Erratum

Journal of Plankton Research 12, 1295–1314, 1990 The abstract was incorrectly printed. The correct version isgiven below. The fine- to coarse-scale distribution patterns of tuna larvaein the east Indian Ocean were investigated by a combinationof continuous transect sampling using surface tows and randomsampling using double oblique tows. Thunnus maccoyii was themost abundant species, reaching densities near patch centresof 22 m^–3 in surface tows, which is 25 times greater thanthe highest previous records for tuna larvae. Patches of T maccoyiilarvae in areas of high abundance appeared to be 5–15km in diameter. Smaller patches in areas of low abundance wereusually composed of older larvae Lloyd's index of patchinesswas consistently high for all tuna species, ranging from 3.0to 5.2 for T.maccoyii. There was no change in the index whentow distance was doubled to 1200 m, which suggests that thedominant patch size was somewhat larger than the larger samplinginterval. Sampling larvae at the same site 4 days apart resultedin estimates of abundance that differed by an order of magnitude.Abundance estimated from a single station would depend largelyon what day the station was occupied and where the sample wastaken in relation to a patch.     

玫烟色拟青霉对小菜蛾致病力的时间-剂量-死亡率模型模拟

小菜蛾Plutella xylostella是我国南方十字花科蔬菜上的重要害虫,已对田间常用的化学杀虫剂产生了严重的抗性。为寻找有效的小菜蛾生物防治措施,本实验研究了一株分离自家白蚁的玫烟色拟青霉Paecilomyces fumosoroseus (SCAU-PFCF01)对小菜蛾2～4龄幼虫的致病力。实验采用浸液法,供试浓度为1×10³、1×10⁴、1×10⁵、1×10⁶和1×10⁷个孢子/mL。结果表明：随玫烟色拟青霉孢子浓度的升高,小菜蛾的感病死亡率增加,在浓度为1×10⁷ /mL时,小菜蛾2、3和4龄幼虫的累计死亡率分别为96%、85%和80%。玫烟色拟青霉对小菜蛾各龄幼虫的致病力与供试龄期有关,其感病的敏感顺序为2龄、3龄和4龄。用时间 剂量 死亡率模型（time-dose-mortality model,TDM）对各龄幼虫的致病力数据进行模拟,所建模型均顺利通过Hosmer-Lemeshow拟合异质性检验,表明模型拟合良好,并由模型估计出了该菌株对小菜蛾各龄幼虫的致死剂量与致死时间。2龄幼虫接种后第7天、3龄幼虫接种后第5天、4龄幼虫接种后第4天的LC₅₀估计值分别为1.17×10⁴、1.44×10⁴和5.21×10⁴ /mL,LC₉₀估计值分别为1.98×10⁶、3.82×10⁷和1.29×10⁸ /mL。玫烟色拟青霉对小菜蛾幼虫的致死时间与浓度相关,供试各龄幼虫的LT₅₀值随着孢子悬浮液浓度的增加而递减,在1×10⁵～1×10⁷ /mL的范围内,2龄幼虫的LT₅₀值从3.16天降低到1.72天,3龄幼虫的LT₅₀从3.21天降低到1.83天,4龄幼虫的LT₅₀从3.69天降低到2.04天。即2龄幼虫致死所需的时间最短,其次为3龄幼虫,4龄幼虫致死所需的时间最长。结果显示了该株玫烟色拟青霉在小菜蛾的生物防治中具较强的应用潜力。     

Functional characterization of the neuronal-specific K-Cl cotransporter: implications for [K+]o regulation

