Hydrotropism in roots: sensing of a gradient in water potential by the root cap

Roots of the agravitropic pea (Pisum sativum L.) mutant, ageotropum, responded to a gradient in water potential as small as 0.5 MPa by growing toward the higher water potential. This positive response occurred when a sorbitol-containing agar block was unilaterally applied to the root cap but not when applied to the elongation region. Unilateral application of higher concentrations of sorbitol to the elongation region caused root curvature toward the sorbitol source, presumably because of growth reduction on the water-stressed side. The control blocks of plain agar applied to either the root cap or the elongation region did not cause significant curvature of the roots. These results demonstrate that hydrotropism in roots occurs following perception of a gradient in water potential by the root cap.     

Effects of lanthanum on rhythmic and nyctinastic leaflet movements in Albizzia lophantha

The involvement of extracellular calcium in rhythmic and nyctinasticmovement oi Albizzia lophantha Benth. leaflets has been studiedby testing the effect of LaCl₃ and its interaction with thephytochrome control of these movements. A 2h pulse of LaCl₃(10–50 mM) promotes a loss of rhythmicity, leaving leafletsin an open position, and also overrides the phase shift causedby phytochrome. A 2 h pulse of LaCl₃ (1 mM) decreases the amplitudeof rhythmic oscillations but does not promote arhythmicity normodify the phase shift caused by red light. The red light pulseabolishes the damping effect of 1 mM La³⁺. LaCl₃ inhibits nyctinasticclosure and decreases the phytochrome control of nyctinasticclosure. A subsequent supply of CaCl₂ (10 to 100 mM) does notreverse La³⁺ (10 mM) inhibition of closure. Light-induced openingis independent of LaCl, but rhythmic opening in darkness showsdifferent responses to La³⁺ depending on the time at which La³⁺is applied. Data suggest that extracellular calcium is requiredfor the closure mechanism and for the expression of rhythmicmovement. It could also be involved in the phytochrome transductionpathway and/or in the linking steps between phytochrome andthe circadian clock. Key words: Albizzia lophantha, calcium, circadian rhythm, lanthanum, phytochrome     

Hydrotropism in abscisic acid,wavy, and gravitropic mutants of Arabidopsis thaliana

We have developed experimental systems to study hydrotropism in seedling roots of Arabidopsis thaliana (L.) Heynh. Arabidopsis roots showed a strong curvature in response to a moisture gradient, established by applying 1% agar and a saturated solution of KCl or K(2)CO(3) in a closed chamber. In this system, the hydrotropic response overcame the gravitropic response. Hydrotropic curvature commenced within 30 min and reached 80-100 degrees within 24 h of hydrostimulation. When 1% agar and agar containing 1 MPa sorbitol were placed side-by-side in humid air, a water potential gradient formed at the border between the two media. Although the gradient changed with time, it still elicited a hydrotropic response in Arabidopsis roots. The roots curved away from 0.5-1.5 MPa of sorbitol agar. Various Arabidopsis mutants were tested for their hydrotropic response. Roots of aba1-1 and abi2-1 mutants were less sensitive to hydrotropic stimulation. Addition of abscisic acid restored the normal hydrotropic response in aba1-1 roots. In comparison, mutants that exhibit a reduced response to gravity and auxin, axr1-3 and axr2-1, showed a hydrotropic response greater than that of the wild type. Wavy mutants, wav2-1 and wav3-1, showed increased sensitivity to the induction of hydrotropism by the moisture gradient. These results suggest that auxin plays divergent roles in hydrotropism and gravitropism, and that abscisic acid plays a positive role in hydrotropism. Furthermore, hydrotropism and the wavy response may share part of a common molecular pathway controlling the directional growth of roots.     

The effects of asymmetric calcium application on primary root curvature of ageotropum pea mutant.

Recent studies indicate that roots of ageotropum seedlings can be used to study the hydrotropic response of roots independent of physiological events related to the gravity response of roots. There is evidence that Ca2+ ions are important in both the gravitropic and hydrotropic response of roots. In this study, we have compared three fully graviresponsive pea cultivars and the ageotropum mutant with regard to: 1) general root anatomy, 2) the effects of unilateral Ca application to both the root cap and DEZ region on root curvature, and 4) effects of unilateral application of EGTA to the DEZ region.     

