Project Summary This purpose of this supplement is to enable the continuation of research that was hindered by limited research operations due to the COVID-19 pandemic and to assist in my transition to an independent PI position. The results obtained from this supplement will enhance my competitiveness for faculty positions. I will gain further training from my mentor on crucial career skills. In particular, our weekly meetings will include discussion of relevant skills such as interviewing, negotiating as well as establishing and maintaining an independent lab. The supplement will also aid in networking to identify potential collaborators and job opportunities, as this critical career advancement activity was severely hindered by COVID-19. I will attend the UT Austin molecular biosciences postdoctoral association weekly professional development seminars. With funds I will also attend relevant meetings, in virtual formats and ideally, as social restrictions ease, in person. These meetings will include RSA, the Society for Neuroscience and the International Zebrafish Meeting. My preliminary data demonstrates ethanol interacts with vangl2 during tangential FBMN migration. Tangential FBMN migration fails to occur in both control and ethanol-exposed vangl2 mutants. While control wildtype (WT) and vangl2 heterozygotes develop normally, ethanol induces tangential FBMN migration defects that are dose and time dependent. Strikingly, many ethanol-exposed WTs and vangl2 heterozygotes phenocopy the mutant. Notably, these defects are observed more often in ethanol-exposed vangl2 heterozygotes compared to WTs. Remarkably, FBMN migration defects are induced by as low as 0.25% v/v ethanol. Similarly, I have found that ethanol interacts with another member of the Wnt/PCP pathway, pk1b, during FBMN migration. I hypothesize that a combined genetic and ethanol-mediated attenuation of planar cell polarity results in aberrant FBMN migration behaviors. In Aim 1 of this supplement, I will use live time-lapsed confocal analyses to assess the migratory behaviors of FBMNs in ethanol-exposed vangl2, pk1b, celsr2, fzd3a and scrib1 heterozygous embryos using the isl1:eGFP;ubi:nlsmApple and gfap:mCherry transgenic lines. In Aim 2 of this supplement, I will characterize vangl2-ethanol interactions underlying neuronal migration defects. I will use confocal analyses to assess hindbrain patterning in ethanol-exposed vangl2 heterozygotes using hoxb1a:RFP, egr2b:GFP, and mafba:GFP transgenics. I will generate the Vangl2:EGFP and Celsr1:RFP transgenes using CRISPR/Cas9 mediated homology directed repair to examine planar cell polarity acquisition and use the actb2:Mmu.Arl13b-GFP transgenes to examine planar cell polarity functionality in vangl2 heterozygotes to determine how ethanol interacts with the Wnt/PCP pathway. In Aim 3 of this supplement, I will map and functionally characterize ethanol-sensitive zebrafish mutants with FBMN migration defects that are identified through an o...