PubMed Health⌕ Search

Biomedical subjects

D L Yudilevich

Publications and source records attributed to D L Yudilevich.

At least 37 records · Page 2Linked to original sources
PubMed1987

Regulation of amino acid influx and efflux at the basolateral plasma membrane of the salivary gland epithelium: effects of parasympathetic nerve stimulation.

Basolateral amino acid transport systems in the salivary epithelium of resting and secreting cat submandibular glands were characterized by means of a rapid paired-tracer dilution technique. Amino acid uptake was measured by comparison of venous tracer concentration profiles for a labeled amino acid and D-mannitol (an extracellular tracer of similar size) following an intra-arterial bolus injection of both radioactive molecules. Unidirectional uptake of 21 amino acids, dopamine, noradrenaline, and serotonin was quantified in non-secreting glands. During 8 Hz parasympathetic nerve stimulation, significant epithelial uptakes were measured for L-[3H] alanine and L-[3H] phenylalanine, but less than 0.2% of the injected amino acid was recovered in the collected saliva. In non-secreting glands, cross-inhibition studies of L-[3H] alanine, L-[3H] phenylalanine, and L-[3H] lysine uptake by unlabeled amino acid competitors and detailed kinetic influx experiments indicated that epithelial uptake was mediated by three distinct parallel transport systems: ASC (short-chain neutral), L (branched-chain and aromatic neutral), and y+ (cationic). Rapid metabolism of alanine was inhibited by aminooxyacetate, and the metabolic uncoupler dinitrophenol selectively accelerated the efflux of transported large neutral amino acids and L-lysine. Concurrent autoradiographic experiments suggest that transport sites for small and large neutral amino acids are localized in the basolateral plasma membrane of acinar, demilunar, and striated ductal cells.

Amino Acids↗
PubMed1986

Autoradiographic localization of transported neutral amino acids in epithelia of cat submandibular and sublingual salivary glands.

Light-microscopic autoradiography was used to localize the cellular sites for neutral amino acid uptake in submandibular and sublingual salivary gland epithelia. The vasculature of isolated glands was perfused for 3-5 min with either L-(3-3H)serine or L-(4-3H)phenylalanine and then fixed by perfusion with buffered glutaraldehyde. In the submandibular gland the small neutral amino acid L-serine and the aromatic amino acid L-phenylalanine were localized to central acinar cells, demilunar cells and ductal cells. In the sublingual gland silver grains associated with each of these tritiated amino acids were localized to central acinar and ductal cells. Perfusion of both submandibular and sublingual glands with unlabelled L-serine (25 mM) or L-phenylalanine (30 mM) resulted in a significant decrease in the silver grain density associated with each labelled amino acid. The absence of silver grains in the lumina of acinar and ductal cells and the presence of tight junctions near the apical surface of the epithelium strongly suggest that the initial uptake of these amino acids was mediated by basolateral plasma membrane carriers.

Amino Acids↗
PubMed1986

Evidence of saturable uptake mechanisms at maternal and fetal sides of the perfused human placenta by rapid paired-tracer dilution: studies with calcium and choline.