Induction of Curvature in Maize Roots by Calcium or by Thigmostimulation : Role of the Postmitotic Isodiametric Growth Zone

We examined the response of primary roots of maize (Zea mays L. cv Merit) to unilateral application of calcium with particular attention to the site of application, the dependence on growth rate, and possible contributions of thigmotropic stimulation during application. Unilateral application of agar to the root cap induced negative curvature whether or not the agar contained calcium. This apparent thigmotropic response was enhanced by including calcium in the agar. Curvature away from objects applied unilaterally to the extreme root tip occurred both in intact and detipped roots. When agar containing calcium chloride was applied to one side of the postmitotic isodiametric growth zone (a region between the apical meristem and the elongation zone), the root curved toward the side of application. This response could not be induced by plain agar. We conclude that curvature away from calcium applied to the root tip results from a thigmotropic response to stimulation during application. In contrast, curvature toward calcium applied to the postmitotic isodiametric growth zone results from direct calcium-induced inhibition of growth.     

Characterization of hydrotropism: the timing of perception and signal movement from the root cap in the agravitropic pea mutant ageotropum

In this study, ageotropum pea mutant was used to determine the threshold time for perception of an osmotic stimulation in the root cap and the time requirement for transduction and transmission of the hydrotropic signal from the root cap to the elongation region. The threshold time for the perception of an osmotic stimulation was compared to current estimates of threshold times for graviperception in roots. The time required for transduction and transmission in the hydrotropic response of ageotropum was compared to the time requirement in the gravity response of Alaska pea roots. We determined that threshold time for perception of an osmotic stimulation in the root cap is very rapid, occurring in less than 2 min following the application of sorbitol to the root cap. Furthermore, a single 5 min exposure of sorbitol to the root cap fully induced a hydrotropic response. We also found that transduction and transmission of an osmotic stimulus requires 90-120 min for movement from the root cap to more basal tissues involved in differential growth leading to root curvature. The very rapid threshold time for perception of root hydrotropism is similar to those times reported for root gravitropism. However, the time required for the transduction and transmission of an osmotic stimulation from the root cap is significantly longer than the time required in gravitropism. These results suggest that there must exist some differences between root hydrotropism and gravitropism in either the rate or mechanisms of transduction and transmission of the tropistic signal from the root cap.     

Induction of hydrotropism in clinorotated seedling roots of Alaska pea,Pisum sativum L.

Roots of the agravitropic pea (Pisum sativum L.) mutantageotropum show positive hydrotropism, whereas roots of Alaska peas are hydrotropically almost non-responsive. When the gravitropic response was nullified by rotation on clinostats, however, roots of Alaska peas showed unequivocal positive hydrotropism in response to a water potential gradlent. These results suggest that roots of Alaska peas possess normal ability to respond hydrotropically and their weak hydrotropic response results from a counteracting effect of gravitropism.     

Peak power output is maintained in rabbit psoas and rat soleus single muscle fibers when CTP replaces ATP

The chemomechanicalcoupling mechanism in striated muscle contraction was examined bychanging the nucleotide substrate from ATP to CTP. Maximum shorteningvelocity [extrapolation to zero force from force-velocity relation(V_max) andslope of slack test plots (V₀)], maximumisometric force (P_o), power, andthe curvature of the force-velocity curve[a/P_o(dimensionless parameter inversely related to the curvature)] weredetermined during maximumCa²⁺-activated isotoniccontractions of fibers from fast rabbit psoas and slow rat soleusmuscles by using 0.2 mM MgATP, 4 mM MgATP, 4 mM MgCTP, or 10 mM MgCTPas the nucleotide substrate. In addition to a decrease in the maximumCa²⁺-activated force in both fibertypes, a change from 4 mM ATP to 10 mM CTP resulted in a decrease inV_max in psoasfibers from 3.26 to 1.87 muscle length/s. In soleus fibers,V_max was reduced from 1.94 to 0.90 muscle length/s by this change in nucleotide. Surprisingly, peak power was unaffected in either fiber type by thechange in nucleotide as the result of a three- to fourfold decrease inthe curvature of the force-velocity relationship. The results areinterpreted in terms of the Huxley model of muscle contraction as anincrease in f₁and g₁ coupled toa decrease in g₂(where f₁ is therate of cross-bridge attachment and g₁ andg₂ are rates ofdetachment) when CTP replaces ATP. This adequately accounts for theobserved changes in P_o,a/P_o,and V_max.However, the two-state Huxley model does not explicitly reveal thecross-bridge transitions that determine curvature of the force-velocityrelationship. We hypothesize that a nucleotide-sensitive transitionamong strong-binding cross-bridge states followingP_i release, but before the release of the nucleotide diphosphate, underlies the alterations ina/P_o reported here.

