Distribution and characterization of the opioid octapeptide met5-enkephalin-arg6-gly7-leu8 in the gastrointestinal tract of the rat

Summary The distribution and characterization of the opioid octapeptide met⁵-enkephalin-arg⁶-gly⁷-leu⁸ (met⁵-enk-arg⁶-gly⁷-leu⁸) within the gastrointestinal tract of the rat has been determined by immunohistochemistry and radioimmunoassay by use of a newly developed antibody to met⁵-enk-arg⁶-gly⁷-leu⁸. With both techniques, met⁵-enk-arg⁶-gly⁷-leu⁸-immunoreactivity (met⁵-enk-arg⁶-gly⁷-leu⁸IR) was detected in all regions of the gastrointestinal (GI) tract except the esophagus. The highest concentration of immunoreactive met⁵-enk-arg⁶-gly⁷-leu⁸ was observed in the colon, while intermediate concentrations were found in the stomach, duodenum, jejunum, and ileum. Immunostained somata were observed chiefly in the myenteric plexus; immunostained processes were present primarily in the myenteric plexus and the circular muscle layer. This distribution pattern is similar to that previously observed with antiserum to met⁵-enkephalin-arg⁶-phe⁷ (met⁵-enk-arg⁶phe⁷). Chromatographic analysis of met⁵-enk-arg⁶-gly⁷leu⁸-immunoreactive peptides extracted from the GI tract revealed the presence of an immunoreactive peptide of high molecular weight which accounted for approximately three-quarters of met⁵-enk-arg⁶-gly⁷-leu⁸-IR in both stomach and colon. These findings suggest a role for peptides related to the octapeptide met⁵-enk-arg⁶-gly⁷-leu⁸ in the regulation of GI function.     

Reversible stimulation of the Na+/K+ pump by monensin in cultures of vascular smooth muscle

Two ionophores, monensin and salinomycin, increased total cell Na⁺ and ouabain-sensitive ⁸⁶Rb⁺ uptake in cultures of smooth muscle cells from rat aorta. Monensin was used to produced graded increases in cell Na⁺ in order to assess the Na⁺ dependence of the Na⁺/K⁺ pump in the intact cell. The relationship between internal Na⁺ and ouabain-sensitive ⁸⁶Rb⁺ uptake was hyperbolic (K¹_{Na = 3 mM)}. Monensin did not stimulate ⁸⁶Rb⁺ uptake in the absence of external Na⁺. Loading the cells with Na⁺ by exposing cultures to a K⁺-free medium for 3 hr maximally increased cell Na⁺ and ouabain-sensitive ⁸⁶Rb⁺ uptake to the same extent as monensin. Total cell Na⁺ and pump activity in monensin-treated cells returned to the initial values after removing the ionophore. Monensin was then able to increase total cell Na⁺ and ouabain-sensitive ⁸⁶Rb⁺ uptake to the same extent as the initial treatment with the ionophore.     

Dual emission of Ce3+,Mn2+‐coactivated Ca3YNa(PO4)3F via energy transfer: a single component white/yellow‐emitting phosphor

A series of Ce³⁺,Mn²⁺‐coactivated Ca₃YNa(PO₄)₃F phosphors were synthesized via a traditional solid‐state reaction under a reductive atmosphere. X‐Ray powder diffraction was used to confirm that the crystal structure and diffraction peaks of Ce³⁺/Mn²⁺‐doped samples matched well with the standard data. A spectral overlap between the emission band of Ce³⁺ and the excitation band of Mn²⁺ suggested the occurrence of energy transfer from Ce³⁺ to Mn²⁺. With increasing Mn²⁺ content, the emission intensities and lifetime values of the Ce³⁺ emission for Ca₃YNa(PO₄)₃F:Ce³⁺,Mn²⁺ phosphors linearly decrease, whereas the energy transfer efficiencies gradually increase to 89.35%. By adjusting the relative concentrations of Ce³⁺ and Mn²⁺, the emission hues are tuned from blue to white and eventually to yellow. These results suggest that Ca₃YNa(PO₄)₃F:Ce³⁺,Mn²⁺ phosphors have promising application as white‐emitting phosphors for near‐ultraviolet light‐emitting diodes.     

