Development of transport mechanisms for sulphate and iodide in immature choroid plexus

The time-course of development of sulphate and iodide transport mechanisms in choroid plexus was studied by measuring uptake of [³⁵S]sulphate and [¹²⁵I]iodide from an incubating medium by isolated choroid plexuses of foetal and newborn rabbits and cats. Sulphate uptake by choroid plexus was poorly developed in rabbit foetuses just before term, but highly developed in newborn animals. Iodide uptake was already well developed in the most immature foetuses studied.     

Transport du sulfate à travers la double membrane limitante, ou enveloppe, des chloroplastes d'épinard

Evidence is presented for low rates of carriermediated uptake of sulphate, thiosulphate and sulphite into the stroma of the C3 plant Spinacia oleracea. Uptake of sulphate in the dark was followed using two techniques (1) uptake of sulphate [³⁵S] as determined by silicon oil centrifugal filtration and (2) uptake as indicated by inhibition of CO₂-dependant O₂ evolution rates after addition of sulphate.Sulphate, thiosulphate and sulphite were transported across the envelope leading to an accumulation in the chloroplasts. Sulphate transport had saturation kinetics of the Michaelis-Menten type (Vmax : 25 μmoles . mg⁻¹ chl . h⁻¹ at 22°C ; Km : 2.5 mM). The rate of transport for sulphate was not influenced either by illumination or pH change in the external medium. Phosphate was a competitive inhibitor of sulphate uptake by chloroplasts (Ki : 0.7 mM, fig. 1). The rate of transport for phosphate appeared to be much higher than for sulphate. When the chloroplasts were pre-loaded with labelled sulphate, radioactivity was rapidly released after addition of phosphate into the external medium. Consequently, the transport of sulphate occurs by a strict counter-exchange : for each molecule of sulphate entering the chloroplast, one molecule of phosphate leaves the stroma, and vice-versa.The uptake of sulphate by isolated intact chloroplasts exchanging for internal free phosphate induced a lower rate of photophosphorylation, which in turn inhibited CO₂-dependent O₂ evolution.The presence, on the inner membrane of the chloroplast envelope, of a specific sulphate carrier, distinct from the phosphate translocator, is discussed.     

Thyroxine transport in choroid plexus

The role of the choroid plexus in thyroid hormone transport between body and brain, suggested by strong synthesis and secretion of transthyretin in this tissue, was investigated in in vitro and in vivo systems. Rat choroid plexus pieces incubated in vitro were found to accumulate thyroid hormones from surrounding medium in a non-saturable process. At equilibrium, the ratio of thyroid hormone concentration in choroid plexus pieces to that in medium decreased upon increasing the concentration of transthyretin in the medium. Fluorescence quenching of fluorophores located at different depths in liposome membranes showed maximal hormone accumulation in the middle of the phospholipid bilayer. Partition coefficients of thyroxine and triiodothyronine between lipid and aqueous phase were about 20,000. After intravenous injection of 125I-labeled thyroid hormones, choroid plexus and parts of the brain steadily accumulated 125I-thyroxine, but not [125I]triiodothyronine, for many hours. The accumulation of 125I-thyroxine in choroid plexus preceded that in brain. The amount of 125I-thyroxine in non-brain tissues and the [125I]triiodothyronine content of all tissues decreased steadily beginning immediately after injection. A model is proposed for thyroxine transport from the bloodstream into cerebrospinal fluid based on partitioning of thyroxine between choroid plexus and surrounding fluids and binding of thyroxine to transthyretin newly synthesized and secreted by choroid plexus.     

VITAMIN B6 TRANSPORT IN THE CENTRAL NERVOUS SYSTEM: IN VITRO STUDIES

Abstract— The transport into and release of tritium labeled vitamin B₆ ([³H]B₆) from rabbit brain slices and isolated choroid plexuses were studied. In vitro, both brain slices and choroid plexus concentrated [³H]B₆ by an energy dependent uptake system when [³H]pyridoxine (PIN) was added to the incubation medium. Most of the [³H] within the tissues was phosphorylated [³H]B₆. In each tissue, the nonphosphorylated vitamers inhibited the uptake of [³H]PIN from the medium significantly more than the phosphorylated vitamers. The concentrations of the nonphosphorylated B₆ vitamers necessary to inhibit brain and choroid plexus uptake of [³H]PIN from the medium by 50% were approx 0.4 μm and 5–10μm respectively after a 30 min incubation. Both brain slices and choroid plexus readily released (46 and 56% respectively in 30 min) previously accumulated [³H]B₆ into artificial CSF. However, brain slices released only nonphosphorylated [³H]B₆, whereas the choroid plexus released predominantly phosphorylated [³H]B₆. Addition of unlabeled PIN to the release media significantly increased the percentage of [³H]B₆ released by both brain slices and choroid plexus. The results of these in vitro studies provide evidence that: (1) both brain slices and chloroid plexus possess specific uptake and release mechanisms for B₆, and (2) these mechanisms tend to regulate intracellular B₆ levels. These studies also suggest that the choroid plexus serves as a locus for the transfer of B₆ from blood to CSF and is the source of most of the phosphorylated B₆ in CSF.     

