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   

Notch2 Transduction by Feline Leukemia Virus in a Naturally Infected Cat     

Shinya WATANABE  Jumpei ITO  Takuya BABA  Takahiro HIRATSUKA  Kyohei KUSE  Haruyo OCHI  Yukari ANAI  Masaharu HISASUE  Hajime TSUJIMOTO  Kazuo NISHIGAKI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2014,76(4):553-557

  相似文献   

Urinary free metanephrines measurement in dogs with adrenal gland diseases using a new simple liquid chromatography tandem mass spectrometry method     

Noboru SASAKI  Yoshinori IKENAKA  Yumiko INOUE  Takahiro ICHISE  Noriyuki NAGATA  Mayumi ISHIZUKA  Syouta MM NAKAYAMA  Kensuke NAKAMURA  Mitsuyoshi TAKIGUCHI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(4):648

Measurement of urinary metanephrines in spot samples is used for the diagnosis of canine pheochromocytoma (PC). We describe a simple analytical method based on liquid chromatography tandem mass spectrometry (LC-MS/MS) for measuring free metanephrine (MN) and normetanephrine (NMN) in spot urine samples. Using the developed method, we evaluated the stability of urinary free-MN and free-NMN at various storing conditions. In addition, we assessed the feasibility of urinary free-MN and -NMN measurement for diagnosing PC. Urine samples were mixed with stable isotope internal standards and thereafter purified by ultrafiltration. The purified samples were analyzed by LC-MS/MS in the multiple reaction monitoring mode after separation on a multimode octa decyl silyl column. The coefficient of variation of free-MN and -NMN measurement was 7.6% and 5.5%, respectively. The linearity range was 0.5–10 µg/l for both analytes. Degradation was less than 10% for both analytes under any of the storage conditions. The median free-NMN ratio to creatinine of 9 PC dogs (595, range 144–47,961) was significantly higher (P<0.05) than that of 13 dogs with hypercortisolism (125, range 52–224) or 15 healthy dogs (85, range 50–117). The developed method is simple and may not require acidification of spot urine. The results of this preliminary retrospective study suggest that the measurement of urinary free metanephrines is a promising tool for diagnosing canine PC.  相似文献   

Histopathological changes in the pancreas due to decreased pancreatic blood flow in a canine tachycardia-induced cardiomyopathy model     

Aritada YOSHIMURA  Takahiro OHMORI  Kokoro ITOU  Ryo ISHI  Yuri MATSUMURA  Yuhei WADA  Miori KISHIMOTO  Tomoko IWANAGA  Naoki MIURA  Kazuhiko SUZUKI  Ryuji FUKUSHIMA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2021,83(5):780

In dogs, pancreatic acinar cell injury is thought to be caused by decreased pancreatic blood flow due to heart failure. In previous our report, it demonstrated that decreased heart function causes a significant decrease in pancreatic blood flow in heart failure dog model caused by rapid ventricular pacing (RVP). However, the types of histopathological changes remain unclear. We aimed to verify the types of histopathological changes occurring in the pancreatic tissue due to decreased heart function. After RVP for 4 weeks, atrophy of pancreatic acinar cells, characterized by a decrease in zymogen granules, was observed in all areas of the pancreas. In conclusion, the result of this study suggests that attention should be paid to ischemia/hypoperfusion injury in the pancreas.  相似文献   

Development of a rapid method for identifying asparagus super-males using, N-(4-chloro-2-trifluoromethylphenyl)-N'- propoxyacetamidine to induce flowering   总被引：2，自引：0，他引：2  

Takahiro Sonoda  Hajime Iwamura  Atsuko Uragami  Masayuki Ohwada 《Euphytica》2003,129(2):169-174

