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161.
Peptidergic neurons of the crab,Cardisoma carnifex,in defined culture maintain characteristic morphologies under a variety of conditions 总被引:5,自引:0,他引:5
Summary Peptidergic neurons dissociated from the neurosecretory cell group, the X-organ, of adult crabs (Cardisoma carnifex) show immediate outgrowth on unconditioned plastic dishes in defined medium. Most of the neurons can be categorized as small cells, branchers or veilers. A fourth type, superlarge, found occasionally, has a soma diameter greater than 40 m and multipolar outgrowth. We report here the effects on morphology that follow alterations of the standard defined culturing conditions. The three common types of neurons are present when cells are grown in crab saline or saline with l-glutamine and glucose (saline medium). Changes of pH between 7.0 to 7.9 have no effect. Osmolarity changes cause transient varicosities in small cells. In some veilers, pits rapidly appear in the veil and then disappear within 35 min. In cultures at 26° C instead of 22° C, veilers extend processes from the initial veil in a pattern similar to branchers, and the processes of adjacent veilers sometimes form appositions. Culturing in higher [K+]o medium ([K+]o=15–110 mM; standard=11 mM) has no long-term effect, but growth is arrested by [K+]o greater than 30 mM. Cultures were also grown in media in which [Ca2+]o ranged from 0.1 M to 26 mM (standard=13 mM). Outgrowth occured from all neuronal types in all [Ca2+]o tested. Thus, the expression of different outgrowth morphologies occurs under a wide variety of culturing conditions. 相似文献
162.
C. F. C. Bonting H. W. van Veen A. Taverne G. J. J. Kortstee A. J. B. Zehnder 《Archives of microbiology》1992,158(2):139-144
The response of Acinetobacter strain 210A to low phosphate concentrations was investigated in P- or C-limited chemostat cultures. The organism accumulated poly--hydroxybutyric acid under P-deprivation, at phosphate concentrations ranging from 0.1 to 0.7 mM. The amount of biomass was proportional to the phosphate concentration in the medium and no polyphosphate was formed. When shifting a culture from P- to C-limitation phosphate was accumulated as polyphosphate. No poly--hydroxybutyrate could be detected in these cells. The amount of polyphosphate in the cell showed a hysteresis. When cultures were shifted from low to high phosphate concentrations, polyphosphate reached a maximum of about 60 mg P per gram of dry weight at about 3 times excess phosphate (ca. 2.5 mM Pi). It decreased to 45 mg P per gram dry weight at approximately 5 times the phosphate needed for growth (ca. 3.5 mM Pi). In the reverse case (high to low) polyphosphate did never exceed 45 mg P per gram dry weight. The specific activities of alkaline phosphatase and the phosphate uptake system were induced at residual Pi concentrations below the detection limit (<10 M). The specific uptake rate followed also a hysteresis. The specific activities of polyphosphatase and polyphosphate: AMP phosphotransferase increased when polyphosphate formation was possible.Abbreviations HPP
High polymeric polyphosphates
- PHB
Poly--hydroxybutyric acid
- PPn
Polyphosphate
- PQQ
Pyrrolo-quinoline quinone
- U
1 mol product formed · min-1 相似文献
163.
Toshihiro Mitaka Gerald L. Sattler Henry C. Pitot Yohichi Mochizuki 《Virchows Archiv. B, Cell pathology including molecular pathology》1992,62(1):329-335
Phenotypes of the cells developing into small colonies after days of primary culture of adult rat hepatocytes in serum-free
modified Dulbecco Modified Eagles’ medium containing 10 mM nicotinamide and 10 ng/ml epidermal growth factor were analyzed
immunocytochemically, cytochemically and ultrastructurally. Albumin, cytokeratin 8 and 18 were seen by immunocytochemical
techniques in the cells of the small colonies at Day 6. Transferrin, α-antitrypsin, ceruloplasmin, and haptoglobin, proteins
secreted by mature hepatocytes, were faintly stained in these cells as was α-fetoprotein. These proteins were secreted into
the culture medium as evidenced by immunoblot analysis. γ-Glutamyltransferase, alkaline phosphatase and glucose 6-phosphatase
were not present in the cells of the small colonies as well as the surrounding hepatocytes at Day 6 of culture. In addition,
ultrastructural examinations of the cells in the small colonies indicated that these cells not only had many characteristic
mitochondria and desmosomes, but also a few small peroxisomes. Such cells, even after 20 days in culture were proliferating,
as evidenced by the intranuclear presence of the proliferating cell nuclear antigen. The potential relation of these cells
to hepatocytes which may serve as the principal reserve for replicating hepatocytes is discussed. 相似文献
164.
