Figure 4. Comprehensive analysis of individual neurons in a central complex sublineage through twin-spot MARCM.

Six central complex neurons (a), nc82 staining for revealing relative positions to these neurons (a-b) and a schematic drawing of these neurons (c). Twin-spot MARCM analyses on this central complex sublineage reveals birth-order of these neurons (d-h and o). The first four neurons (d-g) extend into the noduli with the projections to discrete subcompartments, including the dorsal noduli (d), anterior ventral noduli (e), posterior ventral noduli (f) and middle noduli (g). The last two neurons (h and o) innervate the ellipsoid body. Neuronal morphology alternates between narrow and elaborate neurites (d, f and h vs. e, g and o) in the lateral accessory lobe. A third-born chinmo neuron (3rd^4th; i), determined by its reference wild-type Nb clone (p), has the morphology of wild-type fourth-born neuron. This 3rd^4th born neuron innervates the middle noduli that normally is innervated by the 4th born neuron (arrowheads in i, p and w). The neurite pattern in the lateral accessory lobe of this 3rd^4th born neuron is also transformed from narrow to elaborate (triple arrows in f, g and i). In contrast, the reverse twin-spot MARCM analysis shows that the second-born wild-type single-cell clone is associated with a chinmo Nb clone (j, q and x). Compared to the projection to the posterior ventral noduli in the wild-type Nb clone (arrow in l), the same morphology is not present in the chinmo Nb clone (arrow in q), confirming the transformation of the third-born neuron to the wild-type fourth-born neuron morphology. Asterisk indicates cell body position.