The neuronal K-Cl cotransporter isoform (KCC2) was functionallyexpressed in human embryonic kidney (HEK-293) cell lines. Two stablytransfected HEK-293 cell lines were prepared: one expressing anepitope-tagged KCC2 (KCC2-22T) and another expressing theunaltered KCC2 (KCC2-9). The KCC2-22T cells produced aglycoprotein of ~150 kDa that was absent from HEK-293 control cells.The ⁸⁶Rb influx in both cell lineswas significantly greater than untransfected control HEK-293 cells. TheKCC2-9 cells displayed a constitutively active⁸⁶Rb influx that could beincreased further by 1 mMN-ethylmaleimide (NEM) but not by cellswelling. Both furosemide [inhibition constant (K_i) ~25µM] and bumetanide (K_i~55 µM) inhibited the NEM-stimulated ⁸⁶Rb influx in the KCC2-9cells. This diuretic-sensitive⁸⁶Rb influx in theKCC2-9 cells, operationally defined as KCC2 mediated, required external Clbut not external Na⁺ and exhibiteda high apparent affinity for externalRb⁺(K⁺)[Michaelis constant(K_m) = 5.2 ± 0.9 (SE) mM; n = 5] but alow apparent affinity for externalCl(K_m >50 mM). Onthe basis of thermodynamic considerations as well as the unique kineticproperties of the KCC2 isoform, it is hypothesized that KCC2 may servea dual function in neurons: 1) themaintenance of low intracellularCl concentration so as toallow Cl influx vialigand-gated Cl channelsand 2) the buffering of externalK⁺ concentration([K⁺]_o) in the brain.

     

脱氧鬼臼毒素对美洲大蠊的毒力及几种酶系的影响

采用药膜法测试了脱氧鬼臼毒素对美洲大蠊Periplaneta americana初孵若虫的触杀活性,并测定了其对成虫中枢神经系统乙酰胆碱酯酶（AChE）和腺苷三磷酸酶(ATPase)离体活性的影响。结果表明：脱氧鬼臼毒素对美洲大蠊初孵若虫具有较强的毒杀活性,在接触时间24、48、72和96 h 的LC₅₀分别为26.26、4.68、1.51和0.62 μg/cm²;其对AChE没有明显影响; 对Na⁺ -K⁺ -ATPase有明显抑制作用,并存在浓度 效应关系,IC₅₀为44.9 μmol/L; 对Ca²⁺-Mg²⁺-ATPase表现出低剂量激活,高剂量抑制的现象。结果提示美洲大蠊的AChE不是脱氧鬼臼毒素靶标,而ATPase可能是脱氧鬼臼毒素的重要靶标之一。     

自同安钮夜蛾幼虫分离的一种核型多角体病毒

对在林间导致同安钮夜蛾Anua indiscriminate Moore幼虫死亡的一种病原物进行了分离和鉴定,并对该病原物进行了室内毒力测定。结果表明：该病原物为核型多角体病毒, 定名为同安钮夜蛾核型多角体病毒(AiNPV)。其对同安钮夜蛾幼虫的致死浓度LC₅₀和LC₉₀分别为3.4×10⁴和3.8×10⁷ PIB/mL。     

Estimating chlorophyll a abundance from the 'phytoplankton colour' recorded by the Continuous Plankton Recorder survey: validation with simultaneous fluorometry

The green colour (measured with reference to standard colourcharts) of sections of the Continuous Plankton Recorder (CPR)filtering silk was compared with estimates of chlorophyll aconcentration derived from a fluorometer mounted on the CPRduring seven tows in the North Sea between February and May1991. After the green colour was assessed, the abundance ofphytoplankton cells on the filtering silks was quantified bymicroscope analysis. Data were collected for 115 10-nautical-milesamples over a total of seven cruises. For these 115 samples,there was only a weak (F_1.113 = 3.8, P = 0.05, r² = 0.03) positiverelationship between the colour of the filtering silk and thechlorophyll a concentration. However, when this comparison wasrestricted to four tows (68 10-nautical-mile samples) wherethe recorded phytoplankton cell abundance on the silks was verylow, there was a highly significant (F_1.66 /,69.1, P < 0.001,r² = 0.51) positive relationship between the silk colour andthe chlorophyll a concentration. By measuring the relative colourintensity of CPR standard colour categories and quantifyingthe individual variation in the assessment of colour, a theoreticalmodel was developed which pedicted that if the silks were colouredin direct proportion to the chlorophyll a concentration in thewater, then the expert r² for the relationship between silkcolour and chlorophyll a concentration would be 0.62. The greencolour recorded by the CPR survey was therefore identified asa quantitative index of chlorophyll a concentration, but onlywhen numbers of phytoplankton cells on the CPR silks are nothigh.     