     

Effects of Osmotic Stress, Ionic Stress and IAA on the Cell-Membrane Resistance of Vigna Hypocotyls

The cell-membrane resistance (R_m) of Vigna hypocotyls was examined,and the effects of osmotic stress, ionic stress and IAA on R_mwere investigated. R_m decreased by 64 to 77% under osmotic stressin the presence of absorbable solutes (40 mM sorbitol, 15 mMKC1, 30 mM sucrose; or 40 mM sorbitol, 15 mM KC1, 30 mM sucroseplus 10^–4 M IAA) or under ionic stress (50 mM NaCl or50 mM KC1). R_m was not changed by perfusion with 10^–4M IAA. Therefore, the hyper-polarizations of the membrane potentialobserved in both cases should be ascribed totally to the activationof the electrogenic proton pump. Although R_m showed an increaseof 1.6 fold when the hypocotyls were subjected to osmotic stress(100 mM sorbitol or 100 mM sorbitol plus 10^–4 M IAA),83.6% or 92.4% of the hyperpolarization of the membrane potential(V_px was also the result of the activation of the pump. Theresults, calculated on the basis of the current source model,support the viewpoint that the hyperpolarization of the cellmembrane potential of Vigna hypocotyls under osmotic stress,ionic stress or in the presence of IAA is an expression of theactivation of the proton pump, and is not caused by an increasein R_m. ¹ Present address: Researchers and Planners of Natural Environment, Yotsugi Bldg. (2F), 1-5-4 Horinouchi, Suginami-Ku, Tokyo,166 Japan ² Present address: Graduate School of Integrated Science, YokohamaCity University, 22-2 Seto, Kanazawa-Ku, Yokohama, 236 Japan (Received February 14, 1991; Accepted July 24, 1991)     

Collection of Gravitropic Effectors from Mucilage of Electrotropically-stimulated Roots of Zea mays L.

We placed agar blocks adjacent to tips of electrotropicallystimulated primary roots of Zea mays. Blocks placed adjacentto the anode-side of the roots for 3 h induced significant curvaturewhen subsequently placed asymmetrically on tips of vertically-orientedroots. Curvature was always toward the side of the root ontowhich the agar block was placed. Agar blocks not contactingroots and blocks placed adjacent to the cathode-side of electrotropicallystimulated roots did not induce significant curvature when placedasymmetrically on tips of vertically-oriented roots. Atomicabsorption spectrophotometry indicated that blocks adjacentto the anode-side of electrotropically-stimulated roots containedsignificantly more calcium than (1) blocks not contacting roots,and (2) blocks contacting the cathode-side of roots. These resultsdemonstrate the presence of a gradient of endogenous Ca in mucilageof electrotropically-stimulated roots (i.e. roots undergoinggravitropic-like curvature). Zea mays, corn, mucilage, root gravitropism, electrotropism     

Characterization of Hexose Transporter for Facilitated Diffusion of the Tonoplast Vesicles from Pear Fruit

Tonoplast vesicles were prepared from the flesh tissue of maturepear fruit. Sugar uptakes into the vesicles determined by twodifferent methods, the membrane and the gel filtration methods,were quite similar. The uptake was highest for glucose and subsequently,in order, for fructose, sucrose and sorbitol. It was not stimulatedby addition of ATP, although the vesicles could create a protongradient. However, the uptakes were significantly inhibitedby p-chloromercuribenzene sulphonate (PCMBS, SH-reagent andinhibitor of sugar transporter). Further, the PCMBS-sensitiveuptakes of glucose and fructose saturated with their increasedconcentrations. Thus, these PCMBS-sensitive uptakes are mediatedby the transporter of facilitated diffusion. The uptakes ofglucose or fructose each had two K_m values. K_m values for glucosewere 0.35 and 18 mM, and those for fructose were 1.6 and 25raM. The uptake of 0.2 mM glucose was inhibited by 2 mM fructoseand that of 2 mM fructose was inhibited by 2 mM glucose, butneither was inhibited by sucrose or sorbitol. O-methyl-glucose(OMG) also inhibited both the glucose and fructose uptakes.Therefore, the same transporter may mediate both glucose andfructose uptakes at lower concentrations; this hexose transportsystem differed from the sucrose and sorbitol transport systems. ¹Research Fellow of the Japan Society for the Promotion of Science. ²Present address: Faculty of Agriculture, Tohoku University,1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, 981 Japan.     

