Transgenic Pigs with Pancreas-specific Expression of Green Fluorescent Protein     

Hitomi MATSUNARI  Toshihiro KOBAYASHI  Masahito WATANABE  Kazuhiro UMEYAMA  Kazuaki NAKANO  Takahiro KANAI  Taisuke MATSUDA  Masaki NAGAYA  Manami HARA  Hiromitsu NAKAUCHI  Hiroshi NAGASHIMA 《The Journal of reproduction and development》2014,60(3):230-237

The development and regeneration of the pancreas is of considerable interest because of the role of these processes in pancreatic diseases, such as diabetes. Here, we sought to develop a large animal model in which the pancreatic cell lineage could be tracked. The pancreatic and duodenal homeobox-1 (Pdx1) gene promoter was conjugated to Venus, a green fluorescent protein, and introduced into 370 in vitro-matured porcine oocytes by intracytoplasmic sperm injection-mediated gene transfer. These oocytes were transferred into four recipient gilts, all of which became pregnant. Three gilts were sacrificed at 47–65 days of gestation, and the fourth was allowed to farrow. Seven of 16 fetuses obtained were transgenic (Tg) and exhibited pancreas-specific green fluorescence. The fourth recipient gilt produced a litter of six piglets, two of which were Tg. The founder Tg offspring matured normally and produced healthy first-generation (G1) progeny. A postweaning autopsy of four 27-day-old G1 Tg piglets confirmed the pancreas-specific Venus expression. Immunostaining of the pancreatic tissue indicated the transgene was expressed in β-cells. Pancreatic islets from Tg pigs were transplanted under the renal capsules of NOD/SCID mice and expressed fluorescence up to one month after transplantation. Tg G1 pigs developed normally and had blood glucose levels within the normal range. Insulin levels before and after sexual maturity were within normal ranges, as were other blood biochemistry parameters, indicating that pancreatic function was normal. We conclude that Pdx1-Venus Tg pigs represent a large animal model suitable for research on pancreatic development/regeneration and diabetes.  相似文献   

Plasma N-Terminal Pro B-Type Natriuretic Peptide Concentrations in Dogs with Pulmonic Stenosis     

Keiya KOBAYASHI  Yasutomo HORI  Syuuichi CHIMURA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2014,76(6):827-831

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Effects of Storage Temperature and Semen Extender on Stored Canine Semen     

Tatsuya HORI  Ryuta YOSHIKUNI  Masanori KOBAYASHI  Eiichi KAWAKAMI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2014,76(2):259-263

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Unpredictable Chronic Stress-Induced Reproductive Suppression Associated with the Decrease of Kisspeptin Immunoreactivity in Male Mice     

Tetsushi HIRANO  Yoshihiro KOBAYASHI  Takuya OMOTEHARA  Atsutoshi TATSUMI  Rie HASHIMOTO  Yuria UMEMURA  Daichi NAGAHARA  Youhei MANTANI  Toshifumi YOKOYAMA  Hiroshi KITAGAWA  Nobuhiko HOSHI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2014,76(9):1201-1208

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Decreased Gene Expressions of Insulin Signal Molecules in Canine Hyperadrenocorticism     

Satoshi NOZAWA  Hitomi ODA  Ran AKIYAMA  Kaori UEDA  Kaori SAEKI  Saori SHONO  Natsuki MARUYAMA  Atsuki MURATA  Hiroyuki TAZAKI  Akihiro MORI  Yutaka MOMOTA  Daigo AZAKAMI  Toshinori SAKO  Katsumi ISHIOKA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2014,76(8):1177-1182

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Multiple high-throughput analyses monitor the response of E. coli to perturbations     

Ishii N  Nakahigashi K  Baba T  Robert M  Soga T  Kanai A  Hirasawa T  Naba M  Hirai K  Hoque A  Ho PY  Kakazu Y  Sugawara K  Igarashi S  Harada S  Masuda T  Sugiyama N  Togashi T  Hasegawa M  Takai Y  Yugi K  Arakawa K  Iwata N  Toya Y  Nakayama Y  Nishioka T  Shimizu K  Mori H  Tomita M 《Science (New York, N.Y.)》2007,316(5824):593-597

Analysis of cellular components at multiple levels of biological information can provide valuable functional insights. We performed multiple high-throughput measurements to study the response of Escherichia coli cells to genetic and environmental perturbations. Analysis of metabolic enzyme gene disruptants revealed unexpectedly small changes in messenger RNA and proteins for most disruptants. Overall, metabolite levels were also stable, reflecting the rerouting of fluxes in the metabolic network. In contrast, E. coli actively regulated enzyme levels to maintain a stable metabolic state in response to changes in growth rate. E. coli thus seems to use complementary strategies that result in a metabolic network robust against perturbations.  相似文献   

Databases for technical aspects of immunohistochemistry: 2021 update     

Takayasu Moroki  Saori Matsuo  Hirofumi Hatakeyama  Seigo Hayashi  Izumi Matsumoto  Satoshi Suzuki  Takashi Kotera  Kazuyoshi Kumagai  Kiyokazu Ozaki 《Journal of toxicologic pathology》2021,34(2):161