重金属Cd2+对棕尾别麻蝇血细胞数量、包囊作用和形态结构的影响

为了揭示重金属对昆虫细胞免疫的影响及其机理,本文采用血细胞计数、台盼蓝染色、延展与包囊率测定, 以及显微与超微结构观察等方法,以经重金属Cd²⁺（浓度为150 μg/g）处理的棕尾别麻蝇Boettcherisca peregrina及其对照为研究对象,分别测定了处理组与对照组血细胞的数量与存活率、延展与包囊率,观察其形态结构,并比较了两组间的差异。结果表明： 棕尾别麻蝇初产幼虫经连续喂饲含Cd²⁺的饲料24,48,72和96 h后,与对照相比其血细胞总数和存活率显著下降,而血细胞的延展率和包囊作用的显著下降则分别出现于取食72和48 h之内,此后则下降不显著;由Cd²⁺处理幼虫发育形成蛹的血细胞包囊作用也显著低于对照。形态结构观察结果表明,Cd²⁺处理对幼虫的原血细胞、浆血细胞、颗粒血细胞和类绛色血细胞的显微形态影响不大,但可导致部分浆血细胞不能产生伪足;但均能导致各类血细胞的超微结构发生不同程度的变化,其中主要变化包括：细胞膜受损或破裂,染色质呈现凝聚, 线粒体和内质网等细胞器明显减少,以及胞质内出现空囊泡。结果说明重金属Cd²⁺对棕尾别麻蝇的血细胞可具有毒害作用。     

Circadian photosynthetic activity of natural marine phytoplankton isolated in a tank

A large (1200 1) seawater sample from the Gulf of Maine waskept under constant temperature and light conditions for a periodof 72 h. Circadian variations were observed in the photosyntheticcapacity (P^B_max) and efficiency (^B); these occurred in phase,and were not related to changes in chlorophyll concentrations.Such observations are consistent with the hypothesis that oscillationsin photosynthetic characteristics of natural phytoplankton aremediated by an endogenous circadian rhythm. ¹ Bigelow Laboratory contribution 87023 and a contribution tothe programs of GIROQ (Groupe interuniversitaire de recherchesoceanographiques du Québec) and of the Maurice-LamontagneInstitute     

家蝇幼虫消减文库的构建及差异表达基因的鉴定

为了鉴定家蝇Musca domestica免疫相关基因,应用抑制性消减杂交技术,构建刺激家蝇幼虫差异表达cDNA消减文库。以大肠杆菌Escherichia coli和金黄色葡萄球菌Staphylococcus aureus诱导12 h的家蝇幼虫与未诱导的家蝇幼虫为消减杂交对象,获得了差异表达基因的cDNA片段,将其与T/A载体连接并转化大肠杆菌DH5α,构建了刺激家蝇幼虫cDNA消减文库。PCR检测发现,文库的阳性克隆中插入的cDNA片段大小在200~1 000 bp之间,随机挑选了161个含大小不等差异片段的克隆进行测序和同源性分析,鉴定了36种蛋白的基因片段,包括抗菌肽、酶、核糖体蛋白、其他功能蛋白以及功能不明的蛋白。用半定量RT-PCR分析了其中6种蛋白基因的表达,结果显示：防御素和攻击素基因在细菌刺激后24 h内明显上调表达,而溶菌酶、酚氧化酶原活化因子、糜蛋白酶和蛋白质合成起始因子基因在细菌刺激后0-4 h内表达受抑制,12 h后上调表达。该研究结果为家蝇免疫相关基因的克隆和家蝇免疫防御机制的探讨奠定了良好的基础。     

