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At least 1,531 records · Page 85Linked to original sources
PubMed1979

Sodium dependence of the nerve growth factor--regulated hexose uptake in chick embryo ganglionic cells.

Embryonic dorsal root ganglionic cells, when incubated in vitro in the absence of nerve growth factor (NGF) undergo a general metabolic degeneration which is preceded by certain changes in permeation properties. Previous studies demonstrated that NGF can rapidly modulate permeation properties which regulate the availability to the cell of an important energy source, glucose. Hexose uptake was determined by measuring the ability of the cells to accumulate [3H]labeled 2-deoxy-D-glucose. The work reported here shows that the NGF-dependent portion (about one-third) of the total specific hexose uptake was also dependent on the presence of Na+, with the apparent uptake constant (Kt) for deoxyglucose varying inversely with an external Na+ concentration of 70-140 mM; Vmax was unaffected in this range. Preincubation of ganglionic cells with 10 mM ouabain for 15-60 min, followed by a pulse with [3H]-deoxyglucose, also resulted in 50-95% reduction of the NGF-sensitive uptake. A similar pretreatment of cells with veratridine gave a 25-50% reduction in uptake. The NGF-controlled hexose uptake was also energy dependent, being diminished 50-95% after a 30-90 min preincubation with 2 mM 2,4-dinitrophenol. Uptake activities for other substrates (alpha-aminoisobutyric acid, uridine) which exhibited NGF regulation were likewise Na+-sensitive. These results indicate that availability of major energy substrates to NGF-dependent dorsal root ganglionic neurons is controlled by sodium gradients across their membranes. It is conceivable that NGF provides for maintenance and development of its target neurons by acting on such sodium gradients and, consequently, regulating the intake of essential nutrients.

Amino Acids↗
PubMed1982

Effect of 4-aminopyridine on transmitter release in synaptosomes.

The effect of 4-aminopyridine (4-AP) on the release of labeled transmitters in mouse brain synaptosomes was studied in a superfusion system. 4-AP at microM concentrations notably stimulated the spontaneous release of labeled GABA and glutamate, and of acetylcholine (ACh) derived from tritiated choline. No effects on the release of labeled alpha-aminoisobutyric acid were observed. The stimulation of GABA and ACh release was dependent on the presence of Ca2+ in the superfusion media, whereas the effect on glutamate release was more variable and no clear Ca2+-dependence was observed. In contrast to these results, 4-AP did not have any effect on the release of the above transmitters stimulated by K+-depolarization in the presence of Ca2+. These results are discussed in terms of the possible participation of Ca2+ in the action of 4-AP on spontaneous transmitter release in isolated nerve endings.

4-Aminopyridine↗
PubMed1984

Neutral amino acid transport by plasma membrane vesicles of the rabbit choroid plexus.

Uptakes of neutral L-amino acids into rabbit choroid plexus apical membrane vesicles were studied using L-[14C]amino acids. Apical membrane vesicles were prepared using Ca2+ precipitation and uptakes of 14C-labeled substrates were measured by a rapid mixing and filtration procedure. Na-dependent, concentrative uptake was observed for proline, histidine and methylaminoisobutyric acid (MeAIB). Phenylalanine and D-glucose uptakes showed no significant Na dependence. Proline uptake was a saturable function of the proline concentration with Jmax 3.5 nmol/(mg min) and Kt 0.3 mM. Competition experiments in the presence of Na indicated that proline and MeAIB are mutually competitive. Proline uptake was also inhibited 25% by phenylalanine, but MeAIB uptake was relatively unaffected. Neither proline nor MeAIB transport was significantly inhibited by glycine. We conclude that proline uptake across rabbit choroid plexus apical membrane vesicles in via an Na-dependent pathway which is shared by MeAIB and, to a minor extent, by phenylalanine, but from which glycine is excluded. Histidine uptake was inhibited by glycine, GABA, phenylalanine, proline and MeAIB. This suggests that histidine may utilize a different pathway in addition to that shared with proline and MeAIB. These transporters should play an active role in the regulation of amino acids in the CSF.

Amino Acids↗
PubMed1987

Local blood flow and vascular permeability of autonomic ganglion-transplants in the brain.

