Salivary ions and neural taste responses in the hamster

Saliva is a chemically complex fluid that bathes oral surfacesand may affect early events in mammalian gustation. We measuredchorda tympani responses to taste stimuli in hamsters (Mesocricetusauratus) while their tongues were adapted to either water, artificialsaliva or natural saliva. Artificial saliva on the tongue loweredneural responses to taste stimuli that were present in the artificialsaliva and to those stimuli that cross-adated with salivarycomponents. Changing from a water-adapted tongue to one soakedwith pilocarpine-stimulated saliva from donor hamsters led tosignificantly smaller responses to NaCl. Responses to sucrose,NH₄Cl and quinine were unaffected. Chemical analysis of hamstersaliva revealed ‘normal’ mammalian levels of K⁺,Ca²⁺ and Mg²⁺, but unexpectedly low levels of Na⁺ and Cl^–.     

Gustatory neural response latencies in the frog

The major rate limiting steps in bullfrog peripheral nerve gustatoryresponse latencies were studied by measuring glossopharyngealnerve multi-unit activity, detecting response onset times, andcalculating rates of stimulus diffusion to receptor cells andsignal propagation along first order neurons. The stimulus deliverytechnique minimized physicochemical and mechanical artifacts,as well as neural responses to mechanical stimulation of thetongue. Neural activity was processed in 10 ms bins. Responseonsets were determined by a criterion that compared the statisticalprobability of the neural events during stimulus liquid presentationswith those during both Ringer's solution presentations afteradaptation to Ringer's and no-stimulus control conditions. Thiscriterion yielded response latencies of 70–110 ms for10 mM CaCl₂, 2 mM quinine hydrochloride, and 10^–5 M and10^–6 M cantharidin or Ringer's, and H₂O. No responsesoccurred during presentations of 10^–7 M cantharidin orRinger's after adaptation to Ringer's, or during the no-stimuluscontrol condition. From the measured latencies and calculatedrates of stimulus diffusion to receptor cells, and signal propagationalong first order neurons, we conclude that taste receptor cellevents and not perireceptor or signal propagatiog events arethe major rate limiting steps in gustatory response latencies.     

The Perception of Saltiness is Eliminated by NaCl Adaptation: Implications for Gustatory Transduction and Coding

The tastes of salts to humans are complex. NaCl is the mostpurely salty of all salts, but even this stimulus tastes sweetat low concentrations and somewhat sour at mid-range intensities.Other salts taste significantly sour or bitter in addition tosalty. Previous studies have shown that the saltiness of simplehalide salts is reduced by adaptation to NaCl, suggesting thata single mechanism might be responsible for the salty tasteof these stimuli. In electrophysiological studies in rodents,the response to NaCl is reduced by application to the tongueof the Na⁺- channel blocker amiloride. Organic Na⁺ salts aremore heavily dependent on this amiloride-sensitive transductioncomponent than NaCl, and are generally less salty and more sour.In order to investigate the relationship between NaCl saltinessand that evoked by other salts, we adapted the tongue to distilledH₂O and to 0.1 M NaCl and obtained direct magnitude estimatesof the taste intensity of 15 organic and inorganic Na⁺, Li⁺,K⁺ and Ca²⁺ salts, matched for total intensity. Subjects dividedthese magnitude estimates among the component taste qualities.Adaptation to NaCl abolished the taste of NaCl and LiCl, andeliminated the saltiness of all other salts. The magnitude estimatesof the bitterness and sourness of many salts increased afterNaCI adaptation. Since recent biophysical data suggest thatadaptation in taste receptors may involve whole-cell mechanisms,we propose that saltiness is reduced by NaCl adaptation becauseit originates in the subset of taste receptors responsive toNaCl. This implies that saltiness is coded within the CNS incells whose receptive fields include the NaCl-sensitive receptorcells and that the degree to which any salt tastes salty isdetermined by its ability to drive these receptors. This modelproposes, for example, that KCl has a salty component becauseit stimulates some of the same receptor cells as NaCl, eventhough the transduction mechanisms for KCl are different thanthose engaged by NaCl. Adaptation to NaCl blocks the saltinessof KCl and other salts because they stimulate NaCl-sensitivereceptor cells. Chem. Senses 20: 545–557, 1995.     