虫酰肼及其衍生物0593对家蚕的毒性及作用机理

为明确虫酰肼衍生物0593对家蚕Bombyx mori的毒性,本研究采用食下毒叶法测定了虫酰肼及其衍生物0593对家蚕的毒性,观察了亚致死浓度下对家蚕生长发育的影响,并测定了虫酰肼及其衍生物0593对家蚕幼虫体内保护酶的影响,对虫酰肼0593的作用机理进行了初步探索。结果表明：虫酰肼及其衍生物0593对2龄家蚕96 h的LC₅₀值分别为1.2863和0.3364 mg·L^-1,属高毒级药剂;虫酰肼及其衍生物0593在亚致死剂量下对家蚕的生长发育有明显的不利性,可使幼虫历期缩短0.5～2 d;处理组眠期体重、全茧量、蛹重和化蛹率与对照相比均显著降低;对4龄幼虫体内多酚氧化酶和几丁质酶也有较明显影响,虫酰肼及其衍生物0593处理家蚕,处理后6 h对体内多酚氧化酶有明显激活作用,12 h后表现出显著的抑制作用;虫酰肼及其衍生物0593对家蚕体内几丁质酶均有激活作用。0593对保护酶的影响较虫酰肼明显。结果提示虫酰肼及其衍生物0593对家蚕毒性高,对其生长发育和保护酶类均有不利性,不适合在桑园及其周边农田使用。     

The search for proteins involved in the formation of crustacean cuticular structures

Summer ichthyoplankton surveys were conducted in the northern Gulf of Mexico from 2007 to 2010 to characterize the distribution and abundance of tuna larvae. Larval assemblages of tunas were comprised of four genera: Thunnus, Auxis, Euthynnus, and Katsuwonus. Thunnus were the most abundant and four species were detected; T. atlanticus [blackfin tuna], T. obesus [bigeye tuna], T. albacares [yellowfin tuna], and T. thynnus [bluefin tuna]. Intra- and inter-annual variability in the distribution and abundance of Thunnus species were observed with higher densities in 2008 and 2009, with a decline in abundance observed in 2010. Distribution and abundance of Thunnus larvae were influenced by physical and chemical conditions of the water mass, notably sea surface temperature and salinity. Distinct species-specific habitat preferences were observed and the location of mesoscale oceanographic features influenced larval abundance with higher densities of T. atlanticus, T. obesus, and T. albacares near anticyclonic (warm core) regions and the Loop Current, while T. thynnus was observed in higher densities near cyclonic (cold core) regions. This study demonstrates that spatial and temporal variability in the location of mesoscale oceanographic features may be important to partitioning nursery habitat among Thunnus species.     

Morphological and Physiological Responses of Echinoderm Larvae to Nutritive Signals

SYNOPSIS. Echinoid larvae have been found previously to developshorter arms as phytoplankton concentrations increase. In thepresent study, the skeletal dimensions of larvae of the seaurchins Lytechinus pictus and Strongylocentrotus purpuratuswere measured after exposure to dissolved organic compoundsin seawater. The presence of glucose or individual amino acids(at 1 or 2 µM), a mixture of 16 different amino acids(100 nM each), or algal exudate resulted in larvae with shorterarms (by 12 to 88%), relative to larvae in seawater with noadditions. Larvae exposed to mixtures of amino acids also hadchanges in the constituents of their internal free amino acidpools (as determined by HPLC). For another echinoid, Dendrasterexcentricus, amino acid transport (T, from 500 nM) by individuallarvae (n = 47) scaled to arm length (L) as follows: T = 2.06L^0.81(0.81 ± 0.26,95% confidence intervals). Mass (M) andmetabolic rate (MR) did not scale in the same manner (as transport)to arm length for larvae of the echinoid Centrostephanus coronatus(MR = 148L^1.51; M = 1.7L^2.01). These characteristics may scaleto arm length independently from transport rate under certainconditions. Larvae of echinoderms respond morphologically to"signals" in their environment that may indicate the availabilityof dissolved and paniculate nutrients. This in turn will haveconsequences for their ability to take up and metabolize thesenutrients. The response may be mediated by surface receptorsfor dissolved organic compounds, or by changes in the sizesof intracellular substrate pools.     