A comparison of regulatory effects of chloride on nitrate uptake, and of nitrate on chloride uptake into Pisum sativum seedlings

Net nitrate uptake, ³⁶ClO^?₃/NO^?₃ influx and ³⁶Cl^? influx into Pisum sativum L. cv. Feltham First seedlings have been examined following growth in culture medium containing different combinations of chloride and nitrate. When young (6 days old) seedlings, that had been grown in the absence of N were used, nitrate accumulation stimulated net nitrate uptake and ³⁶ClO^?₃/NO^?₃ influx (r²= 0.99) while chloride accumulation inhibited nitrate uptake and ³⁶ClO^?₃/NO^?₃ influx (r²= 0.65). When nitrate was provided during growth there was no effect of chloride pretreatment on net nitrate uptake and there was little effect of total [NO^?₃+ Cl^?]_i on ³⁶ClO^?₃/NO^?₃ influx (r²= 0.26). A direct effect of Cl^? on ³⁶ClO^?₃/NO^?₃ influx was only found when seedlings had been starved of N for more prolonged periods (14 days). When moderate chloride was supplied during growth, ³⁶Cl^? influx was insensitive to nitrate or chloride accumulated, but significantly correlated with log_e [NO^?₃+ Cl^?]_i (r²= 0.75). When trace amounts of Cl^? were supplied during growth ³⁶Cl^? influx was inhibited by (a) NO^?₃ in the external medium and (b) Cl^? pretreatment, but was insensitive to NO^?₃ pretreatment. The sensitivity of ³⁶Cl^? influx to external nitrate was not found following Cl^? pretreatment in the absence of nitrate. The possibility that there are two populations of chloride carriers which differ in their sensitivity to external nitrate is discussed. Tentative schematic models to account for the regulation of nitrate and chloride uptake are proposed in the context of current hypotheses for regulation of ion transport and control systems theory.     

Identification of Residues in Domain III of Bacillus thuringiensis Cry1Ac Toxin That Affect Binding and Toxicity

Alanine substitution mutations in the Cry1Ac domain III region, from amino acid residues 503 to 525, were constructed to study the functional role of domain III in the toxicity and receptor binding of the protein to Lymantria dispar, Manduca sexta, and Heliothis virescens. Five sets of alanine block mutants were generated at the residues ⁵⁰³SS⁵⁰⁴, ⁵⁰⁶NNI⁵⁰⁸, ⁵⁰⁹QNR⁵¹¹, ⁵²²ST⁵²³, and ⁵²⁴ST⁵²⁵. Single alanine substitutions were made at the residues ⁵⁰⁹Q, ⁵¹⁰N, ⁵¹¹R, and ⁵¹³Y. All mutant proteins produced stable toxic fragments as judged by trypsin digestion, midgut enzyme digestion, and circular dichroism spectrum analysis. The mutations, ⁵⁰³SS⁵⁰⁴-AA, ⁵⁰⁶NNI⁵⁰⁸-AAA, ⁵²²ST⁵²³-AA, ⁵²⁴ST⁵²⁵-AA, and ⁵¹⁰N-A affected neither the protein’s toxicity nor its binding to brush border membrane vesicles (BBMV) prepared from these insects. Toward L. dispar and M. sexta, the ⁵⁰⁹QNR⁵¹¹-AAA, ⁵⁰⁹Q-A, ⁵¹¹R-A, and ⁵¹³Y-A mutant toxins showed 4- to 10-fold reductions in binding affinities to BBMV, with 2- to 3-fold reductions in toxicity. Toward H. virescens, the ⁵⁰⁹QNR⁵¹¹-AAA, ⁵⁰⁹Q-A, ⁵¹¹R-A, and ⁵¹³Y-mutant toxins showed 8- to 22-fold reductions in binding affinities, but only ⁵⁰⁹QNR⁵¹¹-AAA and ⁵¹¹R-A mutant toxins reduced toxicity by approximately three to four times. In the present study, greater loss in binding affinity relative to toxicity has been observed. These data suggest that the residues ⁵⁰⁹Q, ⁵¹¹R, and ⁵¹³Y in domain III might be only involved in initial binding to the receptor and that the initial binding step becomes rate limiting only when it is reduced more than fivefold.     

