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21.
SUMMARY: A calponin-like protein of 45 kDa was isolated from mussel anterior byssus retractor muscle (ABRM) and its inhibitory effects on actomyosin Mg2+ -ATPase was demonstrated. The 2-D electrophoresis for ABRM myofibrils gave a spot of 45 kDa protein in addition to myofibrillar proteins such as myosin and actin. The 45 kDa protein, which was more basic and showed a slightly higher molecular weight than actin, was isolated by ion-exchange chromatography and subjected to chymotryptic digestion. N-terminal amino acid sequencing of polypeptide fragments produced gave two sequences, ASQKGMTSFGAVRHH and GMDRALISKMGSKYDSGL, both of which showed a high homology to those of vertebrate calponins and invertebrate calponin-related proteins. Furthermore, the 45 kDa protein strongly reacted with commercially available antibody raised against chicken smooth muscle calponin, demonstrating that the mussel ABRM 45 kDa protein is a new member of the calponin family. Then, actomyosin Mg2+ -ATPase activity of ABRM was measured in the presence and absence of the 45 kDa protein. The 45 kDa protein clearly inhibited actomyosin Mg2+ -ATPase activity in a dose-dependent manner as in the case of other vertebrate calponins. These results indicate that the 45 kDa calponin-like protein is involved in the thin filament-associated regulation of molluscan smooth muscle contraction, possibly of a unique contraction called catch. 相似文献
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Ryota Takeuchi Mitsuru Jimbo Fumika Tanimoto Mariko Iijima Hiroshi Yamashita Go Suzuki Saki Harii Yoshikatsu Nakano Ko Yasumoto Shugo Watabe 《Marine drugs》2021,19(3)
Many corals establish symbiosis with Symbiodiniaceae cells from surrounding environments, but very few Symbiodiniaceae cells exist in the water column. Given that the N-acetyl-d-glucosamine-binding lectin ActL attracts Symbiodiniaceae cells, we hypothesized that corals must attract Symbiodiniaceae cells using ActL to acquire them. Anti-ActL antibody inhibited acquisition of Symbiodiniaceae cells, and rearing seawater for juvenile Acropora tenuis contained ActL, suggesting that juvenile A. tenuis discharge ActL to attract these cells. Among eight Symbiodiniaceae cultured strains, ActL attracted NBRC102920 (Symbiodinium tridacnidorum) most strongly followed by CS-161 (Symbiodinium tridacnidorum), CCMP2556 (Durusdinium trenchii), and CCMP1633 (Breviolum sp.); however, it did not attract GTP-A6-Sy (Symbiodinium natans), CCMP421 (Effrenium voratum), FKM0207 (Fugacium sp.), and CS-156 (Fugacium sp.). Juvenile polyps of A. tenuis acquired limited Symbiodiniaceae cell strains, and the number of acquired Symbiodiniaceae cells in a polyp also differed from each other. The number of Symbiodiniaceae cells acquired by juvenile polyps of A. tenuis was correlated with the ActL chemotactic activity. Thus, ActL could be used to attract select Symbiodiniaceae cells and help Symbiodiniaceae cell acquisition in juvenile polyps of A. tenuis, facilitating establishment of symbiosis between A. tenuis and Symbiodiniaceae cells. 相似文献
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Shiro?Itoi Misako?Nakaya Gen?Kaneko Hidehiro?Kondo Keijiro?Sezaki Shugo?WatabeEmail author 《Fisheries Science》2005,71(6):1356-1364
ABSTRACT: Standard molecular techniques, such as sequencing and restriction fragment length polymorphism analysis after polymerase chain reaction (PCR) amplification are relatively complicated, and species identification can take a long time when using such techniques. We established a quick method, using PCR with species-specific TaqMan Minor Groove Binder (MGB) probes based on single nucleotide polymorphism (SNP) to distinguish the two eel species Anguilla japonica and Anguilla anguilla . This method can be used in processed products. Partial sequences of the mitochondrial 16S rRNA gene were compared between A. japonica and A. anguilla to design a primer pair common to both A. japonica and A. anguilla and probes specific to A. japonica and A. anguilla . Different fluorescence intensities were produced in two PCR microtubes each containing A. japonica - and A. anguilla -specific probes for one target sample. We observed the fluorescence intensity of PCR products in microtubes under ultraviolet transillumination, with similar results to those obtained by real-time PCR. Therefore, SNP-based PCR is a powerful tool for identifying materials of processed foods from either A. japonica or A. anguilla . 相似文献
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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
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冀东地区野生大豆愈伤组织诱导及其耐盐性 总被引:5,自引:1,他引:4
以冀东沿海及内地不同条件下生长的14个野生大豆为试验材料,选取下胚轴和子叶作为外植体,在四种不同成分的培养基上诱导愈伤组织,经继代观察愈伤组织的质量,同时,在含不同浓度NaCl(0,0.2%,0.4%,0.6%,0.8%,1.0%)的继代培养基上,观察愈伤组织的耐盐性,试验结果表明;诱导野生大豆愈伤组织的适宜培养基为MS 2,4-D2.0mg/L 6-BA0.5mg/L 蔗糖5% 琼脂0.8%;不同材料之间愈伤组织的耐盐性有明显差异;耐盐性强的可在0.8%NaCl浓度下生存低的只能忍耐0.2%NaCl含量;本试验确定野生大豆愈伤组织耐盐性鉴定适宜的浓度为0.6%左右;盐碱条件下生长的野生大豆一般耐盐性较强,非盐碱环境下生长的野生大豆耐盐性也存在着耐盐性较强的类型,说明生长环境与细胞水平的耐盐性之间并没有完全的对应关系。 相似文献
30.
Nobuhiko Ojima Miyuki Mekuchi Toshinao Ineno Koichi Tamaki Akio Kera Shigeharu Kinoshita Shuichi Asakawa Shugo Watabe 《Fisheries Science》2012,78(5):1051-1057
Rainbow trout Oncorhynchus mykiss is a cold-water aquaculture species, and a thermally selected strain has been developed by multigenerational high-temperature breeding in Japan. We examined the expression of heat-shock proteins as candidates responsible for thermotolerance in rainbow trout using F2 offspring from F1 hybrids produced between thermally selected and normal strains. From F2 offspring, two groups were selected for western blot analysis, namely, low- and high-thermotolerance groups (times to loss of equilibrium were <30 and ??60?min, respectively). We demonstrated that the expression levels of Hsp70, Hsp60, and Hsp40 in tail fin tissues were significantly higher in the individuals with high thermotolerance than in those with low thermotolerance under non-heat-shock conditions. In particular, Hsp70 was expressed only in the individuals with high thermotolerance. These results suggest that Hsp70 is a major protein responsible for conferring thermotolerance in rainbow trout. 相似文献