Protein kinase C{varepsilon} contributes to regulation of the sarcolemmal Na+-K+ pump

A reduction in angiotensinII (ANG II) in vivo by treatment of rabbits with theangiotensin-converting enzyme inhibitor, captopril, increasesNa⁺-K⁺ pump current (I_p)of cardiac myocytes. This increase is abolished by exposure of myocytesto ANG II in vitro. Because ANG II induces translocation of the-isoform of protein kinase C (PKC), we examined whether thisisozyme regulates the pump. We treated rabbits with captopril, isolatedmyocytes, and measured I_p of myocytes voltageclamped with wide-tipped patch pipettes. I_p ofmyocytes from captopril-treated rabbits was larger thanI_p of myocytes from controls. ANG II superfusionof myocytes from captopril-treated rabbits decreasedI_p to levels similar to controls. Inclusion ofPKC-specific blocking peptide in pipette solutions used to perfusethe intracellular compartment abolished the effect of ANG II. Inclusionof RACK, a PKC-specific activating peptide, in pipettesolutions had an effect on I_p that was similarto that of ANG II. There was no additive effect of ANG II andRACK. We conclude that PKC regulates the sarcolemmalNa⁺-K⁺ pump.

     

辣椒碱对小菜蛾的驱避活性及其体内谷胱甘肽-S-转移酶和Na+，K+-ATP酶活性的影响

采用常规生测方法和酶活力测定方法,初步研究了辣椒碱对小菜蛾Plutella xylostella L.的产卵忌避和拒食作用,及其对小菜蛾体内谷胱甘肽-S-转移酶、Na⁺,K⁺-ATP酶活性的影响,以期阐明辣椒碱对害虫的作用机制。结果表明,辣椒碱对小菜蛾表现出较强的产卵忌避活性和拒食活性。在6.25×10⁴ mg/L浓度下,处理24 h辣椒碱对小菜蛾的非选择性产卵忌避率达96.55%,选择性产卵忌避率为84.30%;在相同浓度下,处理48 h辣椒碱对小菜蛾的非选择性拒食率达81.47%,选择性拒食率为69.69%。 另外,经1.25×10⁵ mg/L辣椒碱不同时间处理后,小菜蛾体内的谷胱甘肽-S-转移酶酶活力和Na⁺,K⁺-ATP酶活力与对照相比均产生了波动,处理18 h时小菜蛾体内GSTs活力最高,为152.01 U·mg^-1pro·min^-1,处理1 h时小菜蛾体内Na⁺,K⁺-ATP酶活力最高,为19.99 U·mg^-1pro·min^-1。结果说明辣椒碱能够影响小菜蛾产卵和取食行为,并且对其体内的酶系也产生了影响。     

Dietary cholesterol alters Na+/K+ selectivity at intracellular Na+/K+ pump sites in cardiac myocytes

A modest diet-induced increase in serum cholesterol in rabbits increases the sensitivity of the sarcolemmal Na⁺/K⁺ pump to intracellular Na⁺, whereas a large increase in cholesterol levels decreases the sensitivity to Na⁺. To examine the mechanisms, we isolated cardiac myocytes from controls and from rabbits with diet-induced increases in serum cholesterol. The myocytes were voltage clamped with the use of patch pipettes that contained osmotically balanced solutions with Na⁺ in a concentration of 10 mM and K⁺ in concentrations ([K⁺]_pip) ranging from 0 to 140 mM. There was no effect of dietary cholesterol on electrogenic Na⁺/K⁺ current (I_p) when pipette solutions were K⁺ free. A modest increase in serum cholesterol caused a [K⁺]_pip-dependent increase in I_p, whereas a large increase caused a [K⁺]_pip-dependent decrease in I_p. Modeling suggested that pump stimulation with a modest increase in serum cholesterol can be explained by a decrease in the microscopic association constant K_K describing the backward reaction E₁ + 2K⁺ E₂(K⁺)₂, whereas pump inhibition with a large increase in serum cholesterol can be explained by an increase in K_K. Because hypercholesterolemia upregulates angiotensin II receptors and because angiotensin II regulates the Na⁺/K⁺ pump in cardiac myocytes in a [K⁺]_pip-dependent manner, we blocked angiotensin synthesis or angiotensin II receptors in vivo in cholesterol-fed rabbits. This abolished cholesterol-induced pump inhibition. Because the -isoform of protein kinase C (PKC) mediates effects of angiotensin II on the pump, we included specific PKC-blocking peptide in patch pipette filling solutions. The peptide reversed cholesterol-induced pump inhibition. partial reactions; protein kinase C; angiotensin converting enzyme inhibitors; arteriosclerosis; insulin resistance     

