Abstract
The insect renal (Malpighian) tubule has long been a model system for the study of fluid secretion and its neurohormonal control, as well as studies on ion transport mechanisms. To extend these studies beyond the boundaries of classical physiology, a molecular genetic approach together with the ‘omics technologies is required. To achieve this in any vertebrate transporting epithelium remains a daunting task, as the genetic tools available are still relatively unsophisticated. Drosophila melanogaster, however, is an outstanding model organism for molecular genetics. Here we describe a technique for fluid secretion assays in the D. melanogaster equivalent of the kidney nephron. The development of this first physiological assay for a Drosophila epithelium, allowing combined approaches of integrative physiology and functional genomics, has now provided biologists with an entirely new model system, the Drosophila Malpighian tubule, which is utilized in multiple fields as diverse as kidney disease research and development of new modes of pest insect control.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Morgan TH (1910) Sex limited inheritance in Drosophila. Science 32:120–122
Rubin GM, Lewis EB (2000) A brief history of Drosophila’s contributions to genome research. Science 287:2216–2218
Rubin GM, Spradling AC (1983) Vectors for P element-mediated gene transfer in Drosophila. Nucleic Acids Res 11:6341–6351
Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118:401–415
Rosay P et al (1997) Cell-type specific calcium signalling in a Drosophila epithelium. J Cell Sci 110(Pt 15):1683–1692
Davies SA, Terhzaz S (2009) Organellar calcium signalling mechanisms in Drosophila epithelial function. J Exp Biol 212:387–400
Tian L et al (2009) Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators. Nat Methods 6:875–881
Shafer OT et al (2008) Widespread receptivity to neuropeptide PDF throughout the neuronal circadian clock network of Drosophila revealed by real-time cyclic AMP imaging. Neuron 58:223–237
Cabrero P et al (2014) Chloride channels in stellate cells are essential for uniquely high secretion rates in neuropeptide-stimulated Drosophila diuresis. Proc Natl Acad Sci U S A 111:14301–14306
Akerboom J et al (2013) Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics. Front Mol Neurosci 6:2
Efetova M et al (2013) Separate roles of PKA and EPAC in renal function unraveled by the optogenetic control of cAMP levels in vivo. J Cell Sci 126:778–788
Dietzl G et al (2007) A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila. Nature 448:151–156
Gramates L et al (2017) FlyBase at 25: looking to the future. Nucleic Acids Res 45:D663–D671
Chintapalli VR, Wang J, Dow JA (2007) Using FlyAtlas to identify better Drosophila melanogaster models of human disease. Nat Genet 39:715–720
Leader DP, Krause SA, Pandit A, Davies SA, Dow JAT (2018) FlyAtlas 2: a new version of the Drosophila melanogaster expression atlas with RNA-Seq, miRNA-Seq and sex-specific data. Nucleic Acids Res 46:D809–D815
Dow JT, Davies SA (2003) Integrative physiology and functional genomics of epithelial function in a genetic model organism. Physiol Rev 83:687–729
Dow JAT (2012) Drosophila as an experimental organism for functional genomics. In: eLS. John Wiley & Sons Ltd, Chichester
Ugur B, Chen K, Bellen HJ (2016) Drosophila tools and assays for the study of human diseases. Dis Model Mech 9:235–244
Dow JA, Romero MF (2010) Drosophila provides rapid modeling of renal development, function, and disease. Am J Physiol Renal Physiol 299:F1237–F1244
Strange K (2016) Drug discovery in fish, flies, and worms. ILAR J 57:133–143
