title: Comparative proteomic analysis of liver mitochondrial proteins derived from cloned adult pigs reconstructed with Meishan pig fibroblast cells and European pig enucleated oocytes.
abstract: Somatic cell nuclear transfer (SCNT) has been exploited in efforts to clone and propagate valuable animal lineages. However, in many instances, recipient oocytes are obtained from sources independent of donor cell populations. As such, influences of potential nuclear-cytoplasmic incompatibility, post SCNT, are largely unknown. In the present study, alterations in mitochondrial protein levels were investigated in adult SCNT pigs produced by microinjection of Meishan pig fetus fibroblast cells into enucleated matured oocytes (maternal Landrace genetic background). Mitochondrial fractions were prepared from liver samples by mechanical homogenization and differential centrifugation. Liver mitochondria were then subjected to two-dimensional difference gel electrophoresis (2-D DIGE). Protein expression changes were confirmed with a volume ratio greater than 2 fold (P<0.05). 2-D DIGE analysis further revealed differential expression of three proteins between the Meishan (n=3) and Landrace (n=3) breeds. Differential expression patterns of 16 proteins were detected in SCNT pig liver tissue (n=3) when compared with Meishan control samples. However, none of the 16 proteins correlated with the three differentially expressed Meishan and Landrace liver mitochondrial proteins. In summary, alteration of mitochondrial protein expression levels was observed in adult SCNT pigs that did not reflect the breed difference of the recipient oocytes. Comparative proteomic analysis represents an important tool for further studies on SCNT animals.
authors.affiliation: National Agricultural and Food Research Organization (NARO), Institute of Livestock and Grassland Science (NILGS), Ibaraki 305-0901, Japan. kumiko@affrc.go.jp
authors.forename: Kumiko
authors.initials: K
authors.lastname: Takeda
authors.forename: Mariko
authors.initials: M
authors.lastname: Tasai
authors.forename: Masaki
authors.initials: M
authors.lastname: Iwamoto
authors.forename: Mika
authors.initials: M
authors.lastname: Oe
authors.forename: Koichi
authors.initials: K
authors.lastname: Chikuni
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Takahiro
authors.initials: T
authors.lastname: Tagami
authors.forename: Keijiro
authors.initials: K
authors.lastname: Nirasawa
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Hanada
authors.forename: Carl A
authors.initials: CA
authors.lastname: Pinkert
authors.forename: Akira
authors.initials: A
authors.lastname: Onishi
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000378:metabolism*; D017849:Animals, Inbred Strains; D065150:Cellular Reprogramming; D002851:Chromatography, High Pressure Liquid; D030562:Databases, Protein; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D008930:Mitochondria, Liver / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000378:metabolism*; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D013045:Species Specificity; D034421:Sus scrofa / Q000235:genetics / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D057825:Two-Dimensional Difference Gel Electrophoresis
chemical_list: D024101:Mitochondrial Proteins; D010446:Peptide Fragments