Genotype-independent transmission of transgenic fluorophore protein by boar spermatozoa

• Published in: PloS one 2011-11, Vol.6 (11), p.e27563-e27563
• Main Authors: Garrels, Wiebke, Holler, Stephanie, Taylor, Ulrike, Herrmann, Doris, Struckmann, Christina, Klein, Sabine, Barg-Kues, Brigitte, Nowak-Imialek, Monika, Ehling, Christine, Rath, Detlef, Ivics, Zoltán, Niemann, Heiner, Kues, Wilfried A
• Format: Article
• Language: English
• Subjects: Agriculture; Analysis; Animals; Animals, Genetically Modified; Apoptosis; Bacterial Proteins - genetics; Biology; Cell cycle; Cloning; Confocal; DNA Transposable Elements - genetics; Embryo, Mammalian; Estrus; Fertility; Fertility - genetics; Fertility - radiation effects; Fluorescence; Gene expression; Genetic aspects; Genetic engineering; Genomes; Genotype; Germ cells; Light; Luminescent Proteins - genetics; Male; Meiosis; Microscopic analysis; Offspring; Oocytes; Oxidation; Physiology; Proteins; RNA, Messenger - genetics; RNA, Messenger - metabolism; Rodents; Sperm; Spermatozoa; Spermatozoa - metabolism; Spermatozoa - radiation effects; Sus scrofa - embryology; Sus scrofa - genetics; Sus scrofa - physiology; Transcription, Genetic - radiation effects; Transgenes - genetics; Transgenic animals; Transposons; Veterinary Science; X Chromosome - genetics; X Chromosome - radiation effects; Y Chromosome - genetics; Y Chromosome - radiation effects; Y Chromosomes; Zygotes
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0027563

Recently, we generated transposon-transgenic boars (Sus scrofa), which carry three monomeric copies of a fluorophore marker gene. Amazingly, a ubiquitous fluorophore expression in somatic, as well as in germ cells was found. Here, we characterized the prominent fluorophore load in mature spermatozoa of these animals. Sperm samples were analyzed for general fertility parameters, sorted according to X and Y chromosome-bearing sperm fractions, assessed for potential detrimental effects of the reporter, and used for inseminations into estrous sows. Independent of their genotype, all spermatozoa were uniformly fluorescent with a subcellular compartmentalization of the fluorophore protein in postacrosomal sheath, mid piece and tail. Transmission of the fluorophore protein to fertilized oocytes was shown by confocal microscopic analysis of zygotes. The monomeric copies of the transgene segregated during meiosis, rendering a certain fraction of the spermatozoa non-transgenic (about 10% based on analysis of 74 F1 offspring). The genotype-independent transmission of the fluorophore protein by spermatozoa to oocytes represents a non-genetic contribution to the mammalian embryo.