Porcine γ-synuclein: molecular cloning, expression analysis, chromosomal localization and functional expression

  • Pernille Munk Frandsen
  • Lone Bruhn Madsen
  • Christian Bendixen
  • Knud Larsen


The γ-synuclein protein is involved in breast carcinogenesis and has also been implicated in other forms of cancer and in ocular diseases. Furthermore, γ-synuclein is believed to have a role in certain neurodegenerative diseases, such as Parkinson’s disease and Alzheimer’s disease. This work reports the cloning and characterization of the porcine (Sus scrofa) γ-synuclein cDNA (SNCG). The SNCG cDNA was amplified by reverse transcriptase polymerase chain reaction (RT-PCR) using oligonucleotide primers derived from in silico sequences. The porcine SNCG cDNA codes for a protein of 126 amino acids which shows a high similarity to bovine (90%), human (87%) and mouse (83%) γ-synuclein. A genomic clone containing the entire porcine SNCG gene was isolated and its genomic organization determined. The gene is composed of five exons, the general structure being observed to be very similar to that of the human SNCG gene. Expression analysis by quantitative real-time RT-PCR revealed the presence of SNCG transcripts in all examined organs and tissues. Differential expression was observed, with very high levels of SNCG mRNA in fat tissue and high expression levels in spleen, cerebellum, frontal cortex and pituitary gland. Expression analysis also showed that porcine SNCG transcripts could be detected in different brain regions during early stages of embryo development. The porcine SNCG orthologue was mapped to chromosome 14q25–q29. The distribution of recombinant porcine γ-synuclein was studied in three different transfected cell lines and the protein was found to be predominantly localized in the cytoplasm.


Animal models Brain Embryogenesis γ-synuclein Pig SNP 



Phosphate buffered saline


Parkinson’s disease


Single nucleotide polymorphism


Untranslated region



We gratefully acknowledge Dr. Martine Yerle of INRA Toulouse, France for providing the pig-rodent hybrid panel. The authors wish to thank Connie Jakobsen Juhl and Helle Jensen for excellent technical assistance, Dr. Dorothy K. Madsen and Dr. Mark Henryon for critically reading of the manuscript and Dr. Rikke K.K. Vingborg for help with artwork. Our thanks are also due to Dr. Ole Højbjerg for assistance with confocal microscopy. This work was supported by a grant from the Danish Parkinson Association. The sequence of the porcine SNCG cDNA, encoding the g-synuclein protein, the entire genomic sequence of SNCG, and a short SNCG promoter sequence have been submitted to DDBJ/ EMBL/GenBank under the accessions numbers EF104640, EF104639 and EF486511, respectively.


