An anion channel in guinea pig gallbladder epithelial cells is highly permeable to HCO(-)(3)

In guinea pig gallbladder epithelium, a secretion of fluid, secondary to an electrogenic secretion of Cl(-) and HCO(-)(3), is elicited in the presence of a high intracellular concentration of adenosine 3'-5'-cyclic monophosphate (cAMP). The aim of this study was to analyze the effects of secretagogues on the activity of anionic channels in isolated epithelial cells using the patch-clamp technique and measuring the electrical potential difference of the cellular membrane (pd(cm)). In cell-attached configuration, with the microelectrode filled with a solution of N-methylglucamine-Cl, or in inside-out configuration (symmetrical solution), it was possible to demonstrate the presence of an 18-pS Cl(-) channel with linear current/voltage (I/V) relationship and voltage independence; this channel is not activated by cAMP (cell-attached configuration). In inside-out configuration (symmetrical solution), another anionic channel with a conductance of 2.8 pS, voltage independence, and a linear I/V relationship was also identified. This channel was stimulated by cAMP (cell-attached configuration) and by PKA + ATP + cAMP (inside-out configuration). The channel was inhibited by NPPB (10(-5) M), but not by other anionic inhibitors. Measurements of the pd(cm) value suggested that in isolated cells, as in whole tissue, cAMP activates conductance for both Cl(-) and HCO(-)(3). The selectivity of the channel was gluconate < SO(2-)(4) < Cl(-) < Br(-) < I(-) < HCO(-)(3) < SCN(-) and the P(HCO(3))/P(Cl) was 2.6. Some features of the channel resemble those of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel and RT-PCR performed on mRNA from isolated epithelial cells detected the presence of a CFTR homologue mRNA. The results obtained indicate that this channel is responsible for the HCO(-)(3) conductance activated by cAMP.     

Heterogeneity of chloride channels in the apical membrane of isolated mitochondria-rich cells from toad skin

The isolated epithelium of toad skin was disintegrated into single cells by treatment with collagenase and trypsine. Chloride channels of cell-attached and excised inside-out apical membrane-patches of mitochondria-rich cells were studied by the patch-clamp technique. The major population of Cl- channels constituted small 7-pS linear channels in symmetrical solutions (125 mM Cl-). In cell-attached and inside-out patches the single channel i/V-relationship could be described by electrodiffusion of Cl- with a Goldmann-Hodgkin-Katz permeability of, PCl = 1.2 x 10(-14) - 2.6 x 10(-14) cm3. s-1. The channel exhibited voltage-independent activity and could be activated by cAMP. This channel is a likely candidate for mediating the well known cAMP-induced transepithelial Cl- conductance of the amphibian skin epithelium. Another population of Cl- channels exhibited large, highly variable conductances (upper limit conductances, 150-550 pS) and could be activated by membrane depolarization. A group of intermediate-sized Cl(- )-channels included: (a) channels (mean conductance, 30 pS) with linear or slightly outwardly rectifying i/V-relationships and activity occurring in distinct "bursts," (b) channels (conductance-range, 10-27 pS) with marked depolarization-induced activity, and (c) channels with unresolvable kinetics. The variance of current fluctuations of such "noisy" patches exhibited a minimum close to the equilibrium-potential for Cl-. With channels occurring in only 38% of sealed patches and an even lower frequency of voltage-activated channels, the chloride conductance of the apical membrane of mitochondria-rich cells did not match quantitatively that previously estimated from macroscopic Ussing- chamber experiments. From a qualitative point of view, however, we have succeeded in demonstrating the existence of Cl-channels in the apical membrane with features comparable to macroscopic predictions, i.e., activation of channel gating by cAMP and, in a few patches, also by membrane depolarization.     

Ion channels in human endothelial cells.

