Fine surface structure of bovine acrosome-intact and reacted spermatozoa observed by atomic force microscopy     

Saeki K  Sumitomo N  Nagata Y  Kato N  Hosoi Y  Matsumoto K  Iritani A 《The Journal of reproduction and development》2005,51(2):293-298

The atomic force microscope (AFM) provides nanometer resolution, topographic data of the natural surface structure of materials. We studied the topology of the surface structure of bovine sperm heads during the acrosome reaction by AFM. In addition, we numerically analyzed the areas of the median sagittal plane of the sperm heads. Bovine frozen-thawed spermatozoa were washed, capacitated by heparin, and incubated with lysophosphatidylcholine (LPC) to induce the acrosome reaction, smeared on a cover glass, air-dried, and observed with AFM using the dynamic force (tapping) mode. AFM analysis of spermatozoa showed the clear surface structure of acrosomes, equatorial segments, postacrosomal regions and necks. Although AFM images of spermatozoa capacitated by heparin had complete acrosomes, most spermatozoa treated with LPC had no acrosomal caps as shown by AFM. These observations coincided with those obtained by light microscopy after staining with naphthol yellow S and erythrosin B. Furthermore, numerical analysis of AFM images indicated that areas of the median sagittal plane of the anterior portions of acrosome-reacted sperm heads (2679 +/- 616 pixels) were approximately 40% less than those of intact heads (4535 +/- 174 pixels, P<0.05). These results indicate that AFM can usefully observe and numerically analyze the fine surface structures of bovine spermatozoa.  相似文献   

Cloning of bovine MAIL and its mRNA expression in white blood cells of Holstein cows     

Yamaji D  Kitamura H  Kimura K  Matsushita Y  Okada H  Shiina T  Morimatsu M  Saito M 《Veterinary immunology and immunopathology》2004,98(3-4):175-184

Molecule possessing ankyrin-repeats induced by lipopolysaccharide (MAIL) is known as an IkappaB protein induced after administration of bacterial lipopolysaccharide (LPS) to mice. In the present study, we cloned bovine MAIL cDNA and examined its mRNA expression in white blood cells isolated from Holstein cows. Bovine MAIL had more than 80% amino acid identities with murine and human MAILs, highly conserved ankyrin-repeat motifs and PEST-like sequences. Bovine MAIL mRNA was undetectable in isolated peripheral white blood cells, but rapidly induced (<1h) after stimulation by LPS and lipid A in vitro in a dose-dependent manner. The lipid A-induced MAIL mRNA expression was found in polymorphonuclear cells, monocytes/macrophages and total lymphocytes, but not in T-lymphocytes. MAIL mRNA was also induced in vivo in peripheral blood leukocytes of cows after intramammary injection of Escherichia coli derived from coliform mastitis. Thus, bovine MAIL, as rodent MAILs, is induced by inflammatory stimuli in specific immune cells in vitro and in vivo, suggesting a role in inflammatory responses to bacterial infection in cattle.  相似文献   

Molecular cloning, chromosomal location, and biological activity of porcine interleukin-21     

Muneta Y  Kikuma R  Uenishi H  Hoshino T  Yoshihara K  Tanaka M  Hamashima N  Mori Y 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2004,66(3):269-275

A pig interleukin-21 (IL-21) cDNA was successfully cloned and sequenced from porcine peripheral blood lymphocytes (PBL) stimulated with 10 microg/ml concanavalin A (ConA), 10 microg/ml phytohemagglutinin P (PHA), 50 ng/ml phorbol 12-myristate 13-acetate (PMA), and 0.5 microg/ml anti-porcine CD3 antibody for 48 hr. The open reading frame of the porcine IL-21 cDNA is 459 base pairs in length and encodes 152 amino acids. The predicted amino acid sequence of the porcine IL-21 shows 86.2%, 77.7%, and 58.4% identity to the bovine, human, and murine IL-21, respectively. The porcine IL-21 gene was mapped to porcine chromosome 8 (8q22-->q23) by means of fluorescence in situ hybridization and radiation hybrid mapping, where the porcine IL-2 gene had been mapped nearby. The recombinant porcine mature IL-21 expressed by E. coli induced dose-dependent proliferation and IFN-gamma production from a human NK cell line, NK0. The porcine IL-21 identified in this study will be helpful for the enhancement of innate immune responses of pigs.  相似文献   

Developmental competence of porcine blastocysts produced in vitro   总被引：1，自引：0，他引：1  

