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Neil A. Hukriede, Ph.D.
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Assistant Professor
5061 BST3
3501 Fifth Avenue
Pittsburgh, Pennsylvania 15213
Phone: (412) 648-9918
Fax: (412) 383-5918
E-mail: hukriede@pitt.edu
Lab Phone: (412) 648-9483
Lab Website
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Biography
2006 Basil O’Connor Research Award, March of Dimes
2004 Carl W. Gottschalk Research Scholar, American Society of Nephrology
National Research Council Advanced Level Associateship, National Academy of Sciences, National Institutes of Health, 2000-2002
National Research Council Associateship, National Academy of Sciences, National Institutes of Health, 1997-2000
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Research
The vertebrate kidney is a complex homeostatic organ that functions to detoxify blood, maintain ion and water equilibrium, and regulate hormone release. The physiological consequences of abnormal kidney formation or function are frequently fatal, with dialysis and organ transplantation the only long-term treatments for kidney disease. Future strategies to fight kidney disease must rely on a fundamental understanding of the earliest events that lead to the formation of the embryonic kidney. Importantly, kidney structure and function is conserved among vertebrates and current evidence suggests that mechanisms of kidney development are similarly conserved. We exploit the embryological and genetic features of Xenopus and zebrafish embryos to (1) identify the earliest cells (progenitors) that give rise to the pronephric kidney and (2) understand the molecular events that initiate and regulate kidney development during embryogenesis.
The principal focus of my laboratory is delineating the events for specifying kidney-restricted cells from the intermediate mesoderm. One area of study is determining the function of the LIM-homeodomain transcription factor Lim1. In previous work, I demonstrated that Lim1 is involved at multiple steps of gastrulation in Xenopus. Using oligonucleotide-mediated depletion of Lim1 transcripts, it was determined that Lim1 is a transcriptional activator controlling expression of protocadherins, which are involved in cell movements during gastrulation. In addition to the gastrulation defect, loss of Lim1 during kidney organogenesis results in complete loss of the pronephric kidney. Our current thinking is that either Lim1 plays a role in specification of the nephric tissue and is sufficient to induce kidney, or Lim1 is activated in already specified tissue and plays a role during morphogenesis. In order to address these questions we are using depletion and over-expression tools in Xenopus embryo and explant studies.
A second focus of the laboratory is to utilize the genetically amenable zebrafish system to identify genes necessary for kidney differentiation. While mutations that affect the kidney have been reported in several systems and a good amount of data exist on kidney morphogenesis, little is known about the earliest inductive events in the formation of this tissue. In addition, several kidney mutants have previously been identified in zebrafish, but none affect early pronephros development. As part of an in situ-based haploid genetic screen, we isolated several mutants that affect kidney development. Future studies will entail phenotypically characterizing these mutations and identifying the mutated loci.
Finally, chemical screens provide a valuable approach for identifying small molecules that can regulate embryogenesis and control biological processes. With the identification of chemical compounds from specific assays, it is possible to define the temporal inductive events that establish embryonic differentiation pathways. Whole organism chemical screens also allow for rapid validation of chemical compounds by eliminating toxic and non-specific compounds often found in high-throughput screening. The purpose of our chemical screens is to identify compounds that can either abate or enhance the formation of the zebrafish pronephros and compliment genetic screens that have already identified several kidney mutants.
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Selected Publications
- Vogt, A., Cholewinski, A., Shen, X., Nelson, S., Lazo, J.S., Tsang, M. and N.A. Hukriede (2009) Automated image-based phenotypic analysis in zebrafish embryos. Dev. Dynamics, 238: 656-663. | Abstract
- Koide, K., Song, F., de Groh, E.D., Garner, A.L., Mitchell, V.D., Davidson, L.A., and N.A. Hukriede (2008) Scalable and Concise Synthesis of Water-Soluble Dichlorofluorescein Derivatives Displaying Tissue Permeability in Live Zebrafish Embryos. CHEMBIOCHEM, 9 (2), 214-218. | Abstract
- Bai, Q., Garver, J.A., Hukriede, N.A., and E.A. Burton (2007) Generation of a transgenic zebrafish model of Tauopathy using a novel promoter element derived from the zebrafish eno2 gene. Nucleic Acids Res. 35 (19), 6501-16. | Abstract
- Pugacheva EM, Kwon YW, Hukriede NA, Pack S, Flanagan PT, Ahn JC, Park JA, Choi KS, Kim KW, Loukinov D, Dawid IB, Lobanenkov VV (2006) Cloning and characterization of zebrafish CTCF: Developmental expression patterns, regulation of the promoter region, and evolutionary aspects of gene organization. Gene 75, 26-36. | Abstract
- Hong, S., Haldin, C.E., Lawson, N.D., Weinstein, B.M., Dawid, I.B., and N.A. Hukriede (2005) The zebrafish kohtalo/trap230 gene is required for the development of the brain, neural crest and pronephric kidney. Proc Natl Acad Sci U S A. 102(51):18473-8. | Abstract
- Kozlowski, D.J., Whitfield, T.T., Hukriede, N.A., Lam, W.K., and Weinberg E.S. (2005) The zebrafish dog-eared mutation disrupts eya1, a gene required for cell survival and differentiation in the inner ear and lateral line. Dev. Bio. 277, 27-41. | Abstract
- Hukriede, N.A., Tsang, T.E., Habas, R., Khoo, PL., Steiner, K., Weeks, D.L., Tam P.P.L. and Dawid I.B. (2003) Conserved Requirement of Lim1 Function for Cell Movements during Gastrulation. Developmental Cell 4, 83-94. | Abstract
- Chen, L., Segal, D., Hukriede, N.A., Podtelejnikov, A., Bayarsaihan, D., Kennison, J., Ogryzko, V., Dawid, I.B., Westphal H. (2002) Ssdp proteins interact with the LIM-domain binding protein Ldb1 to regulate development. Proc Natl Acad Sci U S A. 99(22):14320-5. | Abstract
- Hukriede, N.A., Fisher, D., Epstein, JA., Joly, L., Tellis, P., Zhou, X., Barbazuk, B., Cox, K., Fenton-Noriega, L., Hersey, C., Miles, J., Sheng, X., Song, A., Waterman, R., Johnson, SL.,. Dawid, IB., Chevrette, M., Zon, LI., McPherson, J. and M. Ekker. (2001) The LN54 radiation hybrid map of zebrafish expressed sequences. Genome Research 11, 2127-2132. | Abstract
- Hukriede, N.A., Joly, L., Tsang, M., Miles, J., Tellis, P., Epstein, J.A., Barbazuk, W.B., Li, F.N., Paw, B., Postlethwait, J.H., Hudson, T.J., Zon, L,I,. McPherson, J.D., Chevrette, M., Dawid, I.B., Johnson, S.L. and M. Ekker (1999) Radiation hybrid mapping of the zebrafish genome. Proc. Natl. Acad. Sci. 96, 9745-9750. | Abstract
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