Ion channels were studied in human endothelial cells from umbilical cord by the patch clamp technique in the cell attached mode. Four different types of ion channels were recorded: i) potassium channel current that rectifies at positive potentials in symmetrical potassium solutions (inward rectifier); ii) low-conductance non-selective cation channel with a permeability ratio K:Na:Ca = 1:0.9:0.2; iii) high-conductance cation-selective channel that is about 100 times more permeable for calcium than for sodium or potassium; iv) high-conductance potassium channel with a permeability ratio K:Na = 1:0.05. The extrapolated reversal potential of the inwardly rectifying current was near to the potassium equilibrium potential. The slope conductance decreased from 27 pS in isotonic KCl solution to 7 pS with 5.4 mmol/l KCl and 140 mmol/l NaCl in the pipette but 140 mmol/l KCl in the bath. The low-conductance non-selective cation channel showed a single-channel conductance of 26 pS with 140 mmol/l Na outside, 28 pS with 140 mmol/l K outside, and rectified in inward direction in the presence of Ca (60 mmol/l Ca, 70 mmol/l Na, 2.7 mmol/l K in the pipette) at negative potentials. The current could be observed with either chloride or aspartate as anion. The high-conductance non-selective channel did not discriminate between Na and K. The single-channel conductance was about 50 pS. The extrapolated reversal potential was more positive than +40 mV (140 K or 140 Na with 5 Ca outside). Both the 26 and 50 pS channel showed a run-down, and they rapidly disappeared in excised patches. The high-conductance potassium channel with a single-channel conductance of 170 pS was observed only rarely. It reversed near the expected potassium equilibrium potential. The 26 pS channel could be stimulated with histamine and thrombin from outside in the cell-attached mode. Both the 26 pS as well as the 50 pS channel can mediate calcium flux into the endothelial cell.     

K+ channel cAMP activated in guinea pig gallbladder epithelial cells

In guinea pig gallbladder epithelial cells, an increase in intracellular cAMP levels elicits the rise of anion channel activity. We investigated by patch-clamp techniques whether K(+) channels were also activated. In a cell-attached configuration and in the presence of theophylline and forskolin or 8-Br-cAMP in the cellular incubation bath, an increase of the open probability (P(o)) values for Ca(2+)-activated K(+) channels with a single-channel conductance of about 160 pS, for inward current, was observed. The increase in P(o) of these channels was also seen in an inside-out configuration and in the presence of PKA, ATP, and cAMP, but not with cAMP alone; phosphorylation did not influence single-channel conductance. In the inside-out configuration, the opioid loperamide (10(-5) M) was able to reduce P(o) when it was present either in the microelectrode filling solution or on the cytoplasmic side. Detection in the epithelial cells by RT-PCR of the mRNA corresponding to the alpha subunit of large-conductance Ca(2+)-activated K(+) channels (BK(Ca)) indicates that this gallbladder channel could belong to the BK family. Immunohistochemistry experiments confirm that these cells express the BK alpha subunit, which is located on the apical membrane. Other K(+) channels with lower conductance (40 pS) were not activated either by 8-Br-cAMP (cell-attached) or by PKA + ATP + cAMP (inside-out). These channels were insensitive to TEA(+) and loperamide. The data demonstrate that under conditions that induce secretion, phosphorylation activates anion channels as well as Ca(2+)-dependent, loperamide-sensitive K(+) channels present on the apical membrane.     

Transpiration rates of the understory annual Impatiens capensis (Balsaminaceae) in response to the autumnal changes in canopy leaf area

Leaf transpiration rates of Impatiens capensis were measured beneath a broadleaved deciduous forest canopy over successive growing seasons using a steady-state porometer. The transpiration measurements, which continued into early autumn, provided a framework for assessing whether I. capensis exhibits stomatal opening in response to the autumnal increase in available direct-beam radiation reaching the forest floor. The deciduous canopy LAI (leaf area index) decreased from a growing season maximum of 3.94 m² m⁻², while the understory I. capensis population located along a stream channel maintained LAI values ranging from 0.58 to 1.05 m² m⁻² late into the growing season. Late morning and early afternoon leaf transpiration rates during the months of June and July averaged about 8 μg cm⁻² s⁻¹, with a mean stomatal conductance of 0.5 cm s⁻¹. In August, leaf transpiration averaged almost 12 μg cm⁻² s⁻¹, with stomatal conductance exceeding 1.5 cm s⁻¹. However, beginning in early to mid-September, before canopy leaf-fall, the persistent green leaves of I. capensis exhibited a sharp decline in transpiration, possibly a result of decreasing vapor pressure deficits or non-lethal physiological damage induced by cold stress. This physiological decline offsets any advantage that could have been gained by the increased exposure to direct-beam radiation after canopy leaf-fall in mid-October. Although green leaf area and seed-bearing capsules may persist until the first frost in October or early November, there is no evidence of stomatal opening suggestive of carbon assimilation for enhanced seed development during this early autumn period. We conclude that the persistent green leaf area of I. capensis fails to exploit the increase in available direct-beam radiation in the final stage of its life cycle.     