Lena Lewin Mats-Olof Mattsson David K. Rassin Åke Sellström 《Neurochemical research》1992,17(4):333-337
The uptake of radioactive -aminobutyric acid (GABA) andd-aspartate and the effect of SKF 89976-A, a non-substrate inhibitor of the GABA transporter, on this uptake have been investigated. Neuronal cultures from eight-day-old chick embryos grown for three or six days in vitro, were used as a model. For comparison, we also used the P2-fraction from rat. Neuronal cultures grown for three and six days expressed high-affinity uptake systems for [3H]GABA and ford-[3H]aspartate with an increasing Vmax during this period. The lipophilic non-substrate GABA uptake inhibitor, SKF 89976-A, inhibited transporter mediated uptake of GABA both in cell cultures from chicken, and in P2-fractions from rat. The results also showed that SKF 89976-A was a poor inhibitor of the uptake ofd-aspartate. We found no non-saturable uptake ofd-aspartate. 相似文献
165.
A continuous-culture approach to the question of inorganic carbon concentration by Synechococcus species 总被引:1,自引:0,他引:1
Abstract Chemostat cultures of Synechococcus PCC7942 were established in steady state over ten generations with inorganic carbon-limiting biomass production. The bicarbonate-concentration process was not significantly induced; RuBisCo activity was increased six-fold with decreasing dissolved inorganic carbon concentration and the presence of the 42-kDa cytoplasmic membrane polypeptide was observed but not implicated in the process. 相似文献
166.
Summary High yields of protoplasts were obtained from leaves of aseptically grown plants and calli originated from different explants, in several cultivars of Cajanus cajan L. The protoplasts divided to form cell clusters in modified KM 8p medium and developed to protocolonies after dilution with liquid Caboche's medium within three to four weeks of culture. The protocolonies proliferated to form green calli on solid Caboche's medium. No shoots or plants were obtained.Abbreviations BAP
6-benzylaminopurine
- NAA
-napthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- Kin
kinetin
- Zea
zeatin
- Adn S
adenine sulphate
- GA 3
gibberellic acid 相似文献
167.
Taina Tiainen 《Plant cell reports》1992,10(12):604-607
Summary The role of ethylene in embryogenesis of cultured potato anthers was studied indirectly by testing various substances known to affect ethylene formation. The reducing agents ascorbic acid and L-cysteine prevented browning of anther cultures and significantly stimulated embryogenesis. Embryogenesis was also promoted by the use of the ethylene inhibitors AgNO3 and n-propyl-gallate and by the polyamines spermidine and putrescine. The use of the ethylene releasing compound ethrel significantly inhibited embryogenesis.Abbreviations MS
Murashige & Skoog
- PVP
polyvinylpyrrolidone
- MW
molecular weight
- ACC
1-aminocyclopropane-1-carboxylic acid
- ethrel
2-chloroethylphosphonic acid (ethephon) 相似文献
168.