Rapid uptake and efflux of 45Ca2+ and [3H]choline at the maternal and fetal interfaces of the syncytiotrophoblast in the dually-perfused human placenta was investigated by application of the single circulation paired-tracer dilution method (Yudilevich, Eaton, Short & Leichtweiss 1979). Cotyledons were perfused with Krebs-bicarbonate containing dextran (30 g/l; MW = 60-70,000) at 20 and 6 ml/min on maternal and fetal sides, respectively. The paired-tracer (test substrate and extracellular marker) technique consisted of an intra-arterial injection of a tracer bolus, followed by venous sampling over 5-6 min. There was a rapid (sec) uptake of 45Ca2+, followed by backflux (efflux into the ipsilateral circulation) which, over 5-6 min, was 59-100% on the fetal side. It was more variable but generally lower on the maternal interface. At 0.1 mM calcium, 45Ca2+ maximal uptake (Umax) was about 53% on the fetal side but on the maternal side it was variable and averaged 17%. At 2.4 mM calcium fetal side Umax was reduced to 40%. However, on the maternal side the effect was not consistent. Unidirectional influx (nmol/min per g) appeared to be not different on the two sides of the placenta. For [3H]choline (in choline-free perfusates) Umax was about 50% and 30% on fetal and maternal sides, respectively; tracer backflux was variable on the maternal side and averaged 50% on the fetal side. [3H]Choline uptake was highly inhibited by either 1.0 mM choline or the specific competitive inhibitor, hemicholinium-3 (0.1 mM). Specific transplacental transfer of 45Ca2+ (i.e. in excess of the extracellular marker) was not significant in either direction. For [3H]choline there was an apparent small excess (about 4%) preferential towards the fetal circulation. These findings in the human placenta are similar to those demonstrated previously in the guinea-pig placenta which suggested the existence of specific transport systems for choline and calcium on both sides of the syncytiotrophoblast.

Biological Transport, Active↗
PubMed1985

Transport of folates at maternal and fetal sides of the placenta: lack of inhibition by methotrexate.

Folate (pteroylglutamate) and methotrexate rapid (seconds) uptake by the trophoblast was investigated from either the maternal or fetal circulations of the isolated dually-perfused guinea-pig placenta. Tissue uptake was measured by using a single-circulation paired-tracer (3H-test and 14C-extracellular marker) technique. [3H]Folate uptakes were 80 and 52% (mean) in perfusates without unlabelled folate, on maternal and fetal sides, respectively. There was negligible 3H-tracer backflux into the circulation up to 6 min probably due to metabolic sequestration. [3H]Methotrexate uptakes were about 85 and 22% on maternal and fetal sides, respectively; however these uptakes were followed by rapid and complete backflux of the label. Specific transplacental transfer of [3H]folate or [3H]methotrexate in either direction was not detectable within 5-6 min. At the brush-border side (maternal) uptake of [3H]folate was highly inhibited by 100 nM unlabelled folate or its reduced form, methyltetrahydrofolate (the main form in plasma); however, equimolar methotrexate (an antifolate chemotherapeutic agent) failed to produce any inhibition of folate uptake. Our findings demonstrate that on both sides of the placenta a high-affinity transport system exists for trophoblast uptake of folate compounds. For methotrexate, either a separate transport system may exist or methotrexate may have a very low affinity for the folate system. These results are distinct from the findings reported in mouse L1210 leukemia cells.

Animals↗
PubMed1985

Evidence for a lactate transport system in the sarcolemmal membrane of the perfused rabbit heart: kinetics of unidirectional influx, carrier specificity and effects of glucagon.

The kinetics and specificity of L-lactate transport into cardiac muscle were studied during a single transit through the isolated perfused rabbit heart using a rapid (15 s) paired-tracer dilution technique. Kinetic experiments revealed that lactate influx was highly stereospecific and saturable with an apparent Kt = 19 +/- 6 mM and a Vmax = 8.4 +/- 1.5 mumol/min per g (mean +/- S.E., n = 14 hearts). At high perfusate concentrations (10 mM), the inhibitors alpha-cyano-4-hydroxycinnamate (Ki = 7.3 mM), pyruvate (Ki = 6.5 mM), acetate (Ki = 19.4 mM) and chloroacetate (Ki = 28 mM) reduced L-lactate influx, and Ki values were estimated assuming a purely competitive interaction of the inhibitors with the monocarboxylate carrier. The monocarboxylic acids [14C]pyruvate and [3H]acetate were themselves transported, and sarcolemmal uptakes of respectively 38 +/- 1% and 70 +/- 8% were measured relative to D-mannitol. Perfusion of hearts for 10-30 min with 0.15 or 1.5 microM glucagon increased myocardial lactate production and simultaneously inhibited tracer uptake of lactate, pyruvate and acetate. It is concluded that a stereospecific lactate transporter exhibiting an affinity for other substituted monocarboxylic acids is operative in the sarcolemmal plasma membrane of the rabbit myocardium.