Low-conductance states of K+ channels in adult mouse skeletal muscle

Single-channel currents were recorded from Ca2+-activated or ATP-sensitive K+ channels in inside-out membrane patches excised from isolated mouse toe muscles. In addition to the closed and fully open configurations, both types of channels may exhibit several intermediate low-conductance states which are clustered near multiples of elementary conductance units. The units are 1/8 or 1/6 of the channel conductance for Ca2+-activated channels and 1/4 or 1/3 for ATP-sensitive channels. Normally, low-conductance states are rare, but they occur more frequently directly after patch excision. An increased probability of low-conductance states of ATP-sensitive K+ channels was also observed in the presence and during washout of the internal channel blocker adenine. The results suggest that Ca2+-activated and ATP-sensitive K+ channels are composed of several membrane pores with strong positive cooperativity among the elementary conductance units.     

Single delayed rectifier channels in frog atrial cells. Effects of beta-adrenergic stimulation.

The patch-clamp technique with two pipettes was used to record single delayed K+ channels (cell-attached electrode) and to control the potential and the composition of the intracellular compartment (whole-cell electrode). With 30 microM cAMP in the cell and physiological potassium concentrations inside and outside the patch, a channel carrying an outward current was characterized. Its open probability was very low and the channel was recorded in only 5% of patches under control conditions. Increasing intracellular cAMP increased the probability of finding a channel in a patch 10-fold. The channel had the characteristics expected of a delayed rectifier channel. The time-course of its ensemble average resembled the whole-cell current in the same cell. The current-voltage relationship exhibited inward rectification, with a slope conductance of 20 pS in the linear portion and a reversal potential close to EK. Both the open- and the closed-time distributions were described by the sum of two exponentials, suggesting a complicated gating scheme involving two closed states and two open states. The beta-adrenergic stimulation did not change the conductance of the channel, but increased its probability of opening.     

Serotonin stimulates a Ca2+ permeant nonspecific cation channel in hepatic endothelial cells.

The contraction of hepatic endothelial cell fenestrae after exposure to serotonin is associated with an increase in intracellular Ca2+ which is derived from extracellular Ca2+, is inhibited by pertussis toxin and is not associated with activation of phosphoinositol turnover or cAMP. Using cell-attached and excised patches in primary cultures of rat hepatic endothelial cells, we identified a serotonin-activated channel with conductance of 26.4+/-2.3 pS. The channel was also permeant to Na+, K+ and Ca++ but not to anions. In cell-attached patch recordings, addition of serotonin to the bath significantly increased channel activity with Ca2+ or Na+ as the charge-carrying ions. This channel provides a mechanism whereby serotonin can raise the cytosolic Ca2+ concentration in hepatic endothelial cells.     

