Extracellular Mg2+Inhibits both Histamine-Stimulated Ca2+-Signaling and Exocytosis in Human Tracheal Secretory Gland Cells

Sebille, Stéphane; Pereira, Manuella; Millot, Jean-Marc; Jacquot, Jacky; Delabroise, A. M.; Arnaud, Maurice J.; Manfait, Michel

doi:10.1006/bbrc.1998.8494

Cited by 7 publications

(3 citation statements)

References 26 publications

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“…Winter flounder mucus calcium and magnesium concentrations, as measured by atomic absorption, were 0.68 mM magnesium and 0.36 mM calcium (data not shown). These values are within the functional range of pleurocidin and comparable to concentrations in human tracheal secretory gland cells (35).…”

Section: Resultssupporting

confidence: 72%

Characterization of a Fish Antimicrobial Peptide: Gene Expression, Subcellular Localization, and Spectrum of Activity

Cole

Darouiche

Legarda

et al. 2000

Antimicrob Agents Chemother

143

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Antimicrobial peptides are proposed to act as the first line of mucosal host defense by exerting broadspectrum microbicidal activity against pathogenic microbes. Pleurocidin, a new 25-residue linear antimicrobial peptide, was recently isolated from the skin secretions of winter flounder (Pleuronectes americanus). The present study identifies the cDNA and gene encoding pleurocidin. The pleurocidin gene comprises four exons. Its upstream region demonstrates consensus binding sequences for transcription factors found in host defense genes in mammals, including sequences identical to the NF-IL6 and alpha and gamma interferon response elements. Pleurocidin is predicted to exist as a 68-residue prepropeptide that undergoes proteolytic cleavage of its amino-terminal signal and carboxy-terminal anionic propiece to form the active, mature peptide. Transmission electron microscopy localized pleurocidin to the mucin granules of skin and intestinal goblet cells. Significant synergy was shown to occur between pleurocidin and D-cycloserine targeting Mycobacterium smegmatis. Pleurocidin was functionally active at physiologic concentrations of magnesium and calcium; however, high concentrations of these divalent cations ablated pleurocidin's activity against a standard test strain, Escherichia coli D31. Pleurocidin was tested against bacterial and fungal clinical isolates and showed broad-spectrum antimicrobial activity. Together, these data support the hypothesis that pleurocidin participates in innate mucosal immunity, and it may prove to be a beneficial therapeutic agent.Increasing evidence suggests that endogenous peptides with antimicrobial properties play an important role in host defense. These peptides possess marked microbicidal activity and have been isolated from a variety of cells of myeloid lineage and mucosal surfaces in most species tested thus far (4,5,8,12). The recent focus has been on mucus-derived peptides and their roles in innate host defense at organism-environment interfaces, such as the integument and the respiratory and digestive epithelia. The multitude of peptides discovered at mucosal surfaces include human ␤-defensin 1 (HBD-1) in urogenital tissues (43) and bronchoalveolar lavage fluid (39), HBD-2 at sites of inflammation (24, 39), cryptdins from the Paneth cells (18), tracheal antimicrobial peptide (TAP) and lingual antimicrobial peptide (LAP) from cows (10, 33), and magainin and PGLa from frogs (46). Each peptide class confers a broad spectrum of antimicrobial activity and cationic charge at physiologic pH. Many peptides show high interspecies cDNA and protein sequence homology, frequently across evolutionarily diverse phyla.We recently discovered pleurocidin, a novel 25-residue linear antimicrobial peptide in the skin mucous secretions of the winter flounder, Pleuronectes americanus (6). The sequence of the mature molecule, GWGSFFKKAAHVGKHVGKAAL THYL, shows sequence homology with the dermaseptin (tree frog) and ceratotoxin (medfly) classes of antimicrobial peptides. Pleurocidin is a highly basic...

