Multiple calcium mobilization pathways in single avian salt gland cells

1990 ◽  
Vol 258 (2) ◽  
pp. C289-C298 ◽  
Author(s):  
E. L. Stuenkel ◽  
S. A. Ernst
Keyword(s):  
Salt Gland ◽  
Transient Increase ◽  
Secretory Cells ◽  
Free Medium ◽  
Subsequent Removal ◽  
Intracellular Pool ◽  
Gland Cells ◽  
Agonist Stimulation ◽  
Induced Changes ◽  
Sustained Increase

Agonist-induced changes in intracellular Ca2+ concentration ([Ca2+]i) in individual secretory cells from the avian salt gland were detailed using dual-wavelength microspectrofluorimetry of the Ca2(+)-sensitive fluorescent probe fura-2. Resting [Ca2+]i averaged 42 +/- 5 nM. Stimulation with the cholinergic agonist carbachol (1 microM) resulted in a rapid increase in [Ca2+]i to 308 +/- 26 nM, which was sustained at a nearly constant elevated level (328 +/- 31 nM) throughout agonist application. In the absence of extracellular Ca2+ or in the presence of an inorganic blocker of Ca2+ entry (Ni2+, 1 mM), only a transient increase in [Ca2+]i occurred on agonist stimulation, whereas subsequent readmission of Ca2+ or washout of Ni2+ reinitiated a sustained increase in [Ca2+]i. The initial transient response results from Ca2+ release from intracellular stores, whereas the sustained phase represents entry of extracellular Ca2+ into the cytoplasm. Repetitive stimulations in Ca2(+)-free medium alternating with Ca2(+)-containing medium were performed to examine the mechanisms involved in refilling of the agonist-sensitive intracellular pool. After depletion of the intracellular pool by stimulation in Ca2(+)-free medium, removal of the agonist and readmission of Ca2+ resulted in a rapid transient increase in [Ca2+]i that could be blocked by Ni2+, La3+, or elevated K+. Subsequent removal of extracellular Ca2+ and restimulation nonetheless showed that complete refilling of the intracellular pool had occurred in each case. These results suggest that two separate Ca2(+)-entry mechanisms, one sensitive to Ni2+, La3+, and elevated K+ and responsible for the agonist-induced increase in [Ca2+]i and one insensitive to the blockers and involved in refilling of the intracellular pool, may exist in salt gland cells. Spontaneous oscillations of [Ca2+]i that are independent of extracellular Ca2+ have also been observed in 10% of the cells. The abolition of the oscillations by depletion of the agonist-sensitive pool suggests this pool as the Ca2+ source for the oscillations.

Agonist-induced frequency modulation of Ca2+ oscillations in salt gland secretory cells

1991 ◽  
Vol 261 (1) ◽  
pp. C177-C184 ◽  
Author(s):  
K. M. Crawford ◽  
E. L. Stuenkel ◽  
S. A. Ernst
Keyword(s):  
Intracellular Calcium ◽  
Frequency Modulation ◽  
Calcium Concentration ◽  
Salt Gland ◽  
Effective Concentration ◽  
Secretory Cells ◽  
Oscillatory Frequency ◽  
Fura 2 ◽  
Dose Dependent ◽  
Sustained Increase