Thymic lymphocytes: II. Phenotypic modifications of thymocytes after concanavalin A stimulation in the presence of interleukin 2: Early modifications of Lyt 1+2+ subset and later proliferation of cells with more mature phenotypes

Cortical thymocytes are devoid of any immune function, as tested by presently available techniques. The ability of this subpopulation to respond to mitogens or antigens in the presence of interleukin 2 (IL-2) produced by activated mature T lymphocytes has been claimed but is still questioned. In an attempt to study the participation of the different thymocyte subsets and especially that of the cortical type, phenotypic modifications were examined during concanavalin A activation in the presence of IL-2. An immunofluorescent double labeling technique with anti-Lyt 1 and anti-Lyt 2 antibodies was used which led to the determination of four different phenotypes: Lyt 1⁺2⁺, Lyt 1⁺2^?, Lyt 1^?2⁺, and Lyt 1^?2^?. Careful analysis of cell viability in culture and expression of the results in absolute numbers of living cells per culture allowed us to follow modifications of small cellular subsets. Cultures of total thymocytes and PNA-agglutinated (enriched in Lyt 1⁺2⁺ cells) and non-PNA-agglutinated cells (enriched in Lyt 1⁺2^?, Lyt 1^?2⁺, and Lyt 1^?2^? cells) were studied. It was shown that thymocyte activation began by early phenotypic modifications which took place within the first 2 hr of culture but only when Con A plus IL-2 were used. These modifications imply the reduction of the Lyt 1⁺2⁺ pool and a compensatory enhancement of Lyt 1^?2⁺ and Lyt 1^?2^? cells, without modification of the total cell number or [³H]thymidine incorporation. These early phenotypic changes are interpreted as the modulation of antigens on the surface of Lyt 1⁺2⁺ cells. The second phase of thymocyte activation implies cell death (essentially Lyt 1⁺2⁺ cells) and cell proliferation. The cells which specifically proliferate in the presence of Con A and IL-2 are Lyt 1⁺2^? and Lyt 1^?2⁺, the latter always being present in greater number. Cell survival and absolute number of Lyt 1⁺2^? and Lyt 1^?2⁺ cells in the activated PNA^?-enriched population are always higher than in total thymocyte and PNA⁺ cells cultures. Thus, if Lyt 1⁺2⁺ cortical thymocytes do not proliferate by themselves, they seem to intervene by providing Lyt 1^?2⁺ cells which proliferate secondarily.     

Serum stability of 67Cu chelates: Comparison with 111In and 57Co

Simple chelates and chelate conjugated monoclonal antibodies labeled with ¹¹¹In, ⁵⁷Co and ⁶⁷Cu demonstrate marked differences in stability when exposed to a serum environment. Among these radiometals, on DTPA, the order of stability is ¹¹¹In ⁵⁷Co ⪢ ⁶⁷Cu. On benzyl-EDTA, the order of stability is ¹¹¹In ⋍ ⁵⁷Co ⪢ ⁶⁷Cu. Among those investigated, the only serum stable ⁶⁷Cu chelate found was ⁶⁷Cu-TETA. The order of stability observed for ⁵⁷Co vs ⁶⁷Cu is contrary to published equilibrium constants. These in vitro studies suggest that the in vivo behavior of metal chelates exposed to a complex molecular environment may not be predicted by classically determined equilibrium constants.     

