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In migrating cells, force production relies essentially on a polarized actomyosin system, whereas the spatial regulation of actomyosin contraction and substrate contact turnover involves a complex cooperation between the microtubule (MT) and the actin filament networks (Goode, B.L., D.G. Drubin, and G. Barnes. 2000. Curr. Opin. Cell Biol., 12:63-71). Targeting and capture of MT plus ends at the cell periphery has been described, but whether or not the minus ends of these MTs are anchored at the centrosome is not known. Here, we show that release of short MTs from the centrosome is frequent in migrating cells and that their transport toward the cell periphery is blocked when dynein activity is impaired. We further show that MT release, but not MT nucleation or polymerization dynamics, is abolished by overexpression of the centrosomal MT-anchoring protein ninein. In addition, a dramatic inhibition of cell migration was observed; but, contrary to cells treated by drugs inhibiting MT dynamics, polarized membrane ruffling activity was not affected in ninein overexpressing cells. We thus propose that the balance between MT minus-end capture and release from the centrosome is critical for efficient cell migration.  相似文献   

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Photoinactivation of PSII is thought to be caused by the excessive light energy that is neither used for photosynthetic electron transport nor dissipated as heat. However, the relationship between the photoinactivation rate and excess energy has not been quantitatively evaluated. Chenopodium album L. plants grown under high-light and high-nitrogen (HL-HN) conditions show higher tolerance to photoinactivation and have higher photosynthetic capacity than the high-light and low-nitrogen (HL-LN)- and low-light and high-nitrogen (LL-HN)-grown plants. The rate of photoinactivation in the LL-HN plants was faster than that in the HL-LN, which was similar to that in the HL-HN plants, while the LL-HN and HL-LN plants had similar photosynthetic capacities [Kato et al. (2002b) Funct. Plant Biol. 29: 787]. We quantified partitioning of light energy between the electron transport and heat dissipation at the light intensities ranging from 300 to 1,800 micromol m(-2) s(-1). The maximum electron transport rate was highest in the HL-HN plants, heat dissipation was greatest in the HL-LN plants, and the excess energy, which was neither consumed for electron transport nor dissipated as heat, was greatest in the LL-HN plants. The first-order rate constant of the PSII photoinactivation was proportional to the magnitude of excess energy, with a single proportional constant for all the plants, irrespective of their growth conditions. Thus the excess energy primarily determines the rate of PSII photoinactivation. A large photosynthetic capacity in the HL-HN plants and a large heat dissipation capacity in the HL-LN plants both contribute to the protection of PSII against photoinactivation.  相似文献   

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The cyanobacterial symbionts in the fern Azolla have generally been ascribed to either the Anabaena or Nostoc genera. By using comparisons of the sequences of the phycocyanin intergenic spacer and a fragment of the 16S rRNA, we found that the cyanobiont from an Azolla belongs to neither of these genera.  相似文献   

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Summary The influence of glutamate on the GABA-activated Cl- conductance was studied in the slowly adapting stretch-receptor neuron and dactylopodite opener muscle fibre of the crayfish (Astacus astacus) using a two-microelectrode and a three-microelectrode voltage clamp, respectively. Glutamate (0.5–1.0 mM) had no effect on the GABA-activated conductance in either preparation. This indicates that the availability of the inhibitory channels for activation of GABA is not influenced by glutamate. The present results are in sharp contrast to those obtained by Franke et al. (J Comp Physiol A 159:591–609, 1986) in experiments on excised membrane patches, which suggested that glutamate is capable of both activating and desensitizing inhibitory postsynaptic channels in the crayfish opener muscle fibre.Abbreviations GABA -aminobutyric acid - GGABA and G GABA p GABA-gated conductance and peak conductance - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulphonic acid - I current - SRN stretch-receptor neuron - Vm and Vl membrane voltage in two- and three-microelectrode voltage clamp, respectively  相似文献   

