COVERSHEET

TITLE: Roles of Zic2 and Zic3 in early development

DEPARTMENT: Department of Zoology

MENTOR: Yevgenya Grinblat

DEPARTMENT: Department of Zoology

MENTOR(2): ______

DEPARTMENT(2): ______

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ABSTRACT

Roles of Zic2 and Zic3 in early development

The Hedgehog (Hh) signaling pathway plays a crucial role in modulating embryonic development. Malfunctions in the vertebrate Hh pathway involving Homolog (Shh) have been linked to cancers including basal cell carcinoma and developmental disorders like . Zic3, a zinc-finger transcription factor, is hypothesized to activate Shh- mediated Hh signaling. This is based on data demonstrating ZIC3 often binds to GLI consensus motif, and Zic3-depleted embryos express lower levels of Shlz. To test this hypothesis, a line ofzebrafish embryos carrying a nonsense mutation of Zic3 was examined for morphology ofthe forebrain and retina. Unexpecte.{lly, no visible defects were found in embryos homozygous for the mutant allele through five days of development, and an expression assay of three Hh pathway (Shh, Hhip, and G/i2a) via in situ hybridization in Zic3 mutants showed normal patterns of Hh target expression. These data suggest Zic3 has redundant functions or compensation is occurring, in comparison to ZicJ-depleted embryos.

In a parallel approach, we are investigating A/xi, a candidate target of Zic2. A/xi is strongly implicated in the development of neural crest (NC) cells, as is the Zic gene family. Zic2 and A/xi promote specification of NC cells. Pericytes, a mural cell type important for vascular development, are one of many cell lineages derived from NC cells. Zic2 mutants exhibit hemorrhage indicative of aberrant vasculogenesis, which may be due to aberrant pericyte formation. To test this hypothesis, immunohistochemistry was used to assay expression of Pdgfi·b, a gene expressed in pericytes, in A/xi morpholino­ injected embryos. Analysis of the dorso-medial midrain-hindbrain boundary found smaller quantities of pericytes within A/xi- depleted embryos than in controls, suggesting A/xi has an impact on migration, differentiation, or a combination of both factors on pericyte development. This study has provided potential insight into Zic gene family function in early development and may enhance understanding of diseases associated with it.

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_5/11/2016______Date

Roles of Zic2 and Zic3 in early development

Dan Fox1

1Deparhnent of Zoology, University of\Visconsin-Madison