title: Programmed cell death is impaired in the developing brain of FMR1 mutants.
abstract: Fragile X syndrome (FXS), due to transcriptional silencing of fragile X mental retardation protein (FMRP), is characterized by excess synaptic connections and impaired dendrite maturation. Programmed cell death (PCD) is critical for synaptogenesis and elimination of aberrant neuronal connections in the developing brain; however, the role of FMRP in PCD is unknown. The aim of this work was to assess the intrinsic apoptosis pathway in the developing brain of Fmr1 mutants. To accomplish this, we evaluated two different Fmr1 mutant strains of 10-day-old male mice compared with appropriate controls. We performed immunohistochemistry for activated caspase-3 and TUNEL assays, quantified the number of neurons in neocortex and hippocampus, determined cytochrome c peroxidase activity, measured the amount of cytochrome c release from forebrain mitochondria, and assessed levels of key pro- and antiapoptotic mediators with immunoblot analysis. Both Fmr1 mutant strains demonstrated decreased apoptosis in neocortex, hippocampus, and basolateral amygdala, impaired cytochrome c and procaspase-9 release from mitochondria despite intact Bax translocation, increased expression of the antiapoptotic protein, BCL-xL, and increased number of neurons. Taken together, the data suggest that PCD is impaired due to increased BCL-xL expression and is associated with excess neurons in the developing brain of FMRP-deficient mice. It is possible that deficient PCD prevents neuron elimination and results in abnormal retention of developmentally transient neurons. Thus, defective PCD may contribute to the excess synaptic connections known to exist in Fmr1 mutants and could play a role in the behavioral phenotype of children with FXS.
authors.affiliation: Division of Anesthesiology and Pain Medicine, Children's National Medical Center, The George Washington University School of Medicine and Health Sciences, Washington, DC 20010, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Cheng
authors.forename: Joshua G
authors.initials: JG
authors.lastname: Corbin
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Levy
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics* / Q000502:physiology*; D051017:Apoptosis Regulatory Proteins / Q000096:biosynthesis / Q000235:genetics; D015153:Blotting, Western; D001921:Brain / Q000254:growth & development* / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D002452:Cell Count; D003578:Cytochrome-c Peroxidase / Q000378:metabolism; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000235:genetics*; D005600:Fragile X Syndrome / Q000235:genetics / Q000473:pathology; D006418:Heme / Q000031:analogs & derivatives / Q000378:metabolism; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation / Q000235:genetics / Q000502:physiology; D009474:Neurons / Q000502:physiology; D051058:Oncogene Protein v-akt / Q000235:genetics / Q000502:physiology; D010766:Phosphorylation; D051025:bcl-Associated Death Protein / Q000235:genetics / Q000502:physiology
chemical_list: D051017:Apoptosis Regulatory Proteins; D053148:Caspase 3; D003578:Cytochrome-c Peroxidase; D051860:Fragile X Messenger Ribonucleoprotein 1; D006418:Heme; D051058:Oncogene Protein v-akt; D051025:bcl-Associated Death Protein; C497684:Fmr1 protein, mouse; C031690:heme C
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