Nitrogen uptake and assimilation in Enteromorpha intestinalis (L.) Link (Chlorophyta): using N to determine preference during simultaneous pulses of nitrate and ammonium

We investigated the ability of Enteromorpha intestinalis (L.) Link to take up pulses of different species of nitrogen simultaneously, as this would be an important mechanism to enhance bloom ability in estuaries. Uptake rates and preference for NH₄⁺ or NO₃⁻ following 1, 3, 6, 9, 12 or 24 h of exposure to either ¹⁵NH₄NO₃ or NH₄¹⁵NO₃ were determined by disappearance of N from the medium. Differences in assimilation rates for NH₄⁺ or NO₃⁻ were quantified by the accumulation of NH₄⁺, NO₃⁻, and atom % ¹⁵N in the algal tissue. NH₄⁺ concentration was reduced more quickly than water NO₃⁻ concentration. Water column NH₄⁺ concentration after the longest time interval was reduced from 300 to 50 μM. Water NO₃⁻ was reduced from 300 to 150 μM. The presence of ¹⁵N or ¹⁴N had no effect on uptake of either NH₄⁺ or NO₃⁻. ¹⁵N was removed from the water at an almost identical rate and magnitude as ¹⁴N. Differences in accumulation of ¹⁵NH₄⁺ and ¹⁵NO₃⁻ in the tissue reflected disappearance from the water; ¹⁵N from NH₄⁺ accumulated faster and reached an atom % twice that of ¹⁵N from NO₃⁻. This outcome suggested that when NH₄⁺ and NO₃⁻ were supplied in equal concentrations, more NH₄⁺ was taken up and assimilated. The ability to take up high concentrations of NH₄⁺, and NO₃⁻ simultaneously is important for bloom-forming species of estuarine macroalgae subject to multiple nutrient species from various sources.     

Vanadate inhibition of the Ca-ATPase from human red cell membranes

1. (1) VO₃⁻ combines with high affinity to the Ca²⁺-ATPase and fully inhibits Ca²⁺-ATPase and Ca²⁺-phosphatase activities. Inhibition is associated with a parallel decrease in the steady-state level of the Ca²⁺-dependent phosphoenzyme.

2. (2) VO₃⁻ blocks hydrolysis of ATP at the catalytic site. The sites for VO₃⁻ also exhibit negative interactions in affinity with the regulatory sites for ATP of the Ca²⁺-ATPase.

3. (3) The sites for VO₃⁻ show positive interactions in affinity with sites for Mg²⁺ and K⁺. This accounts for the dependence on Mg²⁺ and K⁺ of the inhibition by VO₃⁻. Although, with less effectiveness, Na⁺ substitutes for K⁺ whereas Li⁺ does not. The apparent affinities for Mg²⁺ and K⁺ for inhibition by VO₃⁻ seem to be less than those for activation of the Ca²⁺-ATPase.

4. (4) Inhibition by VO₃⁻ is independent of Ca²⁺ at concentrations up to 50 μM. Higher concentrations of Ca²⁺ lead to a progressive release of the inhibitory effect of VO₃⁻.

Photochemistry of Co(EDTA)−-I− System in Aqueous Solutions

The Co(EDTA)⁻ complex in aqueous solution gives rise to a specific interaction with I⁻ ions as evidenced by a new, relatively intense band formed at 290–300 nm. This specific interaction is attributed to the formation of an ion-pair between Co(EDTA)⁻ and I⁻, even though they are like charged ions. Irradiation of this ion-pair in air- equilibrated solutions with 313 nm light, causes the reduction of the Co(EDTA)⁻ to Co(EDTA)²⁻ and the oxidation of the I⁻ ion to I₃⁻. The results obtained are interpreted on the basis of a mechanism in which Co(EDTA)²⁻ and I^· are the primary photoproducts. The I^· radical is then scavenged by I⁻ to yield I₂⁻, which subsequently disproportionates to I₃⁻ and I⁻ and reoxidizes Co(EDTA)²⁻ to Co(EDTA)⁻. At the beginning of the photoreaction, the I₂⁻ decay is equally distributed on the two reaction pathways. It was possible to determine a value of 0.2 ± 0.05 for the photoreaction quantum yield and an efficiency of the primary photochemical step almost unitary. A schematic representation of the energetics of the overall reaction is reported.     

