Cat tracheal gland cells in primary culture

1992 ◽  
Vol 263 (2) ◽  
pp. L264-L275 ◽  
Author(s):  
D. J. Culp ◽  
D. K. Lee ◽  
D. P. Penney ◽  
M. G. Marin
Keyword(s):  
Molecular Weight ◽  
Primary Culture ◽  
High Molecular Weight ◽  
Cultured Cells ◽  
Short Circuit ◽  
Light And Electron Microscopy ◽  
Collagen Gels ◽  
Culture Dish ◽  
Gland Cells ◽  
Secretion Granules

Conditions for the primary culture of isolated cat tracheal gland cells were established. Cells plated onto glutaraldehyde-fixed gels of rat tail collagen grew to confluency after 8 days of culture forming a monolayer of cuboidal cells with ultrastructural characteristics of epithelial cells and immunoreactivity to antikeratins. Cultured cells synthesized and released radiolabeled high-molecular-weight glycoconjugates. Glycoconjugate secretion was increased approximately 10% in response to the cholinergic agonist, carbachol. Secretion of glycoconjugates was unrelated to regulated exocrine secretion, since these cells were devoid of secretion granules as assessed by light and electron microscopy. Confluent cultures also generated a spontaneous potential difference and short-circuit current, which were both inhibited by ouabain and increased by carbachol. This suggested gland cells contribute to fluid secretion by active ion-transport mechanisms. We also plated cells onto unfixed collagen gels that were released from the culture dish at confluency. Cells were columnar with apically oriented secretion granules that stained with alcian blue and for blood group A immunoreactivity. Secretion of radiolabeled high-molecular-weight glycoconjugates was increased 27% by carbachol. These cell culture systems may serve as models to investigate glandular secretory mechanisms and their regulation.

Correlative light and electron microscopy of dissociated immature rat testicular cells undergoing morphogenesis in vitro

Development ◽  
1980 ◽  
Vol 60 (1) ◽  
pp. 283-293
Author(s):  
L. A. Erickson ◽  
J. C. Davis ◽  
P. R. Burton ◽  
J. Snyder
Keyword(s):  
Electron Microscopy ◽  
Primary Culture ◽  
Cell Monolayer ◽  
Light And Electron Microscopy ◽  
Outer Cell ◽  
Culture Dish ◽  
Tubular Aggregates ◽  
Immature Rat ◽  
Testicular Cells

Immature rat testicular cells undergo morphogenesis in primary culture (Davis, 1978). Depending upon the number of dissociated testicular cells added to the culture dish, spherical or tubular aggregates were formed. Spherical aggregates resulted from movement of cells into centers of aggregation and the detachment of these cells from the substratum; on the other hand, tubular aggregates resulted from detachment and retraction of the cell monolayer at certain points along its outer edge. In this investigation, the different methods of formation of aggregates by immature rat testicular cells in primary culture were examined with the scanning electron microscope (SEM). The cell types involved in such morphogenesis and their associations within completely formed structures were examined by transmission electron microscopy (TEM). In addition, the rates of formation of aggregates were established by time-lapse cinemicroscopy. During formation of spherical aggregates, the rate of recruitment of cells into centers of aggregation (0·04± 0·006 μm/min; x±S.E.M., n = 78) was much slower than the rate of cell detachment during formation of tubular aggregates (11·7 ± 1·8 μm/sec; x±S.E.M., n = 110). Although specific roles for each cell type in formation of aggregates have not been determined, the associations of cells within the two types of reformed aggregates appeared to be similar. Myofibroblast cells were located in outer cell layers and Sertoli cells were observed to underlie the layers of myofibroblasts in both types of aggregates. Germinal cells, however, were found on the outer surface of spherical aggregates, but in tubular aggregates they were located on the inner surface. Since spherical and tubular aggregates are formed by different methods, this observation suggests that rearrangement of cells within the aggregates takes place and contributes to the internal morphology of newly-formed aggregates.