多基因联合分析棒束孢属Isaria（Ascomycota,Cordycipitaceae）系统发育关系

对棒束孢属Isaria及近缘属物种开展5基因（nrSSU、nrLSU、tef-1α、rpb1和 rpb2）测序并联合分析,结合GenBank相关类群序列,探讨棒束孢属系统发育关系,最终获得95个菌株、58个明确分类群的2-5基因序列。利用MEGA和MrBayes软件进行多基因聚类分析,结果表明棒束孢属多系起源于虫草菌科中,分3个不同分支。A支主要由Isaria cicadae、I. teniupes、I. coleopterorum、I. fumosorosea和I. cateniannulata等组成;B支包括I. poprawkii、I. locustica、I. javanica、I. amoenerosea和I. cateniobliqua;C支仅有I. farinosa。分支间被Cordyceps militaris、C. ninchukispora、C. pruinosa等隔开。棒束孢在形态上,主要以瓶梗基部膨大、尖端变细及孢子呈链状等特征与其他类群分开,但同时也发现有棒状分生孢子梗和单孢子类型。基于节点的分歧时间预测分析,推测棒束孢属首次分化于70Mya,但棒束孢属主要物种形成却在60-55Mya,且3个分支的棒束孢物种为快速同时形成,而后大多数类群表现遗传稳定。同时发现,与Isaria Clade A较近一支有粉被玛利亚霉Mariannaea pruinosa（C. pruinosa无性型）和蛹草蚧霉Lecanicillium militaris（C. militaris无性型）;与粉棒束孢距离最近一支有Akanthomyces aculeatus（C. tuberculata无性型）和L. attenuatum（C. confragosa无性型）,是两个不同的属征分类群,且相互间遗传距离较近。根据棒束孢属及其近缘种属形态特征的复杂性推测,棒束孢属在快速物种形成中,其近缘类群存在一定程度的丢失和选择性演化。     

Evidence for modulation of cell-to-cell electrical coupling by cAMP in mouse islets of Langerhans

The effects of forskolin on electrical coupling among pancreatic β-cells were studied. Two microelectrodes were used to measure membrane potentials simultaneously in pairs of islet β-cells. Intracellular injection of a current pulse (ΔI) elicited a membrane response ΔV₁ in the injected cell and also a response ΔV₂ in a nearby β-cell confirming the existence of cell-to-cell electrical coupling among islet β-cells. In the presence of glucose (7 mM), application of forskolin evoked a transient depolarization of the membrane and electrical activity suggesting that the drug induced a partial inhibition of the β-cell membrane K⁺ conductance. Concomitant with this depolarization of the membrane there was a marked decrease in β-cell input resistance (ΔV₂/ΔI) suggesting that exposure to forskolin enhanced intercellular coupling. Direct measurements of the coupling ratio ΔV₂/ΔV₁ provided further support to the idea that forskolin enhances electrical coupling among islet cells. Indeed, application of forskolin reversibly increased the coupling ratio. These results suggest that cAMP might be involved in the modulation of electrical coupling among islet β-cells.     

Basolateral Cl channels in primary airway epithelial cultures

Salt and water absorption and secretion across the airway epithelium are important for maintaining the thin film of liquid lining the surface of the airway epithelium. Movement of Cl across the apical membrane involves the CFTR Cl channel; however, conductive pathways for Cl movement across the basolateral membrane have been little studied. Here, we determined the regulation and single-channel properties of the Cl conductance (G(Cl)) in airway surface epithelia using epithelial cultures from human or bovine trachea and freshly isolated ciliated cells from the human nasal epithelium. In Ussing chamber studies, a swelling-activated basolateral G(Cl) was found, which was further stimulated by forskolin and blocked by N-phenylanthranilic acid (DPC) = sucrose > flufenamic acid = niflumic acid = glibenclamide > CdCl(2) = 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) = DIDS = ZnCl(2) > tamoxifen > 4,4'-dinitro-2,2'-stilbene-disulfonate disodium salt (DNDS). In whole cell patch-clamp experiments, three types of G(Cl) were identified: 1) a voltage-activated, DIDS- (but not Cd-) blockable and osmosensitive G(Cl); 2) an inwardly rectifying, hyperpolarization-activated and Cd-sensitive G(Cl); and 3) a forskolin-activated, linear G(Cl), which was insensitive to Cd and DIDS. In cell-attached patch-clamp recordings, the basolateral pole of isolated ciliated cells expressed three types of Cl channels: 1) an outwardly rectifying, swelling-activated Cl channel; 2) a strongly inwardly rectifying Cl channel; and 3) a forskolin-activated, low-conductance channel. We propose that, depending on the driving force for Cl across the apical membrane, basolateral Cl channels confine Cl(-) secretion or support transcellular Cl(-) absorption.     

