Cytoplasmic Channels and Chromatin Migration in the Meiocytes of Arnebia hispidissima (Sieb.) DC

Microsporogenesis was studied in Arnebia hispidissima whichprovided interesting evidence for the occurrence of cytoplasmicchannels and DNA migration between the meiocytes. The intercommunicatingchannels (0.25–1.75 µ in diameter) are consideredto be a prerequisite for chromatin migration and to act as pathwaysbetween the connected meiocytes. They may develop at any stageof meiosis from early prophase to metaphase II and the migrationmay take place at any time following channel formation. Thetendency of chromatin to migrate seemed to be higher in theearly prophasic stages when it moved in the form of ill-definedmasses or clumps. The chances of migration as well as the amountof migrating chromatin is reduced as the meiosis advances. Theamount of chromatin migrating varies from a small portion tothe entire chromatin mass or nucleus. Pollen mother cells withdouble the normal amount of chromatin as well as those withlittle or none were studied as well as cells with increasedor decreased chromosome numbers. In the chromatin mass, uni-or bivalents stretched between the meiocytes or attenuated towardsthe channel, were frequently seen. Occasional semi-asyndesis,multivalent formation, precocity, and nondisjunction was alsonoted. The possibility of channel formation and the induction of chromatinmovement as a consequence of mechanical or procedural defectis ruled out. It is believed that these cytoplasmic channelshave no relation with the plasmodesmata but they arise de novoand that chromatin migration occurs as a natural phenomenon(in some species). The observations are not due to faulty fixation,staining, or squashing.     

The formation of cytoplasmic channels between tapetal cells inZea mays

Summary In this report we show that large cytoplasmic channels form between the tapetal cells ofZea mays (maize) during the period of tapetal cell differentiation. Tapetal cells are connected by plasmodesmata through their cellulosic cell walls prior to the first meiotic division of the meiocytes. As the tapetal cellulose wall is degraded at the onset of meiosis, both plasmodesmata and cytoplasmic channels measuring 50–200 nm are detectable between tapetal cells. By the time the meiotic tetrad is formed, the cytoplasmic channels are well-established and vary in size from 100–400 nm. The channels, with an average diameter of 200–300 nm, persist after the microspores are released from the callose wall and throughout the period of exine development in microsporogenesis. The channels could potentially allow for free exchange of cytoplasm and organelles. As the tapetal cells begin to pull apart and become vacuolate prior to microspore mitosis, the connecting channels are no longer detectable.     

A Cytochemical Study of DNA, RNA, and Protein in the Developing Maize Anther: II. Observations

Changes in acetic-alcohol fixable DNA, RNA, and protein werefollowed in the tapetum, sporogenous tissue, and spores of thedeveloping maize anther using standard cytochemical methodsand microdensitometry. In the tapetum, early nuclear divisionsoccur without prior DNA synthesis, giving a population of IC nuclei. Subsequent synthesis produces the equivalent of 34,000C amounts per pollen sac, 20 times more than is present in thespores before pollen mitosis. The main tapetal RNA synthesisis during the meiotic prophase, with a further period of accumulationin the interval, tetrad to young spores. In the meiocytes, theprincipal accumulation is in the early prophase, with no synthesisduring the meiotic divisions or through the tetrad period. Proteinaccumulation occurs in the tapetum up to mid-meiotic prophase;after this there is a pause, followed by further synthesis frommeiotic metaphase I to the final dissolution of the tissue.In the meiocytes, protein is accumulated through the early prophase;there is no synthesis during the meiotic mitoses or in the tetradperiod, but active accumula-tion occurs in the developing spores. The implications of these observations are discussed in relationto the function of the tapetum.     

New Insights into the Role of the Maize Ameiotic1 Locus

In maize the am1-1 mutant allele results in both the male and female meiocytes undergoing mitosis in place of the meiotic divisions. A second mutant allele am1-praI enables both the male and female meiocytes to proceed to the early zygotene stage of meiotic prophase I before being blocked. Here we report on three new alleles that allow all male meiocytes to undergo mitosis but in female meiocytes approximately one quarter (am1-2), one half (am1-485), or all (am1-489) of them are blocked at an abnormal interphase stage. Previous analysis has shown that am1-praI is dominant to am1-1 in male meiocytes. Cytological analysis of heteroallelic combinations in female meiocytes now indicates a dominance relationship of am1-praI > am1-1 > am1-2/am1-485 > am1-489. The evidence provided by the female phenotypes of the new mutant alleles suggest that, whereas the normal am1 allele is required for the meiocytes to proceed through meiosis, a partially functional allele may be required for their diversion into a mitotic division. The partial or complete blockage of mitosis in female meiocytes carrying the new am1 alleles rules out the possibility that the mitotic division of mutant meiocytes reflects a simple default pathway for cells that cannot initiate meiosis. This locus may have a dual function.     

