218 | Metallomics, 2020, 12, 218--240 This journal is © The Royal Society of Chemistry 2020 Cite this: Metallomics, 2020, 12, 218 Intestinal response to dietary manganese depletion in Drosophila† Johana Va ´ squez-Procopio, a Beatriz Osorio, a Leticia Corte ´ s-Martı ´ nez, b Fidel Herna ´ ndez-Herna ´ ndez, b Oscar Medina-Contreras, c Emmanuel Rı ´ os-Castro, d Aram Comjean, e Fangge Li, e Yanhui Hu, e Stephanie Mohr, e Norbert Perrimon ef and Fanis Missirlis * a Manganese is considered essential for animal growth. Manganese ions serve as cofactors to three mitochondrial enzymes: superoxide dismutase (Sod2), arginase and glutamine synthase, and to glycosyltransferases residing in the Golgi. In Drosophila melanogaster, manganese has also been implicated in the formation of ceramide phosphoethanolamine, the insect’s sphingomyelin analogue, a structural component of cellular membranes. Manganese overload leads to neurodegeneration and toxicity in both humans and Drosophila. Here, we report specific absorption and accumulation of manganese during the first week of adulthood in flies, which correlates with an increase in Sod2 activity during the same period. To test the requirement of dietary manganese for this accumulation, we generated a Drosophila model of manganese deficiency. Due to the lack of manganese- specific chelators, we used chemically defined media to grow the flies and deplete them of the metal. Dietary manganese depletion reduced Sod2 activity. We then examined gene and protein expression changes in the intestines of manganese depleted flies. We found adaptive responses to the presumed loss of known manganese-dependent enzymatic activities: less glutamine synthase activity (amination of glutamate to glutamine) was compensated by 50% reduction in glutaminase (deamination of glutamine to glutamate); less glycosyltransferase activity, predicted to reduce protein glycosylation, was compensated by 30% reduction in lysosomal mannosidases (protein deglycosylating enzymes); less ceramide phosphoethanolamine synthase activity was compensated by 30% reduction in the Drosophila sphingomyeline phospodiesterase, which could catabolize ceramide phosphoethanolamine in flies. Reduced Sod2 activity, predicted to cause superoxide- dependent iron–sulphur cluster damage, resulted in cellular iron misregulation. Significance to metallomics Manganese is considered an essential micronutrient in animals, although clear demonstrations of manganese deficiency are scarce. We describe a manganese deficiency study in Drosophila melanogaster, showing that females transfer the metal to their eggs and that young adults highly and specifically absorb the metal from their diet during the first few days of their life, presumably to support manganese-dependent enzyme activities. Manganese deficient flies offer an experimental model to investigate the physiological roles of manganese. Combining metallomic, transcriptomic and proteomic analysis, we show that intestinal cells compensate for the loss of manganese-dependent biosynthetic activities by downregulating reciprocal catabolic pathways. Introduction How animals discriminate Mn is not understood Manganese (Mn) is considered an essential micronutrient in animals, although clear demonstrations of Mn deficiency are scarce. 1,2 Three mitochondrial enzymes function using Mn ions as cofactors: superoxide dismutase (Sod2), which converts superoxide radicals to hydrogen peroxide and oxygen, glutamine synthase (GS), which adds ammonia to glutamate and arginase (Arg), which subsequently removes the ammonia as urea. 3–5 In the Golgi apparatus, Mn serves as a cofactor to galactosyltransferases. 6–8 a Departamento de Fisiologı ´a, Biofı ´sica y Neurociencias, Centro de Investigacio´n y de Estudios Avanzados, Av. IPN 2508, Mexico City, 07360, Mexico. E-mail: fanis@fisio.cinvestav.mx b Departamento de Infecto´mica y Patoge ´nesis Molecular, Centro de Investigacio´n y de Estudios Avanzados, Av. IPN 2508, Mexico City, 07360, Mexico c Laboratorio de Investigacio´n en Inmunologı ´a y Proteo´mica, Hospital Infantil de Me ´xico Federico Go´mez, Dr Ma ´rquez 162, Mexico City, 06720, Mexico d Unidad de Geno´mica, Proteo´mica y Metabolo´mica, LaNSE, Centro de Investigacio´n y de Estudios Avanzados, Av. IPN 2508, Mexico City, 07360, Mexico e Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA f Howard Hughes Medical Institute, Boston, Massachusetts 02115, USA † Electronic supplementary information (ESI) available. See DOI: 10.1039/ c9mt00218a Received 5th September 2019, Accepted 26th November 2019 DOI: 10.1039/c9mt00218a rsc.li/metallomics Metallomics PAPER Downloaded from https://academic.oup.com/metallomics/article/12/2/218/5961849 by guest on 21 June 2022