Figure 2.
In situ hybridization of an antisense ptth RNA probe to a stage 17 embryo (A) and wandering third instar brain (B). The black circle in A indicates the position of two positive neurons (enlarged in the inset) that correspond to one of the neuron pairs shown in B. (C) No staining is seen in a third instar brain hybridized with a sense control probe. (D) The dendritic arbors and axon terminals (yellow arrow) of the PG neurons are highlighted using ptth-Gal4 to drive expression of a membrane-bound UAS- cd8GFP reporter. (E) Immunolocalization of PTTH using an HA-tagged genomic transgene. The yellow arrow indicates synaptic connections on the prothoracic gland. (F) Immunolocalization of PTTH-HA (green) and the ecdysone biosynthetic enzyme Phm (red) in the prothoracic gland. (G) Axons from the PDF-producing neurons (red) contact the dendritic arbors of PTTH-producing neurons.