Meiotic telomere protein Ndj1p is required for meiosis-specific telomere distribution, bouquet formation and efficient homologue pairing

We have investigated the requirements for NDJ1 in meiotic telomere redistribution and clustering in synchronized cultures of Saccharomyces cerevisiae. On induction of wild-type meiosis, telomeres disperse from premeiotic aggregates over the nuclear periphery, and then cluster near the spindle pole body (bouquet arrangement) before dispersing again. In ndj1Delta meiocytes, telomeres are scattered throughout the nucleus and fail to form perinuclear meiosis-specific distribution patterns, suggesting that Ndj1p may function to tether meiotic telomeres to the nuclear periphery. Since ndj1Delta meiocytes fail to cluster their telomeres at any prophase stage, Ndj1p is the first protein shown to be required for bouquet formation in a synaptic organism. Analysis of homologue pairing by two-color fluorescence in situ hybridization with cosmid probes to regions on III, IX, and XI revealed that disruption of bouquet formation is associated with a significant delay (>2 h) of homologue pairing. An increased and persistent fraction of ndj1Delta meiocytes with Zip1p polycomplexes suggests that chromosome polarization is important for synapsis progression. Thus, our observations support the hypothesis that meiotic telomere clustering contributes to efficient homologue alignment and synaptic pairing. Under naturally occurring conditions, bouquet formation may allow for rapid sporulation and confer a selective advantage.     

Meiotic chromosomes of the spermatocytes ejaculated by bulls

The Sperling and Kaden (1971) approach was used in studies of the bull meiotic chromosomes. The number of meiocytes in the ejaculates of normal bulls ranged from 0.0001 to 0.0114% of the whole set of cellular elements in the ejaculate. Two of the studied bulls had a chromosomal abnormality in somatic cells (60, XX/60, XY mosaic). These bulls displayed no deviation as concerns the quantity of meiocytes in their ejaculates. In two other bulls examined, with cryptorchism and azoospermia, about 0.0017 and 4.75% meiocytes were counted in the ejaculate, respectively. A comparative analysis of ejaculated chromosomes and meiotic chromosomes obtained from the normal testes by biopsy did not reveal any differences in the chromosomal morphology. Various premeiotic and meiotic stages, from spermatogonium A and B to metaphase 1, were identified in the bull ejaculates. This technique may be valuable in cytogenetic diagnosis of meiotic abnormalities in sires.     

Characterization of a novel voltage-dependent outwardly rectifying anion current in Caenorhabditis elegans oocytes

An inwardly rectifying swelling- and meiotic cell cycle-regulated anion current carried by the ClC channel splice variant CLH-3b dominates the whole cell conductance of the Caenorhabditis elegans oocyte. Oocytes also express a novel outwardly rectifying anion current termed I_Cl,OR. We recently identified a worm strain carrying a null allele of the clh-3 gene and utilized oocytes from these animals to characterize I_Cl,OR biophysical properties. The I_Cl,OR channel is strongly voltage dependent. Outward rectification is due to voltage-dependent current activation at depolarized voltages and rapid inactivation at voltages more hyperpolarized than approximately +20 mV. Apparent channel open probability is zero at voltages less than +20 mV. The channel has a 4:1 selectivity for Cl^– over Na⁺ and an anion selectivity sequence of SCN^– > I^– > Br^– > Cl^– > F^–. I_Cl,OR is relatively insensitive to most conventional anion channel inhibitors including DIDS, 4,4'-dinitrostilbene-2,2'-disulfonic acid, 9-anthracenecarboxylic acid, and 5-nitro-2-(3-phenylpropylamino)benzoic acid. However, the current is rapidly inhibited by niflumic acid, metal cations including Gd³⁺, Cd²⁺, and Zn²⁺, and bath acidification. The combined biophysical properties of I_Cl,OR are distinct from those of other anion currents that have been described. During oocyte meiotic maturation, I_Cl,OR activity is rapidly downregulated, suggesting that the channel may play a role in oocyte Cl^– homeostasis, development, cell cycle control, and/or ovulation. chloride channel; ovulation; cell cycle; meiotic maturation     