Nucleoside transport in choroid plexus: Mechanism and specificity

In vitro the transport into and release of [³H]thymidine, [³H]deoxyuridine, and [³H]nitrobenzylthioinosine (NBTI) from the isolated choroid plexus, the anatomical locus of the blood-cerebrospinal fluid barrier, were studied separately. Using the ability of NBTI to inhibit nucleoside efflux from the choroid plexus, the transport of [³H]thymidine and [³H]deoxyuridine into the choroid plexus at 37 °C was measured. Like thymidine, deoxyuridine was transported into the choroid plexus against a concentration gradient by a saturable process that depended on intracellular energy production but not intracellular binding or metabolism. The Michaelis-Menten constants (K_T) for the active transport of thymidine and deoxyuridine into the choroid plexus were 13.6 and 7.2 μM, respectively. Deoxyuridine and adenosine were competitive inhibitors of thymidine transport into the choroid plexus with inhibitor constants (K_I) of 6.8 and 14.5 μM, respectively. [³H]NBTI was also transported into the choroid plexus at 37 °C; unlike [³H]thymidine and [³H]deoxyuridine, the release of [³H]NBTI was not inhibited by NBTI itself. These studies provide evidence that the choroid plexus contains an active nucleoside transport system of low specificity for nucleosides, and a separate, saturable efflux system for nucleosides that is very sensitive to inhibition by NBTI. In vivo these systems transport nucleosides from blood into cerebrospinal fluid.     

Utilization of 35S-Thiosulphate and an appraisal of the role of ATP-sulphurylase in chemolithotrophic Thiobacillus ferrooxidans

Differentially labelled ³⁵S-thiosulphate was taken up by washed cells of Thiobacillus ferrooxidans which were previously grown on thiosulphate. The uptake was proportional to the biomass over the range 0.5–4.0 mg dry wt. of bacteria and showed typical saturation kinetics with an estimated K _m value of 0.5 mM for ³⁵S-thiosulphate. Dithionate and Group VI anions inhibited the uptake, which was under pH control and had a temperature optimum of 50°C. In the absence of thiosulphate, the cells bound ³⁵S-sulphate but the binding did not increase on prolonged incubation and the label could be removed completely by washing with dilute sulphuric acid. Increasing amounts of the label were incorporated from [outer-³⁵S]thiosulphate into cellular materials over a 60-min period, whereas little or no assimilation was observed from either the [inner-³⁵S]thiosulphate or ³⁵S-sulphate. The kinetic properties of the sulphate-activating enzyme ATP_sulphurylase enriched from bacteria grown with either thiosulphate or ferrous-iron were similar although this enzyme has an assimilatory function only when the bacterium is grown with ferrous-iron.Abbreviation APS adenosine-5-sulphatophosphate     

Amino acid transport by choroid plexus in vitro

Choroid plexus from mongrel cats was incubated from 1 to 120 min in artificial cerebrospinal fluid containing α-amino[1-¹⁴C]isobutyric acid. The uptake of α-amino [1-¹⁴C]isobutyric acid occurred against a concentration gradient, was saturable, dependent on metabolic energy, and inhibited by natural amino acids. These results indicate that a transport mechanism is present in choroid plexus which could serve to regulate amino acid concentration in the cerebrospinal fluid of animals.     