基于多组织稳定同位素比值的热带大西洋4种鲨鱼营养生态位分化

不同代谢速率组织间的碳、氮稳定同位素比值(δ¹³C和δ¹⁵N)可以反映生物不同时间尺度的摄食信息,对探讨物种间摄食、栖息地利用和营养生态位的变化具有重要的指示作用。本研究以在大西洋热带海域兼捕的大青鲨、长鳍鲭鲨、拟锥齿鲨和尖吻鲭鲨为对象,通过测定其肌肉、肝脏和血液的δ¹³C和δ¹⁵N值,探讨4种鲨鱼营养生态位分化。结果表明: 与长鳍鲭鲨相比,尖吻鲭鲨、拟锥齿鲨和大青鲨的δ¹⁵N值相似且相对较高;大青鲨与其他鲨鱼存在摄食隔离,表现出独特的营养生态位;尖吻鲭鲨营养生态位宽幅最大,摄食食物种类和(或)栖息环境类型更多样化,其与拟锥齿鲨的营养生态位重叠度最高,说明两种鲨鱼具有潜在的资源竞争关系。尖吻鲭鲨、拟锥齿鲨和大青鲨组织间的δ¹³C、δ¹⁵N差值与其叉长均无显著相关关系,说明3种鲨鱼近期内无明显摄食变化;而长鳍鲭鲨的肝脏、血液和肌肉组织的δ¹⁵N差值与叉长显著相关,说明长鳍鲭鲨在短期内存在摄食变化。肝脏和血液的δ¹³C、δ¹⁵N值的相似性反映了两种组织整合摄食时间周期相近,其较高的代谢速率可以反映相对短时间周期的摄食信息。     

Acetylcholine-activated currents in quail myotubes expressing viral oncogenes

Acetylcholine-activated currents were recorded in cultured myotubes arising from embryonic quail myoblasts transformed by the v-src and v-ras oncogenes. In src-myotubes, the whole cell inward current decayed more slowly than in non-transformed controls. In ras-myotubes, the current had a faster decay and smaller amplitude than in the controls. The single-channel conductance and mean open times recorded from cell-attached patches were similar in transformed and control cells. However, in ras-myotubes the frequency of channel openings strongly decreased with time.

It is concluded that oncogenic tyrosine-specific protein kinase (v-src product) and G-like p21 protein (v-ras product) can induce differential changes in the function of nicotinic ACh receptor, perhaps related to specific biochemical events elicited in the establishment of the transformed state.     

Two types of chloride channels in hen colon epithelial cells identified by patch-clamp experiments

Summary Freshly isolated epithelial cells from hen colon were investigated using the patch-clamp technique. The aim of this investigation was to characterise the cellular conducting site for Cl^- secretion. In cell-attached mode two types of Cl^--channels were found. Both showed distinct outward rectification. The channel types differed in single channel conductances and the marked voltage dependence of the open probabilities. A low conductance Cl^--channel was observed with a mean conductance at negative holding potentials of g_-=9 pS, and of g₊=34 pS at positive potentials. This channel was predominantly open at negative potentials, corresponding to cell hyperpolarization. The second channel type observed had conductances of g_-=35 pS and g₊=77 pS, and showed increasing open probabilities with increasing holding potentials (cell depolarisation). Both channel types were blockable by the Cl^--channel blocker NPPB. These data in combination with previously published transepithelial transport data on hen colon indicate that these channels are the Cl^- secretory sites in colon epithelium.Abbreviations DNSO dimethylsulfoxide - EGTA ethyleneglycol triacetic acid - g₊, g_- single channel conductance at positive and negative voltages - HEPES N-(2-hydroxy-ethyl)piperazine-N-(2-ethane-sulfonic acid) - i single channel current - NMDG N-methyl-d-glucosamine - NPPB 5-hitro-2-(3-phenylpropylamino)-benzoate - P_o open probability - V_p holding potential     