Sonoshita M, Cagan RL (2017) Modeling human cancers in Drosophila. Curr Top Dev Biol 121:287–309
Krench M, Littleton JT (2017) Neurotoxicity pathways in Drosophila models of the polyglutamine disorders. Curr Top Dev Biol 121:201–223
Berridge MJ, Oschman JL (1969) A structural basis for fluid secretion by malpighian tubules. Tissue Cell 1:247–272
Maddrell SHP (1971) The mechanisms of insect excretory systems. Adv Insect Physiol 8:199–331
Dow JAT (2013) In: Chapman RF, Simpson SJ, Douglas AE (eds) The insects, structure and function. Cambridge University Press, Cambridge
Marcelli Malpighii Philosophii & Medici Bononiensis Dissertatio epistolica De Bombyce: Societati Regiae, Londini ad Scientiam Naturalem promovendam institutae, dicata Malpighi, Marcello, 1628–1694. https://encore.lib.gla.ac.uk/iii/encore/record/C_Rb2671014;jsessionid=DD1890B70359EE4BC74ADB74D5041C5B?lang=eng
Maddrell S (2009) Insect homeostasis: past and future. J Exp Biol 212:446–451
Wessing A, Eichelberg D (1978) The genetics and biology of Drosophila, vol 2c. Academic Press, London
Denholm B (2013) Shaping up for action: the path to physiological maturation in the renal tubules of Drosophila. Organogenesis 9:40–54
Beyenbach KW, Skaer H, Dow JA (2010) The developmental, molecular, and transport biology of Malpighian tubules. Annu Rev Entomol 55:351–374
Sozen MA, Armstrong JD, Yang M, Kaiser K, Dow JA (1997) Functional domains are specified to single-cell resolution in a Drosophila epithelium. Proc Natl Acad Sci U S A 94:5207–5212
Dow JA et al (1994) The Malpighian tubules of Drosophila melanogaster: a novel phenotype for studies of fluid secretion and its control. J Exp Biol 197:421–428
Dube K, McDonald DG, O'Donnell MJ (2000) Calcium transport by isolated anterior and posterior Malpighian tubules of Drosophila melanogaster: roles of sequestration and secretion. J Insect Physiol 46:1449–1460
Dube KA, McDonald DG, O'Donnell MJ (2000) Calcium homeostasis in larval and adult Drosophila melanogaster. Arch Insect Biochem Physiol 44:27–39
O’Donnell MJ, Maddrell SH (1995) Fluid reabsorption and ion transport by the lower Malpighian tubules of adult female Drosophila. J Exp Biol 198:1647–1653
Dow JA (2009) Insights into the Malpighian tubule from functional genomics. J Exp Biol 212:435–445
Dow JA (1999) The multifunctional Drosophila melanogaster V-ATPase is encoded by a multigene family. J Bioenerg Biomembr 31:75–83
Allan AK, Du J, Davies SA, Dow JAT (2005) Genome-wide survey of V-ATPase genes in Drosophila reveals a conserved renal phenotype for lethal alleles. Physiol Genomics 22:128–138
Torrie LS et al (2004) Resolution of the insect ouabain paradox. Proc Natl Acad Sci U S A 101:13689–13693
Kaufmann N et al (2005) Developmental expression and biophysical characterization of a Drosophila melanogaster aquaporin. Am J Physiol Cell Physiol 289:C397–C407
Kerr M, Davies SA, Dow JA (2004) Cell-specific manipulation of second messengers; a toolbox for integrative physiology in Drosophila. Curr Biol 14:1468–1474
Wang J et al (2004) Function-informed transcriptome analysis of Drosophila renal tubule. Genome Biol 5:R69
Chintapalli VR et al (2012) Functional correlates of positional and gender-specific renal asymmetry in Drosophila. PLoS One 7:e32577
Davies SA et al (2012) Immune and stress response ‘cross-talk’ in the Drosophila Malpighian tubule. J Insect Physiol 58:488–497
Davies SA et al (2014) Cell signalling mechanisms in stress tolerance. J Exp Biol 217:119–128
Terhzaz S et al (2010) Cell-specific inositol 1,4,5 trisphosphate 3-kinase mediates epithelial cell apoptosis in response to oxidative stress in Drosophila. Cell Signal 22:737–748