Animals↗
PubMed1985

Characterization of choline transport at maternal and fetal interfaces of the perfused guinea-pig placenta.

Unidirectional influx and efflux of choline into the syncytiotrophoblast were investigated from both maternal and fetal circulations of the perfused guinea-pig placenta by using a single-circulation paired-tracer (extracellular reference and test substrate) dilution technique. Cellular uptake of [3H]choline at 0.05 mM was (mean percentage +/- S.E. of mean, n = 14 placentae) 51 +/- 2 and 49 +/- 2, on maternal and fetal sides, respectively. Kinetics of unidirectional influx (0.05-4.0 mM-choline) indicated the existence of saturable and non-saturable components on both sides: on maternal and fetal interfaces the Km (mM) values were respectively, 0.12 and 0.13, the Vmax (mumol min-1 g-1) values, 0.08 and 0.07 and the apparent linear transfer constants (min-1 g-1) 0.11 and 0.12. Efflux of [3H]choline from the placenta back into the ipsilateral circulation (backflux) was generally fast (20-60% in 5-6 min) and asymmetric with the fetal: maternal ratio usually above unity. Transplacental specific choline transfer in the dually perfused placenta, when observed, was small (less than 10% of the injected dose) following tracer injections in either direction based on the 5-6 min collection of the contralateral circulation (at 0.05 mM-choline). Placental retention of [3H]choline at the end of the 5-6 min period was about 25% of the injected dose when the tracers were injected from either circulation. Analogues of choline such as hemicholinium-3, thiamine, ethanolamine and N,N-dimethylethanolamine inhibited choline unidirectional influx, whereas betaine and acetate had no effect. The absence of the normal sodium gradient (perfusate sodium was replaced by Tris or by lithium) did not inhibit choline transport. The metabolic inhibitors dinitrophenol (1.0 mM) and potassium cyanide (1.0 mM) were essentially ineffective (up to 40 min perfusion). The sulphydryl reagent N-ethylmaleimide did not appear to inhibit the influx, in comparison with its effect on [3H]choline backflux which was greatly accelerated, resulting in a dramatic reduction in placental net uptake of the label. Our findings show that choline transport into the placenta is a rapid carrier-mediated process occurring at both maternal and fetal sides of the trophoblast, at physiological blood concentrations. This cellular uptake is possibly related to the synthesis of acetylcholine, which is known to occur in human placental tissue. Specific transplacental transfer of choline was a very slow process under the conditions of our experiments and this contrasted with the observed fast and high uptake into the trophoblast.

Animals↗
PubMed1984

Discrimination of parallel neutral amino acid transport systems in the basolateral membrane of cat salivary epithelium.

Transport of short-chain and long-chain neutral amino acids across the basolateral membrane of the epithelium in the perfused cat salivary gland has been studied using a rapid (less than 30 s) single circulation paired-tracer dilution technique. Amino acid uptake was measured by comparing the venous dilution profiles for a tritiated amino acid and D-[14C]mannitol (extracellular reference) following a bolus intra-arterial injection of a mixture containing both molecules. Unidirectional influx (v) was estimated from the maximal tracer uptake (Umax), the perfusate flow (F) and the perfusate amino acid concentration (Ca): v = [-F . ln (1-Umax)] . Ca. L-alanine influx was saturable and apparently mediated by a single entry system (Km = 0.83 +/- 0.11 mM and Vmax = 655 +/- 32 nmol/min . g). These kinetic constants were considerably lower than our previously reported values for L-phenylalanine: Km = 6.4 mM and Vmax = 1719 nmol/min . g. In cross-inhibition experiments performed over a wide range of concentrations (0.05-24 mM), influx of L-alanine and L-phenylalanine could be further discriminated, since both L-phenylalanine (Ki = 22 mM) and L-alanine (Ki = 19 mM) behaved as poor competitors. Removal of Na+ from the perfusate resulted in a selective inhibition of L-alanine and L-serine influx, whereas influx of the long-chain neutral amino acids L-leucine, L-phenylalanine and L-tryptophan remained unaffected. Although prolonged perfusion of glands with dinitrophenol (0.8 mM for 20-30 min) caused a variable but net inhibition of unidirectional uptake, it markedly enhanced the tracer efflux of L-leucine, L-phenylalanine, L-tyrosine and the basic amino acid L-lysine. It appears that at least two separate neutral amino acid transport systems are operative at the blood-tissue interface of the salivary epithelium: (i) a Na+-dependent alanine-serine-cysteine preferring type of carrier exhibiting a high affinity for amino acids with short, polar or linear side chains and (ii) a Na+-independent leucine preferring type of carrier selective for large neutral amino acids.