  1. 1.
    Uversky VN, Li J, Souillac P, Millett IS, Doniach S, Jakes R, Goedert M, Fink AL (2002) Biophysical properties of the synucleins and their propensities to fibrillate: inhibition of alpha-synuclein assembly by beta- and gamma-synucleins. J Biol Chem 277:11970–11978PubMedCrossRefGoogle Scholar
  2. 2.
    Maroteaux L, Scheller RH (1991) The rat brain synucleins; family of proteins transiently associated with neuronal membrane. Brain Res Mol Brain Res 11:335–343PubMedCrossRefGoogle Scholar
  3. 3.
    Ueda K, Fukushima H, Masliah E, Xia Y, Iwai A, Yoshimoto M, Otero DA, Kondo J, Ihara Y, Saitoh T (1993) Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease. Proc Natl Acad Sci USA 90:11282–11286PubMedCrossRefGoogle Scholar
  4. 4.
    Ueda K, Saitoh T, Mori H (1994) Tissue-dependent alternative splicing of mRNA for NACP, the precursor of non-A beta component of Alzheimer’s disease amyloid. Biochem Biophys Res Commun 205:1366–1372PubMedCrossRefGoogle Scholar
  5. 5.
    Jakes R, Spillantini MG, Goedert M (1994) Identification of two distinct synucleins from human brain. FEBS Lett 345:27–32PubMedCrossRefGoogle Scholar
  6. 6.
    Iwai A, Masliah E, Yoshimoto M, Ge N, Flanagan L, de Silva HA, Kittel A, Saitoh T (1995) The precursor protein of non-A beta component of Alzheimer’s disease amyloid is a presynaptic protein of the central nervous system. Neuron 14:467–475PubMedCrossRefGoogle Scholar
  7. 7.
    Lavedan C, Leroy E, Dehejia A, Buchholtz S, Dutra A, Nussbaum RL, Polymeropoulos MH (1998) Identification, localization and characterization of the human gamma-synuclein gene. Hum Genet 103:106–112PubMedCrossRefGoogle Scholar
  8. 8.
    Ninkina NN, Alimova-Kost MV, Paterson JW, Delaney L, Cohen BB, Imreh S, Gnuchev NV, Davies AM, Buchman VL (1998) Organization, expression and polymorphism of the human persyn gene. Hum Mol Genet 7:1417–1424PubMedCrossRefGoogle Scholar
  9. 9.
    Ji H, Liu YE, Jia T, Wang M, Liu J, Xiao G, Joseph BK, Rosen C, Shi YE (1997) Identification of a breast cancer-specific gene, BCSG1, by direct differential cDNA sequencing. Cancer Res 57:759–764PubMedGoogle Scholar
  10. 10.
    Surgucheva I, McMahon B, Surguchov A (2006) gamma-Synuclein has a dynamic intracellular localization. Cell Motil Cytoskeleton 63:447–458PubMedCrossRefGoogle Scholar
  11. 11.
    Buchman VL, Adu J, Pinon LG, Ninkina NN, Davies AM (1998) Persyn, a member of the synuclein family, influences neurofilament network integrity. Nat Neurosci 1:101–103PubMedCrossRefGoogle Scholar
  12. 12.
    Galvin JE, Giasson B, Hurtig HI, Lee VM, Trojanowski JQ (2000) Neurodegeneration with brain iron accumulation, type 1 is characterized by alpha-, beta-, and gamma-synuclein neuropathology. Am J Pathol 157:361–368PubMedGoogle Scholar
  13. 13.
    Bruening W, Giasson BI, Klein-Szanto AJ, Lee VM, Trojanowski JQ, Godwin AK (2000) Synucleins are expressed in the majority of breast and ovarian carcinomas and in preneoplastic lesions of the ovary. Cancer 88:2154–2163PubMedCrossRefGoogle Scholar
  14. 14.
    Gupta A, Godwin AK, Vanderveer L, Lu A, Liu J (2003) Hypomethylation of the synuclein gamma gene CpG island promotes its aberrant expression in breast carcinoma and ovarian carcinoma. Cancer Res 63:664–673PubMedGoogle Scholar
  15. 15.
    Wu K, Weng Z, Tao Q, Lin G, Wu X, Qian H, Zhang Y, Ding X, Jiang Y, Shi YE (2003) Stage-specific expression of breast cancer-specific gene gamma-synuclein. Cancer Epidemiol Biomarkers Prev 12:920–925PubMedGoogle Scholar
  16. 16.
    Inaba S, Li C, Shi YE, Song DQ, Jiang JD, Liu J (2005) Synuclein gamma inhibits the mitotic checkpoint function and promotes chromosomal instability of breast cancer cells. Breast Cancer Res Treat 94:25–35PubMedCrossRefGoogle Scholar
  17. 17.
    Bjerre D, Madsen LB, Bendixen C, Larsen K (2006) Porcine parkin: molecular cloning of PARK2 cDNA, expression analysis, and identification of a splicing variant. Biochem Biophys Res Commun 347:803–813PubMedCrossRefGoogle Scholar
  18. 18.
    Yerle M, Echard G, Robic A, Mairal A, Dubut-Fontana C, Riquet J, Pinton P, Milan D, Lahbib-Mansais Y, Gellin J (1996) A somatic cell hybrid panel for pig regional gene mapping characterized by molecular cytogenetics. Cytogenet Cell Genet 73:194–202PubMedCrossRefGoogle Scholar
  19. 19.
    Pronin AN, Morris AJ, Surguchov A, Benovic JL (2000) Synucleins are a novel class of substrates for G protein-coupled receptor kinases. J Biol Chem 275:26515–26522PubMedCrossRefGoogle Scholar
  20. 20.
    Lu A, Gupta A, Li C, Ahlborn TE, Ma Y, Shi EY, Liu J (2001) Molecular mechanisms for aberrant expression of the human breast cancer specific gene 1 in breast cancer cells: control of transcription by DNA methylation and intronic sequences. Oncogene 20:5173–5185PubMedCrossRefGoogle Scholar
  21. 21.
    Lu A, Li Q, Liu J (2006) Regulatory mechanisms for abnormal expression of the human breast cancer specific gene 1 in breast cancer cells. Sci China Life Sci 49:403–408CrossRefGoogle Scholar
  22. 22.
    Madsen LB, Thomsen B, Larsen K, Bendixen C, Holm IE, Fredholm M, Jørgensen AL, Nielsen AL (2007) Molecular characterization and temporal expression profiling of presenilins in the developing porcine brain. BMC Neurosci 8:72PubMedCrossRefGoogle Scholar
  23. 23.
    Buchman VL, Hunter HJ, Pinon LG, Thompson J, Privalova EM, Ninkina NN, Davies AM (1998) Persyn, a member of the synuclein family, has a distinct pattern of expression in the developing nervous system. J Neurosci 18:9335–9341PubMedGoogle Scholar
  24. 24.
    Specht CG, Tigaret CM, Rast GF, Thalhammer A, Rudhard Y, Schoepfer R (2005) Subcellular localisation of recombinant alpha- and gamma-synuclein. Mol Cell Neurosci 28:326–334PubMedCrossRefGoogle Scholar
  25. 25.
    Surguchov A, Surgucheva I, Solessio E, Baehr W (1999) Synoretin—A new protein belonging to the synuclein family. Mol Cell Neurosci 13:95–103PubMedCrossRefGoogle Scholar
  26. 26.
    Surgucheva I, McMahon B, Surguchov A (2006) gamma-Synuclein has a dynamic intracellular localization. Cell Motil Cytoskeleton 63:447–458PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Pernille Munk Frandsen
    • 1
  • Lone Bruhn Madsen
    • 1
  • Christian Bendixen
    • 1
  • Knud Larsen
    • 1
  1. 1.Department of Genetics and Biotechnology, Faculty of Agricultural SciencesUniversity of AarhusTjeleDenmark

Personalised recommendations