Complete Genome Sequencing and Comparative Genomic Analysis of Helicobacter Apodemus Isolated From the Wild Korean Striped Field Mouse (Apodemus agrarius) for Potential Pathogenicity.
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ABSTRACT:
SUBMITTER: Kim J
PROVIDER: S-EPMC6090156 | biostudies-literature | 2018
REPOSITORIES: biostudies-literature
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Complete Genome Sequencing and Comparative Genomic Analysis of <i>Helicobacter Apodemus</i> Isolated From the Wild Korean Striped Field Mouse <i>(Apodemus agrarius</i>) for Potential Pathogenicity.
Kim Junhyung J Kim Woohyun W An Jae-Uk JU Suh Jun Gyo JG Seong Je Kyung JK Jeon Bo-Young BY Cho Seongbeom S
Frontiers in pharmacology 20180731
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Project description:Adult ascarid worms from the field mice, Apodemus agrarius, were observed with a light and scanning electron microscope, and molecularly analized with 18S rRNA gene. In the scanning electron microscope, 3 prominent labia were present in the anterior end of male and female worms, but the interlabia and gubernaculum were absent. Scanning electron micrographs showed cervical alae as vestigial organs that looked like a slightly uplifted superficial sewing stitch. Total 6 pairs of post-cloacal papillae were observed on the tail of the male worms. The tail of female worms was blunt and conical shape with a spine-like structure, mucron. The eggs were sub-globular, coated with the albuminous layer and 73 by 82 μm in average size. The superficial pits of T. apodemi egg (mean 8.6×6.7 μm) are obviously bigger than those of Toxocara spp. The partial sequence of 18S rRNA showed the sequence homology of Toxocara canis (99.6%), Toxocara cati (99.4%), Toxascaris leonina (99.4%), and Toxocara vitulorum (99.2%). Conclusively, it was confirmed that ascarid nematodes, Toxocara apodemi, recovered from striped field mice in Korea are taxonomically conspecific relationship with genus Toxocara and genetic divergence from other Toxocara species.
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Project description:BackgroundIn rodents, the cell surface complement regulatory protein CD46 is expressed solely on the spermatozoal acrosome membrane. Ablation of the CD46 gene is associated with a faster acrosome reaction. Sperm from Apodemus flavicollis (yellow-necked field mice), A. microps (pygmy field mice) and A. sylvaticus (European wood mice) fail to express CD46 protein and exhibit a more rapid acrosome reaction rate than Mus (house mice) or BALB/c mice. A. agrarius (striped field mice) belong to a different Apodemus subgenus and have pronounced promiscuity and large relative testis size. The aim of this study was to determine whether A. agrarius sperm fail to express CD46 protein and, if so, whether A. agrarius have a faster acrosome reaction than Mus.MethodsReverse transcription polymerase chain reaction (RT-PCR) was used to assess whether A. agrarius transcribe testicular CD46 mRNA. RT-PCR was supplemented with 3'- and 5'-rapid amplification of cDNA ends to determine the complete nucleotide sequence of A. agrarius CD46. Fluorescence microscopy was used to assess whether CD46 protein is expressed by A. agrarius sperm. The acrosome status of A. agrarius sperm was calculated over time by immunocytochemistry using peanut agglutinin lectin.ResultsWe demonstrate that A. agrarius mice transcribe two unique alternatively spliced testicular CD46 mRNA transcripts, both lacking exon 7, which differ from those described previously in other Apodemus species. The larger A. agrarius CD46 transcript has an insert between exons 10 and 11 which, if translated, would result in a novel cytoplasmic tail. In addition, A. agrarius CD46 transcripts have an extended AU-rich 3'-untranslated region (UTR) and a truncated 5'-UTR, resulting in failure to express spermatozoal CD46 protein. We show that A. agrarius has a significantly faster spontaneous acrosome reaction rate than A. sylvaticus and Mus.ConclusionAbsence of CD46 protein expression is associated with acrosomal instability in rodents. A. agrarius mice express novel CD46 transcripts, resulting in the trade of spermatozoal CD46 protein expression for a rapid acrosome reaction rate, in common with other species of field mice. This provides a strategy to increase competitive sperm advantage for individuals, leading to faster fertilisation in this highly promiscuous genus.
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Project description:BackgroundHeligmosomoides polygyrus is a widespread gastro-intestinal nematode infecting wild Apodemus (wood mice) throughout Europe. Using molecular and morphological evidence, we review the status of Heligmosomoides from Apodemus agrarius in Poland previously considered to be an outlying clade of H. polygyrus, to further resolve the status of the laboratory model species, H. bakeri.MethodsMorphological analysis of the male bursa and the synlophe, and molecular analyses of concatenated nuclear (28S rDNA, ITS1 and ITS2) and mitochondrial (CO1 and cytb) genes, of Heligmosomoides collected from Apodemus agrarius from two sites in Poland and comparison with related heligmosomids from voles and mice in Eurasia.ResultsHeligmosomoides neopolygyrus, a heligmosomid nematode from Apodemus species from China and Japan, is recognised for the first time in western Europe infecting Apodemus agrarius in Poland. It can be distinguished from H. polygyrus by the filiform externo-dorsal rays of the male copulatory bursa and the small, equally distributed longitudinal crêtes on the body. Specimens from A. agrarius are 20% different at ribosomal (ITS1 and ITS2) nuclear loci, and 10% different at the mitochondrial cytb locus from H. polygyrus, and in phylogenetic analyses group with the vole-infecting genus Heligmosomum.ConclusionsDespite morphological similarity, H. neopolygyrus is only distantly related to H. polygyrus from western European Apodemus, and may be more closely related to vole-infecting taxa. It was brought into Europe by the recent rapid migration of the host mice. Inclusion of H. neopolygyrus in phylogenies makes it clear that Heligmosomoides is paraphyletic, with the pika-infecting Ohbayashinema and the vole-infecting Heligmosomum nesting within it. Clarification of the European status of H. neopolygyrus also allows H. bakeri, the laboratory model species, to be seen as a terminal sister clade to H. polygyrus, rather than as an internal clade of the latter taxon.
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