Oscillations in intracellular calcium concentration ([Ca2+]i) induced by the acetylcholine analogue carbachol (CCh) were characterized by microspectrofluorimetry of fura-2 in single secretory cells from the avian salt gland. The frequency of oscillations increased in graded fashion with [CCh] between 25 nM (2.7 +/- 0.6 min-1) and 250 nM (11.8 +/- 1.4 min-1), whereas the amplitude of the spikes was independent of [CCh]. An interperiod return to prestimulatory [Ca2+]i was generally seen only at very low (25 nM) CCh. Between 50 and 250 nM CCh, oscillations were associated with sustained elevated [Ca2+]i levels. The amplitude of the oscillatory spikes was found not to exceed that of initial spikes arising from prestimulatory [Ca2+]i, despite the dose-dependent [effective concentration at 50% (EC50) = 200 nM CCh] sustained rise in [Ca2+]i. At 1 microM CCh, oscillations gave way to a maximal sustained increase in [Ca2+]i. Reduction of [Ca2+]o to 1.5 microM during an oscillatory train or blockage of Ca2+ influx with Ni+ resulted in a reduction in sustained Ca2+i levels and in frequency, but not amplitude, of oscillations. A relationship between the sustained partial rise in [Ca2+]i derived from Ca2+ influx and the oscillatory frequency at a given [CCh] was further indicated by the lower frequency (P less than 0.01) of the early spikes in a train when interspike [Ca2+]i initially returned to near-basal levels. In some cells, oscillations were slow enough (less than 2 min-1) to resolve an interperiod of elevated baseline [Ca2+]i, showing that the latter can occur independent of the repetitive Ca2+ spikes. (ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Trypsin-induced increase in cyclic AMP concentration in rat thymocytes An effect independent of calcium and calmodulin

1986 ◽  
Vol 239 (3) ◽  
pp. 603-607 ◽  
Author(s):  
J Segal
Keyword(s):  
Cyclic Amp ◽  
Free Medium ◽  
Standard Medium ◽  
Intracellular Pool ◽  
Calmodulin Inhibitor ◽  
Related Increase ◽  
In Cells

Trypsin produces a dose-related increase in cellular cyclic AMP concentration in rat thymocytes [Shneyour, Patt & Trainin (1976) J. Immunol. 117, 2143-2149; Segal & Ingbar (1983) Clin. Res. 31, 277A]. In the present study, I examined whether this effect of trypsin requires Ca2+ and whether it is modified by calmodulin. In fresh thymocytes suspended in standard medium (containing 1 mM-Ca2+), trypsin produced a concentration-dependent increase in cytoplasmic free Ca2+ concentration, which was evident at a concentration of 50 micrograms of trypsin/ml and reached maximal values at about 1 mg/ml. This effect of trypsin was very prompt in onset, almost immediate, and reached maximal values within 2-3 min. But in cells suspended in essentially Ca2+-free medium (6 nM free Ca2+), trypsin had no effect on cytoplasmic free Ca2+ concentration, which indicates that trypsin acted by increasing Ca2+ uptake rather than Ca2+ release from an intracellular pool. However, the increase in thymocyte cyclic AMP concentration produced by trypsin was independent of extracellular Ca2+ and was not influenced by calmodulin, because it was the same in the presence or absence of Ca2+ and was not changed by the calmodulin inhibitor trifluoperazine. I therefore suggest that in rat thymocytes the trypsin-induced increase in cyclic AMP concentration does not require Ca2+ and is not influenced by calmodulin.

Biogenesis of Plasma Membranes in Salt Glands of Salt-Stressed Domestic Ducklings: Localization of Acyltransferase Activity

1972 ◽  
Vol 11 (3) ◽  
pp. 855-873
Author(s):  
A. M. LEVINE ◽  
JOAN A. HIGGINS ◽  
R. J. BARRNETT
Keyword(s):  
Plasma Membrane ◽  
Plasma Membranes ◽  
Specific Activity ◽  
Salt Gland ◽  
Secretory Cells ◽  
Salt Glands ◽  
Acyltransferase Activity ◽  
Structural Localization ◽  
Differentiated Cells ◽  
Golgi Membranes