Calcium and Rhizodermal Differentiation in Primary Maize Roots

Rhizodermal differentiation of maize (Zea mays L. cv. LG 11)roots cultured in humid air was influenced by a pretreatmentfor 2 h in CaCl₂ or CaSO₄ solutions. This increased the numberof hair-producing roots and the density of hairs. Ethylene glycol-bis-(ß-aminoethylether)N,N'-tetraacetic acid (EGTA) was inhibitory. Root hairsemerged in the part of the cell nearer to the tip. Trichoblastswere shorter and elongated more slowly than atrichoblasts. Theelongation of the lower part of the trichoblast was less thanthat of the upper part. Key words: Cell length, cell number, hair position     

Correlations between Gravitropic Curvature and Auxin Movement across Gravistimulated Roots of Zea mays

We compared the kinetics of auxin redistribution across the caps of primary roots of 2-day-old maize (Zea mays, cv Merit) seedlings with the time course of gravitropic curvature. [³H] indoleacetic acid was applied to one side of the cap in an agar donor and radioactivity moving across the cap was collected in an agar receiver applied to the opposite side. Upon gravistimulation the roots first curved upward slightly, then returned to the horizontal and began curving downward, reaching a final angle of about 67°. Movement of label across the caps of gravistimulated roots was asymmetric with preferential downward movement (ratio downward/upward = ca. 1.6, radioactivity collected during the 90 min following beginning of gravistimulation). There was a close correlation between the development of asymmetric auxin movement across the root cap and the rate of curvature, with both values increasing to a maximum and then declining as the roots approached the final angle of curvature. In roots preadapted to gravity (alternate brief stimulation on opposite flanks over a period of 1 hour) the initial phase of upward curvature was eliminated and downward bending began earlier than for controls. The correlation between asymmetric auxin movement and the kinetics of curvature also held in comparisons between control and preadapted roots. Both downward auxin transport asymmetry and downward curvature occurred earlier in preadapted roots than in controls. These findings are consistent with suggestions that the root cap is not only the site of perception but also the location of the initial redistribution of effectors that ultimately leads to curvature.     

玉米初生根向水性诱导优化试验研究

为了研究湿度梯度对根系向水性反应的影响,采用Takahashi and Scott于1993年创建的方法,设置以下3个试验:1)向水性诱导物不同倾斜角试验;2)根系距向水性诱导物不同距离试验;3)根尖距底部饱和K2CO3溶液不同距离试验。同时,还研究了根长和根系延伸速率对根系向水性弯曲的影响。结果表明,用饱和K2CO3溶液控制湿度时根系的向水性弯曲度明显大于纯水。随着诱导物倾斜角的增大,向水性弯曲增强。与距诱导物3 mm和6 mm相比,根系直接接触诱导物时表现出最大的向水性反应。与根尖距底部盐溶液6 cm相比,相距4 cm时向水性弯曲度增大,这些与根尖周围的湿度梯度增大有关。当根长为1.0、1.5、2.0、2.5、3.0 cm时,短根比长根表现出更大的向水性反应,这可能与其较慢的延伸速率为根系对湿度梯度的反应提供了更充足的时间有关。为了验证这个假说,用相同长度的根系、通过控制不同温度进行试验,结果表明根系的向水性弯曲随温度升高而降低。可见,玉米初生根的向水性反应受环境和根系发育阶段两方面影响。当根系相距诱导物较近、根系周围的湿度梯度较大时,根系向水性反应更强。而且,具有较小延伸速率根系的向水性反应更大。考虑到干旱条件下根系伸长慢、且土壤中湿度梯度大,因而可以认为干旱条件下根系的向水性生长在玉米吸收水分中有重要作用。同时,对根系向水性诱导方法的优化有助于其生理机制的进一步研究。     

Studies on the change of the respiratory system in roots in relation to the growth of plants II. Activity of iron-containing oxidases in roots of cereal crops with the aging of plants