Strontium induces murine keratinocyte differentiation in vitro in the presence of serum and calcium

Primary mouse keratinocytes in culture are induced to terminally differentiate by increasing extracellular Ca²⁺ concentrations (Ca_O) from 0.05 mM to ≥ 0.1 mM. The addition of Sr²⁺ (≥ 2.5 mM) to medium containing 0.05 mM Ca²⁺ induces focal stratification and terminal differentiation, which are similar to that found after increasing the Ca_O to 0.12 mM. Sr²⁺ in 0.05 mM Ca²⁺ medium induces the expression of the differentiation-specific keratins, keratin 1 (K1), keratin 10 (K10), and the granular cell marker, filaggrin, as determined by both immunoblotting and immunofluorescence. Sr²⁺ induces the expression of those differentiation markers in a dose dependent manner, with an optimal concentration of 5 mM. In the absence of Ca²⁺ in the medium, the Sr²⁺ effects are reduced, and Sr²⁺ is ineffective when both Ca²⁺ and serum are deleted from the medium. Sr²⁺ treatment increases the ratio of fluorescence intensity of the intracellular Ca²⁺ sensitive probe, fura-2, indicating an associated rise in the level of intracellular free Ca²⁺ and/or Sr²⁺. At doses sufficient to induce differentiation, Sr²⁺ also increases the level of inositol phosphates in primary keratinocytes within 30 min. The uptake curves of ⁸⁵Sr²⁺ by primary keratinocytes are similar to those of ⁴⁵Ca²⁺. At low concentrations, the initial uptake of both ⁴⁵Ca²⁺ and ⁸⁵Sr²⁺ reaches a plateau within 1 hr; at higher concentrations, the uptake of both ⁴⁵Ca²⁺ and ⁸⁵Sr²⁺ increases continuously for 12 hr. In keratinocytes pre-equilibrated with ⁴⁵Ca²⁺ in 0.05 mM Ca²⁺ medium, Sr²⁺ causes an increase of ⁴⁵Ca²⁺ uptake, which is dependent on the presence of serum. These results suggest that Sr²⁺ utilizes the same signalling pathway as Ca²⁺ to induce keratinocyte terminal differentiation and that Ca²⁺ may be required to exert these effects. © 1993 Wiley-Liss, Inc.     

CHLORIDE AND SODIUM UPTAKE POTENTIAL OVER AN ENTIRE ROTATION OF POPULUS IRRIGATED WITH LANDFILL LEACHATE

There is a need for information about the response of Populus genotypes to repeated application of high-salinity water and nutrient sources throughout an entire rotation. We have combined establishment biomass and uptake data with mid- and full-rotation growth data to project potential chloride (Cl^?) and sodium (Na⁺) uptake for 2- to 11-year-old Populus in the north central United States. Our objectives were to identify potential levels of uptake as the trees developed and stages of plantation development that are conducive to variable application rates of high-salinity irrigation. The projected cumulative uptake of Cl^? and Na⁺ during mid-rotation plantation development was stable 2 to 3 years after planting but increased steadily from year 3 to 6. Year six cumulative uptake ranged from 22 to 175 kg Cl^? ha^?1 and 8 to 74 kg Na⁺ ha^?1, while annual uptake ranged from 8 to 54 kg Cl^? ha^?1 yr^?1 and 3 to 23 kg Na⁺ ha^?1 yr^?1. Full-rotation uptake was greatest from 4 to 9 years (Cl^?) and 4 to 8 years (Na⁺), with maximum levels of Cl^? (32 kg ha^?1 yr^?1) and Na⁺ (13 kg ha^?1 yr^?1) occurring in year six. The relative uptake potential of Cl^? and Na⁺ at peak accumulation (year six) was 2.7 times greater than at the end of the rotation.     