Development of Nuclear Vacuoles in Sugar Beet Male Meiocytes

The formation, development and disappearance of nuclear vacuolesin three genetically different lines of Beta vulgaris L. arepresented. Nuclear vacuoles may be generated in zygotene bythe nuclear envelope, providing the membrane necessary for theformation of vacuoles within the nucleus. During subsequentstages of meiotic prophase nuclear vacuole volume increasesrapidly, peaking in pachytene. In diplotene/diakinesis totalnuclear vacuole volume falls gradually, and vacuoles eventuallydisappear in metaphase I. Maximum nuclear vacuole size in pachytenecoincides with a notable increase in nuclear surface. We thereforesuggest that nuclear vacuoles do not perform a mechanical functionin the contraction of the karyoplasm, but instead act as compartmentsin which products of nuclear metabolism may be stored, and subsequentlyparticipate in the elimination of nuclear products to the cytoplasm.These processes may be related to changes occurring preparatoryto the transition from sporophyte to gametophyte generation. Beta vulgaris L, sugar beet, meiocytes, nuclear vacuoles, ultrastructure     

The IJsselmeer and its phytoplankton--with special attention to the suitability of the lake as a habitat for Oscillatoria agardhii Gom

The IJsselmeer (surface area 1200 km², mean depth 4.5 m, residencetime 0.4 year, phosphorus load 7 g m^–2 year^–1) isa very important conservation area. Regular summer bloomingof Oscillatoria spp. can depreciate this value, so the boundsof possibility of this kind of blooming have been investigated.Therefore samples were taken along the shore and in the openwater from 1974 to 1982, continuous temperature profile measurementswere made in the same period in the middle of the lake and insitu primary production was measured in 1976 and 1977. The phytoplanktonconsists of green algae throughout the year, diatoms in spring,and blooms of Microcystis aeruginosa in summer. Blooming ofOscillatoria agardhii Gom occurs regularly in summer along theFrisian shore. In 1976, however, a heavy bloom of this algaoccurred in the whole lake. Comparing the IJsselmeer with shallowerOscillatoria-lakes in the Netherlands distinct differences arepresent concerning biomass, chlorophyll a content, relativevolume of the euphotic zone and light-dark cycle. Not only thelarger depth and extensiveness of the IJsselmeer are unfavourablefactors for Oscillatoria, but also the separation by land reclamationof many of the shallow littoral regions from the main body ofthe lake. High temperature and microstratification are neededto develop a bloom in the whole lake.     

Development, growth and egg production of Centropages abdominalis in the eastern subarctic Pacific

Centropages abdominalis is a neritic, omnivorous, temporallyabundant copepod present throughout the subarctic Pacific andits marginal seas. The two main objectives of this study wereto determine how temperature influences the development of C.abdominalis and whether growth rates of in situ populationsmay be limited by available food. At 6.9°C, median developmenttime from eggs laid to 50% adults was 42 days and the averageweight-specific somatic growth rate was 0.17 day^–1. At4.6°C, median development time to adult was 59 days (projected)and growth rate averaged 0.08 day^–1, suggesting that 4.6°Cmay be approaching the lower temperature for development andgrowth in this species. The functional relationship betweendevelopment time and temperature was established over the temperaturerange in which this species occurs. The in situ adult growthrates between 10 and 13°C averaged 0.14 day^–1 andwere generally lower than the laboratory-reared juvenile growthrates, which may indicate that adult C. abdominalis are foodlimited in the field during summer and autumn.     

Production primaire en baie de Concarneau. Relations algues-bacteries et filtration differentielle

A study of the phytoplankton in Concarneau Bay (Bretagne, France)was carried out on 6–8 June, 1978. Autotrophy was measuredby the ¹⁴C method and heterotrophic activity was estimated by¹⁴C labelled glucose and protein hydrolysate uptake. The measurementswere made on the whole of the sample as well as on the bacterialfraction, isolated by size-fractionation on Nuclepore 3 µmfilters. Moreover, a study of the relationships between naturalphytoplanktonic and bacterial populations was possible becauseof cell counts. The phytoplankton shows coastal characteristicsand diatoms are dominant. The distribution of the genus Chaetoceros,Nitzschia and Leptocylindrus seems to have an influence on therelationship between algae and bacteria so that environmentalconditions not only interfere directly at the bacterial levelon heterotrophic activity, but also through the specific compositionand abundance of the phytoplankton.     