Kikuchi K 《The Journal of reproduction and development》2004,50(1):21-28

The establishment of in vitro embryo production (IVP) system in pigs enables us to generate viable embryos with a quality equal to that of in vivo derived embryos. This technology contributes not only to developments in reproductive physiology and agriculture but also to biotechnologies for producing cloned or genetically modified pigs. The birth of piglets from in vitro matured and fertilized embryos at the two- to 4-cell stage was first achieved about 10 years ago, but it was only quite recently that piglets were produced after the transfer of IVP blastocysts. This improvement to the blastocyst stage of the in vitro culture system after in vitro maturation and fertilization can be expected to play a part in the development of an advanced IVP system. Here, we discuss the developmental ability of porcine embryos produced by our improved IVP system and the utilization of this technique in the new biotechnology.  相似文献   

Bovine hepatocyte growth factor and its receptor c-Met: cDNA cloning and expression analysis in the mammary gland     

Yamaji D  Kimura K  Watanabe A  Kon Y  Iwanaga T  Soliman MM  Ahmed MM  Saito M 《Domestic animal endocrinology》2006,30(3):239-246

Hepatocyte growth factor/scatter factor (HGF/SF) is a pleiotropic cytokine that plays a crucial role in the embryonic and postnatal development of various organs including the mammary gland. We cloned bovine HGF and its c-Met receptor cDNAs, and examined their expression during mammary gland development in dairy cows. The 2.5-kbp HGF cDNA clone contained a 2190 bp open reading frame coding a 730 amino acid protein, while the 4.8-kbp c-Met cDNA clone contained a 4152 bp open reading frame coding a 1384 amino acid protein. The bovine HGF and c-Met sequences exhibited more than 87% identity with those of other mammals. RT-PCR analysis revealed ubiquitous expression of both HGF and c-Met mRNAs in various bovine tissues tested. HGF mRNA was detected only in the inactive stage of bovine mammary gland development and not in the developing, lactating, and involuting stages, while c-Met mRNA was detected in the inactive and involuting stages. Immunohistochemical analysis demonstrated that the c-Met protein was found on mammary epithelial cells in the inactive, developing, and involuting stages, and on myoepithelial cells in all stages. These results suggest pivotal roles of HGF and c-Met in the development of bovine mammary gland.  相似文献   

Magnetic resonance imaging of alveolar echinococcosis experimentally induced in the rat lung     

Asanuma T  Kawahara T  Inanami O  Nakao M  Nakaya K  Ito A  Takiguchi M  Hashimoto A  Kuwabara M 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2006,68(1):15-20

Pulmonary alveolar echinococcosis (AE) caused by the metacestode of Echinococcus multilocularis is a lethal zoonosis and is a lesion secondarily induced by hematogenous dissemination from hepatic AE lesions. In the present study, a hematogenous pulmonary AE model was experimentally induced in rats by the injection of echinococcal larval tissue homogenate to the tail vein, and then the pathological and diagnostic aspects of pulmonary AE were examined by magnetic resonance imaging (MRI). Histological primary, mature and degenerated AE lesions were observed 5, 18 and 50 weeks after injection, respectively. These lesions were discriminated as signal-void, hypointense and hyperintense regions in T1-weighted MRI (T1WI), respectively. The change in signal intensity in T1WI might reflect the content of proteinaceous fluid as a result of AE cyst degeneration. Western blot analysis of sera with antibodies of two epitopes (Em18 and Em16) of E. multilocularis provided evidence for AE infection in the early stage. T1WI in combination with Western blot analysis could possibility become definitive and early signs of hematogenous pulmonary AE infection.  相似文献   

In vitro and in vivo developmental ability of oocytes derived from porcine primordial follicles xenografted into nude mice     

Kikuchi K  Kaneko H  Nakai M  Noguchi J  Ozawa M  Ohnuma K  Kashiwazaki N 《The Journal of reproduction and development》2006,52(1):51-57

