Poster Session II
pH Regulatory Protein Nhe2 Localization in Neuroblasts & Abundance within Neuroblasts (type 1 & 2) During Mitosis and Interphase
Project Type
Poster
Faculty Mentor’s Full Name
Professor Beverly Piggott
Faculty Mentor’s Department
Division of Biological Sciences
Abstract / Artist's Statement
pH regulation is essential for neuronal excitability as different pH states dramatically affect ion channel function. Considerably less is known about how pH dynamics might shape brain development. Our preliminary data has discovered a novel role for pH regulatory proteins, Na+/H+ Exchangers (Nhe) in neural development. pH regulatory proteins are important for brain development as their human mutations are linked to developmental disorders like Christianson’s syndrome. While humans have 9 of these proteins, Drosophila melanogaster have 3, making them a straightforward model to investigate the subcellular roles of Nhe proteins in brain development. However, it remains unclear where the 3 Nhe proteins localize. Preliminary experiments suggest localization of Nhe2 at the plasma membrane in type II neuroblasts. Building on previously gathered data, I aim to both localize & quantify proliferation of Nhe2 in neuroblasts (type I & II) as well as localize & quantify abundance of Nhe2 during mitosis and interphase. To mark specific neuroblasts lineages, I will be utilizing the UAS/GAL4 system within control or endogenously tagged Nhe2-10x-MYC fly stocks that were generated via CRISPR. Immunohistochemistry will be used to identify cell types and for localization of Nhe2 within neuroblasts. Mammalian homologs of Nhe2 have been shown to localize to the cell cortex to facilitate mitotic cell swelling. To determine if Nhe2 displays a conserved role in cell swelling, I will examine Nhe2 expression of neuroblasts in mitosis identified by Phosphohistone3 (pH3) staining compared to NB in interphase that lack pH3. Additionally, I will measure NB volume at these stages to link NB localization to cell size. I hypothesize that Nhe2 will localize primarily to the plasma membrane of both type I & II neuroblasts during mitosis to facilitate mitotic cell swelling but be internalized during interphase (pH3- NB). Data collection is ongoing, with project completion projected in early April. Understanding mechanisms of pH regulatory proteins during neural development may help further elucidate how cells shift pH to facilitate developmental signaling and cell behavior. In addition, this work will resolve the relationship between disordered pH and developmental disorders such as Christianson’s syndrome & Parkinson’s, helping aid future research for a cure.
Category
Life Sciences
pH Regulatory Protein Nhe2 Localization in Neuroblasts & Abundance within Neuroblasts (type 1 & 2) During Mitosis and Interphase
UC South Ballroom
pH regulation is essential for neuronal excitability as different pH states dramatically affect ion channel function. Considerably less is known about how pH dynamics might shape brain development. Our preliminary data has discovered a novel role for pH regulatory proteins, Na+/H+ Exchangers (Nhe) in neural development. pH regulatory proteins are important for brain development as their human mutations are linked to developmental disorders like Christianson’s syndrome. While humans have 9 of these proteins, Drosophila melanogaster have 3, making them a straightforward model to investigate the subcellular roles of Nhe proteins in brain development. However, it remains unclear where the 3 Nhe proteins localize. Preliminary experiments suggest localization of Nhe2 at the plasma membrane in type II neuroblasts. Building on previously gathered data, I aim to both localize & quantify proliferation of Nhe2 in neuroblasts (type I & II) as well as localize & quantify abundance of Nhe2 during mitosis and interphase. To mark specific neuroblasts lineages, I will be utilizing the UAS/GAL4 system within control or endogenously tagged Nhe2-10x-MYC fly stocks that were generated via CRISPR. Immunohistochemistry will be used to identify cell types and for localization of Nhe2 within neuroblasts. Mammalian homologs of Nhe2 have been shown to localize to the cell cortex to facilitate mitotic cell swelling. To determine if Nhe2 displays a conserved role in cell swelling, I will examine Nhe2 expression of neuroblasts in mitosis identified by Phosphohistone3 (pH3) staining compared to NB in interphase that lack pH3. Additionally, I will measure NB volume at these stages to link NB localization to cell size. I hypothesize that Nhe2 will localize primarily to the plasma membrane of both type I & II neuroblasts during mitosis to facilitate mitotic cell swelling but be internalized during interphase (pH3- NB). Data collection is ongoing, with project completion projected in early April. Understanding mechanisms of pH regulatory proteins during neural development may help further elucidate how cells shift pH to facilitate developmental signaling and cell behavior. In addition, this work will resolve the relationship between disordered pH and developmental disorders such as Christianson’s syndrome & Parkinson’s, helping aid future research for a cure.