An NTP-Binding Protein That Cross with a Ras-Specific Antibody in the Myceia of Neurospora crassa

A Ras-related NTP-binding protein was partially purified froma membrane fraction derived from the mycelia of Neurospora crassa.[-³²P]ATP and [-³²P]GTP were incubated with mem brane and solublefractions which were then irradiated with UV light to inducecrosslinking of tightly bound nucleotides. After SDS-polyacrylamidegel electrophoresis, blotting onto a nitrocellulose filter andautoradiography it was apparent that most of the proteins thatbound [-³²P]-GTP also bound [-³²P]ATP. Pretreatment of the membranefraction with Ras-specific antibody effectively blocked thebinding of [-³²P]ATP and [-³²P]GTP to several ATP-GTP-bindingproteins. The band of a protein with a molecular weight of 26kDa on the SDS-polyacrylamide gel cross-reacted strongly withthe Ras-specific antibody. The protein was extracted from thegel and further purified by repeated gel electrophoresis. Thepurified protein bound [-³²P]ATP, [-³²P]-GTP, [-³²P]CTP and[-³²P]UTP at 1.6x10^– M and was autophosphorylated in thepresence of [-³²P]ATP and [-³²P]GTP at 1.7x10 M. Pretreatmentof the protein with Ras-specific antibody partially blockedthe autophosphorylation in the presence of these nucleotides.The binding of [-³²P]ATP to the NTP-binding protein was blockedby addition of ATP at 10^–4–10^–3 M. ATP ata concentration of 10^–4 M prevented the binding of [-³²P]to a greater extent than did GTP at the same concentration.Binding of [-³²P]CTP and [-³²P]UTP to the protein was also observed. (Received October 7, 1991; Accepted July 14, 1992)     

Temperature, food and mate limitation of copepod reproductive rates: separating the effects of multiple hypotheses

Temperature, food and the availability of mates may all limitrates of egg production is freshwater diaptomids. We have usedthe reproductive phases of Diaptomus paliidus to develop anindex of food limitation (f) and an index of mate limitation(M), each of which responds independently of temperature. Theresponse of the food limitation index to high and low temperaturesand high and low food concentrations was examined in a two-wayfactorial experiment in the laboratory. The index was highlyresponsive to a change in food concentration, stable duringa change in temperature alone at the high food level and responsiveto the synergistic interaction of food and temperature effectswhen both factors were limiting. Laboratory data indicate thatthe f index should not be biased by mate limitation unless maledensities are 6.7 x 10^–l0 males 1^–1 or less. Themate limitation index was highly responsive to mate availability,and ranged from a low of 9–19 in the presence of abundantmales to 100 in tbe absence of males. Applications of the fand M indices to a natural population of D. paliidus over 6weeks indicated that both food and mate availability were limitingthe reproductive rates of the copepods during the sampling period. ¹Present address: Institute of Animal Resource Ecology, Universityof British Columbia, Vancouver, Canada V6T 1W5     

Cell Potentials and Turgor Pressures in Epidermal Cells of Tradescantia and Commelina

Cell membrane potentials have been measured both in epidermalstrips and intact leaf sections of Tradescantia virginiana andCommelina communis, and in epidermal cells over green and overalbino mesophyll cells of T. albiflora var. albovittata. Membranepotentials (_cell) in strips were considerably lower than thosein intact sections and were insensitive to light and to theabsence or presence of calcium. Their response to external cationlevels was indifferent to ionic species. However, in intactleaf sections incubated with calcium present, membrane potentialsresponded to K⁺ levels but not to Na⁺. were more negative thancells in epidermal strips, and responded to changes in illumination. Long-term recordings of _cell and vacuolar K⁺ levels in T. virginianaduring stomatal closure suggest that the fluctuations of _cellwere unrelated to K⁺ movement (which we could not detect) andthus probably to stomatal movement as well. Turgor pressures measured in epidermal cells of intact leafsections of T. virginiana were found to be of the same magnitudeas those previously reported for epidermal strips. It is concludedthat epidermal cells maintain their solute contents during strippingwithout the involvement of an electrophysiological transportsystem. With the possible exception of lateral subsidiary cells,there was no evidence suggesting that ordinary epidermal cellsare capable of osmotic adjustment even when additional KCI wassupplied in the osmoticum. Absolute turgor levels in intactleaf sections kept at constant external KCI were unrelated tosteady state _cell.     