The choroid plexus removes beta-amyloid from brain cerebrospinal fluid

beta-Amyloid (Abeta) concentration in the cerebrospinal fluid (CSF) of the brain may be regulated by the choroid plexus, which forms a barrier between blood and brain CSF. Abeta uptake from CSF was determined as its volume of distribution (V(D)) into isolated rat choroid plexus tissue. The V(D) of [125I]Abeta1-40 was corrected by subtraction of the V(D) of [14C]sucrose, a marker for extracellular space and diffusion. Abeta uptake into choroid plexus was time and temperature dependent. Uptake of [125I]Abeta was saturable. Abeta uptake was not affected by addition of transthyretin or apolipoprotein E3. In studies with primary culture monolayers of choroidal epithelial cells in Transwells, Abeta permeability across cells, corrected by [(14)C]sucrose, was greater from the CSF-facing membrane than from the blood-facing membrane. Similarly, cellular accumulation of [125I]Abeta was concentrative from both directions and was greater from the CSF-facing membrane, suggesting a bias for efflux. Overall, these results suggest the choroid plexus selectively cleanses Abeta from the CSF by an undetermined mechanism(s), potentially reducing Abeta from normal brains and the brains of Alzheimer's disease patients.     

NIACIN AND NIACINAMIDE ACCUMULATION BY RABBIT BRAIN SLICES AND CHOROID PLEXUS IN VITRO

The transport into and release of¹⁴C-labeled niacin and niacinamide from rabbit brain slices and isolated choroid plexuses were studied. In vitro, both brain slices and choroid plexus concentrated ¹⁴C by specific, energy-dependent mechanisms when [¹⁴C]niacinamide was added to the incubation medium. The saturable accumulation velocities, which were linear for 30 min, depended, in part, on incorporation of the [¹⁴C]niacinamide into NAD. The X_T and Y_max for ¹⁴C accumulation with [¹⁴C]niacinamide in the medium by brain slices and choroid plexus were 0.80 μM and 1.45 μmolkg^?1 (30 min)^?1, and 0.23 μM and 18.6 μmol kg^?1 (30 min)^?1 respectively. In vitro, the choroid plexus, unlike brain slices, vigorously concentrated ¹⁴C by a separate, specific energy-dependent process when ¹⁴C niacin was added to the incubation medium. The saturable accumulation velocity, which was linear for 30 min, depended completely on the metabolism of [¹⁴C]niacin. The K_T and Y_max for¹⁴C accumulation by choroid plexus with [¹⁴C]niacin in the medium were 18.1 μM and 439 μmol kg^?1 (30 min)^?1 respectively. Whether preincubated in [¹⁴C]niacin or [¹⁴C]niacinamide, choroid plexus released predominantly [¹⁴C]niacinamide.     

TRANSPORT OF LYSINE FROM CEREBROSPINAL FLUID OF THE CAT

—The clearance from cerebrospinal fluid of l-[¹⁴C]lysine and l-[³H]arginine was measured during ventriculo-cisternal perfusions of anaesthetized cats. Increasing in the perfusate the concentration of unlabelled l-lysine produced a gradual reduction in clearance of the labelled amino acids without altering the uptake of l-[¹⁴C]lysine by the choroid plexus. Net transport of l-lysine out of cerebrospinal fluid occurred by saturable and non-saturable components. The saturable component satisfied Michaelis-Menton kinetics, while the behaviour of the non-saturable component was consistent with diffusion. A V_max of 0·017 μmol/min and an affinity constant (k_t) of 0·83 mm were estimated. The clearance of l-lysine was unaffected by the addition to the perfusate of high concentrations of selected neutral amino acids, but was stimulated by the presence of l-cystine. Conversely, a high concentration of l-lysine did not affect the clearance of glycine or cycloleucine. The dibasic amino acids appear to be removed from cerebrospinal fluid by a relatively specific, mediated transport system which may serve to regulate their concentrations in the cerebrospinal fluid.     

Protein synthesis and transport by the rat choroid plexus and ependyma

Summary Light (LM-ARG) and electron microscope (EM-ARG) autoradiographs were prepared from immature rat choroid plexus and ependyma at 5, 10, 30, and 60 min and 16 h following intraperitoneal administration of [³H]- labeled amino acid mixtures. Intracellular protein synthesis and transport were ascertained in lateral and fourth ventricle choroid plexus epithelium by quantitative EM-ARG at the several post-injection intervals. ARG were also prepared from choroid plexuses cultured for one day, pulse labeled for one hour and reincubated for various periods in nonradioactive media. Significant labeling of both attached and free apical protrusions (blebs) was observed in both choroid plexus and ependyma in vivo and in choroid plexus in vitro. This phenomenon was interpreted as a physiologically significant mechanism for protein transport (apocrine secretion) by epithelia into the cerebrospinal fluid (CSF).This study was supported in part by N.I.H. Research Grant NS 12906     

Forebrain binding sites for atrial natriuretic factor: Alterations in spontaneously hypertensive (SHR) rats

