Neurons but not glial cells show reciprocal imprinting of sense and antisense transcripts of Ube3a.

The human UBE3A gene shows brain-specific partial imprinting, and lack of a maternally inherited allele causes Angelman syndrome (AS), which is characterized by neurobehavioral anomalies. In several AS model mice, imprinted Ube3a expression is detected predominantly in the hippocampus, cerebellar Purkinje cells and the olfactory bulb. Therefore, imprinting of mouse Ube3a is thought to be region-specific with different levels of silencing of the paternal Ube3a allele in different brain regions. To determine cell types of imprinted Ube3a expression, we analyzed its imprinting status in embryonic brain cells by using primary cortical cell cultures. RT-PCR and immunofluorescence were performed to determine the allelic expression of the gene. The Ube3a gene encodes two RNA transcripts in the brain, sense and antisense. The sense transcript was expressed maternally in neurons but biallelically in glial cells in the embryonic brain, whereas the antisense transcript was expressed only in neurons and only from the paternal allele. Our data present evidence of brain cell type-specific imprinting, i.e. neuron-specific imprinting of Ube3a in primary brain cell cultures. Reciprocal imprinting of sense and antisense transcripts present only in neurons suggests that the neuron-specific imprinting mechanism is related to the lineage determination of neural stem cells.

[1]  Gordon Sato,et al.  Cell Culture in the Neurosciences , 2011, Current Topics in Neurobiology.

[2]  D. Ledbetter,et al.  Allele-specific expression analysis by RNA-FISH demonstrates preferential maternal expression of UBE3A and imprint maintenance within 15q11- q13 duplications. , 2002, Human molecular genetics.

[3]  G. Holmes,et al.  Neurobehavioral and Electroencephalographic Abnormalities in Ube3a Maternal-Deficient Mice , 2002, Neurobiology of Disease.

[4]  D. J. Driscoll,et al.  Distinct phenotypes distinguish the molecular classes of Angelman syndrome , 2001, Journal of medical genetics.

[5]  A. Hüttenhofer,et al.  The IC-SNURF-SNRPN transcript serves as a host for multiple small nucleolar RNA species and as an antisense RNA for UBE3A. , 2001, Human molecular genetics.

[6]  Stormy J. Chamberlain,et al.  The Prader-Willi syndrome imprinting center activates the paternally expressed murine Ube3a antisense transcript but represses paternal Ube3a. , 2001, Genomics.

[7]  S. Goderie,et al.  Timing of CNS Cell Generation A Programmed Sequence of Neuron and Glial Cell Production from Isolated Murine Cortical Stem Cells , 2000, Neuron.

[8]  T. Jessell,et al.  Progression from Extrinsic to Intrinsic Signaling in Cell Fate Specification A View from the Nervous System , 1999, Cell.

[9]  S. Tilghman The Sins of the Fathers and Mothers Genomic Imprinting in Mammalian Development , 1999, Cell.

[10]  Gregor Eichele,et al.  Mutation of the Angelman Ubiquitin Ligase in Mice Causes Increased Cytoplasmic p53 and Deficits of Contextual Learning and Long-Term Potentiation , 1998, Neuron.

[11]  Carlos Cardoso,et al.  An imprinted antisense RNA overlaps UBE3A and a second maternally expressed transcript , 1998, Nature Genetics.

[12]  G. Brewer,et al.  Electrophysiology of embryonic, adult and aged rat hippocampal neurons in serum-free culture , 1998, Journal of Neuroscience Methods.

[13]  J. Sutcliffe,et al.  Imprinted expression of the murine Angelman syndrome gene, Ube3a, in hippocampal and Purkinje neurons , 1997, Nature Genetics.

[14]  A. Hoffman,et al.  Imprinting of the Angelman syndrome gene, UBE3A, is restricted to brain , 1997, Nature Genetics.

[15]  C. Rougeulle,et al.  The Angelman syndrome candidate gene, UBE3AIE6-AP, is imprinted in brain , 1997, Nature Genetics.

[16]  M. Lalande,et al.  UBE3A/E6-AP mutations cause Angelman syndrome , 1996, Nature Genetics.

[17]  G. Brewer Serum‐free B27/neurobasal medium supports differentiated growth of neurons from the striatum, substantia nigra, septum, cerebral cortex, cerebellum, and dentate gyrus , 1995, Journal of neuroscience research.

[18]  P. Glenister,et al.  Ubiquitous expression and imprinting of Snrpn in the mouse , 1995, Mammalian Genome.

[19]  Bernhard Horsthemke,et al.  Inherited microdeletions in the Angelman and Prader–Willi syndromes define an imprinting centre on human chromosome 15 , 1995, Nature Genetics.

[20]  M. Scheffner,et al.  A family of proteins structurally and functionally related to the E6-AP ubiquitin-protein ligase. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Martin Scheffner,et al.  Protein ubiquitination involving an E1–E2–E3 enzyme ubiquitin thioester cascade , 1995, Nature.

[22]  D. Ledbetter,et al.  Imprinting analysis of three genes in the Prader-Willi/Angelman region: SNRPN, E6-associated protein, and PAR-2 (D15S225E). , 1994, Human molecular genetics.

[23]  G. Brewer,et al.  Optimized survival of hippocampal neurons in B27‐supplemented neurobasal™, a new serum‐free medium combination , 1993, Journal of neuroscience research.

[24]  B. Horsthemke,et al.  Evaluation of potential models for imprinted and nonimprinted components of human chromosome 15q11-q13 syndromes by fine-structure homology mapping in the mouse. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[25]  M. Scheffner,et al.  A cellular protein mediates association of p53 with the E6 oncoprotein of human papillomavirus types 16 or 18. , 1991, The EMBO journal.

[26]  M. Pembrey,et al.  Uniparental paternal disomy in Angelman's syndrome , 1991, The Lancet.

[27]  B. Cattanach,et al.  Autosomal and X-chromosome imprinting. , 1990, Development (Cambridge, England). Supplement.

[28]  J. Knoll,et al.  On the parental origin of the deletion in Angelman syndrome , 1989, Human Genetics.

[29]  M. Surani,et al.  Development of reconstituted mouse eggs suggests imprinting of the genome during gametogenesis , 1984, Nature.

[30]  Ping Fang,et al.  De novo truncating mutations in E6-AP ubiquitin-protein ligase gene (UBE3A) in Angelman syndrome , 1997, Nature Genetics.

[31]  S. Weber [Developmental neurology]. , 1992, Kinderkrankenschwester : Organ der Sektion Kinderkrankenpflege.