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Masahiro Miyazaki Yasunori Suzuki Munehiro Oda Akira Kawai Liyan Bai Jiro Sato 《In vitro cellular & developmental biology. Plant》1989,25(9):839-848
Summary For serum-free primary culture of adult rat hepatocytes, a synthetic medium DM-160 and rat-tail collagen were selected for
the basal medium and for the culture substratum, respectively. Barbiturates, such as phenobarbital and 1-ethyl-5-isobutylbarbiturate,
efficiently supported survival of hepatocytes and maintained their morphologic features at lower concentrations under the
serum-free conditions than under the serum-supplemented conditions. However, the hepatocyte survival rates under the serum-free
conditions were lower than those under the serum-supplemented conditions in the presence or absence of barbiturates. Supplementation
of the basal medium with a combination of five groups of factors (5Fs), such as eight amino acids (Ala, Arg, Gly, Ile, Met,
Phe, Pro, and Trp), two unsaturated fatty acids (linoleate and oleate), a protease inhibitor (aprotinin), three vitamins (A,
C, and E), and five trace elements (Mn, Fe, Cu, Zn, and Se), improved the hepatocyte survival under the serum-free conditions
in the presence or absence of barbiturates. In other words, the serum could be completely substituted by the 5Fs. Hepatocyte
cultures maintained in the 5Fs-suppelemented basal medium showed excellent induction of tyrosine aminotransferase activity
in response to dexamethasone in the presence or absence of barbiturates. The efficiency of the 5Fs-supplemented basal medium
for maintaining hepatocytes was not inferior to those of other media in common use with hepatocytes, such as Williams' medium
E and Waymouth's medium MB-752/1. In conclusion, maintenance of functional hepatocytes in serum-free primary culture could
be improved by use of the new medium preparation (the 5Fs-supplemented DM-160) in the presence of barbiturates.
This work was supported by a grant no. 61771923 from the Ministry of Education, Science and Culture of Japan. 相似文献
3.
Integration of Agrobacterium T-DNA into a tobacco chromosome: Possible involvement of DNA homology between T-DNA and plant DNA 总被引:13,自引:0,他引:13
Shogo Matsumoto Yukihiro Ito Tsuyoshi Hosoi Yosuke Takahashi Yasunori Machida 《Molecular & general genetics : MGG》1990,224(3):309-316
Summary We established tobacco tumour cell lines from crown galls induced by Agrobacterium. Restriction fragments containing T-DNA/plant DNA junctions were cloned from one of the cell lines, which has a single copy of the T-DNA in a unique region of its genome. We also isolated a DNA fragment that contained the integration target site from nontransformed tobacco cells. Nucleotide sequence analyses showed that the right and left breakpoints of the T-DNA mapped ca. 7.3 kb internal to the right 25 by border and ca. 350 by internal to the left border respectively. When the nucleotide sequences around these breakpoints were compared with the sequence of the target, significant homology was seen between the region adjacent to the integration target site and both external regions of the T-DNA breakpoints. In addition, a short stretch of plant DNA in the vicinity of the integration site was deleted. This deletion seems to have been promoted by homologous recombination between short repeated sequences that were present on both sides of the deleted stretch. Minor rearrangements, which included base substitutions, insertions and deletions, also took place around the integration site in the plant DNA. These results, together with previously reported results showing that in some cases sequences homologous to those in T-DNA are present in plant DNA regions adjacent to left recombinational junctions, indicate that sequence homology between the incoming T-DNA and the plant chromosomal DNA has an important function in T-DNA integration. The homology may promote close association of both termini of a T-DNA molecule on a target sequence; then TDNA may in some cases be integrated by a mechanism at least in part analogous to homologous recombination.Shogo Matsumoto is on leave from Biochemical Research Institute, Nippon Menard Cosmetic Co., Ltd, Ogaki, Gifu-ken 503, Japan 相似文献
4.
Prosaposin Facilitates Sciatic Nerve Regeneration In Vivo 总被引:3,自引:0,他引:3
Yasunori Kotani Seiji Matsuda Masahiro Sakanaka Keiji Kondoh Shu-ichi Ueno Akira Sano 《Journal of neurochemistry》1996,66(5):2019-2025
Abstract: Prosaposin, a multifunctional protein, is the precursor of saposins, which activate sphingolipid hydrolases. In addition to acting as a precursor for saposins, prosaposin has been shown to rescue hippocampal CA1 neurons from lethal ischemic damage in vivo and to promote neurite extension of neuroblastoma cells in vitro. Here we show that prosaposin, when added to a collagen-filled nerve guide after sciatic nerve transection in guinea pigs, increased dramatically the number of regenerating nerve fibers within the guide. To identify the target neurons of prosaposin during peripheral nerve regeneration, we determined the degree of atrophy and chromatolysis of neurons in the spinal anterior horn and dorsal root ganglia on the prosaposin-treated and untreated side. The effect of prosaposin on large spinal neurons and small neurons of the dorsal root ganglion was more conspicuous. Subsequent immunohistochemistry demonstrated that the atrophy of cholinergic large neurons in the anterior horn is prevented to significant extent by prosaposin treatment. These findings suggest that prosaposin promotes peripheral nerve regeneration by acting on α-motor neurons in the anterior horn and on small sensory neurons in the dorsal root ganglion. The present study raises the possibility of using prosaposin as a tool for the treatment of peripheral nerve injuries. 相似文献
5.