The purpose of this study was to determine whether the blood vessels of transplanted neural tissue retain their functional characteristics. Quantitative autoradiography was used to measure local blood flow (F) with iodoantipyrine and the blood-to-tissue transfer constant (K) with alpha-aminoisobutyric acid in superior cervical ganglion (SCG) allografted to the surface of ventricle IV and into the cerebellum of the same rat. The F of the intraparenchymal grafts was slightly lower than that of the intraventricular grafts; F decreased between 1 and 4 weeks in SCG grafts at both sites. The permeability-surface area (PS) product of the microvessels and extraction fraction of AIB were calculated from these results and indicated restricted transvascular passage of the amino acid in both the in situ and grafted SCG. Surface area (S) and average length (L) of the microvessels were determined morphometrically and their permeability (P) was calculated from these data. Although K and PS decreased in the grafts compared to in situ SCG, a comparable decrease in S indicated that P was similar for the microvessels of both in situ and 1-week-old SCG transplants: 3.5-4.3 x 10(-6) cm/s. Between 1 and 4 weeks after transplantation, the P of the microvessels decreased to approximately 1.6-2.3 x 10(-6) cm/s without any change in S. Thus, the blood vessels of SCG grafts within or upon the brain initially retain the functional attributes of in situ SCG microvessels, but the average permeability of the graft microvessels decreases to approximately one half of the initial value by 4 weeks after transplantation.

Alkaline Phosphatase↗
PubMed1992

Efferent microvascular responses to electrical stimulation of the area postrema in rats.

As part of its role to transduce blood-borne and afferent neural stimuli to the brain, the area postrema conducts efferent projections monosynaptically to individual nuclei of the medulla oblongata and pons. We hypothesized that electrical activation of the area postrema would mimic this transduction process and couple microvascular responses in efferent sites to local increases in tissue metabolism reported previously. We used quantitative autoradiographic techniques and image analysis to measure capillary transfer constants for [14C]alpha-aminoisobutyric acid (AIB, a small, neutral amino acid) and blood flow (iodo[14C]antipyrine) in individual brainstem structures of anesthetized rats. The area postrema was stimulated electrically by means of a monopolar microelectrode positioned stereotaxically 100 microns deep in the dorsocentral aspect of the organ. There were no significant effects of stimulation on [14C]AIB influx or blood flow in control hindbrain structures where postremal projections are sparse or absent--the spinal trigeminal nucleus, reticular formation, or cerebellar vermis. Stimulation of the area postrema produced equivalent increases in transcapillary influx of [14C]AIB and capillary blood flow in the nucleus of the solitary tract, dorsal motor nuclei of the vagus nerves, ventrolateral medullary C1 region, locus coeruleus, dorsal tegmental nuclei, and lateral parabrachial nuclei. Formation of ratios interrelating rates of [14C]AIB influx and blood flow with previously assessed values of tissue glucose metabolism indicated that these measures increased proportionately during postremal stimulation. Such proportional increases in capillary [14C]AIB transfer and blood flow during tissue activation by area postrema stimulation are consistent with interpretation that the increase in blood flow resulted from recruitment of unused surface area in the capillary networks of individual efferent nuclei.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Effects of the lipophilic anticancer drug teniposide (VM-26) on membrane transport.

The epipodophyllotoxin glucopyranosides have previously been shown to interact with membrane lipids and to alter the activity of several lipid-embedded membrane proteins. To determine if these agents are acting as general membrane perturbants, we have further examined their effects on membrane processes in Ehrlich ascites tumor cells. [3H]VM-26 and [3H]VP-16 were taken up rapidly and concentrated within the cells in proportion to their lipophilicity. Neither agent was found to have any significant effect on the influx of L-[3H]leucine or alpha-[3H]aminoisobutyric acid. Likewise, these drugs had no significant effects on the hexose transporter. The nucleoside transporter, which is structurally and functionally similar to the hexose transporter, was dramatically affected, however. VM-26 was a non-competitive inhibitor of equilibrium-exchange influx of cytosine arabinoside in Ehrlich cells with a Ki of 15 microM. Equilibrium-exchange influx increased with temperature in control cells (Q10 = 2) but not in VM-26-treated cells; thus, VM-26 was a more potent inhibitor at higher temperatures. VM-26 also significantly reduced zero-trans influx in Ehrlich, P388, L5178Y, and ML-1 cells, and these effects were immediate in onset. VM-26 inhibited high-affinity binding of the nucleoside transport inhibitor nitrobenzylmercaptopurine riboside (NBMPR), but VM-26 enhanced non-specific NBMPR binding to Ehrlich cells. The apparent specificity of the epipodophyllotoxins for the nucleoside transporter is discussed.

Amino Acids↗
PubMed1991

Gas chromatography method for the separation of amino acids enantiomers in plasma and urine. Application in a case of short bowel syndrome.

Urinary amino acids were isolated from the urine of healthy controls and a patient with a short bowel syndrome. Following derivatization with isopropyl alcohol/HCl and trifluoroacetic anhydride the amino acid enantiomers were separated by gas chromatography on a Chirasil-L-Val column. All subjects excreted D-alanine (10-30% of total Ala). The percentage D-alanine was higher in the patient with the short bowel syndrome. The excretion of D-alanine did not correlate with the D-lactate excretion. An intestinal origin for the D-amino acids is the most probable explanation.

Adolescent↗