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• W1000656469 abstract "Decision making is vital for Drosophila melanogaster to find food or avoid hazards. When offering wild type flies ethanol enriched food and food without ethanol, flies prefer 5% ethanol containing food (Ogueta et al., 2010). This behavior is caused by olfactory stimuli (Schneider et al., 2012). When the odor information is processed, the decision to approach one odor source has to be converted in movement. In addition, flies tend to climb up the vials after they have been shaken down which is known as negative geotaxis (Kamikouchi et al., 2009). Walking speed measured in negative geotaxis assays can be used to analyze locomotion behavior (Strauss and Heisenberg, 1993). The neurotransmitter serotonin (5-HT) modulates olfactory processing in antennal lobe of Drosophila (Dacks et al., 2007). Increased serotonin level by feeding 5-HTP, the serotonin precursor, also causes reduced locomotion activity in flies (Yuan et al., 2006). The role serotonin plays in ethanol preference has not been analyzed. In addition, it is not clear whether serotonin involved in negative geotaxis locomotion.To dissect the role of serotonin in odor evoked ethanol preference, the function of the key regulator in serotonin signaling‒the serotonin transporter (SERT) in olfactory ethanol preference was analyzed. The serotonin transporter removes serotonin from synaptic cleft via reuptake it into the pre-synaptic neuron and therefore terminates the action of serotonin in the synaptic cleft. Even though different dSERT mutants have different transcript level, western blot showed that dSERT protein levels are severely reduced in all dSERT mutants. The loss of SERT expression is correlated with changes in locomotion since dSERT16 mutants fail to perform climbing task and also dSERT18 showed impaired negative geotaxis climbing. dSERT mutants were tested for odor evoked ethanol preference. The dSERT16 mutants could not decide for either food odors or ethanol containing food odor. These results suggested that serotonin is a negative regulator, as increased serotonin levels lead to decreased climbing ability and loss of ethanol odor preference. To confirm the accurate role of serotonin signaling in odor evoked ethanol preference, a dominant-negative version of the serotonin transporter unable to bind serotonin was expressed in different serotonergic neurons in the fly brain to increase serotonin signaling. Expression of this modified transporter in TPH-GAL4 driven neurons indeed caused a reduction of ethanol preference. That is due to prolonged 5-HT signaling, since a similar phenotype was observed when flies were fed with the serotonin precursor 5-HTP resulting in increased 5-HT levels (Schlager, 2013). Locomotion did not contribute to the reduce preference, since TPH-GAL4/UAS-SERTDN flies behaved normal in anti-geotaxis climbing. These results indicate that increased serotonin level suppresses ethanol preference and a subset of serotonergic neurons driven by TPH-GAL4 is required for ethanol odor induced behavior. When disturbing dSERT function in SERT3-GAL4 dependent serotonergic neurons a decreased preference to ethanol was also recorded. However, these flies exhibit robust ability in climbing. A subset of six serotonergic neurons was found in IP, LP1 and SE1 clusters. Four common serotonergic neurons in IP and LP1 clusters were targeted after compared neuronal expression pattern of SERT3-GAL4 with TPH-GAL4. Therefore, ethanol preference is modulated by four serotonergic neurons from IP and LP1 clusters in the brain. Surprisingly expression of UAS-SERTDN in TRH-GAL4 dependent neurons which covered 83% of serotonergic neurons in CNS does not alter ethanol preference. Beside the same neurons found in TPH-GAL4 and SERT3-GAL4 drivers, additional serotonergic neurons in CSD, DP and abdominal ganglia were detected. This data suggests another opposing serotonergic neuronal circuit exists to modulate ethanol preference. To verify that preference changes were not due to the strength of different GAL4 expression, UAS-SERTDN was expressed simultaneously in SERT3-GAL4 and TRH-GAL4 driven neurons. Thereby no change in preference was detected. Same result was observed by expressing UAS-SERTDN in SERT3-GAL4/RN2-E-GAL4 driver. However, those flies showed defects in negative geotaxis climbing. RN2-E-GAL4 drives CSD neuron in the brain and a cluster in the abdominal ganglia. Serotonergic cells in CSD cluster and abdominal ganglia are involved in modulating ethanol preference and climbing. In conclusion, dSERT participates in the modulation of odor evoked preference and negative geotaxis climbing. Serotonin acts as a negative modulator in ethanol preference. Increased serotonin level leads to decreased ethanol preference and four putative serotonergic neurons in IP and LP1 clusters are responsible for this behavior. The preference change is not due to movement ability. Another opposing serotonergic circuit is also involved in regulating ethanol odor evoked ethanol preference in Drosophila melanogaster." @default.
• W1000656469 created "2016-06-24" @default.
• W1000656469 creator A5051746234 @default.
• W1000656469 date "2013-12-04" @default.
• W1000656469 modified "2023-09-24" @default.
• W1000656469 title "Two opposing serotonergic neuronal circuits modulate ethanol preference of Drosophila melanogaster" @default.
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