In response to salt water stress there is a marked increase in the plasma membranes of the epithelial secretory cells of the salt glands of domestic ducklings. In the present study, the fine-structural localization of the acyltransferases involved in synthesis of phospholipids has been investigated in this tissue during this increased biogenesis of plasma membranes. The specific activity of the acyltransferases of the salt gland rose in response to salt stress, and this preceded the rapid increase in weight and cellular differentiation. After the weight increase of the gland became established, the specific activity of the acyltransferases declined, but the total activity remained constant. Salt gland tissue fixed in a mixture of glutaraldehyde and formaldehyde retained 35% of the acyltransferase activity of unfixed tissue. Cytochemical studies of the localization of acyltransferase activity in fixed and unfixed salt gland showed reaction product associated only with the lamellar membranes of the Golgi complex. This localization occurred in partially differentiated cells from salt-stressed glands to the greatest extent; and to only a small extent in cells of control tissue from unstressed salt glands. Omission of substrates resulted in absence of reaction product in association with the Golgi membranes. In addition, vesicles having limiting membranes morphologically similar to the plasma membrane occurred between the Golgi region and the plasma membrane in the partially differentiated cells. The phospholipid component of the plasma membrane appears therefore to be synthesized in association with the Golgi membranes and the membrane packaged at this site from which it moves in the form of vesicles to fuse with the pre-existing plasma membrane.

Mucus glycoconjugate secretion in cool and hypertonic solutions

1997 ◽  
Vol 272 (6) ◽  
pp. L1121-L1125 ◽  
Author(s):  
T. M. Dwyer ◽  
J. M. Farley
Keyword(s):  
Average Rate ◽  
Transient Increase ◽  
Productive Cough ◽  
Submucosal Gland ◽  
Basal Secretion ◽  
Cold Air ◽  
Dry Air ◽  
Gland Cells ◽  
Hypertonic Solutions ◽  
Exercise Induced

For sensitive individuals, exercise-induced asthma is triggered by cold and dry air and is often accompanied by a productive cough. In this study, we determined whether cold solutions and/or solutions of increased tonicity directly caused an increase in glycoconjugate (GC) secretion. To test this, we used isolated swine tracheal submucosal gland cells (TSGCs) and measured the rate of GC secretion at 37 and 32 degrees C in isotonic solutions and in solutions made hypertonic by 30 mosM. TSGCs were isolated under conditions that minimized the rate of GC secretion and were perfused with medium 199 equilibrated with 5% CO2 to a pH of 7.4. A lectin-based assay was used to specifically detect GC present in each 2-min fraction of the perfusate. Basal secretion was 3.1-fold greater at 32 degrees C (n = 3) than at 37 degrees C (n = 4; P < 0.05). At 37 degrees C, increasing perfusate osmolarity by 30 mosM increased the average rate of secretion by 41 +/- 11% (n = 4; P < 0.05); return to isotonic perfusate caused a 4.5 +/- 1.8-fold transient increase in secretion (n = 4; P < 0.05) that was complete within 10 min. At 32 degrees C, changing tonicity of the perfusate had no significant effect but returning to isotonic perfusate caused a 2.3 +/- 0.7-fold transient increase in secretion (n = 3; P < 0.05). Thus key stimuli that trigger obstruction of airflow (cold and increased osmolarity) can also directly stimulate GC secretion in the airway. Such increased secretions may contribute to the productive cough observed in some individuals in response to cold air.

Calcium signaling in cultured human and rat duodenal enterocytes

1998 ◽  
Vol 275 (2) ◽  
pp. G296-G304 ◽  
Author(s):  
Catherine S. Chew ◽  
Bengt Säfsten ◽  
Gunnar Flemström
Keyword(s):  
Extracellular Matrix ◽  
Calcium Signaling ◽  
Primary Culture ◽  
Calcium Concentration ◽  
Duodenal Mucosa ◽  
Transient Increase ◽  
Active Movement ◽  
Culture Model ◽  
Columnar Cells ◽  
Sustained Increase

