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Fig. 5 | BMC Developmental Biology

Fig. 5

From: Developmental nicotine exposure affects larval brain size and the adult dopaminergic system of Drosophila melanogaster

Fig. 5

Developmental nicotine exposure decreases the number of TH+ neurons in the PPM3 cluster. Flies were reared on control or 0.3 mg/ml nicotine food. Adult brains were dissected, immunostained, mounted and imaged on a confocal microscope. a, d Schematic of the approximate position of the adult dopaminergic clusters. a Anterior clusters shown, d posterior clusters shown. b, c Maximum projection images of the anterior region of a brain from an adult fly that was reared in control food. The PAM and PAL dopaminergic clusters are visible. e, f are maximum projection images of the posterior region of the same brain. The PPM1, PPM2, PPM3, PPL1, PPL2ab and PPL2c are visible. c, f Anti-tyrosine hydroxylase (TH) was used as marker for dopaminergic neurons. b, e Anti-bruchpilot (BRP) was used as background staining for the whole brain. Merged images of the BRP and TH channels. Scale bar 50 μM. g Average number of TH+ neurons counted in the PPM3 cluster of flies reared in control food (black bars) or nicotine food (red bars). h, i Representative images of the PPM3 cluster from flies reared in control (h) or nicotine food (i). The number of TH+ neurons in the PPM3 cluster was reduced in brains from nicotine-exposed flies. Sample size was n = 20 brain hemispheres for control and n = 34 brain hemispheres for nicotine from n ≥ 5 independent experiments. Student’s t-test was used to compare the control versus the nicotine condition

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