Identifying Components of Vg1 Anchoring Complex

RNA localization is important in developing asymmetry throughout an oocyte by restricting the location of proteins and when they are translated and are later expressed. My experiments focused on understanding Vegetal1 (Vg1) RNA anchoring by identifying a protein involved in anchoring of Vg1 RNA to the vegetal cortex in Xenopus laevis. Our hypothesis is that the protein Huntingtin is one of the proteins involved in anchoring. Huntingtin protein in Xenopus laevis is homologous to the already sequenced Huntingtin in Xenopus Tropicalis. Immunofluorescence will be used to image the localization of specific proteins in the oocyte. Specifically, we looked at the candidate protein Huntingtin. We microinjected it’s fluorescently labeled RNA to simulate the localization of Vg1 RNA.  We will use the reference proteins tubulin and actin to find where the candidate proteins are located during RNA localization. The reference proteins inside of oocytes will be treated with drugs cytochalasin B and nocodazole to inhibit the protein functions. Currently our results confirm that Huntingtin protein from Xenopus Tropicalis is co-localized in Xenopus laevis where Vg1 RNA would be, even after using the drugs and it remains likely that the protein is involved in anchoring. We will later microinject the sequenced Huntingtin protein obtained from Xenopus laevis to further examine the role of Vg1 RNA anchoring in addition to microinjecting Alexa RNA to fluoresce where the Vg1 RNA would be present.