Differential binding of 3H-imipramine and 3H-mianserin in rat cerebral cortex

Drug competition profiles, effect of raphé lesion, and sodium dependency of the binding of two antidepressant drugs ³H-imipramine and ³H-mianserin to rat cerebral cortex homogenate were compared to examine whether the drugs bound to a common “antidepressant receptor.” Of the neurotransmitters tested, only serotonin displaced binding of both ³H-imipramine and ³H-mianserin. ³H-mianserin binding was potently displaced by serotonin S₂ antagonists and exhibited a profile similar to that of ³H-spiperone binding. In the presence of the serotonin S₂ antagonist spiperone, antihistamines (H₁) potently displaced ³H-mianserin binding. ³H-Imipramine binding was displaced potently by serotonin uptake inhibitors. The order of potency of serotonergic drugs in displacing ³H-imipramine binding was not similar to their order in displacing ³H-spiperone or ³H-serotonin binding. Prior midbrain raphé lesions greatly decreased the binding of ³H-imipramine but did not alter binding of ³H-mianserin. Binding of ³H-imipramine but not ³H-mianserin was sodium dependent. These results show that ³H-imipramine and ³H-mianserin bind to different receptors. ³H-Imipramine binds to a presynaptic serotonin receptor which is probably related to a serotonin uptake recognition site, the binding of which is sodium dependent. ³H-Mianserin binds to postsynaptic receptors, possibly both serotonin S₂ and histamine H₁ receptors, the binding of which is sodium independent.     

Characterization and Location of Met5-Enkephalin-Arg6-Phe7 Stored in Various Rat Brain Regions

A specific antiserum against met⁵-enkepha-lin-arg⁶-phe⁷ was raised and used to study the distribution and characterization of met⁵-enkephalin-arg⁶-phe⁷-like immunoreactive material in rat brains by radioimmunoassay and immunohistochemical procedures. The antiserum appears to be directed to the COOH-terminus of the peptide, as it fails to cross-react with met⁵-enkeph-alin, met³-enkephalin-arg⁶, met⁵-enkephalin-arg⁶-arg⁷, met⁶-enkephalin-lys⁶, and leu-enkephalin. However, it cross-reacts with phe-met-arg-phe by about 10% and with phe-met-arg-phe-NH₂ to an insignificant degree. The highest content of met⁵-enkephalin-arg⁶-phe⁷ was found in the striatum, which contains a dense network of immunoreactive varicose fibers and terminals, as well as immunoreac tive cell bodies. The met⁵-enkephalin-arg⁶-phe⁷ in striatum can be released in a Ca²⁺-dependent manner by a depolarizing concentration of KC1, raising the possibility of a neu-roregulatory role for met⁵-enkephalin-arg⁶-phe⁷. Characterization of the immunoreactive material by gel filtration and high pressure liquid chromatography revealed the presence of multiple forms of immunoreactive material in some brain regions.     

Sodium and potassium interactions with Na+-ATPase of inside-out membrane vesicles from high-K+ and low-K+ sheep red cells

Na⁺-ATPase of high-K⁺ and low-K⁺ sheep red cells was examined with respect to the sidedness of Na⁺ and K⁺ effects, using inside-out membrane vesicles and very low ATP concentrations (?2 μM). With varying amounts of Na⁺ in the medium, i.e., at the cytoplasmic surface, Na_cyt⁺, the activation curves show that high-K⁺ Na⁺-ATPase has a higher affinity for Na_cyt⁺ compared to low-K⁺. The apparent affinity for Na_cyt⁺ is also increased by increasing the ATP concentrations in high-K⁺ but not low-K⁺. With Na_cyt⁺ present, Na⁺-ATPase is stimulated by intravesicular Na⁺, i.e., Na⁺ at the originally external surface, Na_ext⁺, to a greater extent in low-K⁺ than high-K⁺. Intravesicular K⁺ (K_ext⁺) activates Na⁺-ATPase in high-K⁺ but not in low-K⁺ vesicles and extravesicular K⁺ (K_cyt⁺) inhibits low-K⁺ but not high-K⁺ Na⁺-ATPase. Thus, the genetic difference between high-K⁺ and low-K⁺ is expressed as differences in apparent affinities for both Na⁺ and K⁺ and these differences are evident at both cytoplasmic and external membrane surfaces.     

