The biochemical nature of the cell periphery of the salt gland secretory cells of fresh and salt water adapted mallard ducks

1974 ◽  
Vol 150 (2) ◽  
Author(s):  
B.J. Martin ◽  
CharlesW. Philpott
Keyword(s):  
Salt Water ◽  
Salt Gland ◽  
Secretory Cells ◽  
Cell Periphery

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.

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.

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.

Salt stress increases abundance and glycosylation of CFTR localized at apical surfaces of salt gland secretory cells

1994 ◽  
Vol 267 (4) ◽  
pp. C990-C1001 ◽  
Author(s):  
S. A. Ernst ◽  
K. M. Crawford ◽  
M. A. Post ◽  
J. A. Cohn
Keyword(s):  
Salt Stress ◽  
Single Cells ◽  
Salt Gland ◽  
Cell Protein ◽  
Secretory Cells ◽  
Salt Glands ◽  
Cl Secretion ◽  
Peptide Antibody ◽  
Nacl Secretion ◽  
Apical Membranes

Osmotic stress elicits hypertonic NaCl secretion and promotes structural and biochemical differentiation in avian salt glands. In addition to cholinergic control, Cl- secretion is stimulated by vasoactive intestinal peptide (VIP), suggesting that the cystic fibrosis transmembrane conductance regulator (CFTR) may be present and that its expression may be regulated by chronic salt stress. Anion efflux, assayed by 6-methoxy-N-(3-sulfopropyl)quinolinium fluorescence changes in single cells, was stimulated by VIP or 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate. Immunoblots with a COOH-terminal peptide antibody to human CFTR revealed approximately 170- and approximately 180-kDa bands in lysates from control and salt-stressed glands, respectively. Both variants reduced to approximately 140 kDa after N-glycanase digestion and gave identical tryptic phosphopeptide maps after immunoprecipitation and phosphorylation by protein kinase A. CFTR was localized to apical membranes by immunofluorescence and, additionally, to subapical vesicles by immunoelectron microscopy. Salt stress induced an approximately twofold increase in CFTR abundance/cell protein (approximately 5-fold/cell) and intensified apical membrane immunofluorescence. For comparison, Na+ pump expression increased approximately fourfold per cell protein with little change in actin. Thus differentiation induced by salt stress is accompanied by alteration in CFTR abundance and glycosylation. Upregulation of CFTR likely contributes to increased efficiency of Cl- secretion.

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)

Scanning and Transmission Electron Microscopy of Isolated Cells from the Avian Salt Gland

1977 ◽  
Vol 35 ◽  
pp. 488-489
Author(s):  
Ian G. Thompson
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Physiological Stress ◽  
Single Cells ◽  
Salt Gland ◽  
Secretory Cells ◽  
Surface Coat ◽  
Isolated Cells ◽  
Structural Differentiation ◽  
Transmission Electron

With the advent of new techniques for isolating single cells for biochemical and physiological investigation, an important consideration is the morphological integrity of these cells after dissociation from the intact tissue. Do isolated cells retain the degree of structural differentiation that is apparent in vivo? The principal secretory cells of the avian salt gland are an example of cells that are highly differentiated in form under conditions of physiological stress. This report describes the ultrastructure of dissociated salt gland cells as visualized with the scanning and transmission electron microscope.The dissociation procedure employed here was the same as that applied to the exocrine pancreas. For transmission electron microscopy the cell suspension was centrifuged and the resultant pellet prefixed in cacodylate buffered 3% glutaraldehyde- 1% paraformaldehyde, postfixed in unbuffered 1% osmium tetroxide, and embedded in epon-araldite. An assessment of the cell surface coat following enzymatic dissociation was facilitated by the inclusion of ruthenium red (500 ppm) in both the aldehyde and osmium fixation steps.

Plasma Membrane Biogenesis

1974 ◽  
Vol 32 ◽  
pp. 312-313
Author(s):  
Russell J. Barrnett
Keyword(s):  
Plasma Membrane ◽  
Trigeminal Nerve ◽  
Salt Gland ◽  
Ultrastructural Localization ◽  
Secretory Cells ◽  
Membrane Biogenesis ◽  
Cell Plasma ◽  
Microsome Fraction ◽  
Myelin Fraction

This report presents two examples of plasma membrane biogenesis in which the synthesis and assembly of components, phospholipid and enzyme proteins, were studies by a combination of biochemistry, cytochemistry and electron microscopy. These were: the proliferation of Schwann cell plasma membrane during the process of myelination of the trigeminal nerve in neonatal rats and amplification of the plasma membrane at the lateral and basal borders of secretory cells of the ducklings' salt gland as a result of salt stress.In the study concerning myelination a method for the ultrastructural localization of acyltransferase activities (the first two steps in phospholipid synthesis) was applied to the developing rat trigeminal nerve. Determination of acyltransferase levels in the nerve indicated that a peak of activity occurs at the 8th day after birth with gradual declines of activity up to 15 days. This peak coincided with the peak of a-glycerophosphate incorporation into phospholipids in the microsome fraction of the nerve: wheras, no incorporation was noted in the myelin fraction.

