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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, mmp 14 a 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.

Hyaloid vasculature and mmp2 activity play a role during optic fissure fusion in zebrafish