With the aim of sharing information about the technical aspects of immunohistochemistry (IHC) and facilitating the selection of suitable antibodies for histopathological examination, this technical report describes the results of a questionnaire distributed during the period of 2018 to 2019 among members of the Conference on Experimental Animal Histopathology. Additionally, it describes the immunological properties and supplier details (clone, supplier, catalog number, species reactivity, etc.) as well as the IHC staining conditions (fixing solution, fixing time, embedding, antigen retrieval method, antibody dilution, incubation time, incubation temperature, positive control tissue, blocking condition, secondary antibody information, etc.) for a total of 509 primary antibodies (comprising 220 different types). These survey results were an update on the contents reported by CEAH in 2017.  相似文献   

Three-dimensional live imaging of bovine embryos by optical coherence tomography     

Yasumitsu MASUDA  Ryo HASEBE  Yasushi KUROMI  Masayoshi KOBAYASHI  Misaki IWAMOTO  Mitsugu HISHINUMA  Tetsuya OHBAYASHI  Ryo NISHIMURA 《The Journal of reproduction and development》2021,67(2):149

While embryo transfer (ET) is widely practiced, many of the transferred embryos fail to develop in cattle. To establish a more effective method for selecting bovine embryos for ET, here we quantified morphological parameters of living embryos using three-dimensional (3D) images non-invasively captured by optical coherence tomography (OCT). Seven Japanese Black embryos produced by in vitro fertilization that had reached the expanded blastocyst stage after 7 days of culture were transferred after imaged by OCT. Twenty-two parameters, including thickness and volumes of the inner cell mass, trophectoderm, and zona pellucida, and volumes of blastocoel and whole embryo, were quantified from 3D images. Four of the seven recipients became pregnant. We suggest that these 22 parameters can be potentially employed to evaluate the quality of bovine embryos before ET.  相似文献   

Differentially methylated CpG sites related to fertility in Japanese Black bull spermatozoa: epigenetic biomarker candidates to predict sire conception rate     

Kumiko TAKEDA  Eiji KOBAYASHI  Kazuko OGATA  Akira IMAI  Shinya SATO  Hiromichi ADACHI  Yoichiro HOSHINO  Kagetomo NISHINO  Masahiro INOUE  Masahiro KANEDA  Shinya WATANABE 《The Journal of reproduction and development》2021,67(2):99

For semen suppliers, predicting the low fertility of service bull candidates before artificial insemination would help prevent economic loss; however, predicting bull fertility through in vitro assessment of semen is yet to be established. In the present study, we focused on the methylated CpG sites of sperm nuclear DNA and examined methylation levels to screen new biomarkers for predicting bull fertility. In frozen-thawed semen samples collected from Japanese Black bulls, for which the sire conception rate (SCR) was recorded, the methylation level of each CpG site was analyzed using human methylation microarray. According to regression analysis, 143 CpG sites related to SCR were significantly differentially methylated. Whole genome bisulfite sequence data were obtained from three semen samples and the differentially methylated regions (DMRs) that included the target CpG sites selected by human methylation microarray were confirmed. Using combined bisulfite restriction analysis, fertility-related methylation changes were detected in 10 DMRs. With the exception of one DMR, the methylation levels of these DMRs were significantly different between groups with high fertility (> 50%) and low fertility (< 40%). From multiple regression analysis of methylation levels and SCR, three DMRs were selected that could effectively predict bull fertility. We suggest that these fertility-related differences in spermatozoal methylation levels could be new epigenetic biomarkers for predicting bull fertility.  相似文献   

Simple detection method of powerful antiradical compounds in the raw extract of plants and its application for the identification of antiradical plant constituents     

Masuda T  Inaba Y  Maekawa T  Takeda Y  Yamaguchi H  Nakamoto K  Kuninaga H  Nishizato S  Nonaka A 《Journal of agricultural and food chemistry》2003,51(7):1831-1838

A simple detection method for a powerful radical scavenging compound in a mixture containing a large variety of compounds, such as the raw extract of edible plants, was developed using 1,1-diphenyl-2-picrylhydrazyl (DPPH) as the radical reagent. The method was established on the basis of the features of the typical chain-breaking antioxidation reaction mechanism, which suggests that the radical scavenging antioxidant should be converted to other stable nonradical compounds during the reaction. This method requires only a simple HPLC instrument, and the disappearance or decrease in the peak intensity, which is induced by the addition of DPPH. This change is monitored by the HPLC to detect the powerful radical scavenger from the complex mixture. The method was applied to the detection and identification of the most powerful antiradical compound in the extracts of three antioxidatively active plant extracts (Psidium guajava, Citrus depressa, and Hypericum chinense). The radical scavenging efficiency of a newly identified compound from H. chinense was also compared with that of Trolox and catechin using the method.  相似文献