Junction Complexes between Sieve Tubes in the Secondary Phloem of Myrtaceae

Junction complexes of unusual structure form between neighbouringsieve tubes in the secondary phloem of Eucalyptus species. Thick-walledribs support thin-walled ‘sieve areas’. In longitudinalsections the structures have a ‘concertina’- likeappearance. They are relatively large, up to 0.2 mm in length.Electron micrographs confirmed that the structures consistedof thin-walled areas perforated with pores, supported by muchthicker ribs. The structures provide a vast surface area fortransfer of metabolites between sieve tubes compared with thatof lateral wall sieve areas of other plants. Hydrolysis of parenchymacell walls occurs during the development of the junction complexes.The structures are only found when sieve tubes are in closeproximity and it is the redifferentiation and partitioning ofintervening parenchyma cells which result in junction complexformation. A survey for the presence of the structures in thephloem of other genera in the family Myrtaceae was made andthey were found in Tristania and Angophora but were not observedin Acmena and Metrosideros. Eucalyptus, sieve tubes, lateral walls, ultrastructure     

Three stages of meiotic homologous chromosome pairing in wheat: cognition, alignment and synapsis

 Chromosome painting enabled the study of homologous chromosome behaviour prior to and during meiosis. Total genomic DNA from rye, used as a probe for in situ hybridization, identified the rye chromosome arm in a wheat-rye translocation line (T5AS·5RL) at meiotic prophase and the preceding interphase. Accurate staging of the development of the meiocytes was attained by parallel studies of chromatin morphology, nucleolar behaviour and synaptonemal complex formation in electron microscopy thin sections and silver-stained surface spreads. Three stages of pairing were identified for the large cereal genomes that are organized in a Rabl configuration: first, cognition occurs during the long interphase before leptotene, bringing the homologous chromosome domains into close proximity and possibly starting at the centromere; second, homologous chromosome segments align at late leptotene; and third, zygotene synapsis initiates near the telomere, although it was also observed to occur near the centromere. A pairing model is proposed for wheat, with a genome size of 17000 Mbp, that shows prallels to and notable differences from yeast and mammalian models of meiosis. Received: 25 January 1997 / Revision accepted: 14 July 1997     

Functional characterization of voltage-gated K+ channels in mouse pulmonary artery smooth muscle cells

Mice are useful animal models to study pathogenic mechanisms involved in pulmonary vascular disease. Altered expression and function of voltage-gated K⁺ (K_V) channels in pulmonary artery smooth muscle cells (PASMCs) have been implicated in the development of pulmonary arterial hypertension. K_V currents (I_K(V)) in mouse PASMCs have not been comprehensively characterized. The main focus of this study was to determine the biophysical and pharmacological properties of I_K(V) in freshly dissociated mouse PASMCs with the patch-clamp technique. Three distinct whole cell I_K(V) were identified based on the kinetics of activation and inactivation: rapidly activating and noninactivating currents (in 58% of the cells tested), rapidly activating and slowly inactivating currents (23%), and slowly activating and noninactivating currents (17%). Of the cells that demonstrated the rapidly activating noninactivating current, 69% showed I_K(V) inhibition with 4-aminopyridine (4-AP), while 31% were unaffected. Whole cell I_K(V) were very sensitive to tetraethylammonium (TEA), as 1 mM TEA decreased the current amplitude by 32% while it took 10 mM 4-AP to decrease I_K(V) by a similar amount (37%). Contribution of Ca²⁺-activated K⁺ (K_Ca) channels to whole cell I_K(V) was minimal, as neither pharmacological inhibition with charybdotoxin or iberiotoxin nor perfusion with Ca²⁺-free solution had an effect on the whole cell I_K(V). Steady-state activation and inactivation curves revealed a window K⁺ current between –40 and –10 mV with a peak at –31.5 mV. Single-channel recordings revealed large-, intermediate-, and small-amplitude currents, with an averaged slope conductance of 119.4 ± 2.7, 79.8 ± 2.8, 46.0 ± 2.2, and 23.6 ± 0.6 pS, respectively. These studies provide detailed electrophysiological and pharmacological profiles of the native K_V currents in mouse PASMCs. K_V channels     