In order to shorten the term for the identification of asparagus super-maleplants, a method usingN-(4-chloro-2-trifluoromethylphenyl)-N'-pro-poxyacetamidine (AM12) toinduce flowering was developed. This method is conducted as follows: seedsfrom andromonoecious flowers are treated with AM12 to induce flowersto form on the seedlings, the new male flowers are crossed with normalfemale flowers, and the progeny seeds are again treated with AM12 toidentify super-male plants from the sex ratio of the seedlings.Asparagus officinalis cv. `UC157' seeds were treated with AM12.The seedlings were induced to flower at a high frequency in 25 days. Thesex ratio of the plants was 1:1 and the male flowers had the pollengermination ability. One male flower induced by AM12 was crossed withfemale plant. This cross produced many progeny seeds. The seeds weretreated again with AM12, and induced to flower. Two super-male plantswere found among the progeny of andromonoecious flowers of the all-malecultivar `Gijnlim' within six months by this method. This method thusshortens the time for identifying super-males. Since the female flowers werefertile as well, AM12 treatment would also be effective for cross breeding.  相似文献   

Role of urine in the spawning of female rose bitterling,Rhodeus ocellatus ocellatus     

Takahiro Matsubara 《Fish physiology and biochemistry》1994,13(5):399-405

This study clarified the spawning mechanism of female rose bitterling, Rhodeus ocellatus ocellatus, with special emphasis on the physical role of urine. Ovulated females ingested a significantly greater quantity of water by drinking than non-ovulated fish. The body weight of ovulated females increased about 1.3% while it decreased 1.0% in non-ovulated females. Urine volume in the urinary bladder increased rapidly before spawning, synchronized with the spawning cycle. On the other hand, little urine remained in specimens which had completed spawning. Histological observation demonstrated that the oviduct and the ureter joined at the proximal part of the ovipositor. On the basis of these anatomical and physiological results, it is proposed that urine plays the physical role of pushing the ovulated eggs through the elongated ovipositor during spawning.  相似文献   

Polypolar spindle formation during first cell cycle in rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> embryos after heat-shock treatment     

Xuelian Zhang  Takahiro Asami  Hiroshi Onozato 《Fisheries Science》2007,73(6):1325-1331

ABSTRACT:   Heat shock has been used to inhibit cleavage for the induction of monogenic diploids or tetraploids in animals, but usually the success rate is low. Heat-shocked rainbow trout Oncorhynchus mykiss embryos were used in this histological study to clarify the causes of this low success rate. Embryos treated with hydrostatic pressure were used for comparison. After heat shock had disorganized the spindles, polypolar (tripolar or tetrapolar) spindles in addition to bipolar spindles were often reassembled soon after treatment. The embryos then completed tripolar or tetrapolar division at the first mitosis, and directly turned into three- or four-cell embryos as a result of the first cleavage. During the second mitosis, a monopolar spindle was formed in each blastomere of four-cell embryos and approximately 60% of three-cell embryos. In the remaining three-cell embryos, two of the three blastomeres formed a monopolar spindle, and the third one formed a bipolar spindle. The formation of polypoles is assumed to be caused by insufficient disorganization of daughter centrioles and splitting from the mother centriole by heat shock. Polypolar division is considered to be the cause of aneuploidy and the low success rate of chromosome set doubling. In the case of hydrostatic pressure treatment, the regenerated spindles were bipolar in almost all embryos.  相似文献   

Assemblages of vertical migratory mesopelagic fish in the transitional region of the western North Pacific   总被引：5，自引：0，他引：5  

Chiyuki Sassa †  Kouichi Kawaguchi  Takahiro Kinoshita  & Chikako Watanabe 《Fisheries Oceanography》2002,11(4):193-204

Assemblages of vertical migratory mesopelagic fish are described for the nighttime upper 20 m layer of the transitional and adjacent waters in the western North Pacific. Twenty-three mesopelagic fish species belonging to six families and 16 genera were collected during June 1996. Family Myctophidae was most speciose, represented by 17 species, followed by Sternoptychidae (two spp.) and Bathylagidae (one spp.). Six assemblages are recognized based on their species composition: Subarctic, Northern Transition, Southern Transition, Subtropical, Slope Water and Seamount Assemblages. These assemblages are also characterized by size composition and biomass of the catches. The distributions of the Subarctic, Northern Transition, Southern Transition and Subtropical Assemblages are approximately defined by the Kuroshio and Oyashio fronts and the Subarctic Boundary. Slope Water and Seamount Assemblages are closely related to the slope water around the shelf edge off Japan and waters around the Shatsky Rise and the Emperor Seamount chain, respectively. The importance of the Southern Transition Assemblage area between the Kuroshio front and Subarctic Boundary as spawning and nursery grounds of many subarctic, transitional and subtropical fishes is elucidated, based on the geographical distribution patterns of their larvae and juveniles.  相似文献   

