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201.
Differential Requirements of Sodium for Coupling of Cannabinoid Receptors to Adenylyl Cyclase in Rat Brain Membranes 总被引:1,自引:0,他引:1
Abstract: Sodium is generally required for optimal inhibition of adenylyl cyclase by Gl/o-coupled receptors. Canna-binoids bind to specific receptors that act like other members of the Gl/o-coupled receptor superfamily to inhibit adenylyl cyclase. However, assay of cannabinoid inhibition of adenylyl cyclase in rat cerebellar membranes revealed that concentrations of NaCI ranging from 0 to 150 mM had no effect on agonist inhibition. This lack of effect of sodium was not unique to cannabinoid receptors, because the same results were observed using baclofen as an agonist for GABAB receptors in cerebellar membranes. The lack of sodium dependence was region-specific, because assay of cannabinoid and opioid inhibition of adenylyl cyclase in striatum revealed an expected sodium dependence, with 50 mM NaCI providing maximal inhibition levels by both sets of agonists. This difference in sodium requirements between these two regions was maintained at the G protein level, because agonist-stimulated low Km GTPase activity was maximal at 50 mM NaCI in striatal membranes, but was maximal in the absence of NaCI in cerebellar membranes. Assay of [3H]WIN 55212–2 binding in cerebellar membranes revealed that the binding of this labeled agonist was sensitive to sodium and guanine nucleotides like other Gl/o-coupled receptors, because both NaCI and the nonhydrolyzable GTP analogue Gpp(NH)p significantly inhibited binding. These results suggest that differences in receptor-G protein coupling exist for cannabinoid receptors between these two brain regions. 相似文献
202.
Gordon Y. K. Ng †Brian F. O'Dowd ‡Mirelle Caron ‡Michael Dennis §Mark R. Brann † Susan R. George 《Journal of neurochemistry》1994,63(5):1589-1595
Abstract: We have expressed and biochemically characterized the human D2long (D2L ) dopamine receptor isoform using the baculovirus/Sf9 cell system. The expressed receptor bound ligands with a pharmacological profile similar to that reported for neuronal and cloned D2L receptors expressed in mammalian cell lines. Dopamine binding to D2L receptor was sensitive to guanine nucleotides, indicating receptor coupling to endogenous G proteins. A D2L receptor-specific antibody identified two major protein species at ∼44 kDa and at ∼93 kDa in immunoblots, suggesting the presence of D2L receptor monomers and dimers. Both species were purified by immunoprecipitation from digitonin-solubilized preparation of cells expressing D2L receptor prelabeled with 32 Pi or [3 H]-palmitate. These results constitute the first direct evidence for D2L receptor phosphorylation and palmitoylation. 相似文献
203.
204.
Robert G. Gregerson Susan S. Miller Mary Petrowski J. Stephen Gantt Carroll P. Vance 《Plant molecular biology》1994,25(3):387-399
Genomic clones encoding two isozymes of aspartate aminotransferase (AAT) were isolated from an alfalfa genomic library and their DNA sequences were determined. The AAT1 gene contains 12 exons that encode a cytosolic protein expressed at similar levels in roots, stems and nodules. In nodules, the amount of AAT1 mRNA was similar at all stages of development, and was slightly reduced in nodules incapable of fixing nitrogen. The AAT1 mRNA is polyadenylated at multiple sites differing by more than 250 bp. The AAT2 gene contains 11 exons, with 5 introns located in positions identical to those found in animal AAT genes, and encodes a plastid-localized isozyme. The AAT2 mRNA is polyadenylated at a very limited range of sites. The transit peptide of AAT2 is encoded by the first two and part of the third exon. AAT2 mRNA is much more abundant in nodules than in other organs, and increases dramatically during the course of nodule development. Unlike AAT1, expression of AAT2 is significantly reduced in nodules incapable of fixing nitrogen. Phylogenetic analysis of deduced AAT proteins revealed 4 separate but related groups of AAT proteins; the animal cytosolic AATs, the plant cytosolic AATs, the plant plastid AATs, and the mitochondrial AATs. 相似文献
205.
