排序方式: 共有29条查询结果,搜索用时 15 毫秒
21.
Participation of plasma membrane proteins in the formation of tight junction by cultured epithelial cells
下载免费PDF全文
![点击此处可从《The Journal of cell biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Measurements of the transepithelial electrical resistance correlated with freeze-fracture observations have been used to study the process of tight junction formation under various experimental conditions in monolayers of the canine kidney epithelial cell line MDCK. Cells derived from previously confluent cultures and plated immediately after trypsin- EDTA dissociation develop a resistance that reaches its maximum value of several hundred ohms-cm(2) after approximately 24 h and falls to a steady-state value of 80-150 ohms- cm(2) by 48 h. The rise in resistance and the development of tight junctions can be completely and reversibly prevented by the addition of 10 μg/ml cycloheximide at the time of plating, but not when this inhibitor is added more than 10 h after planting. Thus tight junction formation consists of separable synthetic and assembly phases. These two phases can also be dissociated and the requirement for protein synthesis after plating eliminated if, following trypsinization, the cells are maintained in spinner culture for 24 h before plating. The requirement for protein synthesis is restored, however, if cells maintained in spinner culture are treated with trypsin before plating. Actinomycin D prevents development of resistance only in monolayers formed from cells derived from sparse rather than confluent cultures, but new mRNA synthesis is not required if cells obtained from sparse cultures are maintained for 24 h in spinner culture before plating. Once a steady-state resistance has been reached, its maintenance does not require either mRNA or protein synthesis; in fact, inhibition of protein synthesis causes a rise in the resistance over a 30-h period. Following treatments that disrupt the junctions in steady- state monolayers recovery of resistance also does not require protein synthesis. These observations suggest that proteins are involved in tight junction formation. Such proteins, which do not turn over rapidly under steady-state conditions, are destroyed by trypsinization and can be resynthesized in the absence of stable cell-cell or cell-substratum contact. Messenger RNA coding for proteins involved in tight junction formation is stable except when cells are sparsely plated, and can also be synthesized without intercellular contacts or cell-substratum attachment. 相似文献
22.
A comparison of two methods for constructing evolutionary distances from a weighted contribution of transition and transversion differences 总被引:3,自引:3,他引:0
Since the initial work of Jukes and Cantor (1969), a number of procedures
have been developed to estimate the expected number of nucleotide
substitutions corresponding to a given observed level of nucleotide
differentiation assuming particular evolutionary models. Unlike the
proportion of different sites, the expected number of substitutions that
would have occurred grows linearly with time and therefore has had great
appeal as an evolutionary distance. Recently, however, a number of authors
have tried to develop improved statistical approaches for generating and
evaluating evolutionary distances (Schoniger and von Haeseler 1993;
Goldstein and Polock 1994; Tajima and Takezaki 1994). These studies clearly
show that the estimated number of nucleotide substitutions is generally not
the best estimator for use in reconstruction of phylogenetic relationships.
The reason for this is that there is often a large error associated with
the estimation of this number. Therefore, even though its expectation is
correct (i.e., on average the expected number of substitutions is
proportional to time- -but see Tajima 1993), it is not expected to be as
useful as estimators designed to have a lower variance.
相似文献
23.
WJ Richardson DD van der Voort E Wilson JE Moore Jr. 《Molecular & cellular biomechanics : MCB》2013,10(3):245-265
Non-uniform stress and strain fields are prevalent in many tissues in vivo, and often exacerbated by disease or injury. These mechanical gradients potentially play a role in contributing to pathological conditions, presenting a need for experimental tools to allow investigation of cell behavior within non-uniformly stimulated environments. Herein, we employ two in vitro cell-stretching devices (one previously published; one newly presented) capable of subjecting cells to cyclic, non-uniform stretches upon the surface of either a circular elastomeric membrane or a cylindrical PDMS tube. After 24 hours of cyclic stretch, 10T1/2 cells on both devices showed marked changes in long-axis orientation, with tendencies to align parallel to the direction of minimal deformation. The degree of this response varied depending on location within the stretch gradients. These results demonstrated the feasibility of conducting cell mechanobiology investigations with the two novel devices, while also highlighting the experimental capabilities of non-uniform mechanical environments for these types of studies. Such capabilities include robust data collection for developing mechanobiological dose-response curves, signal threshold identification, and potential spatial targeting for drug delivery. 相似文献
24.
Vittori S Mishra RC Dal Ben D Kachare D Lambertucci C Volpini R Cristalli G 《Nucleosides, nucleotides & nucleic acids》2007,26(10-12):1439-1442
In this article, we report on the synthesis of 2',3',5'-tri-O-acetyl-2-amino-1-deazaadenosine and of 2',3',5'-tri-O-acetyl-2-amino-N(6)-cyclopentyl-1-deazaadenosine, which are very versatile intermediates for the preparation of 2-substituted 1-deazaadenosine derivatives. The two synthesized compounds showed to be quite unstable, with the N(6)-substituted derivatives being less stable than the N(6)-unsubstituted counterpart, according to the calculated HOMO-LUMO energy gap. Stability studies were performed through HPLC-MS analysis. 相似文献
25.