Piermarini PM, Esquivel CJ, Denton S (2017) Malpighian tubules as novel targets for mosquito control. Int J Environ Res Public Health 14. https://doi.org/10.3390/ijerph14020111
Dow JA, Davies SA (2006) The Malpighian tubule: rapid insights from post-genomic biology. J Insect Physiol 52:365–378
Ianowski JP, O’Donnell MJ (2004) Basolateral ion transport mechanisms during fluid secretion by Drosophila Malpighian tubules: Na+ recycling, Na+:K+:2Cl− cotransport and Cl− conductance. J Exp Biol 207:2599–2609
Linton SM, O’Donnell MJ (1999) Contributions of K+:Cl− cotransport and Na+/K+-ATPase to basolateral ion transport in malpighian tubules of Drosophila melanogaster. J Exp Biol 202:1561–1570
Maddrell SH, Overton JA (1988) Stimulation of sodium transport and fluid secretion by ouabain in an insect malpighian tubule. J Exp Biol 137:265–276
Davies SA et al (1996) Analysis and inactivation of vha55, the gene encoding the vacuolar ATPase B-subunit in Drosophila melanogaster reveals a larval lethal phenotype. J Biol Chem 271:30677–30684
Karet FE et al (1999) Mutations in the gene encoding B1 subunit of H+-ATPase cause renal tubular acidosis with sensorineural deafness. Nat Genet 21:84–90
Kamleh MA, Hobani Y, Dow JA, Zheng L, Watson DG (2009) Towards a platform for the metabonomic profiling of different strains of Drosophila melanogaster using liquid chromatography-Fourier transform mass spectrometry. FEBS J 276:6798–6809
O’Donnell MJ (2009) Too much of a good thing: how insects cope with excess ions or toxins in the diet. J Exp Biol 212:363–372
Dow JA (2007) Integrative physiology, functional genomics and the phenotype gap: a guide for comparative physiologists. J Exp Biol 210:1632–1640
Tardif G, Murnik M (1975) Frequency-dependent sexual selection among wild-type strains of Drosophila melanogaster. Behav Genet 5:373–379
Ramsay J (1954) Active transport of water by the Malpighian tubules of the stick insect, Dixippus morosus (Orthoptera, Phasmidae). J Exp Biol 31:104–113
Chintapalli VR, Wang J, Herzyk P, Davies SA, Dow JA (2013) Data-mining the FlyAtlas online resource to identify core functional motifs across transporting epithelia. BMC Genomics 14:518
Du J et al (2006) The SzA mutations of the B subunit of the Drosophila vacuolar H+ ATPase identify conserved residues essential for function in fly and yeast. J Cell Sci 119:2542–2551
Maddrell SH, O’Donnell MJ (1992) Insect Malpighian tubules: v-ATPase action in ion and fluid transport. J Exp Biol 172:417–429
Coast GM, Webster SG, Schegg KM, Tobe SS, Schooley DA (2001) The Drosophila melanogaster homologue of an insect calcitonin-like diuretic peptide stimulates V-ATPase activity in fruit fly Malpighian tubules. J Exp Biol 204:1795–1804
Giannakou ME, Dow JA (2001) Characterization of the Drosophila melanogaster alkali-metal/proton exchanger (NHE) gene family. J Exp Biol 204:3703–3716
Davies SA, Day JP (2006) cGMP signalling in a transporting epithelium. Biochem Soc Trans 34:512–514
Coast G (2007) The endocrine control of salt balance in insects. Gen Comp Endocrinol 152:332–338
Blumenthal EM (2003) Regulation of chloride permeability by endogenously produced tyramine in the Drosophila Malpighian tubule. Am J Physiol Cell Physiol 284:C718–C728
Wu Y, Schellinger JN, Huang CL, Rodan AR (2014) Hypotonicity stimulates potassium flux through the WNK-SPAK/OSR1 kinase cascade and the Ncc69 sodium-potassium-2-chloride cotransporter in the Drosophila renal tubule. J Biol Chem 289:26131–26142
O’Donnell MJ, Ianowski JP, Linton SM, Rheault MR (2003) Inorganic and organic anion transport by insect renal epithelia. Biochim Biophys Acta 1618:194–206