Susan J. Vannucci 《Journal of neurochemistry》1994,62(1):240-246
Abstract: Two glucose transport proteins, GLUT1 and GLUT3, have been detected in brain. GLUT1 is concentrated in the endothelial cells of the blood-brain barrier and may be present in neurons and glia; GLUT3 is probably the major neuronal glucose transporter. Of the few studies of glucose transport in the immature brain, none has quantified GLUTS. This study used membrane isolation and immunoblotting techniques to examine the developmental expression of GLUT1 and GLUT3 in four forebrain regions, cerebral microvessels, and choroid plexus, from rats 1–30 days postnatally as compared with adults. The GLUT1 level in whole brain samples was low for 14 days, doubled by 21 days, and doubled again to attain adult levels by 30 days; there was no regional variation. The GLUT3 level in these samples was low during the first postnatal week, increased steadily to adult levels by 21–30 days, and demonstrated regional specificity. The concentration of GLUT1 in microvessels increased steadily after the first postnatal week; the GLUT1 level in choroid plexus was high at birth, decreased at 1 week, and then returned to near fetal levels. GLUT3 was not found in microvessels or choroid plexus. This study indicates that both GLUT1 and GLUT3 are developmentally regulated in rat brain: GLUT1 appears to relate to the nutrient supply and overall growth of the brain, whereas GLUT3 more closely relates to functional activity and neuronal maturation. 相似文献
206.
207.
Ralph A. Nixon Susan E. Lewis Marc Mercken Ram K. Sihag 《Neurochemical research》1994,19(11):1445-1453
Newly synthesized neurofilament proteins become highly phosphorylated within axons. Within 2 days after intravitreously injecting normal adult mice with [32P]orthophosphate, we observed that neurofilaments along the entire length of optic axons were radiolabeled by a soluble32P-carrier that was axonally transported faster than neurofilaments.32P-incorporation into neurofilament proteins synthesized at the time of injection was comparatively low and minimally influenced the labeling pattern along axons.32P-incorporation into axonal neurofilaments was considerably higher in the middle region of the optic axons. This characteristic non-uniform distribution of radiolabel remained nearly unchanged for at least 22 days. During this interval, less than 10% of the total32P-labeled neurofilaments redistributed from the optic nerve to the optic tract. By contrast, newly synthesized neurofilaments were selectively pulse-labeled in ganglion cell bodies by intravitreous injection of [35S]methionine and about 60% of this pool translocated by slow axoplasmic transport to the optic tract during the same time interval. These findings indicate that the steady-state or resident pool of neurofilaments in axons is not identical to the newly synthesized neurofilament pool, the major portion of which moves at the slowest rate of axoplasmic transport. Taken together with earlier studies, these results support the idea that, depending in part on their phosphorylation state, transported neurofilaments can interact for short or very long periods with a stationary but dynamic neurofilament lattice in axons.Special issue dedicated to Dr. Sidney Ochs. 相似文献
208.