ERRATUM

Delete "In contrast, under 5% CO₂, the cells grew even at 40µM NaCl (Fig. 3), and the growth rate at 0.5–10mM NaCl during the linear-growth period was 0.24 µg Chl?(mlculture)^–1?h^–1"     

EFFECTS OF CHLORAMPHENICOL AND KINETIN ON UPTAKE AND INCORPORATION OF AMINO ACIDS BY TOBACCO LEAF DISKS

The effects of chloramphenicol and kinetin on uptake and incorporationof ³⁵S-methionine and some ₁₄C-amino acids have been investigatedin leaf-disks of Nicotiana rustica in light and dark. Chloramphenicolin a concentration of 1 mg per ml inhibits the uptake of aminoacids from 30 to 60 per cent compared with the water control.The incorporation of amino acids into bulk protein is stronglyinhibited in light (40 to 70 per cent), but only to a smalldegree in dark (10 to 20 per cent), as revealed also by ¹⁴CO₂-photosynthesisof the disks and following treatment with chloramphenicol indark. The stimulating effect of kinetin on uptake and incorporationof amino acids is dependent upon its concentration (10^–5to 10^–6 M ; but 10^–4 M solution inhibits stronglyboth uptake and incorporation). The stimulation seems to influencemore incorporation than uptake processes. Possible interactionsof chloramphenicol and kinetin in the protein metabolism oftobacco leaves have been discussed. (Received April 27, 1964; )     

PLASMALEMMA POTENTIAL IN NITELLA

A considerable part of the response of the vacuolar potentialof Nitella flexilis to the change of external KCl, NaCl, RbCl,LiCl, or CaCl₂ concentration is caused by the response of thecell wall (a cation exchanger) to the external medium. The potentialswere measured on the internodes whose cell sap was exchangedfor simple salt solutions. The potential difference across theplasmalemma which is the internal potential measured againstthe cell wall phase changes largely with the change in concentrationof the external KCl, but also more or less with that of theexternal NaCl, LiCl or RbCl. CaCl₂ depolarizes the plasmalemmapotential by about 50 mv when the concentration is increasedfrom 10^–5 M to 10^–3 M, and hyperpolarizes it againby about 40 mv from 10^–3 M to 10^–1 M leaving thelevel of the peak of the action potential almost unchanged. ¹This work was supported by Research Grants from the Ministryof Education of Japan     

Studies on the effect of certain enzymic poisons on the metabolism of storage organs III. Uptake and utilization of sucrose by radish root slices in presence of iodoacetate

Iodoacetate greatly retarded the uptake of sucrose and slightlyaffected its inversion by radish root slices. Carbohydrate content of the samples decreased substantiallyboth in water and in iodoacetate. Feeding with sucrose led tomarked accumulation of carbohydrates and supplemental additionof iodoacetate induced less accumulation of carbohydrates. Iodoacetate caused exudation of nitrogen fractions into theculture media. Protein synthesis via amino acids seems to beoperative in iodoacetate treated slices. It is also suggestedthat nitrate-N, in presence of sucrose, is converted into peptidesand proteins. Addition of iodoacetate to sucrose media inhibitedthis pathway of protein synthesis. Both sucrose and iodoacetate (4 x 10^–4M) stimulated theCO₂ output whereas 25 x 10^–4M iodoacetate did not changethe CO₂ output when compared with that of controls. Sucroseand iodoacetate (4 x 10^–4 M) when joined together maskedthe accelerating effect of each other. ¹Present address: Department of Botany, Faculty of Science,University of A'in Shams, Abbassia, Cairo, Egypt, U. A. R. (Received November 6, 1968; )     

Studies on CN--insensitive respiration in plant roots III. Induction of CN--insensitive respiration with low concentrations of respiratory inhibitors