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Lysosomal beta-glucuronidase shows a dual localization in mouse liver, where a significant fraction is retained in the endoplasmic reticulum (ER) by interaction with an ER-resident carboxyl esterase called egasyn. This interaction of mouse egasyn (mEg) with murine beta-glucuronidase (mGUSB) involves binding of the C-terminal 8 residues of the mGUSB to the carboxylesterase active site of the mEg. We isolated the recombinant human homologue of the mouse egasyn cDNA and found that it too binds human beta-glucuronidase (hGUSB). However, the binding appears not to involve the active site of the human egasyn (hEg) and does not involve the C-terminal 18 amino acids of hGUSB. The full-length cDNA encoding hEg was isolated from a human liver cDNA library using full-length mEg cDNA as a probe. The 1941-bp cDNA differs by only a few bases from two previously reported cDNAs for human liver carboxylesterase, allowing the anti-human carboxylesterase antiserum to be used for immunoprecipitation of human egasyn. The cDNA expressed bis-p-nitrophenyl phosphate (BPNP)-inhibitable esterase activity in COS cells. When expressed in COS cells, it is localized to the ER. The intracellular hEg coimmunoprecipitated with full-length hGUSB and with a truncated hGUSB missing the C-terminal 18-amino-acid residue when extracts of COS cells expressing both proteins were treated with anti-hGUSB antibody. It did not coimmunoprecipitate with mGUSB from extracts of coexpressing COS cells. Unlike mEg, hEg was not released from the hEg-GUSB complex with BPNP. Thus, hEg resembles mEg in that it binds hGUSB. However, it differs from mEg in that (i) it does not appear to use the esterase active site for binding since treatment with BPNP did not release hEg from hGUSB and (ii) it does not use the C terminus of GUSB for binding, since a C-terminal truncated hGUSB (the C-terminal 18 amino acids are removed) bound as well as nontruncated hGUSB. Evidence is presented that an internal segment of 51 amino acids between 228 and 279 residues contributes to binding of hGUSB by hEg.  相似文献   

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Protein tyrosine phosphorylation has been implicated in several aspects of neurite outgrowth regulation. To address specific roles in early neuronal morphogenesis, hippocampal neurons in culture were treated with the tyrosine phosphatase inhibitor orthovanadate. This treatment completely suppressed axon formation, yet enhanced formation of minor neurites. The inhibition of axonogenesis was dose dependent and occurred in parallel with a marked increase in cellular phosphotyrosine immunoreactivity, which was especially concentrated within neuritic growth cones and showed partial colocalization with f-actin. Both the blockade of axonogenesis and the elevation of phosphotyrosine were completely reversible. An additional and unexpected effect of orthovanadate was the appearance of many binucleate neurons. Immunoblotting experiments using a phosphotyrosine-specific antibody revealed an orthovanadate-induced reversible hyperphosphorylation of several protein bands, especially of two at 115 and 125 kD. These data suggest a potentially important role for tyrosine phosphatases and their phosphoprotein substrates in axonogenesis. © 1998 John Wiley & Sons, Inc. J Neurobiol 35: 17–28, 1998  相似文献   

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Brandt KD  Smith GN  Myers SL 《Biorheology》2004,41(3-4):493-502
We previously reported that intraarticular injections of hyaluronan (HA), administered prophylactically to dogs in whom knee osteoarthritis had been induced by transection of the anterior cruicate ligament, did not significantly modify the intraarticular pathology but decreased the proteogylcan concentration of the articular cartilage by as much as 30%. Because the cartilage proteoglycan concentration is directly related to the stiffness of the tissue, these results raised the possibility that intraarticular HA therapy could exacerbate OA. In the present study, using a different HA formulation, with a longer interval between intraarticular HA injection and examination of joint tissues, we found that neither prophylactic nor therapeutic administration of HA had an effect on the severity of OA pathology, the magnitude of vertical ground reaction forces generated by the unstable hind limb (a surrogate for joint pain), or the cartilage proteoglycan concentration. The data suggest that the suppression of proteoglycan synthesis induced by HA is temporary and fully reversible and that HA injections do not result in overloading of the OA extremity. A significant correlation was noted between the severity of chondropathy and the magnitude of the vertical ground reaction forces generated by the unstable limb.  相似文献   

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Weeks et al. (2006) have reported their inability to find a cline in the frequencies of the major Thr-Gly encoding length variant alleles of the period gene in Drosophila melanogaster in Eastern Australia. This is in contrast to a study by Sawyer et al. (2006), who found a cline on this continent from samples collected in 1993. Weeks et al. then cast doubt on the validity of a robust cline found for these variants in Europe by Costa et al. (1992), criticizing their molecular techniques and sampling methods. We show how these claims are unjustified, and reveal a number of potential problems in their own methodology. Finally by reanalysing the subset of their data which they state is more reliable, we suggest that their results from Australia may be reasonably consistent with our own.  相似文献   