Binding sites for atrial natriuretic factor (ANF-28) were studied in forebrain areas of spontaneously hypertensive (SHR) and Wistar Kyoto (WKY) normotensive male rats by quantitative autoradiography. The maximum binding capacity of [¹²⁵I]ANF-28 was significantly reduced in the subfornical organ and choroid plexus of 4 and 14 week old SHR rats compared to age-matched WKY controls. In contrast, the affinity constant for [¹²⁵I]ANF-28 binding was elevated in the choroid plexus of 14 week old SHR rats. These findings indicate that marked reductions in the number of ANF-28 binding sites occur in weanling SHRs as well as in adult SHRs with elevated arterial blood pressures. Thus, these persistant reductions in forebrain ANF-28 binding sites in SHR rats may contribute to the development and maintenance of this form of experimental hypertension.     

A radiotracer method to study efflux transport of iodide liberated from thyroid hormones via deiodination metabolism in the brain

AimsThyroid hormones (TH) play an important role in the development and functional maintenance of the central nervous system. The purpose of this study was to develop a radiotracer method for studying the in vivo efflux transport of iodide liberated by the TH metabolism in the brain. The rationale of our method is as follows: a radioiodinated compound can enter the brain and rapidly release iodide in situ; the iodide efflux rate can be estimated from the clearance of brain radioactivity after disappearance of the iodinated compound.Main methods6-[¹²⁵I]Iodo-9-pentylpurine ([¹²⁵I]9Pe6IP) was designed to enter the brain and release ¹²⁵I^? by the reaction with glutathione and synthesized from the corresponding bromo derivative in a Br/¹²⁵I exchange reaction. The brain kinetics of radioactivity and radioactive metabolites were investigated after intravenous injection of [¹²⁵I]9Pe6IP into mice. The iodide efflux rate was estimated in mice pretreated with perchlorate, an inhibitor of iodide transport from the brain.Key findingsHigh brain uptake (5.3% injected dose/g) was observed at 1 min, and almost complete conversion of [¹²⁵I]9Pe6IP to ¹²⁵I^? occurred 10 min after injection. The ¹²⁵I^? uptake from the blood was negligible. ¹²⁵I^? was eliminated from the brain along a single-exponential curve with a half-life of 6.0 min. Furthermore, dose-dependent inhibition of ¹²⁵I^? efflux was observed in mice pretreated with perchlorate.SignificanceWe conclude that 9Pe6IP labeled with ¹²⁴I (positron emitter) or ¹²³I (single-photon emitter) may be useful for studying the in vivo efflux transport of iodide in the brain using nuclear medicine imaging devices.     

Endocytosis and degradation of serglycin in liver sinusoidal endothelial cells

We have previously reported that liver sinusoidal endothelial cells (LSECs) are responsible for the clearance of monocyte chondroitin sulfate proteoglycan serglycin from the circulation (øynebråten et al.(2000) J. Leukocyte Biol. 67; 183–188). The aim of the present study was to investigate the kinetics of degradation of endocytosed serglycin in primary cultures of LSECs. The final degradation products of serglycin labelled biosynthetically in the glycosaminoglycan (GAG) chains with [³H] in the acetyl groups of N-acetyl galactosamine residues, [¹⁴C] in the pyranose rings, or [³⁵S] in the sulfate groups were identified as[³H]-acetate, [¹⁴C]-lactate and [³⁵S]-sulfate. Comparison of the rate of release of degradation products from the cells after endocytosis of serglycin labelled chemically with ¹²⁵I in the tyrosine residues, or biosynthetically with [³⁵S] or [³H] in the sulfate or acetyl groups, respectively, showed that ¹²⁵I appeared more rapidly in the medium than [³⁵S]-sulfate and [³H]-acetate. Judging from the speed of appearance of free ¹²⁵I both intracellularly and in the medium, the core protein is degraded considerably more rapidly than the GAG chains.Desulfation of the GAG chains starts after the GAG chains are released from the core protein. Generation of lactate and acetate as the final products from degradation of the carbon skeleton of the GAG chains indicates that catabolism of endocytosed macromolecules in LSECs proceeds anaerobically.     

