Table 1.

Summary of Volado immunoreactivity inDrosophila larvae

Stage	Number of larvae	% VOL(+) in CNS neurophil	% Vol(+) at NMJ	Number of hemisegments scored	% NMJs VOL(+)	NMJ	% VOL (+)
2nd instar (24–30 hr AH)	12	67	42	81	2.3	6 /7	3.7
						12 /13	14.8
3rd instar (∼96 hr AH)	47	100	94	420	5.1	4	2.9
						6 /7	4.5
						12 /13	22.6

VOL immunoreactivity (visualized with DAB reaction) is summarized for wild-type second and third instar larval preparations. Multiple classes of NMJs were examined and scored in 8–10 hemisegments (A2–A6) per animal, including longitudinal (muscles 6/7, 12/13, 3, 4), oblique (muscles 15/16/17), and transverse (muscles 8, 21, 22) fibers. Only strong, localized bouton expression (as illustrated in Figs.1A, 2B,C) was scored as positive. Columns 3 and 4 list percentages of larvae with VOL-immunpositive (VOL(+)) synaptic boutons in the CNS or at NMJs, respectively. Columns 5 and 6 list the total number of hemisegments examined, and the overall percentage of VOL(+) NMJs of all the NMJs scored in each age group, respectively. Column 7 lists the frequency of VOL(+) expression at three specific NMJs; muscles 6/7 and 12/13 were scored as single units. All other classes of NMJs not shown had 2%–5% frequency of VOL(+) expression.