Halberg KA et al (2016) The cell adhesion molecule Fasciclin2 regulates brush border length and organization in Drosophila renal tubules. Nat Commun 7:11266
Wu Y, Baum M, Huang CL, Rodan AR (2015) Two inwardly rectifying potassium channels, Irk1 and Irk2, play redundant roles in Drosophila renal tubule function. Am J Physiol Regul Integr Comp Physiol 309:R747–R756
Rodan AR, Baum M, Huang CL (2012) The Drosophila NKCC Ncc69 is required for normal renal tubule function. Am J Physiol Cell Physiol 303:C883–C894
Day JP et al (2008) Identification of two partners from the bacterial kef exchanger family for the apical plasma membrane V-ATPase of Metazoa. J Cell Sci 121:2612–2619
Terhzaz S, Cabrero P, Chintapalli VR, Davies SA, Dow JAT (2010) Mislocalization of mitochondria and compromised renal function and oxidative stress resistance in Drosophila SesB mutants. Physiol Genomics 41:33–41
MacPherson MR, Lohmann SM, Davies SA (2004) Analysis of Drosophila cGMP-dependent protein kinases and assessment of their in vivo roles by targeted expression in a renal transporting epithelium. J Biol Chem 279:40026–40034
MacPherson MR et al (2004) The dg2 (for) gene confers a renal phenotype in Drosophila by modulation of cGMP-specific phosphodiesterase. J Exp Biol 207:2769–2776
Ruka KA, Miller AP, Blumenthal EM (2013) Inhibition of diuretic stimulation of an insect secretory epithelium by a cGMP-dependent protein kinase. Am J Physiol Renal Physiol 304:F1210–F1216
Broderick KE et al (2003) Interactions between epithelial nitric oxide signaling and phosphodiesterase activity in Drosophila. Am J Physiol Cell Physiol 285:C1207–C1218
Pollock VP et al (2003) NorpA and itpr mutants reveal roles for phospholipase C and inositol (1,4,5)- trisphosphate receptor in Drosophila melanogaster renal function. J Exp Biol 206:901–911
MacPherson MR et al (2005) Transient receptor potential-like channels are essential for calcium signaling and fluid transport in a Drosophila epithelium. Genetics 169:1541–1552
Terhzaz S et al (2012) Mechanism and function of Drosophila capa GPCR: a desiccation stress-responsive receptor with functional homology to human neuromedinU receptor. PLoS One 7:e29897
Cannell E et al (2016) The corticotropin-releasing factor-like diuretic hormone 44 (DH) and kinin neuropeptides modulate desiccation and starvation tolerance in Drosophila melanogaster. Peptides 80:96–107
Zandawala M, Marley R, Davies SA, Nassel DR (2018) Characterization of a set of abdominal neuroendocrine cells that regulate stress physiology using colocalized diuretic peptides in Drosophila. Cell Mol Life Sci 75:1099–1115
Blumenthal EM (2009) Isoform- and cell-specific function of tyrosine decarboxylase in the Drosophila Malpighian tubule. J Exp Biol 212:3802–3809
Ashburner M (1989) Drosophila: A laboratory handbook. Cold Spring Harbor Laboratory: 1331
Maddrell SHP (1991) BioEssays 13(7):357
Acknowledgments
Extensive support by the Biotechnology and Biological Sciences Research Council UK to the corresponding authors has been instrumental in developing the new fields of D. melanogaster integrative physiology and functional genomics.
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Davies, SA. et al. (2019). Epithelial Function in the Drosophila Malpighian Tubule: An In Vivo Renal Model. In: Vainio, S. (eds) Kidney Organogenesis. Methods in Molecular Biology, vol 1926. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9021-4_17
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9021-4_17
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9020-7
Online ISBN: 978-1-4939-9021-4
eBook Packages: Springer Protocols