Induction of CN^–-insensitive respiration with low concentrationsof respiratory inhibitors was studied. If roots were treatedwith 10^–3 M CN^– for 96 hr, the plants died, whilethose treated with 10^–4 M CN^– showed healthy growth. O₂ uptake in untreated rice and wheat roots showed a negativeresponse to 10^–2 M CN^– to a considerable extent.On the other hand, pretreatment with 10^–4 M CN^–for more than 6 hr did not greatly affect respiratory rate,but made respiration insensitive to 10^–2 M CN^–.A similar induction of CN^–-insensitivity was also broughtabout with 10^–4 and 10^–3 M H₂S and 10^–4 MNaN₃. (Received July 6, 1971; )     

Light-induced changes in membrane potential in Spirogyra

Spirogyra cells exhibited changes in membrane potential whenthey were exposed to light. Cells made chloroplast-free didnot show any light-induced potential change (LPC) upon illuminationwith white light and also monochromatic red (680 nm) and farred (720 nm) light. LPC was observed when the cell containedonly a small fragment of chloroplast, whether the cell had anucleus or not. The magnitude of LPC depended on the amountof chloroplast in the cell. DCMU at 10^–5 M, CCCP at 10^–5 M and DNP at 10^–4M at pH 5.5 suppressed LPC, while CCCP at 1–5 ? 10^–6M, NH₄Cl at 5 ? 10^–2 M and DNP at 10^–4 M at pH 7.0stimulated LPC. PMS at 10^–4 M stimulated LPC and couldinduce LPC which was completely inhibited by DCMU. These factssuggest that LPC is related to noncyclic and cyclic electronflows. The influences of light and dark conditions and various metabolicinhibitors (DCMU, DNP, CCCP, NH₄Cl) on ATP level have been investigated.No significant difference in the ATP level was observed betweencells in the light and dark. DNP at 10^–4 M (pH 5.5) andCCCP at 5 ? 10^–6 M decreased the ATP level significantly,while DCMU and NH₄Cl only slightly. Good correlation was notfound between the total ATP level and LPC in Spirogyra. LPC occurred even when the external medium contained only asingle salt such as KCl, NaCl or CaSO₄. LPC was also recorded in chloroplasts in situ and in vitro.The mode of LPC of chloroplasts was quite different from thatof the cell. On illumination, the chloroplast potential changedvery rapidly and transiently in the positive direction thenrecovered spontaneously to almost the original potential level. Possible causes of LPC are discussed in relation to the electrogenicion pump. ¹ Present address: Department of Botany, Faculty of Science,University of Tokyo, Hongo, Bunkyo, Tokyo 113, Japan. (Received November 9, 1977; )     

The Effect of Selected Growth-Promoting and Growth-Inhibiting Substances on the Extension of Segments of Etiolated Tomato Seedling Hypocotyls

The effects of a number of growth-promoting and growth-inhibitingsubstances, including two fungal toxins, were studied on theextension of segments of etiolated tomato seedling hypocotyls.The bioassay was sensitive to small quantities of NaF, coumarinand ₂, ₄-DNP and inhibition was observed at all concentrations.₂, ₄-DNP or Iodoacetate stimulated growth at concentrationsbetween 1? 10^–4 and 5 ? 10^–6M. or 1 ? 10^–6and 1 ? 10^–7M. respectively. Inhibitor experiments inbuffered nutrient solution were approximately 10 per cent. moresensitive than those in deionized water. By means of paper partition chromatography small quantitiesof two fungal toxins, fusaric and alternaric acid were chromatographedand bioassayed. The effect of fussric acid (5, n-butyI picolinicacid) on hypocoty1 growth was detected at concentrations aslow as 1 ? 10^–5M. Experiments with recongnized growth-promoting substances showedthat Kinetin inhibited growth at concentrations up to 1 ?10^–8M.in both light and dark. IAA inhibited growth up to 1 ? 10^–6M.At 1 ? 10^–7 and 1 ? 10^–8 only small increases occurredwith IAA and the effect of light was negligible. Gibberellicacid (GA₂)stimulated growth at concentrations from 10^–3to 10^–7M. and significant increases up to 17 per cent.were recorded in the light. Since the light induced inhibitionwas only partly restored, the existence of some other naturallight sensitive growth substance is suggested. The value ofthe bioassay as a method for estimating natural growth-inhibitingand growth-promoting substances is discussed.     