入侵植物五爪金龙根系深色有隔内生真菌的侵染特征

深色有隔内生真菌（DSE）广泛侵染华南地区外来入侵植物五爪金龙。显微观察发现五爪金龙根系具有大量的泡囊,其最显著的特征是大量脂质的富集和被分解成小脂质体,并通过透明或深色菌丝转移,说明了五爪金龙和DSE之间进行碳转移。富含脂质的泡囊可产生透明菌丝和自我复制,说明泡囊具有繁殖功能。富含脂质的泡囊和透明菌丝等非典型结构没有可分辨的或具透明的细胞壁,是活跃的真菌结构。透明菌丝和富含脂质的泡囊主要定殖在生理活性强的根组织中。深色有隔菌丝和微菌核是DSE的典型结构,主要定殖在相对不活跃的根组织中。DSE定殖于五爪金龙根的维管柱、皮层、表皮细胞和表面,同时延伸到土壤,在宿主植物和土壤之间形成连续的系统的真菌网络。根据DSE的多态性质、庞大的真菌网络和脂质的富集及其分布,探讨了DSE和入侵植物五爪金龙潜在的互惠共生关系。     

Cyclic adenosine monophosphate regulates calcium channels in the plasma membrane of Arabidopsis leaf guard and mesophyll cells

The effect of cAMP on Ca(2+)-permeable channels from Arabidopsis thaliana leaf guard cell and mesophyll cell protoplasts was studied using the patch clamp technique. In the whole cell configuration, dibutyryl cAMP was found to increase a hyperpolarization-activated Ba(2+) conductance (I(Ba)). The increase of I(Ba) was blocked by the addition of GdCl(3). In excised outside-out patches, the addition of dibutyryl cAMP consistently activated a channel with particularly fast gating kinetics. Current/voltage analyses indicated a single channel conductance of approximately 13 picosiemens. In patches where we measured some channel activity prior to cAMP application, the data suggest that cAMP enhances channel activity without affecting the single channel conductance. The cAMP activation of these channels was reversible upon washout. The results obtained with excised patches indicate that the cAMP-activated I(Ba) seen in the whole cell configuration could be explained by a direct effect of cAMP on the Ca(2+) channel itself or a close entity to the channel. This work represents the first demonstration using patch clamp analysis of the presence in plant cell membranes of an ion channel directly activated by cAMP.     

Heteromeric channel formation and Ca-free media reduce the toxic effect of the weaver Kir 3.2 allele

weaver mice have a severe hypoplasia of the cerebellum with an almost complete loss of the midline granule cells. Recent genetic studies of weaver mice have identified a mutation resulting in an amino acid substitution (G156S) in the pore of the inwardly rectifying potassium channel subunit K_ir 3.2. When expressed in Xenopus oocytes the weaver mutation alters channel selectivity from a potassium-selective to a nonspecific cation-selective pore. In this study we confirm by cell-attached patch-clamp recording that the mutation produces a non-selective cation channel. We also demonstrate that the cell death induced by weaver expression may be prevented by elimination of calcium from the extracellular solution as well as by coexpression with the wild-type K_ir 3.2 allele, or other members of the K_ir 3.0 subfamily. These results suggest that the weaver defect in K_ir 3.2 may cause cerebellar cell death by cell swelling and calcium overload. Cells which express the weaver subunit, but which normally survive, may do so because of heteromeric subunit assembly with wild-type subunits of the K_ir 3.0 subfamily.     

G-proteins mediate intestinal chloride channel activation.

