Abstract
This chapter is intended to highlight aspects of the serotonin system in the brain of the fruit fly, Drosophila melanogaster, while providing detailed methodology for visualizing neurons with immunofluorescence and confocal microscopy. These techniques can be easily translated to the use of immunofluorescence and confocal microscopy for visualization of cells and circuits using any reporter or fluorescent antibody system in the fly brain. Further, many of these techniques are also applicable to immunohistochemistry, immunofluorescence microscopy, and confocal microscopy in general to multiple types of samples like mammalian brain tissue sections and cultured cells.
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References
Waddell S, Quinn WG (2001) Flies, genes, and learning. Annu Rev Neurosci 24:1283–1309
Davis RL (2004) Olfactory learning. Neuron 44:31–48
Nichols CD (2006) Drosophila melanogaster neurobiology, neuropharmacology, and how the fly can inform central nervous system drug discovery. Pharmacol Ther 112:677–700
Pandey UB, Nichols CD (2011) Human disease models in Drosophila melanogaster and the role of the fly in therapeutic drug discovery. Pharmacol Rev 63:411–436
Becnel J, Johnson O, Majeed ZR, Tran V, Yu B, Roth BL, Cooper RL, Kerut EK, Nichols CD (2013) DREADDs in Drosophila: a pharmacogenetic approach for controlling behavior, neuronal signaling, and physiology in the fly. Cell Rep 4:1049–1059
Saudou F, Boschert U, Amlaiky N, Plassat JL, Hen R (1992) A family of Drosophila serotonin receptors with distinct intracellular signalling properties and expression patterns. EMBO J 11:7–17
Witz P, Amlaiky N, Plassat JL, Maroteaux L, Borrelli E, Hen R (1990) Cloning and characterization of a Drosophila serotonin receptor that activates adenylate cyclase. Proc Natl Acad Sci U S A 87:8940–8944
Colas J, Launay J, Kellermann O, Rosay P, Maroteaux L (1995) Drosophila 5-HT2 serotonin receptor: coexpression with fushi-tarazu during segmentation. Proc Natl Acad Sci U S A 92:5441–5445
Yuan Q, Lin F, Zheng X, Sehgal A (2005) Serotonin modulates circadian entrainment in Drosophila. Neuron 47:115–127
Nichols CD (2007) 5-HT2 receptors in Drosophila are expressed in the brain and modulate aspects of circadian behaviors. Dev Neurobiol 67:752–763
Becnel J, Johnson O, Luo J, Nässel DR, Nichols CD (2011) The serotonin 5-HT7Dro receptor is expressed in the brain of Drosophila, and is essential for normal courtship and mating. PLoS One 6:e20800
Luo J, Becnel J, Nichols CD, Nässel DR (2012) Insulin-producing cells in the brain of adult Drosophila are regulated by the serotonin 5-HT1A receptor. Cell Mol Life Sci 69:471–484
Gasque G, Conway S, Huang J, Rao Y, Vosshall LB (2013) Small molecule drug screening in Drosophila identifies the 5HT2A receptor as a feeding modulation target. Sci Rep 3:srep02120
Thamm M, Rolke D, Jordan N, Balfanz S, Schiffer C, Baumann A, Blenau W (2013) Function and distribution of 5-HT2 receptors in the honeybee (Apis mellifera). PLoS One 8:e82407
Okusawa S, Kohsaka H, Nose A (2014) Serotonin and downstream leucokinin neurons modulate larval turning behavior in Drosophila. J Neurosci 34:2544–2558
Nichols CD, Ronesi J, Pratt W, Sanders-Bush E (2002) Hallucinogens and Drosophila: linking serotonin receptor activation to behavior. Neuroscience 115:979–984
Johnson O, Becnel J, Nichols CD (2009) Serotonin 5-HT(2) and 5-HT(1A)-like receptors differentially modulate aggressive behaviors in Drosophila melanogaster. Neuroscience 158:1292–1300
Alekseyenko OV, Lee C, Kravitz EA (2010) Targeted manipulation of serotonergic neurotransmission affects the escalation of aggression in adult male Drosophila melanogaster. PLoS One 5:e10806
Johnson O, Becnel J, Nichols CD (2011) Serotonin receptor activity is necessary for olfactory learning and memory in Drosophila melanogaster. Neuroscience 192:372–381
Sitaraman D, LaFerriere H, Birman S, Zars T (2012) Serotonin is critical for rewarded olfactory short-term memory in Drosophila. J Neurogenet 26:238–244
Sykes PA, Condron BG (2005) Development and sensitivity to serotonin of Drosophila serotonergic varicosities in the central nervous system. Dev Biol 286:207–216
Neckameyer WS (2010) A trophic role for serotonin in the development of a simple feeding circuit. Dev Neurosci 32:217–237
Majeed ZR, Nichols CD, Cooper RL (2013) 5-HT stimulation of heart rate in Drosophila does not act through cAMP as revealed by pharmacogenetics. J Appl Physiol 115:1656–1665
Park JH, Helfrich-Forster G, Liu L, Rosbash M, Hall JC (2000) Differential regulation of circadian pacemaker output by separate clock genes in Drosophila. Proc Natl Acad Sci U S A 97:3608–3613
Venken KJT, Simpson JH, Bellen HJ (2011) Genetic manipulation of genes and cells in the nervous system of the fruit fly. Neuron 72:202–230
Hampel S, Chung P, McKellar CE, Hall D, Looger LL, Simpson JH (2011) Drosophila Brainbow: a recombinase-based fluorescence labeling technique to subdivide neural expression patterns. Nat Methods 8:253–259
Acknowledgments
This work was supported by R01MH083689 (C.D.N.) and P20GM103514. The authors would like to thank Prof. Dick Nässel for advice.
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Nichols, C.D., Sherman, K.J. (2015). Visualization of the Serotonin System in Drosophila Brain: Immunofluorescence and Confocal Microscopy. In: Blenau, W., Baumann, A. (eds) Serotonin Receptor Technologies. Neuromethods, vol 95. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2187-4_10
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DOI: https://doi.org/10.1007/978-1-4939-2187-4_10
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