Thyroid hormone inhibits biliary growth in bile duct-ligated rats by PLC/IP(3)/Ca(2+)-dependent downregulation of SRC/ERK1/2

The role of the thyroid hormone agonist 3,3',5 L-tri-iodothyronine (T3) on cholangiocytes is unknown. We evaluated the in vivo and in vitro effects of T3 on cholangiocyte proliferation of bile duct-ligated (BDL) rats. We assessed the expression of ₁-, ₂-, ₁-, and ₂-thyroid hormone receptors (THRs) by immunohistochemistry in liver sections from normal and BDL rats. BDL rats were treated with T3 (38.4 µg/day) or vehicle for 1 wk. We evaluated 1) biliary mass and apoptosis in liver sections and 2) proliferation in cholangiocytes. Serum-free T3 levels were measured by chemiluminescence. Purified BDL cholangiocytes were treated with 0.2% BSA or T3 (1 µM) in the absence/presence of U-73122 (PLC inhibitor) or BAPTA/AM (intracellular Ca²⁺ chelator) before measurement of PCNA protein expression by immunoblots. The in vitro effects of T3 (1 µM) on 1) cAMP, IP₃, and Ca²⁺ levels and 2) the phosphorylation of Src Tyr139 and Tyr530 (that, together, regulate Src activity) and ERK1/2 of BDL cholangiocytes were also evaluated. ₁-, ₂-, ₁-, and ₂-THRs were expressed by bile ducts of normal and BDL rats. In vivo, T3 decreased cholangiocyte proliferation of BDL rats. In vitro, T3 inhibition of PCNA protein expression was blocked by U-73122 and BAPTA/AM. Furthermore, T3 1) increased IP₃ and Ca²⁺ levels and 2) decreased Src and ERK1/2 phosphorylation of BDL cholangiocytes. T3 inhibits cholangiocyte proliferation of BDL rats by PLC/IP₃/Ca²⁺-dependent decreased phosphorylation of Src/ERK1/2. Activation of the intracellular signals triggered by T3 may modulate the excess of cholangiocyte proliferation in liver diseases. cholestasis; cholangiopathies; hyperplasia; intrahepatic biliary epithelium; mitosis     

Vibrio cholerae ACE stimulates Ca(2+)-dependent Cl(-)/HCO(3)(-) secretion in T84 cells in vitro

ACE, accessory cholera enterotoxin, the thirdenterotoxin in Vibrio cholerae, has been reported toincrease short-circuit current (I_sc) in rabbitileum and to cause fluid secretion in ligated rabbit ileal loops. Westudied the ACE-induced change in I_sc andpotential difference (PD) in T84 monolayers mounted in modified Ussingchambers, an in vitro model of a Cl secretory cell. ACEadded to the apical surface alone stimulated a rapid increase inI_sc and PD that was concentration dependent andimmediately reversed when the toxin was removed. Ion replacement studies established that the current was dependent on Cland HCO₃. ACE acted synergistically with theCa²⁺-dependent acetylcholine analog, carbachol, tostimulate secretion in T84 monolayers. In contrast, the secretoryresponse to cAMP or cGMP agonists was not enhanced by ACE. TheACE-stimulated secretion was dependent on extracellular andintracellular Ca²⁺ but was not associated with an increasein intracellular cyclic nucleotides. We conclude that the mechanism ofsecretion by ACE involves Ca²⁺ as a second messenger andthat this toxin stimulates a novel Ca²⁺-dependent synergy.