The localization of several GTP-binding regulatory proteins in teh apical membrane of intestinal epithelial cells has prompted us to investigate a possible role for G-proteins as modulators of apical Cl- channels. In membrane vesicles isolated from rat small intestine or human HT29-cl.19A colon carcinoma cells, the entrapment of guanosine 5'-O-(3-thiophosphate (GTP gamma S) led to a large increase in Cl- conductance, as evidenced by an increased 125I- uptake and faster SPQ quenching. The enhancement was observed in the presence, but not in the absence of the K+ ionophore valinomycin, indicating that the increased Cl- permeability is not secondary to the opening of K+ channels. The effect of GTP gamma S was counteracted by guanosine 5'-O-(2-thiophosphate (GDP beta S) and appeared to be independent of cytosolic messengers, including ATP, cAMP, and Ca2+, suggesting that protein phosphorylation and/or phospholipase C activation is not involved. Patch clamp analysis of apical membrane patches of HT29-cl.19A colonocytes revealed a GTP gamma S-activated, inwardly rectifying, anion-selective channel with a unitary conductance of 20 +/- 4 pS. No spontaneous channel openings were observed in the absence of GTP gamma S, while the open time probability (Po) increases dramatically to 0.81 +/- 0.09 upon addition with GTP gamma S. Since the electrophysiological characteristics and regulatory properties of this channel are markedly different from those of the more widely studied cAMP/protein kinase A-operated channel, we propose the existence of a separate Cl(-)-selective ion channel in the apical border of intestinal epithelial cells. Our results suggest an alternative regulatory pathway in transepithelial salt transport and a possible site for anomalous channel regulation as observed in cystic fibrosis patients.     

中国马兜铃属和关木通属(马兜铃科)概览

马兜铃属(广义) (Aristolochia sensu lato)具有花单被、花萼管状、合蕊柱、子房下位、中轴胎座、胚珠多数、蒴果等主要特征, 广布于全世界的热带、亚热带和温带地区, 约有600种, 是马兜铃科中种类最多的属。依据Flora of China, 我国有本属植物45种, 其中33种为中国特有。近些年, 国内大量的新类群被相继报道, 特别是云南、广西两地, 而另一些类群则得到了确认、恢复、重新发表或修订。最近, 基于形态和分子证据, 关木通属(Isotrema)因其花萼管急剧弯曲; 合蕊柱3裂; 雄蕊6, 成对与合蕊柱裂片对生; 蒴果由上而下开裂等区别特征而从广义马兜铃属中被分出独立成属。本文基于大量的野外调查、标本鉴定、数码照片考证和相关文献的仔细研究, 重新梳理了中国马兜铃属和关木通属的种类及分布情况, 确认现阶段中国马兜铃属17种和关木通属58种1亚种, 并一一记述同时编制了最新的检索表。其中, 柔叶关木通(I. mollis)、线叶关木通(I. neolongifolia)的种级地位得到重新确认, 并提供了图版。探讨了优贵马兜铃(A. gentilis)、川滇马兜铃(A. chuandianensis)和纤细马兜铃(A. gracillima)的关系, 昆明关木通(I. kunmingense)与波氏关木通(A. bonatii)的关系, 以及卵叶关木通(I. ovatifolium)、葫芦叶关木通(I. cucurbitoides)、西藏关木通(I. griffithii)、过石珠(I. versicolor)、大别山关木通(I. dabieshanensis)等复合群和袋形关木通(I. saccata)等物种存在的问题。     

Large-conductance calcium-activated anion channel characteristics in neuroblastoma cells

Large-conductance anion channel characteristics were investigated in neuroblastoma cells (N2A) by using different configurations of the patch-clamp technique. In excised patches, the channel was induced by depolarising potentials in 90% of experiments, had a conductance of 340 pS in symmetrical 135 mmol/l NaCl and exhibited the typical bell-shape activity. Neither the channel induction nor the channel activity was affected by rising the Ca2+ concentration on the cytopasmic side of membranes. In cell-attached configuration the maximal channel activity was shifted towards more positive potentials in comparison to that of excised patches and an increase in intracellular Ca2+, obtained by extracellular application of the Ca2+-ionophore A23187 in the presence of 0.2 micromol/l Ca2+, induced single-channel currents in 80% of patches compared to 31% of cell-attached experiments showing channel activity in normal conditions. In turn, application of 2 micromol/l Ca2+ induced channel activity in 100% of patches. The reversal potential of the channel in cell-attached patches was around -10 mV as the resting potential of cells eliciting channel activity. For cells where channel activity was not detected in cell-attached mode, the resting potential was around -45 mV. Channel activity could be restored in most whole-cell recordings in the presence of 2 micromol/l or more intracellular Ca2+ concentrations. The Ca2+-induction and the relation between channel activity and cell resting potential seem to suggest a role of the large-conductance anion channel in resting potential modulation during some basic functions of the neuroblastoma cell proliferation.