2,4-Dinitrophenol↗
PubMed1984

Rapid transcapillary exchange and unidirectional neuronal uptake of noradrenaline in the perfused rabbit heart.

Capillary permeability and cellular uptake of noradrenaline by the isolated artificially perfused rabbit heart was measured using rapid (less than 30 s) single-circulation tracer-dilution techniques. In a single coronary circulation capillary extractions of L-[14C]noradrenaline and D-[3H]mannitol (extracellular reference) relative to an intravascular marker, 125I-labelled albumin, were similar and above 60%. The 'apparent' volume of distribution for tracer noradrenaline was 2.5-fold larger than that measured for D-mannitol (0.32 ml g-1) suggesting cellular uptake of the amine. Unidirectional noradrenaline uptake was estimated by directly comparing coronary sinus dilution profiles of L-[3H]noradrenaline and D-[14C]mannitol. Michaelis-Menten saturation kinetics based on a single-entry system were determined (Km = 2.8 +/- 1.5 microM, Vmax = 2.1 +/- 0.5 nmol min-1 g-1, n = 4) by perfusing hearts with varying concentrations of L-noradrenaline (1-10 microM). Various known inhibitors of noradrenaline uptake were investigated to determine whether uptake was mediated by neuronal (uptake1) and/or extraneuronal (uptake2) mechanisms. Desipramine (5 microM), imipramine (5 microM) and metaraminol (2 microM) resulted in a 66-94% inhibition of noradrenaline influx. In comparison, the steroids, 17 beta-oestradiol (1 microM) and corticosterone (10 microM), and the noradrenaline metabolite normetanephrine (5 microM) caused virtually no inhibitory effects. The beta-adrenergic antagonist propranolol (5 microM) was also relatively ineffective. These results together with the kinetic constants estimated suggest that the rapid noradrenaline uptake reflects transport into adrenergic neurones lying in the coronary interstitium. The high resolution of this paired-tracer dilution technique has permitted a 'non-invasive' study of neuronal uptake mechanisms and its application may be of clinical value.

Animals↗
PubMed1984

Characteristics of a cationic amino acid transport system in the basolateral membrane of the cat salivary epithelium.

The specificity and kinetics of L-lysine influx across the basolateral surface of the cat salivary epithelium have been investigated in the perfused cat submandibular gland using a high-resolution, paired-tracer dilution technique. L-lysine influx was measured at several different perfusate concentrations (0.05-2.5 mM) and was found to be saturable. A Michaelis-Menten analysis based on a single entry site gave a Km of 0.49 +/- 0.08 mM and a Vmax of 231 +/- 20 nmol/min X g. The uptake of L-lysine was highly stereospecific and markedly inhibited by L-arginine (0.25-2.5 mM). The inhibitor constant (Ki) was 0.23 mM, suggesting that the carrier had a greater affinity for L-arginine than L-lysine. When the inhibitory effects of L-histidine (0.5-10 mM) were examined the Ki, estimated at 10 mM, was 4.6 mM. Nine other neutral amino acids (L-alanine, L-serine, L-cysteine, glycine, L-proline, L-homoserine, L-leucine, L-phenylalanine and L-glutamine), and an acidic amino acid (L-aspartate) were also tested at 10 mM and, although several caused inhibition, the Ki was always at least 20 times higher than the measured Km for L-lysine. It is concluded the carrier is highly specific for the L-form of the basic amino acids. The sodium dependence of L-lysine influx was investigated over a range of L-lysine concentrations (0.05-1 mM), and total removal of sodium from the perfusate had no effect on L-lysine influx. In the presence of sodium, L-homoserine, an amino acid not normally present in animal tissues, inhibited L-lysine influx (Ki = 13 mM). This inhibition was not observed in the absence of sodium, and contrasts with the observation that the inhibitory action of L-histidine was sodium independent. The present data suggest that a specific cationic amino acid transport system is operative in the basolateral membrane of the cat salivary epithelium. The properties of this system appear to be similar to the system y+ which has been described in several other cell types.

