The NICHD Zebrafish Core
- Benjamin Feldman, PhD, Staff Scientist, Director of the NICHD Zebrafish Core
The NICHD Zebrafish Core was established in 2012 with the goal of providing its clients with consultation, access to equipment and reagents, and service in the area of zebrafish genetics. NICHD investigators as well as investigators from other NIH institutes and from outside the NIH are its clientele. The oversight committee for the Core comprises Harold Burgess, Ajay Chitnis, and Brant Weinstein. The Core's activities consist of:
- oversight and support of client-specific projects,
- custom generation of genetic zebrafish models,
- troubleshooting of new methodologies with promising application in zebrafish,
- maintenance and improvement of equipment and infrastructure, and
- service and educational outreach.
Oversight and support of client-specific projects
Over 2022–2023, Feldman engaged in research projects with four laboratories and directed the closure of the Core.
Dever Lab (NICHD)
Translation of Distinct RNA Populations by the eukaryotic initiation factors 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 Genes. 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 trained, and co-supervised two postbaccalaureate students through this ambitious project to systematically knock-out each zebrafish alpha and beta tubulin isotype in F0 embryos and determine how their absence affects early development. To date, this project has documented essential embryonic phenotypes for more than five tubulin genes.
Golden Lab (NIDDK)
The Core previously generated a targeted amino acid alteration in the cacna1c gene. This year Feldman found that recessive mutants have profound developmental anomalies. Andy Golden, who was also collaborating with Harry Burgess, tragically passed away during the current reporting period. But characterization of this phenotype by Feldman and members of the Burgess 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 with 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
Strategies for CRISPR-Cas9–based homology-directed repair (HDR)
Over the past several years, Feldman and NICHD Zebrafish Core Staff explored several approaches to generating zebrafish lines with amino-acid substitutions cognate to human disease alleles of interest and generated three such alleles in house: atp7a, cacna1c and satb1. This past year, Feldman worked to devise a less labor-intensive pipeline that will feature outsourcing of CRISPR–based design and reagent steps to In Vivo Biosciences, followed by microinjection and allele recovery in-house via high-throughput sequencing of extruded gametes from candidate carriers.
Cryopreservation and in vitro fertilization of zebrafish sperm
Over the last year, Feldman, assisted by Felicia Benoit, has continued to focus on improving quality control measures to ensure viability of cryopreserved zebrafish lines and minimize variability in viability. This year, they developed an approach of pre-assessing the number and activity of sperm from individual males and only cryopreserving when yields exceeding two million active sperm are obtained.
Service
ACUC Membership
Feldman has served on the NICHD ACUC since 2015 and continued in this capacity this year, taking on the role of Alternate Chairperson, meeting monthly to evaluate and decide upon animal-study proposals, renewals and amendments, and ad hoc issues relevant to animal welfare.
Collaborators
- Harold Burgess, PhD, Section on Behavioral Neurogenetics, NICHD, Bethesda, MD
- Thomas Dever, PhD, Section on Protein Biosynthesis, NICHD, Bethesda, MD
- Andy Golden, PhD, Laboratory of Biochemistry and Genetics, NIDDK, Bethesda, MD
- Claudia Kemper, PhD, Laboratory for Complement and Inflammation Research, NHLBI, Bethesda, MD
- Kenneth Olivier, MPH, MD, Laboratory of Chronic Airway Infection, NHLBI, Bethesda, MD
- Daniel Sackett, PhD, Division of Basic and Translational Biophysics, NICHD, Bethesda, MD
Contact
For more information, email bfeldman@mail.nih.gov.