Distribution of iron-containing oxidases in aging nodal rootsof rice and wheat was studied. Activities of cytochrome c oxidase(1.9.3.1 [EC] , cytochrome c : O₂ oxidoreductase), catalase (1.11.1.6 [EC] ,H₂O₂: H₂O₂ oxidoreductase) and peroxidase (1.11.1.7 [EC] , donor:H₂O₂ oxidoreductase) in wheat roots were comparatively higherthan were those in rice roots at corresponding stages. Cytochromec oxidase in roots remained active throughout the lives of therice and wheat crops. In rice roots, catalase seemed to playa distinct role around the panicle formation stage. Decay ofcatalase activity took place earlier than did that of peroxidaseand cytochrome c oxidase activities. In wheat roots similarenzyme activity changes were not observed. Data may suggestthat the high activity of iron containing oxidases at the panicleformation stage (I) may be chiefly due to catalase activityin rice roots. ¹Paper presented at the 14th Annual Meeting of the Society ofthe Science of Soil and Manure, Japan (1968). (Received November 21, 1968; )     

Role of extracellular [Ca2+] in fatigue of isolated mammalian skeletal muscle

The possiblerole of altered extracellular Ca²⁺concentration([Ca²⁺]_o)in skeletal muscle fatigue was tested on isolated slow-twitch soleusand fast-twitch extensor digitorum longus muscles of the mouse. Thefollowing findings were made. 1) Achange from the control solution (1.3 mM[Ca²⁺]_o)to 10 mM[Ca²⁺]_o,or to nominally Ca²⁺-freesolutions, had little effect on tetanic force in nonfatigued muscle.2) Almost complete restoration oftetanic force was induced by 10 mM[Ca²⁺]_oin severely K⁺-depressed muscle(extracellular K⁺ concentration of10-12 mM). This effect was attributed to a 5-mV reversal of theK⁺-induced depolarization andsubsequent restoration of ability to generate action potentials(inferred by using the twitch force-stimulation strength relationship).3) Tetanic force depressed bylowered extracellular Na⁺concentration (40 mM) was further reduced with 10 mM[Ca²⁺]_o.4) Tetanic force loss at elevatedextracellular K⁺ concentration (8 mM) and lowered extracellular Na⁺concentration (100 mM) was partially reversed with 10 mM[Ca²⁺]_oor markedly exacerbated with low[Ca²⁺]_o.5) Fatigue induced by using repeatedtetani in soleus was attenuated at 10 mM[Ca²⁺]_o(due to increased resting and evoked forces) and exacerbated at low[Ca²⁺]_o.These combined results suggest, first, that raised[Ca²⁺]_oprotects against fatigue rather than inducing it and, second, that aconsiderable depletion of[Ca²⁺]_oin the transverse tubules may contribute to fatigue.

     

Effect of EGTA on Nyctinastic Closure Mediated by Phytochrome in Albizzia lophantha

The role of extracellular calcium on nyctinastic closure ofAlbizzia lophantha leaflets has been studied by testing theeffect of ethyleneglycol-bis-(ß-aminoethylether)-N,N,N',N'-tetraaceticacid (EGTA) and its reversibility by calcium. EGTA (1 and 5mM) causes an inhibition of nyctinastic closure and at a concentrationof 1 mM EGTA it decreases the difference between the effectof red light (R) and far-red light (FR) irradiation on leafletclosure. A simultaneous or subsequent supply of CaCl₂ (5 or10 mM) reverses EGTA (5 mM) inhibition on closure as well ascausing an additional promotion of closure. We suggest that external calcium could play a dual role in nyctinasticclosure. Phytochrome control of leaflet closure probably needsexternal Ca²⁺ and, in addition, Ca²⁺ could regulate the closuremechanism by controlling ionic fluxes through the plasma membranein pulvinular motor cells. (Received June 9, 1989; Accepted November 27, 1989)     

