We investigated the in
vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage-
and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of
healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons
containing recombinant human basic fibroblast growth factor (bFGF; 100
ng/ml). The viability of the bFGF-treated cells was
assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time
RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial
markers. Western blotting and immunocytochemical analysis for the neuronal markers were
performed to evaluate the protein expression and localization. The Ca2+
mobilization of the cells was evaluated using the Ca2+ indicator Fluo3 to
monitor Ca2+ influx. To investigate the mechanism of bFGF-induced neuronal
differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide
3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the
maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal
marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore,
in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase
in intracellular Ca2+ levels. Each inhibitor significantly attenuated the
bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF
contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive
neuron-like cells and may lead to the development of new cell-based treatments for
neuronal diseases. 相似文献
Idiopathic polyarthritis (IPA) is a very common inflammatory arthropathy in the dog.
Canine IPA is diagnosed mainly by detecting increased number of leukocytes in the synovial
fluid (SF), which is easily influenced by glucocorticoid therapy. We obtained 31 SF
samples from 24 IPA dogs prior to (n=19) and/or after
(n=12) 1 to 10 weeks of glucocorticoid therapy. The SF total protein
concentrations of IPA dogs were significantly higher than those of dogs with non-arthritis
diseases (n=34) and healthy controls (n=10). Our data
revealed that the SF total protein concentrations are not influenced by several weeks of
glucocorticoid therapy. Hence, the SF total protein concentration is applicable as a
diagnostic marker of canine IPA even when the patients are receiving glucocorticoid
therapy. 相似文献
Somatostatin receptor 2 (SSTR2) is a negative regulator of cell proliferation in human
breast cancer. Since there is little information about SSTR2 in canine mammary gland tumor
(MGT), we clarified its distribution and expression level in normal mammary gland, benign
MGT and malignant MGT. SSTR2 expression determined by immunohistochemical staining was
observed in the cytoplasm of luminal epithelial cells. The intensity was negatively
correlated with malignancy: normal tissues and some of the benign tumors had the highest
levels, while the malignant tumors had little or no SSTR2 expression. As for the Western
blotting, SSTR2 protein level in benign tumors was significantly lower than the normal
mammary gland. On the other hand, SSTR2 protein levels in two of three malignant tumors
were higher than the other groups. These results suggest that SSTR2 expression alters
according to the malignancy of canine MGT. 相似文献
Transgenic mice are essential research tools in developmental biology studies. The 2A peptide allows multiple genes to be
expressed simultaneously at comparable levels in somatic cells, but there are no reports of it being used successfully in
germ cells. We constructed a Cre/loxP-based conditional vector containing the 2A peptide to significantly enhance the
expression of a reporter and target gene from a constitutive promoter in oocytes. Mice with a transgene insertion containing
the chicken β-actin promoter, floxed EGFP-polyA cassette, mCherry reporter, 2A peptide and target gene DNA methyltransferase
3A2 (Dnmt3a2) were crossed with TNAP- or Vasa-Cre mice to produce offspring, in which mCherry and DNMT3A2
proteins were highly expressed in oocytes upon Cre-mediated removal of EGFP-polyA. This novel transgenic mouse line based on
the 2A expression system can serve as a useful tool for examining gene function during oogenesis. 相似文献
ABSTRACT: It is important to clarify trophic dynamics in marine ecosystems for management of the fishing ground. Organic carbon sources and trophic position of pelagic fishes in the coastal waters of the south-eastern Izu Peninsula, Japan, were examined on the basis of carbon and nitrogen stable isotope distributions. The δ13C of the fishes was mostly distributed from −19 to −16‰ for nektonic fishes (13 species of adults and immatures) and planktonic fishes (10 species of larvae and juveniles), close to the δ13C values of particulate organic matter and planktonic decapods. These δ13C signatures for the inhabitants of the water column were in contrast with the high δ13C values (mainly −16 to −13‰) for demersal fishes of Scorpaeniformes and benthic polychaetes collected in the surf zone. These results indicate that nektonic and planktonic fishes depend on phytoplankton for carbon supply. The δ15N signatures suggest that the trophic position ranged 3.1–4.5 for the nektonic fishes and 2.9–3.7 for the planktonic fishes, premised on trophic level 3 for larval Japanese anchovy Engraulis japonicus . Thus, planktivorous fishes should be mainly assigned to trophic levels 3 and 4 in this area. 相似文献
Paddy and Water Environment - In this article, we examined the value of the traditional Japanese flood control measure that uses a discontinuous levee (kasumitei). By avoiding the use of... 相似文献
Although fish roes (or the egg-laden ovary) are seafood products with high nutritional value and are considered abundant in vitamins including vitamin B12, nevertheless, the detailed properties of vitamin B12 have not been fully elucidated in fish roe products. Vitamin B12 content was determined using C18-reversed-phase high-performance liquid chromatography after purification of samples from immunoaffinity columns. Salmon egg-laden ovary products (sujiko), salmon roe products (ikura), dried mullet roe products (bottarga), and white sturgeon roe products (caviar) were found to contain substantial levels of vitamin B12 (more than approximately 15 µg/100 g wet weight). Interestingly, high levels of vitamin B12 per 100 g wet weight have been measured in pollack roe products (tarako) and flying fish roe products (tobiko). Liquid chromatography–electrospray-ionization/tandem mass spectrometry analysis revealed that vitamin B12 was the predominant corrinoid in the evaluated fish roe products examined, while no inactive corrinoid compounds were identified. These results suggest that commercially available fish roe products could be an important source of vitamin B12 for use as a dietary product for humans.
General (GSI) and specific (SSI) selection indices were constructed for three open-mating populations of sugarcane (Saccharum
spp.) seedlings and their ratoon stages. Though specific indices were found to be the most effective in improving mean brix
yield in their own source populations, but the GSI was better to SSI in other populations as indicated by higher mean values
and more number of superior clones at selection and evaluation stages. This indicated the wider applicability of the GSI over
SSI. Inspite of better expected genetic advance through GSI in ratoon crop, the actual gain in brix yield attained, through
GSI, was better in seedlings.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
In this study, we used Herbaspirillum sp. B501gfp1 (B501gfp1), an isolate from wild rice, to investigate the interaction between a non-host nitrogen-fixing endophytic bacterium and micropropagated sugarcane plants under aseptic condition. Two Japanese sugarcane plants (Saccharum sp.) cultivars (cvs) NiF8 and Ni15 were inoculated using B501gfp1 in two inoculum doses of 108 and 102 bacterial-cells-per-milliliter suspension. The results showed that bacterial cells colonized both the root and stem tissues, and colonization was apparent in the intercellular spaces. Higher bacterial numbers were detected in plant tissues inoculated with the higher inoculum concentration treatment. Bacterial numbers also varied between the two cultivars, with the higher values determined in cv Ni15. This study provides evidence that Herbaspirillum sp. B501gfp1, a rice isolate, could colonize sugarcane tissues, suggesting non-specificity of host plant among endophytes. 相似文献