A novel collagen-glycosaminoglycan (C-GAG) substrate was developed to overcome the optical opacity of a HATF nitrocellulose substrate and to provide a more physiological permeable substrate for cocultured Sertoli and spermatogenic cells. Cocultures were prepared on optically transparent C-GAG discs attached to a polyester mesh to facilitate handling. Sertoli cells displayed a cuboidal-to-columnar shape; a large number of spermatogonia and primary spermatocytes connected by intercellular bridges were associated with basolateral and apical surfaces of Sertoli cells up to 12 days after plating. Rat Sertoli-spermatogenic cell cocultures have been used for testing the effect of toxicants on rat spermatogenesis in vitro. In our initial studies, we tested the effects of the toxicant gossypol on spermatogenic cells cocultured with Sertoli cells on nonpermeable (plastic) and permeable substrates (HATF nitrocellulose) under both standard culture conditions and during perifusion after achieving a continuous electrical-resistant cell monolayer. A selective mitochondrial structural damage was observed in spermatogenic cells (spermatogonia and spermatocytes) but not in the coexisting Sertoli cells. This damage was time- (15–60 min) and dose-dependent (0.1–10µM) and developed more rapidly under perifusion conditions. Similar mitochondrial damage was reported in the intact animal but required higher concentrations (mg) and longer administration time (months) for detection. Studies are in progress to evaluate the effect of additional toxic chemical agents on functional properties of Sertoli and spermatogenic cells in cocultures prepared on various classes of C-GAG substrates.Abbreviations C-GAG,
collagen type I-glycosaminoglycans
- C-C6S,
collagen type I-chondroitin-6-sulfate
- C-H,
collagen type I-heparin 相似文献
169.
The organotypic culture of human skin keratinocytes and fibroblasts to achieve form and function 总被引:6,自引:0,他引:6
Dr. Nancy L. Parenteau Patrick Bilbo Cynthia J. M. Nolte Valerie S. Mason Mireille Rosenberg 《Cytotechnology》1992,9(1-3):163-171
We describe an organotypic model of human skin comprised of a stratified layer of human epidermal keratinocytes and dermal
fibroblasts within a contracted collagen lattice. Feasible and reproducible production of the skin construct has required
the use of traditional as well as specialized culture techniques. The configuration of the construct has been engineered to
maintain polarity and permit extended culture at the air-liquid interface. Morphological, biochemical and kinetic parameters
were assessed and functional assays were performed to determine the degree of similarity to human skin. Light and ultrastructural
morphology of the epidermis closely resembled human skin. The immunocytochemical localization of a number of differentiation
markers and extracellular matrix proteins was also similar to human skin. Kinetic data showed a transition of the epidermal
layer to a morein vivo-like growth rate during the development of the construct at the air-liquid interface. The barrier properties of the construct
also increased with time reaching a permeability to water of less than 2%·h after approximately 2 weeks at the air-liquid
interface which is still on average 30-fold more water-permeable than normal human skin. The construct is currently used forin vitro research and testing and is also being tested in clinical applications. 相似文献
170.
The interactive effect of low P supply (0, 10, 20 and 40 M) and plant age on nodule number, mass and functioning (ureide analysis technique), vegetative growth and pod production were investigated in glasshouse-grown nodulated cowpea (Vigna unguiculata L.cv. Kausband) in sand culture. Compared with 40 M P, P stress (0 M P) or very low (10 M P) supply markedly impaired nodulation, allantoin and amino-N concentrations and weight of N solutes in xylem exudates. Consequently, P stress reduced top growth and pod yields by 48 and 90%, respectively. N solutes in xylem exudates and total plant N assayed by Kjeldahl technique (as estimates of N2 fixation) responded similarly to P supply. However, the relative ureide index [(ureide-N/ureide N+amino-N)×100] remained constant (99%), irrespective of P supply, indicating the plants' complete dependency on symbiosis for growth, without implying that growth was markedly increased by N2 fixation. Although P concentrations in plant tops, roots and nodules increased with P supply, N concentrations in these plant tissues were unaffected by P supply. The concentrations of N and P in the nodules were 2–2 1/2 times higher than in plant tops. P application interacted strongly with plant age, with the largest P effect evidently achieved at the early podding stage. The significance and implications of these results are discussed. 相似文献