Effect of Hypoxia on Na+-K+-Cl− Cotransport in Cultured Brain Capillary Endothelial Cells of the Rat

Abstract: The effect of hypoxia on Na⁺,K⁺-ATPase and Na⁺-K⁺-Cl^? cotransport activity in cultured rat brain capillary endothelial cells (RBECs) was investigated by measuring ⁸⁶Rb⁺ uptake as a tracer for K⁺. RBECs expressed both Na⁺,K⁺-ATPase and Na⁺-K⁺-Cl^? cotransport activity (4.6 and 5.5 nmol/mg of protein/min, respectively). Hypoxia (24 h) decreased cellular ATP content by 43.5% and reduced Na⁺,K⁺-ATPase activity by 38.9%, whereas it significantly increased Na⁺-K⁺-Cl^? cotransport activity by 49.1% in RBECs. To clarify further the mechanism responsible for these observations, the effect of oligomycin-induced ATP depletion on these ion transport systems was examined. Exposure of RBECs to oligomycin led to a time-dependent decrease of cellular ATP content (by ～65%) along with a complete inhibition of Na⁺,K⁺-ATPase and a coordinated increase of Na⁺-K⁺-Cl^? cotransport activity (up to 100% above control values). Oligomycin augmentation of Na⁺-K⁺-Cl^? cotransport activity was not observed in the presence of 2-deoxy-d -glucose (a competitive inhibitor of glucose transport and glycolysis) or in the absence of glucose. These results strongly suggest that under hypoxic conditions when Na⁺,K⁺-ATPase activity is reduced, RBECs have the ability to increase K⁺ uptake through Na⁺-K⁺-Cl^? cotransport.     

Responses of soybean to ammonium and nitrate supplied in combination to the whole root system or separately in a split-root system

To address the questions of whether allocation of carbohydrates to roots is influenced by ionic form of nitrogen absorbed and whether allocation of carbohydrates to roots in turn influences proportionality between NH₄⁺ and NO₃^? uptake from mixed sources, NH₄⁺ and NO₃^? were supplied separately to halves of a split-root hydroponic system and were supplied in combination to a whole-root system. Dry matter accumulation in the split-root system was 18% less in the NH₄⁺-fed axis than in the NO₃^?-fed axis. This, however, does not indicate that partitioning of carbohydrate between the two axes was different. Most of the reduction in dry matter accumulation in the NH₄⁺-fed axis can be accounted for by the retransport of CH₂O equivalents from the root back to the shoot with amino acids produced by NH₄⁺ assimilation. Uptake of NH₄⁺ or NO₃^? by the respective halves of the split-root system was proportional to the estimated allocation of carbohydrate to that half. When NH₄⁺ and NO₃^? were supplied to separate halves of the split-root system, the cumulative NH₄⁺ to NO₃^? uptake ratio was 0.81. When supplied in combination to the whole-root system, the cumulative NH₄⁺ to NO₃^? uptake ratio was 1.67. Thus, while the shoot may affect total nitrogen uptake through the export of carbohydrates to roots, the shoot (common for halves of the split-root system) apparently does not exert a direct effect on proportionality of NH₄⁺ and NO₃^? uptake by roots. For whole roots supplied with both NH₄⁺ and NO₃^?, the restriction in uptake of NO₃^? may involve a stimulation of NO₃^? efflux rather than an inhibition of NO₃^? influx. While only the net uptake of NH₄⁺ and NO₃^? was measured by ion chromatography, monitoring at approximately hourly intervals during the first 3 days of treatment revealed irregularly occurring intervals of both depletion (net influx) and enrichment (net efflux) in solutions. In the case of NH₄⁺, numbers of net efflux events were similar (21 to 24 out of 65 sequential sampling intervals) whether NH₄⁺ was supplied with NO₃^? to whole-root systems or separately to an axis of the split-root system. In the case of NO₃^?, however, the number of net efflux events increased from 8 when NO₃^? was supplied to a separate axis of the split-root system to between 19 and 24 when NO₃^? was supplied with NH₄⁺ to whole-root systems.     