Susan M. House 《Oecologia》1993,96(4):555-561
Pollination success in female trees was determined for a population of Neolitsea dealbata (R. Br.) Merr., a locally abundant dioecious tree pollinated by small, unspecialized insects in northern Queensland rain forest, Australia. The population consisted of a clustered group of trees with a mean male-to-female distance of 4.5 m and more isolated individuals, including females more than 90 m away from the nearest pollen source. A map of all reproductive trees was produced to determine accurate male-to-female distances. The size of the pollen source available to females was defined as a function of the distance to the nearest ten male trees and their sizes (male neighbourhood index). The rate of pollen movement to females was measured by counting pollen tubes (and the number of tubes per style) in female trees 6 days after the commencement of population flowering. The pollination rate decreased steeply to less than half when the nearest male was only 6.5 m away. Although pollen reached a female 330 m away from the nearest pollen source, only 10% of receptive flowers had been pollinated. The short flowering period (2–3 weeks) combined with the the slow rate of pollen movement means that a large proportion of flowers in isolated trees are unpollinated, confirming an earlier finding that isolated females set fewer fruits than gregarious females. The reliability of pollen transfer to females was determined by quantifying insects and their pollen loads trapped at female trees with a range of male neighbourhood indices. Quantities of insects and pollen were significantly correlated with the size of the male neighbourhood index, indicating a strong density-dependent response by vectors to flowering. Pollen was also collected from insect visitors to non-flowering trees. Females with large male neighbourhood indices received more pollen than non-flowering trees with equivalent male neighbourhood indices. However, when the male neighbourhood indices were small for both female and non-flowering trees, the changces of pollinators encountering female and non-flowering trees were similar, suggesting random movements of pollinators in sparse-flowering sub-populations. The dioecious breeding system, brief, synchronous flowering period, clustered population structure and random, opportunistic foraging behaviour of vectors interacted in a way that reduced reproduction in relatively isolated trees. These results demonstrate a mechanism for differential breeding success between trees in natural populations and emphasize the possible impact of logging regimes on pollen flow between trees. Large interconspecific distances in species-rich environments may have been a factor in the selection for synchronous flowering between trees in outcrossing tree species with generalist insect pollinators. 相似文献
209.
The temporal changes in extracellular enzyme activities in freshwater microbial biofilms were examined in two contrasting
river sites in North Wales over a 12 month period. Sites were a first order, unshaded oligotrophic upland stream (Nant Waen)
and a fourth order, mildly eutrophic river with riparian tree cover (River Clywedog). When algal populations were low, biofilms
of the more eutrophic site supported greater enzyme activities and higher population densities than the oligotrophic site.
Composition, concentration and origin of substrates available to the respective biofilm communities influenced extracellular
processing patterns. Reduction in algal populations depressed total and extracellular activities in biofilms from the first
order site, suggesting that biofilm communities here were maintained by in situ primary production. Biofilms from Nant Waen were often found to contain higher extracellular activities per cell than the
more eutrophic River Clywedog biofilms, which might represent the enhanced ability of an oligotrophic biofilm to accumulate
extracellular enzymes. In contrast, light and darkgrown River Clywedog biofilms were not enzymatically distinct, inferring
a less important role for biofilm phototrophs. Some evidence was found for increased reliance on allochthonous substrates
in the River Clywedog for biofilm maintenance. 相似文献
210.
Joseph H. Sellin Roland DeSoignie Susan Burlingame 《The Journal of membrane biology》1993,136(2):147-158
Short-chain fatty acids (SCFAs) are the predominant luminal anion in the mammalian colon. Although they are rapidly absorbed
in vivo, little is known about the mechanisms of transepithelial transport in vitro. Previous studies have suggested that
SCFA transport may be linked to Na absorption or an anion exchange mechanism. We compared the transport of propionate under
short-circuit conditions in rabbit proximal and distal colon to determine whether there were segmental differences, how SCFAs
may be linked to either Na absorption or anion transport, and whether SCFAs, as weak electrolytes, may be affected by transepithelial
pH gradients. In distal colon, propionate transport was not significantly altered by stimulation of electrogenic Na absorption,
epinephrine or Cl removal. However, a modest transepithelial pH gradient (luminal 6.8/serosal 7.4) stimulated propionate absorption.
In proximal colon, propionate transport was significantly altered by manuevers that either stimulated (lowered [Na] in the
bathing media) or inhibited (theophylline) apical Na−H exchange. Neither Cl removal, nor the anion exchange inhibitor DIDS,
nor a transepithelial bicarbonate gradient, altered propionate transport. A transepithelial pH gradient inhibited propionate
secretion, but not in a manner entirely consistent with the effect of pH on the distribution of a weak electrolyte. These
results suggest that there is significant segmental heterogeneity in colonic SCFA transport; that transepithelial propionate
fluxes are altered by changes in pH or electroneutral Na absorption (Na−H exchange), but not by chloride removal, bicarbonate
gradients or electrogenic Na absorption. Regulation of SCFA transport may be an important factor in the physiology of colonic
fluid balance. 相似文献