FRUM and En are co-expressed in a subset of neurons throughout development. The expression patterns of FRUM and En were analyzed by immunohistochemistry using anti-FRUM (green) and anti-En (magenta) in early pupae through adults and the number of co-expressing neurons counted (Table 1). (A-B) Doubly-labeled 50 hr APF pupal CNS, anti-FRUM (A) and anti-En (B) are expressed and co-expressed (C) throughout the brain and ventral nerve cord. (D) Schematic of En/FRUM co-expression. Pink circles represent En-positive and black-rimmed white circles represent En/FRUM neurons in the brain (box i), T1 midline (arrow, iii), T1-T2-T3 medial cells (arrowheads, ii, iv, v, respectively), and abdominal ganglion (vi). (E-G) In optical image overlays of regions i, iii, and vi (noted in panel D), En/FRUM co-expression appears white inside cells of the brain (E), T1 ganglion (F), and abdominal ganglion (G). (H-J) In the anterior mediolateral brain (Box i) from a 27 hr APF pupa (H), a 50 hr APF pupa (I) and an adult (J), En/FRUM are co-expressed (arrowheads) at all stages. (K-M) In the VNC (arrowheads, ii and iii) from a 27 hr APF pupa (K), a 50 hr APF pupa (L) and an adult (M), FRUM/En are co-expressed (arrowheads) in midline neurons of T1 and medial neurons in T1-T3 at all stages. Images are confocal z-stacks through the entire CNS (A, B, C) or stacks of a subset of z-slices (E-M). Size bar = 100 μm (A) for panels A and B, 20 μm (E, H, K) for panels E-M.
Latham et al. BMC Neuroscience 2013 14:57 doi:10.1186/1471-2202-14-57