The significance of apical K+ channels in mudpuppy feeding behavior

Behavioral experiments were conducted to determine the effectsof known potassium channel blockes, minnow extract (a naturalfood substance), and otber taste stimuli on feeding behaviorin the mudpuppy (Necturus maculosus). Groups of 10–15mudpuppies were presented with one or more of the followingchemicals dissolved in gelatin cubes: H₂O (control), NaCl (1.0M; 0.2 M), crude minnow extract (0.1 g/ml), CaCl₂ (1.0 M), citricacid (0.1 M; pH2, 0.01 M; pH 3), quinine HCl (0.1 M), tetraemylammoninmchloride (TEA; 0.1 M), KCl (1.0 M) and an amino acid mixturecontaining L-phenylalanine, L-arginine, L-histidine, L-valine,L-glutamic acid and L-lysinc at concentrations of 0.02 M and0.04 M each. Crude minnow extract was centrifuged and separatedinto components consisting of the following fractions: >500D, >1000 D and >14 000 D, each of which was presentedto mudpuppies in cubes. Results were expressed as the percentageof animals rejecting the cube within 5 min of taking the cubeinto the oral cavity. Statistical analysis using the Fisherexact test indicated that cubes containing chemicals known toblock (CaCl₂, TEA, citric acid and quinine) or permeate (KCl)apical K⁺ channels in mudpuppy taste cells were more aversivethan control cubes, and aversion was concentration-dependentIn contrast, cubes containing minnow extract and componentsof minnow extract < 14 000 D were significantly preferredover control cubes. Cubes containing a mixture of minnow extractand an aversive chemical were significantly less aversive thancubes containing only an aversive chemical. These findings suggestthat activation of the apical K⁺ conductance in mudpuppy tastecells triggers an aversive response, but the aversion can beameliorated by low molecular weight compounds present in minnowextract.     

Localization of absorbed aluminium in pea root and its binding to nucleic acids

Aluminium (Al³⁺) considerably inhibited the root elongationof Alaska pea at concentrations higher than 10^–4 M andpH 4.5. The absorbed Al³⁺ in roots was localized in the epidermisand regions where cells actively divided, such as root tip anddifferentiating lateral root. In cells, Al³⁺ accumulated mainlyin the nuclei and walls. Al³⁺ in nucleic acids increased upto 24 hr after treatment with 10^–3M AlCl₃, and did notdecrease markedly after transfer of 10-hr treated plants towater. Molecular sieving chromatography showed that Al³⁺ innucleic acids associated preferentially with DNA. However, invitro association of Al³⁺ with DNA and RNA was nearly the same.When Al³⁺ and nucleic acid were mixed in vitro, the moleculesof Al³⁺ being adjusted to be equal to those of phosphorus innucleic acid, only 25% of the Al³⁺ associated with nucleic acid.The presence of NaCl up to 0.4 M and MgCl₂ up to 0.05 M increasedthe association of Al³⁺ with DNA. When the phosphorus in DNAwas masked by histone, the association of Al³⁺ with DNA wasconsiderably reduced. (Received September 29, 1975; )     

Inhibition of hamster chorda tympani neural response by copper chloride

Copper chloride was evaluated as a specific inhibitor of neuralresponses to sweet taste stimuli in the goldern hamster (Mesocricetusauratus). The chorda tympani whole-nerve response to taste stimuliwas recorded before and after the tongue was treated for 30s with 0.01, 0.1 and 1 mM CuCl₂. Sweet stimuli [sucrose, fructose,saccharin (calcium salt), D-phenylalanine], which primarilystimulate chorda tympani S fibers, and non-sweet stimuli (NaCl,NH₄Cl) were used. At 0.01 mM, copper chloride had little effect.At 0.10 mM it partially inhibited responses to sucrose and saccharin,but had little effect on responses to D-Phe, fructose, NaCl,NH₄Cl, or a mixture of sucrose plus L-Phe. L-Phe, which hasthe same chelating properties as D-Phe, is not an S-fiber stimulusand likely reduced sucrose inhibiton by chelating the cupricion.Analysis of concentration–response functions revealedthat 0.1 mM copper chloride inhibited the neural response tolow concentrations of sucrose by about 25%, but did not significantlyinhibit high concentrations of surcrose, suggesting competitiveinhibition. In contrast, 0.1 mM CuCl₂ reduced saccharin responsesby 25% throughtout the effective range, suggesting non-competitiveinhibition. Occupation of a saccharide receptor site by coppermay interfere with dimer but not monomer reception and distortthe saccharin receptor site. At 1 mM, CuCl₂ non-competitivelyinhibited responses to sucrose, fructose, saccharin and thenon-sweet NaCl (an N-fiber stimulus), but not NH₄Cl (an H-fiberstimulus). The mechanisms of copper chloride inhibition aredifficult to establish because its effects are weak at concentrationswhere they are specific.     