Toshio Ariga Shama Bhat Takashi Kanda Masanaga Yamawaki Tadashi Tai Yasunori Kushi Takeshi Kasama Shizuo Handa Robert K. Yu 《Glycoconjugate journal》1996,13(2):135-145
We analysed the glycolipid composition of glioma cells (N-370 FG cells), which are derived from a culture of transformed human fetal glial cells. The neutral and acidic glycolipid fractions were isolated by column chromatography on DEAE-Sephadex and analysed by high-performance thin-layer chromatography (HPTLC). The neutral glycolipid fraction contained 1.6 µg of lipid-bound glucose/galactose per mg protein and consisted of GlcCer (11.4% of total neutral glycolipids), GalCer (21.5%), LacCer (21.4%), Gb4 (21.1%), and three unknown neutral glycolipids (23%). These unknown glycolipids were characterized as Lewisx (fucosylneolactonorpentaosyl ceramide; Lex), difucosylneolactonorhexaosyl ceramide (dimeric Lex), and neolactonorhexaosyl ceramide (nLc6) by an HPTLC-overlay method for glycolipids using specific mouse anti-glycolipid antibodies against glycolipid and/or liquid-secondary ion (LSI) mass spectrometry. The ganglioside fraction contained 0.6 µg of lipid-bound sialic acid per mg protein with GD1a as the predominant ganglioside species (83% of the total gangliosides) and GM3, GM2, and GM1 as minor components. Trace amounts of sialyl-Lex and the complex type of sialyl-Lex derivatives were also present. Immunocytochemical studies revealed that GD1a and GalCer were primarily localized on the surface of cell bodies. Interestingly, Lex glycolipids and sialyl-Lex were localized not only on the cell bodies but also on short cell processes. Especially, sialyl-Lex glycolipid was located on the tip of fine cellular processes. The unique localization of the Lex glycolipids suggests that they may be involved in cellular differentiation and initiation of cellular growth in this cell line. 相似文献
6.
Susumu Ikegami Yasunori Ooe Takahiko Shimizu Toshihiko Kasahara Tatsuhiko Tsuruta Masako Kijima Minoru Yoshida Teruhiko Beppu 《Development genes and evolution》1993,202(3):144-151
Summary External application of 10 rig/ml (R)-trichostatin A (TSA), a potent and specific inhibitor of mammalian histone deacetylase, to the embryo of the starfish Asterina pectinifera inhibited development during the early gastrula stage before formation of mesenchyme cells. The TSA-sensitive period was limited to the mid-blastula stage before hatching. The pulse-chase experiment clearly demonstrated that TSA induced an accumulation of acetylated histone species in blastulae through inhibition of historic deacetylation. Similar blockage of development at the early gastrula stage was observed with n-butyrate, which has been known as a weak inhibitor of historic deacetylase. These results suggest an intimate role for historic acetylation-deacetylation equilibria in starfish development.
Correspondence to: S. Ikegami 相似文献
7.
Toyomasu Tomonobu; Yamane Hisakazu; Yamaguchi Isomaro; Murofushi Noboru; Takahashi Nobutaka; Inoue Yasunori 《Plant & cell physiology》1992,33(6):695-701
Four 13-hydroxygibberellins, gibberellin A1 (GA1), 3-epi-GA1,GA19 and GA20 were identified by full-scan GC/MS in extractsof lettuce seedlings (Lactuca sativa L. cv. Grand Rapids). Theresults suggest that the early-13-hydroxylation biosyntheticpathway to GA1 functions in the lettuce seedlings. It was alsofound that GA1 is active per se in the control of hypocotylelongation in lettuce seedlings. To investigate the relationshipbetween control by light of hypocotyl elongation and levelsof endogenous GAs in lettuce, endogenous levels of GAs werequantified by radioimmunoassay in seedlings that had been grownfor 5 days in the dark (5D) and in those that had been grownfor 4 days in the dark and then under white light for 1 day(4D1L). The endogenous level of GA1 in the upper and lower partsof hypocotyls in 5D seedlings was about four times higher thanthat in 4D1L seedlings. The response of explants (hypocotylsegments with cotyledons) from dark-grown seedlings to GA1 isknown to be similar in the dark and under white light when theexplants are treated with inhibitors of the biosynthesis ofGA. Therefore, the above information suggests that the highlevel of GA1 in hypocotyls of dark-grown seedlings is responsiblefor the rapid elongation of hypocotyl, while irradiation bywhite light decreases the endogenous level of GA1 in the hypocotylswith a resultant decrease in the rate of hypocotyl elongation. (Received March 13, 1992; Accepted May 21, 1992) 相似文献
8.
Reorientation of Cortical Microtubules in the Sub-Apical Region during Tuberization in Single-Node Stem Segments of Potato in Culture 总被引:6,自引:0,他引:6
The initial events in tuberization were examined in single-nodestem segments of potato, in which the tuberization was easilyregulated in culture. The addition of 8% sucrose to the culturemedium caused the cessation of elongation of lateral shootsand the swelling of the sub-apical region of each shoot. Swellingwas first induced by lateral cell expansion, which was followedby periclinal cell division. The divided cells then expandedlaterally. The alteration in the direction of growth was accompaniedby the reorientation of arrays of cortical microtubules (MTs),which was monitored by immunofluorescence microscopy. Cellsin the sub-apical region of elongating shoots had prominenttransverse arrays of MTs. The MTs in swelling cells were orientedlongitudinally with respect to the axis of the shoot. Finally,the arrays of MTs became completely disorganized. By contrast,the elongation of lateral shoots continued in GA3-treated segmentsand the cells in the sub-apical region of such shoots retainedconspicuous transverse arrays of MTs during culture, even inthe presence of a high concentration (8%) of sucrose. (Received July 2, 1994; Accepted May 19, 1995) 相似文献
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