Alterations of lymphocyte subpopulations in healthy dogs with aging and in dogs with cancer     

Watabe A  Fukumoto S  Komatsu T  Endo Y  Kadosawa T 《Veterinary immunology and immunopathology》2011,142(3-4):189-200

Changes in an individual's immune status are considered major contributing factors towards the morbidity of cancer and mortality of aging. To evaluate age-related changes in the immune status of dogs, the immunophenotypes (CD3, CD4, CD8 and CD21) of peripheral blood lymphocytes were measured in 160 healthy dogs aged from 1 to 17 years, and in 365 dogs with various tumors and at various stages. In healthy dogs, the absolute numbers of white blood cells, lymphocytes, and CD3(+), CD4(+) and CD21(+) lymphocytes decreased significantly with age. The relative percentages of lymphocytes and CD4(+) cells decreased significantly, while CD8(+) cells increased significantly with age. The CD4:CD8 ratio showed a significant age-related decrease. In contrast, dogs with tumors possessed significantly lower absolute numbers and relative percentages of all lymphocyte phenotypes, while the CD4:CD8 ratio was significantly higher than in age-matched controls. The relative percentages of CD3(+) and CD8(+) lymphocytes were significantly lower in dogs with distant metastases compared with dogs without metastases, and the CD4:CD8 ratio increased with advanced stage. These observations illustrate the significant changes in immune status with age and the presence of marked immunological defects in a large-scale study of dogs with advanced tumors.  相似文献   

Chromosome doubling of Lychnis spp. by in vitro spindle toxin treatment of nodal segments     

Takahiro Nonaka  Eriko Oka  Megumi Asano  Sachiko Kuwayama  Hiromu Tasaki  Dong-Sheng Han  Toshinari Godo  Masaru Nakano 《Scientia Horticulturae》2011

Lychnis (Caryophyllaceae) consists of about 30 species distributed throughout the temperate regions of the Northern Hemisphere, from East Asia to Europe. Many Lychnis spp. have high ornamental value and cultivated as pot or garden plants. In the present study, in vitro chromosome doubling of several Lychnis spp. was examined in order to widen their variability in horticultural traits. Initially effect of various spindle toxin treatments [100, 500 or 1000 mg l⁻¹ colchicine (COL), 10, 20 or 50 mg l⁻¹ oryzalin (ORY), or 1, 5, 10 mg l⁻¹ amiprophos-methyl (APM)] of nodal segments of a triploid genotype of L. senno (3x) was investigated on survival of nodal segments and chromosome doubling in nodal segment-derived plantlets. Significantly higher percentage (75.0%) of surviving segments after spindle toxin treatment was obtained in 10 mg l⁻¹ ORY treatment. Flow cytometry (FCM) analysis of leaf tissues showed that 9.4–13.8% of plantlets, which were derived from 10 to 20 mg l⁻¹ ORY, or 5 mg l⁻¹ APM treatments, were hexaploid (6x) or ploidy chimera (3x + 6x, 4x + 6x, 5x + 6x, 3x + 4x + 6x). The results obtained by FCM analysis were confirmed by chromosome observation in root tip cells. Thus 10 mg l⁻¹ ORY treatment of nodal segments is suitable for in vitro chromosome doubling of triploid L. senno. Efficient chromosome doubling was also achieved in diploid L. fulgens (2x) and L. sieboldii (2x) by treating nodal segments with 10 mg l⁻¹ ORY: 68.9–88.7% of nodal segments survived after ORY treatment, and 24.7–26.5% of plantlets derived from ORY-treated nodal segments were tetraploid (4x) or ploidy chimera (2x + 4x) in both species. These results indicate that the in vitro chromosome doubling method established for triploid L. senno may be applicable to a wide range of Lychnis spp. Tetraploid L. fulgens and L. sieboldii showed a compact plant form, and had thick stems and deep green leaves compared with the diploid mother plants. On the other hand, hexaploid L. senno showed very poor growth and died before flowering.  相似文献