Effect of Cytoplasmic Ca2+ on the Membrane Potential and Membrane Resistance of Chara Plasmalemma

Effects of cytoplasmic Ca²⁺ on the electrical properties ofthe plasma membrane were investigated in tonoplast-free cellsof Chara australis that had been internally perfused with media,containing either 1 mM ATP to fuel the electrogenic pump orhexokinase and glucose to deplete the ATP and stop the pump. In the presence of ATP, cytoplasmic Ca²⁺ up to 2.5?10^–5M did not affect the membrane potential (about -190 mV), butmembrane resistance decreased uniformly with increasing [Ca²⁺]_i.In the absence of ATP, the membrane potential, which was onlyabout -110 mV, was depolarized further by raising [Ca²⁺]_i from1.4?10^–6 to 2.5?10^–5 M. Membrane resistance, whichwas nearly the twofold that of ATP-provided cells, decreasedmarkedly with an increase in [Ca²⁺]_i from zero to 1.38?10^–6M, but showed no change for further increases. Internodal cellsof Nitellopsis obtusa were more sensitive to intracellular Ca²⁺with respect to membrane potential than were those of Charaaustralis, reconfirming the results obtained by Mimura and Tazawa(1983). The effect of cytoplasmic Ca²⁺ on the ATP-dependent H⁺ effluxwas measured. No marked difference in H⁺ effluxes was detectedbetween zero and 2.5?10^–5 M [Ca²⁺]_i; but, at 10^–4M the ATP-dependent H⁺ efflux was almost zero. Ca²⁺ efflux experimentswere done to investigate dependencies on [Ca²⁺]_i and [ATP]_i.The efflux was about 1 pmol cm^–2 s^–1 at all [Ca²⁺]_iconcentrations tested (1.38?10^–6, 2.5?10^–5, 10^–4M).This value is much higher than the influx reported by Hayamaet al. (1979), and this efflux was independent of [ATP]_i. Thepossibility of a Ca²⁺-extruding pump is discussed. ¹ Present address: Botanisches Institut der Universit?t Bonn,Venusbergweg 22, 5300 Bonn, F.R.G. (Received September 22, 1984; Accepted February 19, 1985)     

Purification of NADP-Malic Enzyme and Phosphoenolpyruvate Carboxylase from Sugar Cane Leaves

A procedure is described for the purification of phosphoenolpyruvatecarboxylase (EC 4.1.1.31 [EC] ) and NADP-dependent malic enzyme (EC1.1.1.40 [EC] ) from sugar cane leaves. Each enzyme was purified tohomogeneity as judged by sodium dodecyl sulfate-polyacrylamidegel electro-phoresis, with about 30% yield. Phosphoenolpyruvatecarboxylase was purified 54-fold. A molecular weight of 400,000and a homotetrameric structure were determined for the nativeenzyme. The purified carboxylase had a specific activity of20.0 {diaeresis}mol (mg protein)_–1 min_–1, and wasactivated by glucose-6-phosphate and inhibited by L-malate.K_m values at pH 8.0 for phosphoenolpyruvate and bicarbonatewere 0.25 and O.l0 mM, respectively. NADP-malic enzyme, 356-foldpurified, exhibited a specific activity of 71.2 {diaeresis}mol(mg protein)_–1 min_–1 and was characterized as ahomotetramer with native molecular weight of 250,000. Purifiedmalic enzyme showed an absolute specificity for NADP⁺ and requireda divalent metal ion for activity. K_m values of 0.33 and 0.008mM for L-malate and NADP⁺, respectively, were determined. Thisenzyme was inhibited by several organic acids, including ketoand amino acids; while succinate and citrate increased the enzymeactivity when assayed with 10{diaeresis}M L-malate. The effectsshown by amino acids and by citrate were dependent on pH, beinghigher at pH 8.0 than at pH 7.0. (Received October 26, 1988; Accepted February 3, 1989)     