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Section: Resultssupporting

confidence: 72%

Characterization of a Fish Antimicrobial Peptide: Gene Expression, Subcellular Localization, and Spectrum of Activity

Cole

Darouiche

Legarda

et al. 2000

Antimicrob Agents Chemother

143

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“…Fluorescent analogues of calcium chelators have proven extremely useful in measuring the intracellular calcium concentration of various types of living cells. Fluorescent probes that have been widely used include Indo-1 (June and Rabinovitch, 1994;Sipido and Callewaert, 1995;Jacquemond and Allard, 1998;Sebille et al, 1998;Ju and Allen, 1998;Grynkiewicz et al, 1985), Fura-2 (Xu et al, 1997;Sipido and Callewaert, 1995;Chen et al, 1996;Zicha et al, 1996;Greco et al, 1996;Bhattacharya and Chakrabarti, 1998;Miao et al, 1998;Grynkiewicz et al, 1985), Quin-2 (Tsien and Pozzan, 1989;Deber and Hsu, 1986;Jacob et al, 1987;Nijayaraghavan and Hoskins, 1989), calcium green dextran (Zimprich and Bolsover, 1996;Harris, 1994;Mc-Clellan et al, 1994;Tamm and Terasaki, 1994;Kono et al, 1996;Bi et al, 1995;Carroll et al, 1994;Fetcho and O'Malley, 1995), and chlortetracycline (Tao et al, 1992;Jy and Haynes, 1984. Most experiments of this type are performed at constant temperature (e.g., 37°C).…”

Section: Introductionmentioning

confidence: 99%

Effects of Temperature on Calcium-Sensitive Fluorescent Probes

Oliver

Baker

Fugate

et al. 2000

Biophysical Journal

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The effect of temperature on the binding equilibria of calcium-sensing dyes has been extensively studied, but there are also important temperature-related changes in the photophysics of the dyes that have been largely ignored. We conducted a systematic study of thermal effects on five calcium-sensing dyes under calcium-saturated and calcium-free conditions. Quin-2, chlortetracycline, calcium green dextran, Indo-1, and Fura-2 all show temperature-dependent effects on fluorescence in all or part of the range tested (5-40 degrees C). Specifically, the intensity of the single-wavelength dyes increased at low temperature. The ratiometric dyes, because of variable effects at the two wavelengths, showed, in general, a reduction in the fluorescence ratio as temperature decreased. Changes in viscosity, pH, oxygen quenching, or fluorescence maxima could not fully explain the effects of temperature on fluorescence. The excited-state lifetimes of the dyes were determined, in both the presence and absence of calcium, using multifrequency phase-modulation fluorimetry. In most cases, low temperature led to prolonged fluorescence lifetimes. The increase in lifetimes at reduced temperature is probably largely responsible for the effects of temperature on the physical properties of the calcium-sensing dyes. Clearly, these temperature effects can influence reported calcium concentrations and must therefore be taken into consideration during any investigation involving variable temperatures.

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“…Therefore, multispectral f luorescence imaging provides the ability to determine properties of the local environment in a spatially resolved manner. Confocal microspectrof luorometers have been developed to measure spatially resolved f luorescence spectra 13 and have been used to study drug-target interactions, 13 ion concentrations, 14 and pH levels 15 in living cells. However, these devices are essentially benchtop systems with slow mechanical scanning and offer no solution for evaluating remote in vivo locations.…”

mentioning

confidence: 99%

Multispectral imaging with a confocal microendoscope

Rouse

Gmitro

2000

Opt. Lett.

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The concept of a multispectral confocal microscope for in vivo imaging is introduced. To demonstrate the concept we modified a slit-scan fluorescence confocal microendoscope incorporating a fiber-optic catheter for in vivo imaging to record multispectral images. The system was designed to examine cellular structures during optical biopsy and to exploit the diagnostic information contained within the spectral domain. Preliminary experiments were carried out in phantoms and cell cultures to demonstrate the potential of the technique.

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Extracellular Mg2+Inhibits both Histamine-Stimulated Ca2+-Signaling and Exocytosis in Human Tracheal Secretory Gland Cells

Cited by 7 publications

References 26 publications

Characterization of a Fish Antimicrobial Peptide: Gene Expression, Subcellular Localization, and Spectrum of Activity

Characterization of a Fish Antimicrobial Peptide: Gene Expression, Subcellular Localization, and Spectrum of Activity

Effects of Temperature on Calcium-Sensitive Fluorescent Probes

Multispectral imaging with a confocal microendoscope

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