Amino Acids↗
PubMed1984

Asymmetric calcium influx and efflux at maternal and fetal sides of the guinea-pig placenta: kinetics and specificity.

Unidirectional influx of calcium across maternal and fetal sides of the syncytiotrophoblast was investigated in the guinea-pig placenta by using a rapid (less than 30 s) paired-tracer dilution technique. Experiments were performed in an in situ placenta artificially perfused through the umbilical vessels or in an isolated placenta in which both the maternal and fetal circulations were perfused. At equimolar Ca2+ concentrations, unidirectional calcium influx was always significantly lower on the maternal side than on the fetal side. Saturation kinetics were observed: on the fetal side the estimated Km was 1.8 +/- 0.7 mM and Vmax was 1.66 +/- 0.28 mumol/min X g (mean +/- S.E. of mean) and on the maternal side Km ranged from 0.18 to 1.15 mM and Vmax ranged from 0.12 to 0.59 mumol/min X g. When the inhibition of calcium influx was investigated on the fetal side of the trophoblast by using competing cations, the following sequence was observed: Ba2+ greater than Ca2+ congruent to Ni2+ greater than Sr2+ greater than Mg2+ congruent to Li+. Efflux of 45Ca2+ from the trophoblast into the ipsilateral circulation (backflux) was rapid (20-100% in 6 min) and asymmetric since the fetal:maternal ratio was 1.35 +/- 0.11 (mean +/- S.E. of mean) in the presence of 0.1 mM-Ca2+. In the dually perfused placenta, transplacental transfer (6 min) of 45Ca2+ varied over a wide range (0-80%); however, it was similar to that of the extracellular reference tracer, 22Na+, in either maternal-to-fetal or fetal-to-maternal directions. It is suggested that this is a consequence of the 'leakiness' of the dually perfused placenta since the transplacental transfer of 22Na+ and D-[3H]mannitol (or L-[14C]glucose) measured simultaneously were also variable but similar. Transplacental transfer of 45Ca2+ could not be used to characterize specific calcium-transport mechanisms, whereas highly sensitive trophoblast uptake measurements were provided by the single-circulation, paired-tracer technique. Our findings suggest the presence of a specific carrier-mediated transport system for calcium on both maternal and fetal surfaces of the trophoblast. The asymmetries in unidirectional influx into the trophoblast and rapid backflux indicate a mechanism by which the net transfer of calcium from the maternal to the fetal circulation is maintained in favour of the fetus.

Animals↗
PubMed1982

Unidirectional uptake of L-alanine and L-serine by system ASC in the basolateral membrane of cat salivary epithelium. Selective inhibition of rapid alanine metabolism by aminooxyacetate.

Unidirectional influx of L-alanine and L-serine across the basolateral membrane of cat salivary epithelium was investigated in perfused submandibular glands using a rapid (less than 30 s) paired-tracer dilution technique. (Bustamante, J.C., Mann, G.E. and Yudilevich, D.L. (1981) J. Physiol. 313, 65-79). High uptakes were measured for both neutral amino acids, however, only the uptake of L-alanine was followed by an immediate tracer efflux into the circulation. Aminooxyacetate, an inhibitor of alanine aminotransferase, completely inhibited this efflux and glandular pyruvate production. These results demonstrate that in assessing unidirectional amino acid uptake, the influence of metabolism should be considered even during short intervals of measurement.

