title: Massive excretion of calcium oxalate from late prepupal salivary glands of Drosophila melanogaster demonstrates active nephridial-like anion transport.
abstract: The Drosophila salivary glands (SGs) were well known for the puffing patterns of their polytene chromosomes and so became a tissue of choice to study sequential gene activation by the steroid hormone ecdysone. One well-documented function of these glands is to produce a secretory glue, which is released during pupariation to fix the freshly formed puparia to the substrate. Over the past two decades SGs have been used to address specific aspects of developmentally-regulated programmed cell death (PCD) as it was thought that they are doomed for histolysis and after pupariation are just awaiting their fate. More recently, however, we have shown that for the first 3-4 h after pupariation SGs undergo tremendous endocytosis and vacuolation followed by vacuole neutralization and membrane consolidation. Furthermore, from 8 to 10 h after puparium formation (APF) SGs display massive apocrine secretion of a diverse set of cellular proteins. Here, we show that during the period from 11 to 12 h APF, the prepupal glands are very active in calcium oxalate (CaOx) extrusion that resembles renal or nephridial excretory activity. We provide genetic evidence that Prestin, a Drosophila homologue of the mammalian electrogenic anion exchange carrier SLC26A5, is responsible for the instantaneous production of CaOx by the late prepupal SGs. Its positive regulation by the protein kinases encoded by fray and wnk lead to increased production of CaOx. The formation of CaOx appears to be dependent on the cooperation between Prestin and the vATPase complex as treatment with bafilomycin A1 or concanamycin A abolishes the production of detectable CaOx. These data demonstrate that prepupal SGs remain fully viable, physiologically active and engaged in various cellular activities at least until early pupal period, that is, until moments prior to the execution of PCD.
authors.affiliation: Laboratory of Developmental Genetics, Institute of Experimental Endocrinology, Biomedical Centre, Slovak Academy of Sciences, Dúbravská cesta 9, 84505, Bratislava, Slovakia.
authors.forename: Robert
authors.initials: R
authors.lastname: Farkaš
authors.affiliation: Laboratory of Developmental Genetics, Institute of Experimental Endocrinology, Biomedical Centre, Slovak Academy of Sciences, Dúbravská cesta 9, 84505, Bratislava, Slovakia.|Department of Genetics, Comenius University, Mlynská dolina B-1, 84215, Bratislava, Slovakia.
authors.forename: Ludmila
authors.initials: L
authors.lastname: Pečeňová
authors.affiliation: Laboratory of Developmental Genetics, Institute of Experimental Endocrinology, Biomedical Centre, Slovak Academy of Sciences, Dúbravská cesta 9, 84505, Bratislava, Slovakia.|Department of Genetics, Comenius University, Mlynská dolina B-1, 84215, Bratislava, Slovakia.
authors.forename: Lucia
authors.initials: L
authors.lastname: Mentelová
authors.affiliation: Laboratory of Developmental Genetics, Institute of Experimental Endocrinology, Biomedical Centre, Slovak Academy of Sciences, Dúbravská cesta 9, 84505, Bratislava, Slovakia.
authors.forename: Milan
authors.initials: M
authors.lastname: Beňo
authors.affiliation: Laboratory of Developmental Genetics, Institute of Experimental Endocrinology, Biomedical Centre, Slovak Academy of Sciences, Dúbravská cesta 9, 84505, Bratislava, Slovakia.
authors.forename: Denisa
authors.initials: D
authors.lastname: Beňová-Liszeková
authors.affiliation: Laboratory of Developmental Genetics, Institute of Experimental Endocrinology, Biomedical Centre, Slovak Academy of Sciences, Dúbravská cesta 9, 84505, Bratislava, Slovakia.|Department of Medical Biochemistry, Jessenius Faculty of Medicine, Comenius University, Malá Hora 4, 03601, Martin, Slovakia.
authors.forename: Silvia
authors.initials: S
authors.lastname: Mahmoodová
authors.affiliation: Faculty of Agrobiology, Food and Natural Resources, Czech Agricultural University, Kamýcká 129, 16521, Prague 6, Czech Republic.
authors.forename: Václav
authors.initials: V
authors.lastname: Tejnecký
authors.affiliation: Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Albertov 4, 12800, Prague, Czech Republic.
authors.forename: Otakar
authors.initials: O
authors.lastname: Raška
authors.affiliation: Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Albertov 4, 12800, Prague, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Juda
authors.affiliation: Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Albertov 4, 12800, Prague, Czech Republic.
authors.forename: Silvie
authors.initials: S
authors.lastname: Svidenská
authors.affiliation: Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Albertov 4, 12800, Prague, Czech Republic.
authors.forename: Matúš
authors.initials: M
authors.lastname: Hornáček
authors.affiliation: Department of Biology, University of Nebraska at Omaha, 6001 Dodge Street, Omaha, Nebraska, 68182-0040, USA.
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Chase
authors.affiliation: Institute of Cellular Biology and Pathology, 1st Faculty of Medicine, Charles University in Prague, Albertov 4, 12800, Prague, Czech Republic.
authors.forename: Ivan
authors.initials: I
authors.lastname: Raška
mesh_terms: D000818:Animals; D027321:Anion Transport Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D001693:Biological Transport, Active / Q000502:physiology; D002129:Calcium Oxalate / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D012469:Salivary Glands / Q000378:metabolism*
chemical_list: D027321:Anion Transport Proteins; D029721:Drosophila Proteins; C000600831:prestin protein, Drosophila; D002129:Calcium Oxalate; D017346:Protein Serine-Threonine Kinases; C526279:fray protein, Drosophila
keywords: Drosophila; anion extrusion; calcium oxalate; fruitfly salivary glands; labial nephridia; prestin