Some Basic Psychophysics of Calcium Salt Solutions

Detection thresholds and the taste qualities of suprathresholdconcentrations of calcium salt solutions were assessed. Averagetaste detection thresholds for calcium chloride (CaCl₂), lactate(CaLa), hydroxide, phosphate and gluconate ranged between 8and 50 mM, with no reliable differences among the various salts.Between-subject variability ranged over four orders of magnitudeand reliability coefficients for repeated detection thresholdtests of CaCl₂ averaged r = 0.52. In an odor detection test,subjects could reliably discriminate 100 but not 1 mM CaCl₂and CaLa from water. The taste of suprathreshold concentrations(1–100 mM) of CaCl₂ and CaLa was considered unpleasant.At 1 mM, CaCl₂ solution was rated as 35% bitter, 32% sour, 29%sweet and 4% salty. At higher concentrations the sweet componentdiminished and the salty component increased, so that 100 mMCaCl₂ was rated as 44% bitter, 20% sour, 1% sweet and 35% salty.CaLa solutions were considered to be significantly less bitterand marginally more sour than equimolar CaCl₂ solutions. Thus,the taste of calcium varied with both the form and concentrationof salt tested, but included both sour and bitter components.Saltiness was identified only in high (     

Effects of Calcium on NAD(H)-Glutamate Dehydrogenase from Turnip (Brassica rapa L.) Mitochondria

The effect of Ca²⁺ and ammonia on mitochondrial NADH-glutamatedehydrogenase (GDH: EC 1.4.1.2 [EC] ) isolated from turnip root (Brassicarapa L.) activity was examined. Increasing the ammonia [(NH₄)₂SO₄]concentration led to significant substrate inhibition whichcould be reversed by micromolar levels of Ca²⁺. The sensitivityof the enzyme to ammonia inhibition and its reversal by Ca²⁺was affected by proteolysis. After treatment with various proteases,lower concentrations of Ca²⁺ were capable of fully activatingthe enzyme or overcoming the inhibitory effects of high ammonium,compared to non-treated enzyme. However, the protease-treatedenzyme was still sensitive to ethylene glycol-bis(ß-aminoethylether) N,N,N',N'-tetraacetate (EGTA). In contrast, NADH-GDHactivity was inhibited approx. 30% by organic mercurials (200µm), but the residual activity was not affected by thesubsequent additions of EGTA. NADH-GDH activity could also bestimulated by additions of high concentrations of NaCl (300mM) in the absence of added Ca²⁺. These results suggest thathydrophobic and -SH groups may be involved in the regulationof mitochondrial NADH-GDH activity by Ca²⁺. ² Present address: CSIRO Division of Horticulture, Urrbrae,S.A. 5064, Australia (Received April 18, 1990; Accepted July 23, 1990)     

Neuronal swelling and surface area regulation: elevated intracellular calcium is not a requirement

Neurons aremechanically robust. During prolonged swelling, molluscan neurons cantriple their apparent membrane area. They gain surface area andcapacitance independent of extracellular Ca concentration([Ca]_e), but it isunknown if an increase in intracellular Ca concentration([Ca]_i) isnecessary. If Ca for stimulating exocytosis is unnecessary, it ispossible that swelling-induced membrane tension changes directlytrigger surface area readjustments. If, however, Ca-mediated but nottension-mediated membrane recruitment is responsible for surface areaincreases, swelling neurons should sustain elevated levels of[Ca]_i. The purpose ofthis investigation is to determine if the[Ca]_i in swellingneurons attains levels high enough to promote exocytosis and if anysuch increase is required. Lymnaeaneurons were loaded with the Ca concentration indicator fura 2. Calibration was performed in situ using 4-bromo-A-23187 and Ca-ethyleneglycol-bis(-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA), with free Ca concentration ranging from 0 to 5 µM. Swelling perturbations (medium osmolarity reduced to 25% for 5 min)were done at either a standard[Ca]_e or very low[Ca]_e level (0.9 mM or0.13 µM, respectively). In neither case did the[Ca]_i increase tolevels that drive exocytosis. We also monitored osmomechanically drivenmembrane dynamics [swelling, then formation and reversal ofvacuole-like dilations (VLDs)] with the[Ca]_i clamped below 40 nM via1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). [Ca]_idid not change with swelling, and VLD behavior was unaffected,consistent with tension-driven,[Ca]_i-independent surface area adjustments. In addition, neurons with[Ca]_i clamped at 0.1 µM via an ionophore could produce VLDs. We conclude that, undermechanical stress, neuronal membranes are compliant by virtue ofsurface area regulatory adjustments that operate independent of[Ca]_i. The findingssupport the hypothesis that plasma membrane area is regulated in partby membrane tension.