Acetates↗
PubMed1982

Transport specificity for neutral and basic amino acids at maternal and fetal interfaces of the guinea-pig placenta.

1. The unidirectional influx of amino acids into the guinea-pig syncytiotrophoblast was measured using a single circulation paired-tracer dilution technique which allows separate characterization of both fetal and maternal interfaces. An in situ preparation perfused through the fetal circulation was used to examine the fetal side, while an isolated preparation perfused through both the fetal and maternal circulations was used to study both interfaces simultaneously.2. On the fetal side the maximal uptake (U(max)) determined at tracer concentrations was high for the short-chain neutral amino acid alanine (76%) and the long-chain neutrals, leucine (75%), phenylalanine (90%) and tyrosine (82%) and for the basic amino acid lysine (65%). In contrast, U(max) was negligible for alpha-methylaminoisobutyric acid and taurine, a beta-amino acid.3. The uptake of alanine and phenylalanine on the fetal side was inhibited by both short-chain (alanine, serine, cysteine) and long-chain (phenylalanine, methionine, leucine) neutral amino acids. d-alanine had no effect on l-alanine uptake whereas d-phenylalanine significantly inhibited that of l-phenylalanine. Diaminobutyric acid, lysine and arginine were effective inhibitors of alanine uptake but had no effect on phenylalanine uptake.4. On the maternal side uptake of alanine, phenylalanine and lysine was measured. Over a wide range of concentrations self-inhibition of alanine influx was similar to the cross-inhibition observed with phenylalanine. In contrast, the influx of phenylalanine, which was strongly self-inhibited, was only partially cross-inhibited by alanine.5. Influx of alanine and phenylalanine was measured at various perfusate concentrations and was found to be saturable on both maternal and fetal sides. The data were fitted to a single hyperbola and, on the maternal side, the K(m) for alanine (10.3+/-2.7 mm, mean+/-s.e., n = 3) was three-fold higher than the value measured for phenylalanine (3.1+/-0.8 mm). On the fetal side the K(m) values for alanine (8.4+/-1.4 mm, n = 4) and phenylalanine (11.9+/-1.9 mm, n = 3) were similar.6. The uptake of alanine, phenylalanine and lysine appeared to be highly sodium-dependent accounting for 40-70% of the total influx. However, the inhibited fractions were found to be different on the two sides of the placenta.7. The results of uptake, cross-inhibition and Na(+)-dependency experiments suggest the presence of an alanine-serine-cysteine (ASC) type system and a leucine (L) type system with markedly overlapping specificities at both the fetal and maternal interfaces. Separate kinetic characterization of a two carrier system was not possible under the conditions of these experiments. However, kinetic parameters for the over-all transport of alanine and phenylalanine were measured.

Alanine↗
PubMed1982

Unidirectional uptake of substrates at the blood side of secretory epithelia: stomach, salivary gland, pancreas.

This paper reviews studies of cellular uptake of substrates by the gastric mucosa, salivary gland, and pancreas by single-circulation, multiple tracer dilution techniques. The application of this methodology to secretory organs in vivo has permitted the characterization of transport phenomena at the blood-tissue interface of resting and secreting epithelia. To estimate uptake of a test molecule after an intraarterial injection, its venous concentration profile (30 samples in 1 in) was compared with that of 1) an intravascular marker or 2) a diffusible molecule that remains confined to the extracellular space. Among the molecules investigated were 86Rb, 57Co-labeled cyanocobalamin, 125I-labeled insulin, [3H]ouabain, [3H]dopamine, [3H]norepinephrine, and a wide range of labeled amino acids. High tracer uptake (80%) was measured that could be inhibited by specific unlabeled competitors. Unidirectional influx was saturable and Michaelis-Menten kinetic constants could be estimated. The ultimate objective was to identify various transport systems and/or receptors at the basolateral side of these epithelia. However, nerve terminals in the interstitium could be the major site for the uptake of catecholamines.

