title: Immunolocalization of the vesicular acetylcholine transporter in larval and adult Drosophila neurons.
abstract: Vesicular acetylcholine transporter (VAChT) function is essential for organismal survival, mediating the packaging of acetylcholine (ACh) for exocytotic release. However, its expression pattern in the Drosophila brain has not been fully elucidated. To investigate the localization of VAChT, we developed an antibody against the C terminal region of the protein and we show that this antibody recognizes a 65KDa protein corresponding to VAChT on an immunoblot in both Drosophila head homogenates and in Schneider 2 cells. Further, we report for the first time the expression of VAChT in the antennal lobe and ventral nerve cord of Drosophila larva; and we independently confirm the expression of the protein in mushroom bodies and optic lobes of adult Drosophila. Importantly, we show that VAChT co-localizes with a synaptic vesicle marker in vivo, confirming previous reports of the localization of VAChT to synaptic terminals. Together, these findings help establish the vesicular localization of VAChT in cholinergic neurons in Drosophila and present an important molecular tool with which to dissect the function of the transporter in vivo.
authors.affiliation: Neuroscience Program, Department of Biological Sciences, Delaware State University, 1200 N Dupont Highway, Dover, DE 19901, USA.
authors.forename: Sridhar
authors.initials: S
authors.lastname: Boppana
authors.affiliation: Neuroscience Program, Department of Biological Sciences, Delaware State University, 1200 N Dupont Highway, Dover, DE 19901, USA.
authors.forename: Natalie
authors.initials: N
authors.lastname: Kendall
authors.affiliation: Neuroscience Program, Department of Biological Sciences, Delaware State University, 1200 N Dupont Highway, Dover, DE 19901, USA.
authors.forename: Opeyemi
authors.initials: O
authors.lastname: Akinrinsola
authors.affiliation: Neuroscience Program, Department of Biological Sciences, Delaware State University, 1200 N Dupont Highway, Dover, DE 19901, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: White
authors.affiliation: Neuroscience Program, Department of Biological Sciences, Delaware State University, 1200 N Dupont Highway, Dover, DE 19901, USA.
authors.forename: Krushali
authors.initials: K
authors.lastname: Patel
authors.affiliation: Neuroscience Program, Department of Biological Sciences, Delaware State University, 1200 N Dupont Highway, Dover, DE 19901, USA. Electronic address: hlawal@desu.edu.
authors.forename: Hakeem
authors.initials: H
authors.lastname: Lawal
mesh_terms: D000109:Acetylcholine / Q000378:metabolism; D000375:Aging; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D002799:Cholinergic Fibers / Q000378:metabolism*; D059329:Cholinergic Neurons / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000254:growth & development / Q000378:metabolism*; D007150:Immunohistochemistry / Q000379:methods; D007814:Larva; D024521:Mushroom Bodies / Q000378:metabolism*; D013572:Synaptic Vesicles / Q000378:metabolism*; D050494:Vesicular Acetylcholine Transport Proteins / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D050494:Vesicular Acetylcholine Transport Proteins; D000109:Acetylcholine
keywords: Acetylcholine; Localization; Neurotransmitter; Synaptic vesicle; VAChT