The Effect of Dichlorophenyldimethylurea on Light-Stimulated 22Na+, 42K+ and 86Rb+ Absorption in Different Tissues of Phaseolus vulgaris Leaves

The light-stimulated absorption of ⁸⁶Rb⁺ by Phaseolus vulgaris L. leaf slices was found to be sensitive to dichlorophenyldimethylurea in air as well as in nitrogen, whereas light-stimulated ²²Na⁺ absorption in nitrogen was not sensitive to this inhibitor. The absorption of ²²Na⁺ is not affected by light in air. The absorption of ⁴²K⁺ is enhanced by a dichlorophenyldimethylurea-insensitive light effect under anaerobic conditions and further increased by light in the absence of the inhibitor. Light-enhanced ⁴²K⁺ absorption in air was also inhibited by dichlorophenyldimethylurea. Previous work showed that light-stimulated ⁸⁶Rb⁺ and ⁴²K⁺ absorption by Phaseolus vulgaris leaf slices is restricted to the guard cells. The present results are discussed with reference to the effect of light on stomatal opening.     

Simultaneous Influx and Efflux of Nitrate during Uptake by Perennial Ryegrass

Experiments with intact plants of Lolium perenne previously grown with ¹⁴NO₃⁻ revealed significant efflux of this isotopic species when the plants were transferred to solutions of highly enriched ¹⁵NO₃⁻. The exuded ¹⁴NO₃⁻ was subsequently reabsorbed when the ambient solutions were not replaced. When they were frequently replaced, continual efflux of the ¹⁴NO₃⁻ was observed. Influx of ¹⁵NO₃⁻ was significantly greater than influx of ¹⁴NO₃⁻ from solutions of identical NO₃⁻ concentration. Transferring plants to ¹⁴NO₃⁻ solutions after a six-hour period in ¹⁵NO₃⁻ resulted in efflux of the latter. Presence of Mg²⁺, rather than Ca²⁺, in the ambient ¹⁵NO₃⁻ solution resulted in a decidedly increased rate of ¹⁴NO₃⁻ efflux and a slight but significant increase in ¹⁵NO₃⁻ influx. Accordingly, net NO₃⁻ influx was slightly depressed. A model in accordance with these observations is presented; its essential features include a passive bidirectional pathway, an active uptake mechanism, and a pathway for recycling of endogenous NO₃⁻ within unstirred layers from the passive pathway to the active uptake site.     

Permeability of reconstituted sarcoplasmic reticulum vesicles: Reconstitution of the K+, Na+ channel

Permeability properties of reconstituted rabbit skeletal muscle sarcoplasmic reticulum vesicles were characterized by measuring efflux rates of [³H]inulin, [³H]choline⁺, ⁸⁶Rb⁺, and ²²Na⁺, as well as membrane potential changes using the voltage-sensitive probe, 3,3′-dipentyl-2,2′-oxacarbocyanine. Native vesicles were dissociated with deoxycholate and were reconstituted by dialysis. Energized Ca²⁺ accumulation was partially restored. About $\text{1}\text{2}$ of the reconstituted vesicles were found to be ‘leaky’, i.e., permeable to choline⁺ or Tris⁺ but not to inulin. The remaining reconstituted vesicles were ‘sealed’, i.e., impermeable to choline⁺, Tris⁺ and inulin. Sealed reconstituted vesicles could be further subdivided according to their K⁺, Na⁺ permeability. About $\text{1}\text{2}$, previously designated Type I, were readily permeable to K⁺ and Na⁺, indicating the presence of the K⁺, Na⁺ channel of sarcoplasmic reticulum. The remaining sealed vesicles (Type II) formed a permeability barrier to K⁺ and Na⁺, suggesting that they lacked the K⁺, Na⁺ channel. These studies show that the K⁺, Na⁺ channel of sarcoplasmic reticulum can be solubilized with detergent and reconstituted with retention of activity. Furthermore, our results suggest that part or all of the decreased Ca²⁺-loading efficiency of reconstituted vesicles may be due to the presence of a significant fraction of leaky vesicles.