryos and determine how their absence affects early development. Porter Lab (NICHD): Genetic Dissection and Creation of Human Disease Models of Sterol Metabolism. In previous years the Core used CRISPR-Cas9 technology to create genetic mutant zebrafish lines for the Porter lab in five genes: dhcr7, npc1, npc2, cln3 and ebp - with roles in various steps of cholesterol metabolism. Feldman is in the process of cryopreserving these lines for future use. Stratakis Lab (NICHD): Function of Zebrafish Orthologs to Human Genes Implicated in Disorders of the Pituitary-Adrenal Axis. In previous years, the Core used CRISPR-Cas9 technology to generate zebrafish carrying loss-of-function mutations in four zebrafish orthologs to human genes implicated by the Stratakis lab in human growth anomalies and eight zebrafish orthologs to human adrenal hyperplasia and Cushing disease-associated genes. The core also helped the Stratakis lab generate a precisely edited zebrafish satb1 mutant line with a nonsynonymous AA substitution that is cognate to a human disease-associated mutation of interest. Feldman is in the process of cryopreserving these lines for future use. Golden lab (NIDDK): The Core previously generated a targeted AA-alteration in the cacna1c gene. This year Feldman found that recessive mutants have profound developmental anomalies. Characterization of this phenotype by trainees from the Golden lab is ongoing. Kemper lab (NHLBI): Function of zebrafish rca2.1. The Kemper lab is interested in zebrafish rca2.1s function, because it has certain similarities to human CD46 that are not found in the mouse genome. The Core previously generated two mutant rca2.1 alleles, revealing essential roles in growth and cardiac function. Phenotypic characterization is ongoing. Independent Research by the NICHD Zebrafish Core Cryopreservation and in vitro fertilization of zebrafish sperm. Over the last year, the Core has continued to focus on improving quality control measures to ensure viability of cryopreserved zebrafish lines and minimize variability in viability. This year we developed an approach of pre-assessing the number and activity of sperm from individual males and only cryopreserving when we obtain yields exceeding four million active sperm. Institutional Service: ACUC Membership. Feldman has served on the NICHD ACUC since 2015 and continued in this capacity this year, meeting monthly to evaluate and decide upon animal-study proposals, renewals and amendments and ad hoc issues relevant to animal welfare.