Walton D. Jones edited Results_expression.tex  almost 9 years ago

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To determine the expression pattern of DmCa\textsubscript{v}3 in the adult brain, we first generated a \emph{white\textsuperscript{+}} DmCa\textsubscript{v}3\textsuperscript{Founder} allele in which a $\phi$C31 attP landing site and a floxed \emph{mini-white} marker replaces the first coding exon of DmCa\textsubscript{v}3 via homologous recombination\cite{Huang:2009ei}.  We then removed the \emph{mini-white} marker via Cre-mediated recombination to produce a \emph{white\textsuperscript{-}} DmCa\textsubscript{v}3\textsuperscript{Founder} allele.  Finally, using $\phi$C31-mediated integration, we generated a GFP-tagged DmCa\textsubscript{v}3 knock-in allele, GFP::DmCa\textsubscript{v}3, by inserting the coding sequence for GFP in-frame immediately after the transcriptional start site of the first DmCa\textsubscript{v}3 exon (Fig. \ref{fig:S1}).  In GFP::DmCa\textsubscript{v}3 flies, GFP signal appears broadly across the brain, with the antennal lobes, mushroom body calyces, and the optic lobes all showing strong staining (Fig. \ref{Fig 2}). \ref{fig:2}).  The strongest expression, though, is in the central complex, which comprises the ellipsoid body, fan-shaped body, noduli, and protocerebral bridge (Fig. ).  GFP::DmCa\textsubscript{v}3 channels are also expressed broadly in less structured neuropils including subesophageal zone (SEZ), superior neuropil (SNP), inferior neuropil (INP), ventrolateral neuropil (VLNP) and ventromedial neuropil (VMNP).