Enhancing effects of transition metals on the salt taste responses of single fibers of the frog glossopharyngeal nerve: specificity of and similarities among Ca2+, Mg2+ and Na+ taste responses

Fibers of the frog glossopharyngeal nerve (water fibers) thatare sensitive to water also respond to CaCl₂, MgCl₂ and NaCl.In the present study, interaction among cations (Ca²⁺, Mg²⁺and Na⁺) on taste cell membrane in frogs was studied using transitionmetals (NiCl₂, CoCl₂ and MnCl₂), which themselves are barelyeffective in producing neural response at concentrations below5 mM. Unitary discharges from single water fibers were recordedfrom fungiform papillae with suction electrode. Transition metalions (0.05–5.0 mM) had exclusively enhancing effects onthe responses to 50 mM Ca²⁺, 100 mM Mg²⁺ and 500 mM Na⁺. Theeffects of transition metal ions were always reversible. Therank order of effectiveness of transition metals at 1 mM inthe enhancement of the responses to 50 mM CaCl₂, 100 mM MgCl₂and 500 mM NaCl was NiCl₂ > CoCl² > MnCl₂. The concentrationof transition metal ions effective to enhance salt responsewas almost the same among Ca²⁺, Mg²⁺ and Na⁺ responses. Theresults suggest that a common mechanism is involved in the enhancementof Ca²⁺, Mg²⁺ and Na⁺ taste responses. The enhanced Mg²⁺ responseand the enhanced Na⁺ response were greatly inhibited by theaddition of Ca²⁺ ions, and the enhanced Ca²⁺ response was inhibitedby the addition of Mg²⁺ or Na⁺ ions, suggesting that competitiveantagonism occurs between Ca²⁺ and Mg²⁺ ions and between Ca²⁺and Na⁺ ions in the presence of Ni²⁺ ions. Ni²⁺ ions had a dualeffect on the Ca²⁺ response induced by low concentration (0.1mM) of CaCl₂: enhancement at lower concentrations (0.02–0.1mM) of NiCl₂ and inhibition at higher concentrations (0.5–5mM)of NiCl₂. The present results suggest that transition metalions do not affect the receptor-antagonist complex, but affectonly the receptor-agonist complex.     

Effect of Gibberellic Acid and Ethylene on Peroxidase in Pea Internodes

Ethylene at 5–80 µl l^–1 inhibited elongationand induced swelling in internodes of light-grown normal anddwarf pea plants; GA₃ did not prevent swelling in response toethylene. GA₃ neither inhibited nor enhanced the activity of isoperoxidasesin the internodes, regardless of its effect on their elongation.Ethylene at 80 µl l^–1 enhanced peroxidase in GA₃-untreatedand treated normal and dwarf plants. At 5 µl l^–1,ethylene had only a weak effect on peroxidase activity or none.The enzyme enhancement by ethylene was not related to its effecton cell expansion and seems do be due, at least in part, tochemical injury. Electron microscopy revealed peroxidase activity in the roughER and cell walls, including intercellular spaces. Stainingof walls in ethylene-treated tissues was more pronounced thanin untreated ones. Golgi vesicles did not seem to be involvedin the assembly of the enzyme carbohydrate moiety in ethylene-treatedcells. The peroxidase fraction extracted with 20 mM phosphate buffer,pH 6, and that extracted from wall debris with 1 M NaCl accountedfor 98% of total enzyme activity. Both fractions contained thesame six cathodic isoforms which comprised 85–90% of theiractivity. Electrophoresis did not reveal differences in thequalitative isoenzyme patterns in relation to variety, age,GA₃, or ethylene. The only observed quantitative differenceswere age-dependent. Procedural artefacts during separation of protoplast and wallionically bound peroxidase fractions are discussed.     