Large-conductance calcium-activated potassium channel activity is absent in human and mouse neutrophils and is not required for innate immunity

Large-conductance Ca²⁺-activated K⁺ (BK) channels are reported to be essential for NADPH oxidase-dependent microbial killing and innate immunity in leukocytes. Using human peripheral blood and mouse bone marrow neutrophils, pharmacological targeting, and BK channel gene-deficient (BK^–/–) mice, we stimulated NADPH oxidase activity with 12-O-tetradecanoylphorbol-13-acetate (PMA) and performed patch-clamp recordings on isolated neutrophils. Although PMA stimulated NADPH oxidase activity as assessed by O₂^– and H₂O₂ production, our patch-clamp experiments failed to show PMA-activated BK channel currents in neutrophils. In our studies, PMA induced slowly activating currents, which were insensitive to the BK channel inhibitor iberiotoxin. Instead, the currents were blocked by Zn²⁺, which indicates activation of proton channel currents. BK channels are gated by elevated intracellular Ca²⁺ and membrane depolarization. We did not observe BK channel currents, even during extreme depolarization to +140 mV and after elevation of intracellular Ca²⁺ by N-formyl-L-methionyl-L-leucyl-phenylalanine. As a control, we examined BK channel currents in cerebral and tibial artery smooth muscle cells, which showed characteristic BK channel current pharmacology. Iberiotoxin did not block killing of Staphylococcus aureus or Candida albicans. Moreover, we addressed the role of BK channels in a systemic S. aureus and Yersinia enterocolitica mouse infection model. After 3 and 5 days of infection, we found no differences in the number of bacteria in spleen and kidney between BK^–/– and BK^+/+ mice. In conclusion, our experiments failed to identify functional BK channels in neutrophils. We therefore conclude that BK channels are not essential for innate immunity. killing assay; reactive oxygen species; BK-deficient mice; mice infection     

{Omega}-3 and {omega}-6 polyunsaturated fatty acids block HERG channels

Dietary polyunsaturated fatty acids (PUFAs) have been reported to exhibit antiarrhythmic properties, which have been attributed to their availability to modulate Na⁺, Ca²⁺, and several K⁺ channels. However, their effects on human ether-a-go-go-related gene (HERG) channels are unknown. In this study we have analyzed the effects of arachidonic acid (AA, -6) and docosahexaenoic acid (DHA, -3) on HERG channels stably expressed in Chinese hamster ovary cells by using the whole cell patch-clamp technique. At 10 µM, AA and DHA blocked HERG channels, at the end of 5-s pulses to –10 mV, to a similar extent (37.7 ± 2.4% vs. 50.2 ± 8.1%, n = 7–10, P > 0.05). 5,6,11,14-Eicosatetrayenoic acid, a nonmetabolizable AA analog, induced effects similar to those of AA on HERG current. Both PUFAs shifted the midpoint of activation curves of HERG channels by –5.1 ± 1.8 mV (n = 10, P < 0.05) and –11.2 ± 1.1 mV (n = 7, P < 0.01). Also, AA and DHA shifted the midpoint of inactivation curves by +12.0 ± 3.9 mV (n = 4; P < 0.05) and +15.8 ± 4.3 mV (n = 4; P < 0.05), respectively. DHA and AA accelerated the deactivation kinetics and slowed the inactivation kinetics at potentials positive to +40 mV. Block induced by DHA, but not that produced by AA, was higher when measured after applying a pulse to –120 mV (IO). Finally, both AA and DHA induced a use-dependent inhibition of HERG channels. In summary, block induced by AA and DHA was time, voltage, and use dependent. The results obtained suggest that both PUFAs bind preferentially to the open state of the channel, although an interaction with inactivated HERG channels cannot be ruled out for AA. K⁺ channel; membrane currents; ion channels; arrhythmia; antiarrhythmics     

Pollination Ecology ofChloranthus serratus(Thunb.) Roem. et Schult. andCh. fortunei(A. Gray) Solms-Laub. (Chloranthaceae)