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Many seasonally breeding vertebrate species have an associated reproductive pattern: mating behavior, gonadal activity, and peak circulating androgen levels occur simultaneously. In these species, androgens influence the expression of male mating behavior. Other species have a dissociated reproductive pattern: mating behavior occurs at a different time than peak gonadal activity. In such species, it is hypothesized that mating behavior is not dependent on androgen levels [Crews, D., 1984. Gamete production, sex hormone secretion, and mating behavior uncoupled. Horm. Behav. 18, 22-28]. The salamander Desmognathus ochrophaeus mates in the spring and fall while spermatogenesis occurs during the summer, suggesting that it has a dissociated reproductive pattern and that androgens do not mediate mating behavior. To assess whether mating behavior is regulated by gonadal androgens, we castrated males to reduce endogenous androgens and implanted testosterone propionate (TP) to restore androgen levels. Castrated males mated significantly less than did control males. Castrated males given TP mated as much as control males. Compared to controls, circulating androgen levels (both testosterone (T) and dihydrotestosterone (DHT)) were reduced in castrated males and elevated in castrated males given TP implants. We also found that plasma corticosterone (CORT) levels were strongly and positively correlated with T levels. Together, these data indicate that, although spermatogenesis is dissociated in time from mating behavior, androgens are associated with the expression of mating. Thus, the associated-dissociated dichotomy does not adequately describe the reproductive pattern of D. ochrophaeus. We discuss the limitations of the associated-dissociated framework in clarifying hormone-behavior relationships in reptiles and amphibians.  相似文献   

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Nicotinic acid adenine dinucleotide phosphate (NAADP) has recently been shown to act as a second messenger controlling intracellular Ca2+ responses in mammalian cells. Many questions remain regarding this signaling pathway, including the role of the ryanodine receptor (RyR) in NAADP-induced Ca2+ transients. Furthermore, the exact metabolic pathway responsible for the synthesis of NAADP in vivo has not been determined. Here, we demonstrate that the NAADP mediated Ca2+ release system is present in human myometrial cells. We also demonstrate that human myometrial cells use the NAADP second messenger system to generate intracellular Ca2+ transients in response to histamine. It has been proposed in the past that the NAADP system in mammalian cells is dependent on the presence of functional RyRs. Here, we observed that the histamine-induced Ca2+ transients are dependent on both the NAADP and inositol 1,4,5-trisphosphate signaling pathways but are independent of RyRs. The enzyme CD38 has been shown to catalyze the synthesis of NAADP in vitro by the base-exchange reaction. Furthermore, it has been proposed that this enzyme is responsible for the intracellular generation of NAADP in vivo. Using CD38 knockout mice, we observed that both the basal and histamine stimulated levels of NAADP are independent of CD38 and the base-exchange reaction. Our group is the first to demonstrate that NAADP is a second messenger for histamine-elicited Ca2+ transients in human myometrial cells. Furthermore, the NAADP mediated mechanism in mammalian cells can be independent of RyRs and CD38. Our data provides novel insights into the understanding of the mechanism of action and metabolism of this new second messenger system. cADP ribose; inositol 1,4,5-trisphosphate; endoplasmic reticulum; ryanodine channel; nicotinic acid adenine dinucleotide phosphate; CD38; base-exchange reaction  相似文献   

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A study was designed to evaluate whether the time of onset of puberty and fertility of young ewe lambs would be affected by oocyte pick-up conducted in single or repeated sessions during the first months of lambs' live. Five groups of lambs from the Karagouniko breed were used (A-E each n=12). In group A no treatments were applied (control group) while, laparoscopical follicular aspiration (OPU) was performed early in the third, fourth and fifth month of lambs age (groups C-E, respectively). From the second to fifth month of their age, group B lambs were aspirated four times in monthly intervals. All lambs were weighed at birth, weaning, at second month and monthly thereafter until the eighth month of age. Progesterone priming and ovarian stimulation by serial FSH administrations proceeded each OPU session. To determine onset of puberty blood progesterone concentration was assayed in samples collected initially every week and after the seventh month of age twice weekly. From the seventh month a fertile ram was introduced in each group and oestrous behavior/mating was daily monitored and recorded. Pregnancy diagnosis was carried out by transabdominal ultrasound scanning 55 days after rams' removal. At the fourth and fifth month of age group B lambs were lighter (p<0.05) than controls, but this difference was later equalized. The time of onset of puberty did not differ between groups (p=0.069) and ranged between 224 and 270 days. Some animals (n=15) entered puberty with a full-length luteal phase having progesterone concentration greater than 1ng/ml, while others (n=32) exhibited one or two short luteal phases before luteal length restoration. During the first breeding season 41 animals were fertilized and maintained pregnancy to term, without noticeable differences between groups (p=0.555). During the second breeding season, all ewes were naturally served and lambed at the expected time. It is concluded that OPU in young dairy lambs does not affect the time of onset of puberty, the endocrine profile of the lambs and it does not compromise their future fertility even if it is applied at four successive months.  相似文献   

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