Vertebrate retinal development requires timely and precise fusion of the optic fissure (OF). Failure of this event leads to congenital vision impairment in the form of coloboma. Recent studies have suggested hyaloid vasculature to be involved in OF fusion. In order to examine this link, we analyzed OF fusion and hyaloid vasculogenesis in the zebrafish pax2a noi mutant line. We first determined that pax2a−/− embryos fail to accumulate F-actin in the OF prior to basement membrane (BM) degradation. Furthermore, using 3D and live imaging we observed reduced OF hyaloid vascularization in pax2a−/− embryos. When examining the connection between pax2a loss of function and hyaloid vasculature, we observed significant reduction of talin1 expression, a regulator of hyaloid vasculature. In addition, cranial VEGF expression was found to be reduced in pax2a−/− embryos. Pharmacological inhibition of VEGF signaling phenocopied the pax2a−/− vasculature, F-actin and BM degradation phenotypes. Lastly, we determined that OF associated hyaloid vasculature is a source of mmp2, mmp14a and mmp14b expression and showed that mmp2 is functionally necessary for degradation of OF BM. Taken together we propose a pax2a driven mechanism that ensures proper and timely hyaloid vasculature invasion of the OF in order to facilitate availability of the BM remodeler mmp2.
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This work was supported by faculty start-up funds provided to JKF from the department of Biology, University of Kentucky. MLW was supported by the Lyman T Johnson scholarship from the University of Kentucky. WPP was supported by Brazilian National Council for Scientific and Technological Development (CNPq) under grant number 202970/2014-0.
All data and reagents generated in this manuscript will be freely available to any reasonable request. Sequences of primers used for probe synthesis are found in Table 1.
Weaver, Megan L.; Piedade, Warlen P.; Meshram, Nishita N.; and Famulski, Jakub K., "Hyaloid Vasculature and mmp2 Activity Play a Role during Optic Fissure Fusion in Zebrafish" (2020). Biology Faculty Publications. 189.
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