Flowering and pollination biology ofChloranthus serratusandChloranthusfortuneiwere studied. Flowering took place from early Marchto mid–April inCh. fortunei, and from April to September,the whole growth period, inCh. serratus.The flowering periodof an inflorescence ofCh. serratusaveraged about 8 d and anthesisof a single flower was 5–6 d. Flowers are slightly protogynous.The flower emitted fragrance when the androecium became white.Both species are entomophilous with thrips as exclusive pollinators.Under natural conditions, fruit set occurs mainly as a resultof cross-pollination, but self-pollination and agamospermy mayoccur in some cases. In flowers ofCh. fortuneiandCh. serratus,theincurved androecium, the carpel and the spike axis form a nearlyclosed chamber that contains the anthers and stigma. The developmentof a floral-axial chamber may be an important step towards amore economical and effective pollination system. Floral morphology,pollination biology and fossil evidence suggest that the mainevolutionary trend in the genusChloranthusis towards developmentof ‘closed’ flowers. The fidelity of the relationshipbetweenChloranthusand thrips is regarded as a specialized featureof pollination biology and this relationship may have originatedearly in the evolutionary lineage.Copyright 1999 Annals of BotanyCompany Chloranthaceae,Chloranthus, Thysanoptera,flowering, floral-axial chamber, pollination, evolution trends.     

Phytotoxic Compounds produced by Fusarium equiseti

Culture filtrates of several strains of Fusarium equiseti (Corda)Sacc. were found to be highly phytotoxic. Several phytotoxicsubstances were isolated; the most important of these was acolourless crystalline compound, diacetoxyscirpenol, m.p. 161-2°,C₁₉H₂₆O₇. Though highly phytotoxic in many respects, it markedlystimulates elongation of cress roots at 0.01–0.5 µg./ml.This is not an anti-auxin effect. Diacetoxyscirpenol is at mostonly slightly inhibitory to fungi and bacteria, but is highlytoxic to rats. Two minor metabolic products, m.p. 135-6°and 185-8° respectively, are also described. They, too,were highly phytotoxic, though their spectra of activity weresomewhat different from that of diacetoxyscirpenol. They alsowere highly toxic to rats. Production of these substances isprobably not important in the aetiology of disease symptomsin plants parasitized by F. equiseti, but may be significantin cases of toxicosis of animals fed with grain infected byFusaria. Scirpentriol, a hydrolysis product of diacetoxyscirpenol,is much less phytotoxic but of equal toxicity to rats.     

Cl- secretory effects of EBIO in the rabbit conjunctival epithelium

Experiments were conducted to determine whether the Cl^– secretagogue, 1-ethyl-2-benzimidazolinone (EBIO), stimulates Cl^– transport in the rabbit conjunctival epithelium. For this study, epithelia were isolated in an Ussing-type chamber under short-circuit conditions. The effects of EBIO on the short-circuit current (I_sc) and transepithelial resistance (R_t) were measured under physiological conditions, as well as in experiments with altered electrolyte concentrations. Addition of 0.5 mM EBIO to the apical bath stimulated the control I_sc by 64% and reduced R_t by 21% (P < 0.05; paired data). Under Cl^–-free conditions, I_sc stimulation using EBIO was markedly attenuated. In the presence of an apical-to-basolateral K⁺ gradient and permeabilization of the apical membrane, the majority of the I_sc reflected the transcellular movement of K⁺ via basolateral K⁺ channels. Under these conditions, EBIO in combination with A23187 elicited nearly instantaneous 60–90% increases in I_sc that were sensitive to the calmodulin antagonist calmidazolium and the K⁺ channel blocker tetraethyl ammonium. In the presence of an apical-to-basolateral Cl^– gradient and nystatin permeabilization of the basolateral aspect, EBIO increased the Cl^–-dependent I_sc, an effect prevented by the channel blocker glibenclamide (0.3 mM). The latter compound also was used to determine the proportion of EBIO-evoked unidirectional ³⁶Cl^– fluxes in the presence of the Cl^– gradient that traversed the epithelium transcellularly. Overall, EBIO activated apical Cl^– channels and basolateral K⁺ channels (presumably those that are Ca²⁺ dependent), thereby suggesting that this compound, or related derivatives, may be suitable as topical agents to stimulate fluid transport across the tissue in individuals with lacrimal gland deficiencies. Ussing chamber; short-circuit current; electrolyte transport; chloride secretagogue; potassium conductance; 1-ethyl-2-benzimidazolinone; 1,10-phenanthroline