Development of a monkey model for the study of primate genomic imprinting

A. Fujimoto, Shoukhrat Mitalipov, L. L. Clepper, D. P. Wolf

    Research output: Contribution to journalArticle

    15 Citations (Scopus)

    Abstract

    An understanding of the role of imprinted genes in primate development requires the identification of suitable genetic markers that allow analysis of allele-specific expression and methylation status. Four genes, NDN (Necdin), H19, SNRPN and IGF2, known to be imprinted in mice and humans, were selected for study in rhesus monkeys along with two imprinting centres (ICs) associated with the regulation of H19/IGF2, NDN and SNRPN. GAPD was employed as a non-imprinted control gene. Primers designed to amplify polymorphic regions in these genes and ICs were based on human sequences. Genomic DNA was isolated from peripheral blood leukocytes of 93 rhesus macaques of Indian or Chinese-origin. Sequence analysis of amplicons resulted in the identification of 32 unique SNPs. Country-of-origin related differences in SNP distributions were evident. Since disruptions in imprinted gene expression and associated developmental abnormalities may result from in vitro embryo manipulation, we also examined imprinting in NDN,H19, SNRPN and IGF2 in rhesus monkey infants produced by natural mating or by ICSI. Muscle biopsies followed by RT-PCR and sequence analysis were performed in four heterozygous animals produced by natural mating and all four genes were expressed monoallelically supporting the conclusion that these genes are normally imprinted in monkeys. In the case of ICSI, five informative infants were selected based on parental analysis. Allele-specific studies indicated that the expected uniparental expression patterns were retained in animals produced from manipulated embryos. Moreover, methylation analysis revealed that CpG islands within H19/IGF2 and SNURF/ SNRPN ICs were differentially methylated. The approach described here will allow examination of imprinting in the embryos and embryonic stem cells of the monkey.

    Original languageEnglish (US)
    Pages (from-to)413-422
    Number of pages10
    JournalMolecular Human Reproduction
    Volume11
    Issue number6
    DOIs
    StatePublished - Jun 2005

    Fingerprint

    Genomic Imprinting
    snRNP Core Proteins
    Primates
    Haplorhini
    Macaca mulatta
    Genes
    Intracytoplasmic Sperm Injections
    Embryonic Structures
    Methylation
    Single Nucleotide Polymorphism
    Sequence Analysis
    Alleles
    CpG Islands
    Glyceraldehyde-3-Phosphate Dehydrogenases
    Embryonic Stem Cells
    Genetic Markers
    Leukocytes
    Biopsy
    Gene Expression
    Muscles

    Keywords

    • Development
    • Imprinted genes
    • Methylation
    • Primate
    • Single nucleotide polymorphisms

    ASJC Scopus subject areas

    • Obstetrics and Gynecology
    • Genetics
    • Developmental Biology
    • Embryology
    • Cell Biology

    Cite this

    Development of a monkey model for the study of primate genomic imprinting. / Fujimoto, A.; Mitalipov, Shoukhrat; Clepper, L. L.; Wolf, D. P.

    In: Molecular Human Reproduction, Vol. 11, No. 6, 06.2005, p. 413-422.

    Research output: Contribution to journalArticle

    Fujimoto, A. ; Mitalipov, Shoukhrat ; Clepper, L. L. ; Wolf, D. P. / Development of a monkey model for the study of primate genomic imprinting. In: Molecular Human Reproduction. 2005 ; Vol. 11, No. 6. pp. 413-422.
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