Vagal stimuli increase duodenal mucosal[Formula: see text] secretion and may provide anticipatory protection against acid injury, but duodenal enterocyte (duodenocyte) responses and cholinoceptor selectivity have not been defined. We therefore developed a stable primary culture model of duodenocytes from rats and humans. Brief digestion of scraped rat duodenal mucosa or human biopsies with collagenase/dispase yielded cells that attached to the extracellular matrix Matrigel within a few hours of plating. Columnar cells with villus enterocyte morphology that exhibited spontaneous active movement were evident between 1 and 3 days of culture. Rat duodenocytes loaded with fura 2 responded to carbachol with a transient increase in intracellular calcium concentration ([Ca2+]i), with an apparent EC50 of ∼3 μM. In a first type of signaling pattern, [Ca2+]ireturned to basal or near basal values within 3–5 min. In a second type, observed in cells with enlarged vacuoles characteristic of crypt cell morphology, the initial transient increase was followed by rhythmic oscillations. Human duodenocytes responded with a more sustained increase in [Ca2+]i, and oscillations were not observed. Rat as well as human duodenocytes also responded to CCK-octapeptide but not to vasoactive intestinal polypeptide. Equimolar concentrations (100 nM) of the subtype-independent muscarinic antagonist atropine and the M3 antagonist 4-diphenylacetoxy- N-methylpiperidine methiodide prevented the response to 10 μM carbachol, whereas the M1 antagonist pirenzepine and the M2 antagonists methoctramine and AF-DX 116BS had no effect at similar concentrations. Responses in rat and human duodenocytes were similar. A new agonist-sensitive primary culture model for rat and human duodenocytes has thus been established and the presence of enterocyte CCK and muscarinic M3 receptors demonstrated.

Intracellular Ca2+ signalling in secretory cells.

1997 ◽  
Vol 200 (2) ◽  
pp. 303-314
Author(s):  
T J Shuttleworth
Keyword(s):  
Arachidonic Acid ◽  
Calcium Ion ◽  
Critical Role ◽  
Secretory Activity ◽  
Secretory Cells ◽  
Free Calcium ◽  
Cytosolic Free Calcium ◽  
Maximal Stimulation ◽  
Induced Changes ◽  
Extracellular Environment

The secretion of ions and fluid plays a critical role in a variety of physiological activities that are vital to homeostatic mechanisms in animals. Control of such secretory activity is achieved by a range of neurotransmitters and hormones many of which act intracellularly by generating the second messenger inositol 1,4,5-trisphosphate (InsP3) and increasing cytosolic free calcium ion concentrations ([Ca2+]i). These increases are achieved by a combination of the InsP3-induced release of Ca2+ from specific intracellular stores and the activation of Ca2+ entry from the extracellular environment. The [Ca2+]i signal represents a balance between the adequate activation of components of the secretory mechanism and the avoidance of [Ca2+]i levels that are toxic to the cell. Resting [Ca2+]i is maintained low by the action of Ca2+ pumps on the intracellular stores and plasma membrane, with the result that gradients for Ca2+ movement into the cytosol from either of these two sources are very large and there is considerable potential for achieving rapid increases in [Ca2+]i. Consequently, for successful Ca2+ signalling, it is imperative that these two mechanisms of raising [Ca2+]i (i.e. Ca2+ release and Ca2+ entry) are closely integrated. Current models emphasize the activation of Ca2+ entry as a downstream result of the emptying of the intracellular stores ("capacitative' model). Whilst this may be true for situations of maximal stimulation, recent experiments on the oscillatory [Ca2+]i responses typical of more physiological levels of stimulation indicate a previously unsuspected, independent activation of Ca2+ entry involving arachidonic acid. This arachidonic-acid-activated entry plays a key role, along with InsP3, in inducing the repetitive release of Ca2+ from the stores to produce the [Ca2+]i oscillations. In this way, the two components responsible for the elevation of [Ca2+]i are intimately related and their dual effects closely coordinated, resulting in the finely tuned control of agonist-induced changes in [Ca2+]i.