Characterization of the blue light-induced extracellular alkalinization associated with the monovalent anion uptake by Monorapidium braunii. Competition between NO3 and Cl−

The blue light-elicited monovalent anion-dependent alkalinization of the medium of Monoraphidium braunü (Legnerová, 202–7d) was characterized for the NO-³ and Cl- uptake. The maximal H⁺ uptake rates for these two anions have a similar optimum pH around 8.5, and quite similar K^s values for high (38 üM for Cl- and 35üM for NO-³) and low (320 üM for Cl- and 335 üM for NO-³) affinities. The steady H⁺ uptake associated with the uptake and reduction of NO-³ showed a K^s of 125 üM. which in this alga corresponds to the NO-³ reductase (EC 1.6.6.2) K^m for NO-³. The only and striking difference found in the uptake properties of these anions was the delay time between the switching on of the blue light and the start of the alkalinization, which increased from 10 to 90 s as the initial pH decreased from 8.5 to 6.5 in the presence of NO-³, whereas for Cl- uptake this delay time (10s) did not vary in relation to the initial pH. When the NO-³ concentration in the medium was low (100 üM), the presence of relatively high concentrations of Cl- (3 üM), on the one hand, greatly stimulated the maximal alkalinization rates but, on the other, Cl- severely reduced the steady NO-³-dependent rate of alkalinization. The data indicate that Cl- inhibits competitively NO-³ uptake with a Kⁱ of 750 üM. Moreover, high concentrations of NO-³ (above 5 üM) reduced its own maximal, but not the steady, uptake rates. The above results allow us to propose that most of the components of the individual NO-³ and Cl- transport systems are under identical light control and, as the competition data suggest, that these two anions may be taken up by the same transport system.     

Visualizing Dopamine and Serotonin Transporters in the Human Brain with the Potent Cocaine Analogue [125I]RTI-55: In Vitro Binding and Autoradiographic Characterization

Abstract: The cocaine analogue RTI-55 was evaluated as a probe for in vitro labeling and localization of dopamine and serotonin transporters after death in the human brain. Kinetic, saturation, and competition binding experiments indicated complex interactions of the radioligand with the identification of multiple recognition sites. In membrane binding assays, the association of [¹²⁵I]RTI-55 at 25°C to putamen membranes was monophasic. In contrast, dissociation of [¹²⁵I]RTI-55 occurred in two phases with t_1/2 values of 9.4 and 36.5 min, respectively. Saturation analysis of [¹²⁵I]RTI-55 binding demonstrated two binding sites in the human putamen with K_D values of 0.10 ± 0.02 and 1.81 ± 0.46 nM. The binding of [¹²⁵I]RTI-55 was displaced by a wide range of cocaine analogues and monoamine uptake inhibitors. The rank order of potency demonstrated in competition assays with human putamen membranes indicates that the radioligand labels cocaine recognition sites on the dopamine transporter (mazindol > GBR 12909 > GBR 12935 > paroxetine > nisoxetine > desipramine ≥ fluoxetine > citalopram). In the human occipital cortex, [¹²⁵I]RTI-55 recognized multiple binding sites with K_D values of 0.02 ± 0.01 and 4.18 ± 0.46 nM. The rank order of potency for inhibition of [¹²⁵I]RTI-55 binding to cerebral cortex membranes (paroxetine > citalopram > GBR 12909 ≥ mazindol ≥ nisoxetine > benztropine) suggests that [¹²⁵I]RTI-55 labels the serotonin transporter in the human occipital cortex. Autoradiographic mapping of [¹²⁵I]RTI-55 revealed very high densities of cocaine recognition sites over areas known to be rich in dopaminergic innervation, including the caudate, putamen, and nucleus accumbens. Moderately elevated densities of [¹²⁵I]RTI-55 binding sites were also seen throughout the thalamus, hypothalamus, and substantia nigra. [¹²⁵I]RTI-55 binding sites were prevalent throughout the cerebral cortex and amygdala. In autoradiographic studies, the addition of the selective serotonin transport blocker citalopram completely prevented [¹²⁵I]RTI-55 labeling in the thalamus, hypothalamus, and throughout most of the cerebral cortex. In the presence of citalopram, [¹²⁵I]RTI-55 binding site densities remained elevated over the striatum and substantia nigra, with selective residual labeling also seen in the external segment of the globus pallidus and the lateral nucleus of the amygdala. These results demonstrate that in the human brain, [¹²⁵I]RTI-55 labels multiple recognition sites on dopamine and serotonin transporters.     