The NADP$-specific isocitrate dehydrogenase of Rhodospirillum rubrum

The NADP^$-specific isocitrate dehydrogenase was partially purifiedfrom photosynthetically-grown Rhodospirillum rubrum. The pHoptimum is between 7.5 and 9.0 in phosphate buffer. The apparentKm is 3.1x10^–5 M for isocitrate, 5.1x10^–5 M forNADP^$, 1.7x10^–5 M for manganese, 1.5x10^–4 M formagnesium, and 3.5x10^–3 M for inorganic orthophosphate.Arsenate exerts a slight inhibition. The Q₁₀ between 17.5°Cand 40°C is 1.62, and the energy of activation at 25°Cis 9.74 Kcal/mole. Glyoxylate and oxalacetate cause concertedinhibition of the enzyme activity. Various nucleotides inhibitthe activity. The kinetics of inhibition by ATP was found tobe mixed type with respect to NADP^$ and isocitrate, the K_i valuesbeing 1.17x10^–3 M and 1.10x10^–3 M respectively.The inhibition between ATP and orthophosphate is competitivewith a K_i of 10^–4M. Thiol binding reagents are inhibitory;this inhibition is reversed by cysteine or reduced glutathione. (Received October 1, 1971; )     

Changes in Outward K+ Currents in Response to Two Types of Sweeteners in Sweet Taste Transduction of Gerbil Taste Cells

Using the whole cell patch clamp technique, we measured changesin outward K⁺ currents of gerbil taste cells in response todifferent kinds of sweeteners. Outward K⁺ currents of the tastecell induced by depolarizing pulses were suppressed by sweetstimuli such as 10 mM Na-saccharin. The membrane-permeable analogof cAMP, cpt-cAMP, also decreased outward K⁺ currents. On theother hand, the K+ currents were enhanced by amino acid sweetenerssuch as 10 mM D-tryptophan. The outward K⁺ current was enhancedby external application of Ca²⁺-transporting ionophore, 5 µMionomycin, and intracellular application of 5 µM inositol-1,4,5-trisphosphate(IP₃). The outward K⁺ currents were no longer suppressed by10 mM Na-saccharin containing 20 µM gurmarin, but werestill enhanced by 10 mM D-tryptophan containing 20 µMgurmarin. These results suggest that sweet taste transductionfor one group of sweeteners such as Na-saccharin in gerbilsis concerned with an increase of the intracellular cAMP level,and that the transduction for the other group of sweetenerssuch as D-tryptophan is concerned with an increase of the intracellularIP₃ level which releases Ca²⁺ from the internal stores. Chem.Senses 22: 163–169, 1997.     

Spatial and Temporal Aspects of Growth in the Primary Root of Cotton Seedlings: Effects of NaCl and CaCl2

Seedlings of cotton (Gossypium hirsutum L. cv. Acala SJ-2) weregrown in modified Hoagland nutrient solution with various combinationsof NaCl and CaCl₂. Marking experiments and numerical analysiswere conducted to characterize the spatial and temporal patternsof cotton root growth at varied Na/Ca ratios. At 1 mol m^–3Ca, 150 mol m^–3 NaCl reduced overall root elongation rateto 60% of the control, while increasing Ca to 10 mol m^–3at the same NaCl concentration restored the elongation rateto 80% of the control. Analysis of the spatial distributionof elongation revealed that the presence of 150 mol m^–3NaCl in the medium shortened the growth zone by about 2 mm fromthe approximate 10 mm in the control and also reduced the relativeelemental elongation rate (i.e. the longitudinal strain rate,defined as the derivatives of displacement velocity of a cellularparticle with respect to position on root axis). Supply of 10mol m^–3 Ca at the high salt condition restored partiallythe relative elemental elongation rate, but not the length ofthe growth zone. Compared to the control, the growth trajectoriesshowed that at 1 mol m^–3 CaCl2 it took more time for acellular particle to move through the growth zone at 150 molm^–3 NaCl, while at 10 mol m^–3 CaCl it took lesstime and there was no difference between the NaCl treatments Key words: Gossypium hirsutum, salinity stress, root growth kinematics