Amino Acids↗
PubMed1981

Specificity of neutral amino acid uptake at the basolateral side of the epithelium in the cat salivary gland in situ.

1. Amino acid uptake was measured in resting cat submandibular glands with either a natural blood supply or perfused at constant flow with a Krebs-albumin solution. Following a bolus arterial injection of a 3H-labelled amino acid and D-[14C]mannitol (extracellular reference tracer), the venous effluent was immediately sampled sequentially. The maximal uptake, Umax, from the blood or perfusate was determined from the paired-tracer dilution curves using the expression: uptake % = (1 -- (3H/14C) X 100). 2. In glands with a natural blood supply, Umax values up to 46% were measured for short-chain (serine and alanine) and long-chain (valine, methionine, leucine, isoleucine, 1-amino-cyclopentane cyclopentane carboxylic acid, phenylalanine, tryptophan, tyrosine, histidine and glutamine) neutral amino acids. In contrast, Umax was negligible for amino acids of the imino-glycine group (proline and glycine) and the nonmetabolized amino acids, 2-aminoisobutyric acid (AIB) and methylaminoisobutyric acid (MeAIB). 3. In glands with a natural blood supply addition of an unlabelled amino acid to the tracer injectate reduced Umax for the test acid by up to 80%. The pattern of these interactions suggested the presence of two transport systems for neutral amino acids, one preferring short-chain and the other long-chain amino acids. 4. In glands perfused at constant flow rates with an amino acid-free Krebs-albumin solution high Umax values were measured: L-serine (66%), L-alanine (54%), L-leucine (43%), L-phenylalanine (42%) and L-tyrosine (51%). Only a low uptake was observed for L-proline (8%) and glycine (14%). There was no uptake of methylaminoisobutyric acid which confirms the result obtained in glands with an intact circulation. 5. Saturation of L-phenylalanine influx was observed in perfused glands as the perfusate concentration of unlabelled L-phenylalanine was increased from 0.5 to 20 mmol . 1-1. A Michaelis--Menten analysis based on a single entry system indicated an apparent Km of 6.4 +/- 0.8 mmol . 1-1 and a Vmax of 1719 +/- 94 nmol . min-1g.-1 6. Since the fenestrated capillaries in the salivary gland are readily permeable to the test amino acid and D-mannitol, it is most probable that the amino acid carriers are located in the basolateral side of the epithelium. 7. The use of a paired-tracer dilution technique to measure uptake in a single circulatory passage has enabled a detailed characterization of neutral amino acid transport in the salivary gland and has overcome the limitation of previous studies based on solute transfer from blood to saliva.

Amino Acids↗
PubMed1981

Uptake and asymmetric efflux of amino acids at maternal and fetal sides of placenta.

Unidirectional uptake of eighteen amino acids into the syncytiotrophoblast was measured from both the maternal and fetal circulations of isolated dually perfused guinea pig placentas using a single-circulation, paired-tracer dilution technique. A bolus containing a tritiated amino acid and L-[14C]glucose (extracellular marker) was injected intra-arterially into one circulation, and both venous outflows were sequentially sampled. The maximal cellular uptake (Umax) on the injection side was determined from (1-[3H]/[14C]) values and used to calculate the unidirectional influx. Umax values for neutral and basic amino acids ranged between 15 and 58% and were similar on both sides of the trophoblast. Uptake of the acidic amino acids and taurine was minimal. Amino acid influx from either circulation was followed by rapid tracer backflux and transplacental transfer. Tracer efflux was asymmetric and preferentially directed towards the fetal side. It is suggested that amino acid transport systems are present on both surfaces of the placenta and that net transfer from mother to fetus is the result of asymmetric efflux from the trophoblast.

Amino Acids↗