Primary culture of duck salt gland. I. Morphology of confluent cell layers

1985 ◽  
Vol 249 (1) ◽  
pp. C32-C40 ◽  
Author(s):  
R. J. Lowy ◽  
J. H. Schreiber ◽  
D. C. Dawson ◽  
S. A. Ernst
Keyword(s):  
Electron Microscopy ◽  
Primary Culture ◽  
Plasma Membranes ◽  
Freeze Fracture ◽  
Salt Gland ◽  
Direct Access ◽  
Secretory Cells ◽  
Collagen Gels ◽  
Lateral Membrane ◽  
Confluent Cell

Dissociated avian salt gland secretory cells were maintained in primary culture after plating on hydrated collagen gels. When seeded at 3 X 10(6) cells/cm2, confluent cell sheets formed within 2-3 days, whereas cultures seeded at lower densities formed a complex reticulum of cell aggregates, which remained nonconfluent even after 7 days. Scanning electron microscopy showed that the free surface of 3-day confluent cultures consisted of intermixed convex and flattened cell membranes with prominent junctional boundaries and abundant microvilli. Transmission electron microscopy indicated that these cultures were multilayers of 1-4 cells in thickness. The plasma membranes of the superficial cells were polarized into apical and basolateral regions displaying, respectively, microvilli and interdigitating lateral membrane folds. These membrane domains were separated by shallow occluding junctions, which consisted of both single strands and simple net-like arrays in freeze-fracture images. Underlying epithelial cells retained lateral membrane folds and formed desmosomal contacts with superficial and neighboring cells. These cultures, unlike the intact tissue, allow direct access to the apical and basolateral cell surfaces for electrophysiological analysis of transmural active ion transport.

scholarly journals Dissimilar mechanisms of Ca2+response to bradykinin in different types of juxtamedullary glomerular arterioles

1999 ◽  
Vol 277 (5) ◽  
pp. F697-F705 ◽  
Author(s):  
F. Praddaude ◽  
J. Marchetti ◽  
F. Alhenc-Gelas ◽  
J.-L. Ader
Keyword(s):  
Tyrosine Kinases ◽  
Maximal Response ◽  
Lower Sensitivity ◽  
Free Medium ◽  
Herbimycin A ◽  
Intracellular Calcium Level ◽  
Different Types ◽  
Intracellular Ca ◽  
Induced Changes ◽  
Glomerular Arterioles

Bradykinin (BK)-induced changes in intracellular calcium level ([Ca2+]i) were studied on fura 2-loaded afferent (AA) and efferent glomerular arterioles (EA) microdissected from juxtamedullary renal cortex. A distinction was made between thin and muscular EA. In AA and both types of EA, BK increased [Ca2+]ithrough activation of B2 receptors located only on the endothelium. The responses were not affected by nifedipine (10−6 M) and were smaller in a Ca2+-free medium, providing evidence that BK opens voltage-independent Ca2+ channels and mobilizes intracellular Ca2+. Thin EA differed from AA and muscular EA by a lower sensitivity to BK (EC50 = 6.95 ± 3.81 vs. 0.21 ± 0.08 and 0.18 ± 0.13 nM, respectively; P < 0.05), a higher maximal response (89 ± 5 vs. 57 ± 5 and 44 ± 7 nM; P < 0.001), and a spontaneous return to basal Ca2+ level, even in the presence of BK. Genistein (10−4 M) and herbimycin A (25 × 10−6M), specific inhibitors of tyrosine kinases, inhibited the [Ca2+]iresponses exclusively in AA. Genistein reduced the peak and plateau phases of responses by 69 ± 9 and 82 ± 6%, respectively, in a medium with Ca2+ and the peak by 48 ± 9% in a Ca2+-free medium. Similar reductions were observed with herbimycin A. These results show that dissimilar signal transduction pathways are involved in BK effects on juxtamedullary arterioles and that a tyrosine kinase activity could participate in the regulation of BK effect on AA but not on EA.