We evaluated the developmental ability of oocytes in porcine primordial follicles xenografted into nude mice. Ovarian tissues from 20-day-old piglets, in which most of the follicles were primordial, were transplanted under the kidney capsules of ovariectomized nude mice. Forty-nine to 89 days after grafting (mean +/- SEM, 66.9 +/- 1.9 days; n = 64), the host mice showed the presence of cornified epithelial cells in their vaginal smears for the first time. The mice were then treated with 4 IU of equine chorionic gonadotropin (eCG) 60 days after first detection of vaginal cornification. Oocytes were collected from the host mice 48 h after treatment with eCG, and then matured. The maturation rates, based on the incidence of first polar body, ranged from 25.1% to 42.5%. They were then fertilized in vitro and cultured in vitro for 6 days, or transferred into estrous-synchronized recipients and recovered after 6 days. On Day 6 of culture, 15.4% of the matured oocytes had cleaved to the 2- to 8-cell stage. However, neither the embryos cultured in vitro nor those transferred and recovered developed to advanced embryonic stages, such as morulae or blastocysts. This result suggests that the developmental ability of xenografted oocytes is insufficient, even after in vitro maturation. Further strategies, such as improvement of hormonal treatment for host mice, are required to enable oocytes in xenografted ovarian tissues to acquire the cytoplasmic maturation necessary for embryonic development.  相似文献   

Expression and subcellular localization of GSE protein in germ cells and preimplantation embryos     

Mizuno S  Sono Y  Matsuoka T  Matsumoto K  Saeki K  Hosoi Y  Fukuda A  Morimoto Y  Iritani A 《The Journal of reproduction and development》2006,52(3):429-438

We previously identified a novel gonad-specific expression gene (Gse) and investigated its expression during gametogenesis in the mouse testis and ovary. In this study, we generated a polyclonal antibody to GSE protein and determined the profiles of the protein's expression in germ cells and preimplantation embryos in detail using immunocytochemical and immunofluorescence staining. In a Western blot analysis, the anti-GSE antibody recognized long and short isoforms (approximately 27.6 kDa and 23.1 kDa) of the protein in the mouse testis and the long isoform in the ovary. In the mouse testis, GSE protein was expressed in spermatocytes I in the pachytene stage, round spermatids, and elongated spermatids. In the mouse ovary, the protein was located in the cytoplasm and nucleus of all oocytes regardless of the stage of the ovarian follicles. In preimplantation embryos from the pronuclear to blastocyst stage, however, GSE protein was mainly detected in the nuclei of cells. At the blastocyst stage, the protein was confirmed to have accumulated in the inner cell mass (ICM), whereas it had mostly disappeared from the trophectoderm (TE). These findings suggest that GSE protein may play a role in the establishment of nuclear totipotency and may be associated with early lineage specification.  相似文献   

Genes expressed in tissue-cultured seedlings of mountain laurel (Kalmia latifolia L.) with colonizing Streptomyces padanus AOK30     

Akane Meguro  Kazuhiro Toyoda  Hiroshi Ogiyama  Sachiko Hasegawa  Tomio Nishimura  Hitoshi Kunoh  Tomonori Shiraishi 《Journal of General Plant Pathology》2012,78(5):303-310

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Identification and functional analysis of alleles for productivity in two sets of chromosome segment substitution lines of rice     

Kazuhiro Ujiie  Takayuki Kashiwagi  Ken Ishimaru 《Euphytica》2012,187(3):325-337

Using two sets of chromosome segment substitution lines (CSSLs) of crosses between cvs. ‘Koshihikari’ and ‘Kasalath’ (Ko/Ka) and between ‘Koshihikari’ and ‘Nona Bokra’ (Ko/NB), respectively, we have identified alleles for ten traits related to productivity (e.g., harvest index and biomass) in rice (Oryza sativa L.). A total of 43 chromosome regions affecting traits (CRATs) in Ko/Ka CSSLs and 40 in Ko/NB CSSLs were detected. Among them, ten and 18 CRATs in Ko/Ka and Ko/NB CSSLs, respectively, had positive effects. A CRAT for harvest index (HI) with a positive allele from ‘Kasalath’ on chromosome 6 (tentatively named HI6) increased the HI by 25 % relative to ‘Koshihikari’, raising it to the theoretical upper limit in rice (0.6). Functional analysis using CSSLs with HI6 indicated that HI6 reduced the size of the lower parts of the plant, which is not important for production, while maintaining the size of the other organs related to production (e.g., flag leaf and panicle), resulting in improved nitrogen (N) use efficiency. These results suggest an ‘ideal plant type’ with improved N use efficiency that can sustain higher yields. A CRAT for the SPAD (soil plant analysis development) value, which is a chlorophyll meter value commonly used as an indicator of leaf N content and strongly associated with the source ability of a leaf, with a ‘Nona Bokra’ allele on chromosome 4 increased the value by 13 % relative to ‘Koshihikari’ with no loss of leaf area. These CRATs can be used for the improvement of rice productivity.  相似文献