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Ferritin Is Required in Multiple Tissues during Drosophila melanogaster Development.

González-Morales N, Mendoza-Ortíz MÁ, Blowes LM, Missirlis F, Riesgo-Escovar JR - PLoS ONE (2015)

Bottom Line: Mutations in either gene or deletion of both genes results in a similar set of cuticular embryonic phenotypes, ranging from non-deposition of cuticle to defects associated with germ band retraction, dorsal closure and head involution.Overall, our results are consistent with insect ferritin combining three functions: iron storage, intercellular iron transport, and protection from iron-induced oxidative stress.These functions are required in multiple tissues during Drosophila embryonic development.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Neurobiología del Desarrollo y Neurofisiología, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Campus UNAM Juriquilla, Querétaro, 76230, México.

ABSTRACT
In Drosophila melanogaster, iron is stored in the cellular endomembrane system inside a protein cage formed by 24 ferritin subunits of two types (Fer1HCH and Fer2LCH) in a 1:1 stoichiometry. In larvae, ferritin accumulates in the midgut, hemolymph, garland, pericardial cells and in the nervous system. Here we present analyses of embryonic phenotypes for mutations in Fer1HCH, Fer2LCH and in both genes simultaneously. Mutations in either gene or deletion of both genes results in a similar set of cuticular embryonic phenotypes, ranging from non-deposition of cuticle to defects associated with germ band retraction, dorsal closure and head involution. A fraction of ferritin mutants have embryonic nervous systems with ventral nerve cord disruptions, misguided axonal projections and brain malformations. Ferritin mutants die with ectopic apoptotic events. Furthermore, we show that ferritin maternal contribution, which varies reflecting the mother's iron stores, is used in early development. We also evaluated phenotypes arising from the blockage of COPII transport from the endoplasmic reticulum to the Golgi apparatus, feeding the secretory pathway, plus analysis of ectopically expressed and fluorescently marked Fer1HCH and Fer2LCH. Overall, our results are consistent with insect ferritin combining three functions: iron storage, intercellular iron transport, and protection from iron-induced oxidative stress. These functions are required in multiple tissues during Drosophila embryonic development.

No MeSH data available.


Related in: MedlinePlus

Ferritin genes interact genetically with the DMT1 homolog Mvl.(A) The Mvl97f-LacZ line shows a spatially restricted expression pattern for Mvl, mainly in the head region, the brain, and a segmentally repeated pattern. (B) In a ferritin homozygous mutant background, Mvl97f-LacZ expression increases. Black arrows denote the head region, white asterisk the embryonic brain, red asterisk the ventral nerve cord, and red arrows mark the segmented expression pattern. Introduction of a Mvl97f allele into a ferritin mutant background resulted in the appearance of necrotic patches in the cuticle (C, D). (E) Quantification of the total area covered by LacZ staining in control and ferritin mutant backgrounds, (p<0.0001; T-test, asterisk). (F) Quantification of the number of embryos showing necrotic patches with one or two Mvl97f alleles; statistical difference using a Chi squared test with p<0.0001 is shown by an asterisk.
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pone.0133499.g006: Ferritin genes interact genetically with the DMT1 homolog Mvl.(A) The Mvl97f-LacZ line shows a spatially restricted expression pattern for Mvl, mainly in the head region, the brain, and a segmentally repeated pattern. (B) In a ferritin homozygous mutant background, Mvl97f-LacZ expression increases. Black arrows denote the head region, white asterisk the embryonic brain, red asterisk the ventral nerve cord, and red arrows mark the segmented expression pattern. Introduction of a Mvl97f allele into a ferritin mutant background resulted in the appearance of necrotic patches in the cuticle (C, D). (E) Quantification of the total area covered by LacZ staining in control and ferritin mutant backgrounds, (p<0.0001; T-test, asterisk). (F) Quantification of the number of embryos showing necrotic patches with one or two Mvl97f alleles; statistical difference using a Chi squared test with p<0.0001 is shown by an asterisk.

Mentions: In control embryos, LacZ showed a very restricted pattern of Mvl expression consistent with previous studies [21,24]. However, in a mutant ferritin background, Mvl- driven LacZ is upregulated, its levels increasing as development proceeds (Fig 6A, 6B and 6E). We hypothesize than in the absence of functional ferritin, iron depleted cells upregulate Mvl in an effort to curve iron depletion. Quantitation showed a significant difference of expression (Fig 6E). Significantly, the addition of one copy of the Mvl97f hypomorphic allele into a ferritin-depleted embryo resulted significantly in the appearance of necrotic patches, which were rarely present in Mvl97f or in ferritin homozygous mutants alone (Fig 6C, 6D and 6F).


