title: Inactivation of the hereditary spastic paraplegia-associated Hspd1 gene encoding the Hsp60 chaperone results in early embryonic lethality in mice.
abstract: The mitochondrial Hsp60 chaperonin plays an important role in sustaining cellular viability. Its dysfunction is related to inherited forms of the human diseases spastic paraplegia and hypomyelinating leukodystrophy. However, it is unknown whether the requirement for Hsp60 is neuron specific or whether a complete loss of the protein will impair mammalian development and postnatal survival. In this study, we describe the generation and characterization of a mutant mouse line bearing an inactivating gene-trap insertion in the Hspd1 gene encoding Hsp60. We found that heterozygous mice were born at the expected ratio compared to wild-type mice and displayed no obvious phenotype deficits. Using quantitative reverse transcription PCR, we found significantly decreased levels of the Hspd1 transcript in all of the tissues examined, demonstrating that the inactivation of the Hspd1 gene is efficient. By Western blot analysis, we found that the amount of Hsp60 protein, compared to either cytosolic tubulin or mitochondrial voltage-dependent anion-selective channel protein 1/porin, was decreased as well. The expression of the nearby Hspe1 gene, which encodes the Hsp10 co-chaperonin, was concomitantly down regulated in the liver, and the protein levels in all tissues except the brain were reduced. Homozygous Hspd1 mutant embryos, however, died shortly after implantation (day 6.5 to 7.5 of gestation, Theiler stages 9–10). Our results demonstrate that Hspd1 is an essential gene for early embryonic development in mice, while reducing the amount of Hsp60 by inactivation of one allele of the gene is compatible with survival to term as well as postnatal life.
authors.affiliation: Research Unit for Molecular Medicine, Aarhus University Hospital, Skejby, Denmark. jhc@humgen.au.dk
authors.forename: Jane H
authors.initials: JH
authors.lastname: Christensen
authors.forename: Marit N
authors.initials: MN
authors.lastname: Nielsen
authors.forename: Jakob
authors.initials: J
authors.lastname: Hansen
authors.forename: Annette
authors.initials: A
authors.lastname: Füchtbauer
authors.forename: Ernst-Martin
authors.initials: EM
authors.lastname: Füchtbauer
authors.forename: Mark
authors.initials: M
authors.lastname: West
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Corydon
authors.forename: Niels
authors.initials: N
authors.lastname: Gregersen
authors.forename: Peter
authors.initials: P
authors.lastname: Bross
mesh_terms: D000818:Animals; D018835:Chaperonin 10 / Q000378:metabolism; D018834:Chaperonin 60 / Q000235:genetics* / Q000378:metabolism; D004622:Embryo, Mammalian / Q000473:pathology; D047108:Embryonic Development / Q000235:genetics; D005313:Fetal Death / Q000235:genetics; D005804:Genes, Lethal; D005838:Genotype; D006579:Heterozygote; D051379:Mice; D018345:Mice, Knockout; D008928:Mitochondria / Q000378:metabolism; D016254:Mutagenesis, Insertional; D010641:Phenotype; D018272:Porins / Q000378:metabolism; D015419:Spastic Paraplegia, Hereditary / Q000235:genetics / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism
chemical_list: D018835:Chaperonin 10; D018834:Chaperonin 60; D018272:Porins; D014404:Tubulin
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