Dichotomous development of the organic anion transport protein in liver and choroid plexus

Both adult liver and choroid plexus express the organic aniontransport protein (oatp1) and transport[³⁵S]bromosulfophthalein(BSP). Studies of the developing rat liver reveal that oatp1 mRNA andprotein do not begin to be expressed until 15 days postnatal and are atadult levels by 30 days. Uptake of[³⁵S]BSP follows thesame time course. In contrast, neonatal rat choroid plexus expressesoatp1 mRNA and protein. When quantified on a weight basis, the uptakeof [³⁵S]BSP in choroidplexus is lower in the adult than at earlier stages of development.Although fluorescence confocal microscopy of adult rat choroid plexusshows that oatp is localized to the apical surface, facing thecerebrospinal fluid, this method reveals an intracellular localizationof oatp1 in the neonate. Approximately 12 wk are required for theappearance of the adult pattern of distribution. Changes in thelocalization and activity of oatp1 during development could play animportant role in the pathobiology of maturation of the liver and thecentral nervous system.

     

Lumiflavin and Lumichrome Transport in the Central Nervous System

Abstract: The transport of the lipid-soluble sugarless flavins, [¹⁴C]lumiflavin and [¹⁴C]lumichrome, into and from the isolated choroid plexus and brain slices was studied in vitro. The isolated choroid plexus accumulated both [¹⁴C] flavins by a saturable, energy-requiring process that did not depend on binding or intracellular metabolism of the [¹⁴C] flavins. Both sugar-containing and sugarless flavins, as well as cyclic organic acids, significantly inhibited [¹⁴C]lumiflavin and [¹⁴C]Iumichrome uptake by the isolated choroid plexus. Within 2.5 min, 75% of the [¹⁴C]lumiflavin accumulated by the isolated choroid plexus was released into the medium. Brain slices accumulated [¹⁴C]lumiflavin by a saturable process that did not meet all the criteria for active transport. Ninety-five percent of the [¹⁴C]lumiflavin accumulated by brain slices was released into the medium within 7.5 min. In vivo , 2 h after the intraventricular injection of 6.5 nmol [¹⁴C]lumiflavin, almost all of the [¹⁴C]flavin was cleared from the CNS. Addition of 3.5 μmol FMN to the intraventricular injectate significantly decreased the clearance of [¹⁴C]lumiflavin from the CNS. These studies document that the sugarless flavins are transported by the flavin transport systems in the CNS.     

Biotin transport and metabolism in the central nervous system

The mechanisms by which biotin enters and leaves brain, choroid plexus and cerebrospinal fluid (CSF) were investigated by injecting [³H]biotin either intravenously or intraventricularly into adult rabbits. [³H]biotin, either alone or together with unlabeled biotin was infused at a constant rate into conscious rabbits. At 180 minutes, [³H]biotin had entered CSF, choroid plexus, and brain. In brain, CSF, and plasma, greater than 90% of the nonvolatile³H was associated with [³H]biotin. The addition of 400 mol/kg unlabeled biotin to the infusion syringe decreased the penetration of [³H]biotin into brain and CSF by approximately 70 percent. Two hours after an intraventricular injection, [³H]biotin was cleared from the CSF more rapidly than mannitol and minimal metabolism of the [³H]biotin had occurred in brain. However, 18 hours after an intraventricular injection, approximately 35% of the [³H]biotin remaining in brain had been covalently incorporated into proteins, presumably into carboxylase apoenzymes. These results show that biotin enters CSF and brain by saturable transport systems that do not depend on metabolism of the biotin. However, [³H]biotin is very slowly incorporated covalently into proteins in brain in vivo.     

The effect of temperature on the uptake and loss of anions by seedling roots ofZea mays L

Intact maize seedlings were examined for the uptake and leakage of labelled sulphate and phosphate anions affected by temperature. Control plants, grown at 25 °C were compared from the aspects of uptake capacity and leakage with plants incubated in nutrient solutions cooled to 15 °C and 5 °C, respectively. Short time intervals as well as 1–7 d exposure to cooling were used. Already after 1 h exposure at 5 °C and 5 h cooling at 15 °C and at 5 °C, considerable changes were manifested in anion uptake and leakage. The uptake of³²P declined more than that of³⁵S. So, after a 30 min uptake interval the uptake of³⁵S decreased at 15 °C to 49.84% and at 5 °C to 6.05% comparing with the uptake at 25 °C, while the uptake of³²P declined to 28.64% at 15 °C and to 4.45% at 5 °C. The leakage of both anions was the highest at 25 °C in absolute rates, but relatively most of the uptaken³⁵S and³²P was released at 5 °C. Longer exposure to a chilling temperature of 5 °C (1–7 days) resulted in two patterns of sulphate and phosphate uptake.