Catia Lambertucci Ippolito Antonini Michela Buccioni Diego Dal Ben Dhuldeo D. Kachare Rosaria Volpini Karl-Norbert Klotz Gloria Cristalli 《Bioorganic & medicinal chemistry》2009,17(7):2812-2822
Importance of making available selective adenosine receptor antagonists is boosted by recent findings of adenosine involvement in many CNS dysfunctions. In the present work a series of 8-bromo-9-alkyl adenines are prepared and fully characterized in radioligand binding assays or functional cyclase experiments in respect to their interaction with all the four adenosine receptor subtypes. Results show that the presence of the bromine atom in 8-position of 9-substituted adenines promotes in general the interaction with the adenosine receptors, in particular at the A2A subtype. The present study also demonstrates that adenine derivatives could be a good starting point to obtain selective adenosine A2B receptor antagonists. 相似文献
26.
27.
The expansion of genome sequencing projects has produced accumulating evidence for lateral transfer of genes between prokaryotic
and eukaryotic genomes. However, it remains controversial whether these genes are of functional importance in their recipient
host. Nikoh and Nakabachi, in a recent paper in BMC Biology, take a first step and show that two genes of bacterial origin are highly expressed in the pea aphid Acyrthosiphon pisum. Active gene expression of transferred genes is supported by three other recent studies. Future studies should reveal whether
functional proteins are produced and whether and how these are targeted to the appropriate compartment. We argue that the
transfer of genes between host and symbiont may occasionally be of great evolutionary importance, particularly in the evolution
of the symbiotic interaction itself. 相似文献
28.
Noncontact dipole effects on channel permeation. III. Anomalous proton conductance effects in gramicidin 总被引:3,自引:2,他引:1
下载免费PDF全文
![点击此处可从《Biophysical journal》网站下载免费的PDF全文](/ch/ext_images/free.gif)
LR Phillips CD Cole RJ Hendershot M Cotten TA Cross DD Busath 《Biophysical journal》1999,77(5):2492-2501
Proton transport on water wires, of interest for many problems in membrane biology, is analyzed in side-chain analogs of gramicidin A channels. In symmetrical 0.1 N HCl solutions, fluorination of channel Trp(11), Trp-(13), or Trp(15) side chains is found to inhibit proton transport, and replacement of one or more Trps with Phe enhances proton transport, the opposite of the effects on K(+) transport in lecithin bilayers. The current-voltage relations are superlinear, indicating that some membrane field-dependent process is rate limiting. The interfacial dipole effects are usually assumed to affect the rate of cation translocation across the channel. For proton conductance, however, water reorientation after proton translocation is anticipated to be rate limiting. We propose that the findings reported here are most readily interpreted as the result of dipole-dipole interactions between channel waters and polar side chains or lipid headgroups. In particular, if reorientation of the water column begins with the water nearest the channel exit, this hypothesis explains the negative impact of fluorination and the positive impact of headgroup dipole on proton conductance. 相似文献
29.
The molecular organization of the beta-globin complex of the deer mouse, Peromyscus maniculatus 总被引:1,自引:0,他引:1
Padgett RW; Loeb DD; Snyder LR; Edgell MH; Hutchison CA d 《Molecular biology and evolution》1987,4(1):30-45
Recombinant DNA clones have been isolated that contain 80 kb of the
beta-globin complex from the deer mouse, Peromyscus maniculatus.
Comparisons of this complex with that from the laboratory mouse, Mus
domesticus (with an order 5'-Hbby, Hbb-bhO, Hbb-bhl, Hbb-bh2, Hbb-bh3,
Hbb-bl, Hbb-b2 3') highlight organizational trends in the beta-globin
complex since the two species diverged. Unlike other mammals studied thus
far, the deer mouse possesses three adult genes. Partial sequence analysis
indicates that each of the three adult genes is intact and hence may be
functional. Hybridization of one of the two Mus pseudogenes, Hbb-bh3, to
genomic blots from Peromyscus reveals that it has a homologous counterpart
in Peromyscus. Homologous genes to the two gamma-like Mus genes, Hbb-bhO
and Hbb-bhl, are also found in Peromyscus. The strong hybridization between
the Hbb-bhl genes and significant nucleotide similarity between the Hbb-bhO
genes suggest that both pairs are important for the ontogeny of these mice
although no known product has been identified for the Hbb-bhO genes. The
presence of Hbb-bhO and Hbb-bhl in Peromyscus suggests that the duplication
that created this related gene set occurred before the two lineages
diverged. A single gene for Hbb-y has been isolated from Peromyscus. The
adult region in Peromyscus has undergone significant divergence from the
same region in Mus, having three rather than two adult genes, the
acquisition of at least 15 kb of extra DNA relative to Mus, and possibly
the loss of the Hbb-bh2 pseudogene. The nonadult region of the complex, in
contrast, contains the same set of genes apparently distributed over the
same amount of DNA as in the Mus beta- globin complex. This observation
suggests that the embryonic region of the complex is more evolutionarily
stable than the adult region.
相似文献