Ultrastructural profile of metapleural gland cells of the ant Atta laevigata (F. Smith, 1858) (Formicidae: Attini)

2012 ◽  
Vol 62 (1) ◽  
pp. 1-11 ◽  
Author(s):  
Maria Izabel Camargo-Mathias ◽  
Odair Correa Bueno ◽  
Alexsandro Santana Vieira
Keyword(s):  
Endoplasmic Reticulum ◽  
Squamous Epithelium ◽  
Smooth Endoplasmic Reticulum ◽  
Secretory Cells ◽  
Metapleural Gland ◽  
Atta Laevigata ◽  
Gland Cells ◽  
Transmission Electron ◽  
Secretion Granules ◽  
Myelin Figures

AbstractThe metapleural gland is considered a synapomorphy of ants, and is characterized as a paired structure located at the two postlateral ends of the thorax and secretes substances capable of inhibiting and controlling fungi and bacteria in the fungus garden and inside the nest. This study was aimed at investigating if and which are the ultrastructural differences in the metapleural gland cells of workers (three castes) of Atta laevigata using transmission electron microscopy (TEM). This study revealed the presence of Golgi regions, rough endoplasmic reticulum (lamellar and vesicular shapes), smooth endoplasmic reticulum, mitochondria (elongated, round-shaped), vacuoles, secretion granules with different electron densities, and myelin figures in the cytoplasm of secretory cells, indicating that this gland produces substances composed of proteins, lipids, and polysaccharides (glycogen in major workers). Lipid droplets and secretion granules were found very near to the microvilli, especially in minor workers. The intracellular portion of canaliculi exhibited invaginations that increased the surface area and modified the secretion produced by the secretory cells. In the three castes examined, the gland exhibited a reservoir preceded by a collecting chamber, both lined by a simple squamous epithelium with a cuticular intima. Workers of the three castes of A. laevigata might be involved in the production of secretion mainly composed of proteins with antibiotic properties and, minor workers, may be responsible for producing a wider variety of secretions compared to median and major workers in the colony.

External ATP-induced changes in [Ca2+]i and membrane currents in mammalian atrial myocytes

1991 ◽  
Vol 260 (4) ◽  
pp. C673-C680 ◽  
Author(s):  
Y. Hirano ◽  
S. Abe ◽  
T. Sawanobori ◽  
M. Hiraoka
Keyword(s):  
Digital Imaging ◽  
Calcium Concentration ◽  
Transient Increase ◽  
P2 Receptor ◽  
Atrial Myocytes ◽  
Whole Cell ◽  
Inward Current ◽  
Whole Cell Patch Clamp ◽  
Induced Changes ◽  
Whole Cell Recordings

Fura-2 fluorescent digital-imaging microscopy and whole cell patch-clamp recordings were used to study the effects of externally applied ATP on atrial myocytes isolated from rabbit and guinea pig hearts. Application of 100 microM ATP elicited a transient increase in intracellular calcium concentration ([Ca2+]i), which was not suppressed by theophylline, whereas adenosine and ADP failed to evoke the response. The Ca2+ transients were suppressed by the application of Co2+, Ni2+, or verapamil and by the removal of extracellular Ca2+, indicating that the inflow of external Ca2+ is necessary to evoke the response. The Ca2+ transient was suppressed also by ryanodine, suggesting that the mobilization of intracellular Ca2+ is another important factor. In the whole cell recordings, ATP induced a transient depolarization of the membrane potential due to the activation of a rapidly desensitizing inward current which persisted in the presence of Co2+, Ni2+, verapamil, or ryanodine. These results indicate that in mammalian atrial myocytes, ATP evoked transient increase in [Ca2+]i via P2-receptor, through the release of internally stored Ca2+ associated with the inflow of external Ca2+. This response seemed to be triggered mainly by the influx of Ca2+ through L-type Ca2+ channel activated by membrane depolarization, which was caused by the ATP-induced inward current.

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