Ferritin Is Required in Multiple Tissues during Drosophila melanogaster Development.

González-Morales N, Mendoza-Ortíz MÁ, Blowes LM, Missirlis F, Riesgo-Escovar JR - PLoS ONE (2015)

Ferritin genes interact genetically with the DMT1 homolog Mvl.(A) The Mvl97f-LacZ line shows a spatially restricted expression pattern for Mvl, mainly in the head region, the brain, and a segmentally repeated pattern. (B) In a ferritin homozygous mutant background, Mvl97f-LacZ expression increases. Black arrows denote the head region, white asterisk the embryonic brain, red asterisk the ventral nerve cord, and red arrows mark the segmented expression pattern. Introduction of a Mvl97f allele into a ferritin mutant background resulted in the appearance of necrotic patches in the cuticle (C, D). (E) Quantification of the total area covered by LacZ staining in control and ferritin mutant backgrounds, (p<0.0001; T-test, asterisk). (F) Quantification of the number of embryos showing necrotic patches with one or two Mvl97f alleles; statistical difference using a Chi squared test with p<0.0001 is shown by an asterisk.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4508113&req=5

pone.0133499.g006: Ferritin genes interact genetically with the DMT1 homolog Mvl.(A) The Mvl97f-LacZ line shows a spatially restricted expression pattern for Mvl, mainly in the head region, the brain, and a segmentally repeated pattern. (B) In a ferritin homozygous mutant background, Mvl97f-LacZ expression increases. Black arrows denote the head region, white asterisk the embryonic brain, red asterisk the ventral nerve cord, and red arrows mark the segmented expression pattern. Introduction of a Mvl97f allele into a ferritin mutant background resulted in the appearance of necrotic patches in the cuticle (C, D). (E) Quantification of the total area covered by LacZ staining in control and ferritin mutant backgrounds, (p<0.0001; T-test, asterisk). (F) Quantification of the number of embryos showing necrotic patches with one or two Mvl97f alleles; statistical difference using a Chi squared test with p<0.0001 is shown by an asterisk.
Mentions: In control embryos, LacZ showed a very restricted pattern of Mvl expression consistent with previous studies [21,24]. However, in a mutant ferritin background, Mvl- driven LacZ is upregulated, its levels increasing as development proceeds (Fig 6A, 6B and 6E). We hypothesize than in the absence of functional ferritin, iron depleted cells upregulate Mvl in an effort to curve iron depletion. Quantitation showed a significant difference of expression (Fig 6E). Significantly, the addition of one copy of the Mvl97f hypomorphic allele into a ferritin-depleted embryo resulted significantly in the appearance of necrotic patches, which were rarely present in Mvl97f or in ferritin homozygous mutants alone (Fig 6C, 6D and 6F).

Bottom Line: Mutations in either gene or deletion of both genes results in a similar set of cuticular embryonic phenotypes, ranging from non-deposition of cuticle to defects associated with germ band retraction, dorsal closure and head involution.Overall, our results are consistent with insect ferritin combining three functions: iron storage, intercellular iron transport, and protection from iron-induced oxidative stress.These functions are required in multiple tissues during Drosophila embryonic development.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Neurobiología del Desarrollo y Neurofisiología, Instituto de Neurobiología, Universidad Nacional Autónoma de México, Campus UNAM Juriquilla, Querétaro, 76230, México.

ABSTRACT
In Drosophila melanogaster, iron is stored in the cellular endomembrane system inside a protein cage formed by 24 ferritin subunits of two types (Fer1HCH and Fer2LCH) in a 1:1 stoichiometry. In larvae, ferritin accumulates in the midgut, hemolymph, garland, pericardial cells and in the nervous system. Here we present analyses of embryonic phenotypes for mutations in Fer1HCH, Fer2LCH and in both genes simultaneously. Mutations in either gene or deletion of both genes results in a similar set of cuticular embryonic phenotypes, ranging from non-deposition of cuticle to defects associated with germ band retraction, dorsal closure and head involution. A fraction of ferritin mutants have embryonic nervous systems with ventral nerve cord disruptions, misguided axonal projections and brain malformations. Ferritin mutants die with ectopic apoptotic events. Furthermore, we show that ferritin maternal contribution, which varies reflecting the mother's iron stores, is used in early development. We also evaluated phenotypes arising from the blockage of COPII transport from the endoplasmic reticulum to the Golgi apparatus, feeding the secretory pathway, plus analysis of ectopically expressed and fluorescently marked Fer1HCH and Fer2LCH. Overall, our results are consistent with insect ferritin combining three functions: iron storage, intercellular iron transport, and protection from iron-induced oxidative stress. These functions are required in multiple tissues during Drosophila embryonic development.

No MeSH data available.


Related in: MedlinePlus