title: Neonatal mouse hippocampus: phlebotomy-induced anemia diminishes and treatment with erythropoietin partially rescues mammalian target of rapamycin signaling.
abstract: BackgroundPhlebotomy-induced anemia (PIA) is common in premature infants and affects neurodevelopment. PIA alters hippocampal metabolism in neonatal mice through tissue hypoxia and iron deficiency. The mammalian target of rapamycin (mTOR) pathway senses the status of critical metabolites (e.g., oxygen, iron), thereby regulating hippocampal growth and function. We determined the effect of PIA and recombinant human erythropoietin (rHuEpo) treatment on mTOR signaling and expression of genes related to mTOR pathway functions.MethodsMice receiving an iron-supplemented diet were phlebotomized from postnatal day (P)3 to a target hematocrit of <25% by P7. Half were maintained at <25% until P14; half received rHuEpo from P7 to increase the hematocrit to 25-28%. Hippocampal phosphorylated to total protein ratios of four key mTOR pathway proteins were measured by western blotting at P14 and compared with non-phlebotomized, non-anemic control mice. mRNA levels of genes regulated by mTOR were measured by quantitative PCR.ResultsPIA suppressed phosphorylation of all mTOR proteins. rHuEpo restored AMP-activated protein kinase (AMPK) and AKT status, and partially rescued the mTOR output protein S6K. PIA and rHuEpo treatment also altered the expression of genes regulated by S6K.ConclusionPIA compromises and rHuEpo treatment partially rescues a pathway regulating neuronal DNA transcription, protein translation, and structural complexity.
authors.affiliation: Graduate Program in Neuroscience, Department of Neuroscience, University of Minnesota, Minneapolis, Minnesota.
authors.forename: Diana J
authors.initials: DJ
authors.lastname: Wallin
authors.affiliation: Children's Hospitals and Clinics of Minnesota, St. Paul, Minnesota.
authors.forename: Tara G
authors.initials: TG
authors.lastname: Zamora
authors.affiliation: Division of Neonatology, Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota.
authors.forename: Michelle
authors.initials: M
authors.lastname: Alexander
authors.affiliation: Division of Neonatology, Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota.
authors.forename: Kathleen M
authors.initials: KM
authors.lastname: Ennis
authors.affiliation: Division of Neonatology, Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota.
authors.forename: Phu V
authors.initials: PV
authors.lastname: Tran
authors.affiliation: Division of Neonatology, Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota.
authors.forename: Michael K
authors.initials: MK
authors.lastname: Georgieff
mesh_terms: D000740:Anemia / Q000188:drug therapy* / Q000209:etiology; D000818:Animals; D000831:Animals, Newborn; D004921:Erythropoietin / Q000008:administration & dosage* / Q000627:therapeutic use*; D005260:Female; D006624:Hippocampus / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D018962:Phlebotomy / Q000009:adverse effects*; D011247:Pregnancy; D015398:Signal Transduction; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*
chemical_list: D004921:Erythropoietin; C546843:mTOR protein, mouse; D058570:TOR Serine-Threonine Kinases