Influence of salinity on ultrastructure of the secretory cells of the adenohypophyseal pars distalis in yearling coho salmon (Oncorhynchus kisutch)

1977 ◽  
Vol 55 (1) ◽  
pp. 183-198 ◽  
Author(s):  
Yoshitaka Nagahama ◽  
W. Craig Clarke ◽  
W. S. Hoar
Keyword(s):  
Freshwater Fish ◽  
Fresh Water ◽  
Coho Salmon ◽  
Sea Water ◽  
Salt Water ◽  
Light And Electron Microscopy ◽  
Secretory Cells ◽  
Plasma Sodium ◽  
Pars Distalis ◽  
Freshwater Environment

Six different types of secretory cells were identified by light and electron microscopy in the adenohypophyseal pars distalis of yearling coho salmon acclimated to fresh or salt water. Prolactin cells are markedly more active in the freshwater than the seawater fish; these cells exhibit definite functional activity 3 days after transfer from salt to fresh water, indicating an osmoregulatory role of prolactin in the freshwater environment. Plasma sodium showed a significant decline 6 h after transfer from sea water to fresh water and, even after 1 week, remained lower than in the fully acclimated freshwater fish. Corticotropic (ACTH) cells did not appear cytologically different in freshwater and seawater fish. GH cells, the most prominent cells in the proximal pars distalis, appear more numerous and more granulated in the seawater fish, suggesting an osmoregulatory involvement in young coho salmon. Putative thyrotropic (TSH) and putative gonadotropic cells (GTH) can be distinguished by differences in granulation; only one type of GTH cell is evident with ultrastructural features that differ from those of sexually mature salmon. Stellate, non-granulated cells occur in all regions of the adenohypophysis but more frequently in the prolactin follicles; they are much more prominent in the seawater than freshwater fish.

scholarly journals PRESERVATION OF Na-K-ACTIVATED AND Mg-ACTIVATED ADENOSINE TRIPHOSPHATASE ACTIVITIES OF AVIAN SALT GLAND AND TELEOST GILL WITH FORMALDEHYDE AS FIXATIVE

1970 ◽  
Vol 18 (4) ◽  
pp. 251-263 ◽  
Author(s):  
STEPHEN A. ERNST ◽  
CHARLES W. PHILPOTT
Keyword(s):  
Atpase Activity ◽  
Adenosine Triphosphatase ◽  
Electron Microscopic Examination ◽  
Salt Gland ◽  
Chloride Cells ◽  
Secretory Cells ◽  
Salt Glands ◽  
Electron Microscopic ◽  
Fixed Tissue ◽  
Gill Filaments

The effect of glutaraldehyde and formaldehyde fixation on the level of biochemically demonstrable Na-K-adenosine triphosphatase (Na-K-ATPase) and Mg-ATPase of avian salt glands and teleost gill filaments was studied. Sections, 100-200 µ, prepared with the Smith-Farquhar tissue chopper, were fixed for varying periods, homogenized and assayed for ATPase activity. Fixation of salt gland tissue with 0.5% glutaraldehyde for 40-60 min completely inhibited the Na-K-ATPase activity and reduced the level of Mg-ATPase by 85%. In contrast, fixation with 2 or 3% formaldehyde, prepared from paraformaldehyde, for 60-90 min resulted in a loss of only 30% of the Na-K-ATPase activity and 65% of the Mg-ATPase activity. Similar results were obtained with gill filaments. In addition, Na-K-ATPase of formaldehyde-fixed tissue retained an obligatory requirement for Na+ and K+ and was fully sensitive to ouabain. Electron microscopic examination of formaldehyde-fixed tissue, sectioned with either the tissue chopper or in the cryostat and incubated in the Wachstein-Meisel medium, showed excellent morphologic preservation. Reaction product deposition (presumably due to Mg-ATPase) was associated with the extracellular side of the plasma membrane in the secretory cells of the salt gland and over the mitochondrial matrix of chloride cells present in the gill epithelium.

Cholinergic and adrenergic innervation of lingual salt glands of the estuarine crocodile, Crocodylus porosus

2005 ◽  
Vol 53 (6) ◽  
pp. 345 ◽  
Author(s):  
Craig E. Franklin ◽  
Greg Taylor ◽  
Rebecca L. Cramp
Keyword(s):  
Neural Control ◽  
Salt Water ◽  
Cholinergic Neurons ◽  
Secretory Activity ◽  
Salt Gland ◽  
Adrenergic Innervation ◽  
Chloride Ions ◽  
Adrenergic Nerve ◽  
Salt Glands ◽  
Crocodylus Porosus

Many marine reptiles and birds possess extrarenal salt glands that facilitate the excretion of excess sodium and chloride ions accumulated as a consequence of living in saline environments. Control of the secretory activity of avian salt glands is under neural control, but little information is available on the control of reptilian salt glands. Innervation of the lingual salt glands of the salt water crocodile, Crocodylus porosus, was examined in salt water-acclimated animals using histological methods. Extensive networks of both cholinergic and adrenergic nerve fibres were identified close to salt-secreting lobules and vasculature. The identification of both catecholamine-containing and cholinergic neurons in the salt gland epithelium and close to major blood vessels in the tissue suggests the action of the neurotransmitters on the salt-secreting epithelium itself and the rich vascular network of the lingual salt glands.

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