A probe for flagellar dynein in the mammalian mitotic apparatus

1981 ◽  
Vol 48 (1) ◽  
pp. 241-257
Author(s):  
G.W. Zieve ◽  
J.R. NcIntosh
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Indirect Immunofluorescence ◽  
Mitotic Spindle ◽  
Cultured Cells ◽  
Structural Studies ◽  
Mitotic Apparatus ◽  
Specific Staining ◽  
Bovine Sperm ◽  
Sperm Proteins

An anti-serum has been prepared in rabbits that precipitates high-molecular-weight bovine sperm proteins, including the dyneins. The activity of the serum against the dyneins is demonstrated by the recognition of dynein polypeptides in stained electrophoretic profiles of sperm proteins and in immunoprecipitates of radiolabelled sperm proteins. In addition, the serum stains the sperm flagella when used in indirect immunofluorescence and quantitatively inhibits the motility of demembranated sperm reactivated with ATP. However, the serum has additional anti-sperm activities besides those directed against the dyneins as demonstrated by the staining of the acrosome in indirect immunofluorescence. When used to immunoprecipitate proteins from extracts of cultured cells, the serum precipitates 2 polypeptides; one has a molecular weight higher than the flagellar dyneins, one lower. No specific staining of cultured cells is observed when an affinity-purified anti-dynein fraction IgG is used to stain a variety of cultured cells including bovine fibroblasts. We interpret these data to suggest that flagellar dynein is not a component of the mammalian mitotic spindle and discuss how this conclusion is consistent with recent genetic and structural studies on the mitotic spindle.

Primary culture of flounder renal tubule cells: transepithelial transport

1986 ◽  
Vol 251 (3) ◽  
pp. F424-F432 ◽  
Author(s):  
K. G. Dickman ◽  
J. L. Renfro
Keyword(s):  
Proximal Tubule ◽  
Primary Culture ◽  
Short Circuit ◽  
Transport Processes ◽  
Transepithelial Transport ◽  
Ciliary Activity ◽  
Suitable Model ◽  
Collagen Gels ◽  
Ussing Chambers ◽  
Tubule Cells

Renal proximal tubule cells from the winter flounder (Pseudopleuronectes americanus) were maintained in a functionally differentiated state for up to 16 days in primary culture on floating collagen gels. The cells were confluent after 7-8 days in culture, contracted the collagen gels, and exhibited ciliary activity. Electron microscopy indicated that the cultures were composed of continuous sheets of columnar epithelial cells that had established structural polarity. When mounted in Ussing chambers, the cultures exhibited a small mucosa-negative potential difference (0.6 +/- 0.10 mV) and a low transepithelial resistance (23 +/- 2.3 omega X cm2). Short-circuit current averaged 24 microA/cm2. The cultured epithelium was four times more permeable to Na than to Cl and actively secreted sulfate and p-aminohippuric acid and reabsorbed hexoses. Glucose reabsorption was rheogenic and occurred via a high-affinity (Km = 0.16 mM), low-capacity (Vmax = 5 microA/cm2), phlorizin-sensitive transport system. We concluded that the cultured cells express many of the differentiated properties of the intact flounder proximal tubule and thus provide a suitable model system for studying renal transport processes.

scholarly journals NUCLEOLAR-DERIVED RIBONUCLEIC ACID IN CHROMOSOMES, NUCLEAR SAP, AND CYTOPLASM OF CHIRONOMUS TENTANS SALIVARY GLAND CELLS

1971 ◽  
Vol 51 (2) ◽  
pp. 355-368 ◽  
Author(s):  
U. Ringborg ◽  
L. Rydlander
Keyword(s):  
Molecular Weight ◽  
Salivary Gland ◽  
High Molecular Weight ◽  
Ribosomal Rna ◽  
Chironomus Tentans ◽  
Gland Cells ◽  
Composite Gels ◽  
Salivary Gland Cells

The distribution of monodisperse high molecular weight RNA (38, 30, 28, 23, and 18S RNA) was studied in the salivary gland cells of Chironomus tentans. RNA labeled in vitro and in vivo with tritiated cytidine and uridine was isolated from microdissected nucleoli, chromosomes, nuclear sap, and cytoplasm and analyzed by electrophoresis on agarose-acrylamide composite gels. As shown earlier, the nucleoli contain labeled 38, 30, and 23S RNA. In the chromosomes, labeled 18S RNA was found in addition to the 30 and 23S RNA previously reported. The nuclear sap contains labeled 30 and 18S RNA, and the cytoplasm labeled 28 and 18S RNA. On the basis of the present and earlier analyses, it was concluded that the chromosomal monodisperse high molecular weight RNA fractions (a) show a genuine chromosomal localization and are not due to unspecific contamination, (b) are not artefacts caused by in vitro conditions, but are present also in vivo, and (c) are very likely related to nucleolar and cytoplasmic (pre)ribosomal RNA. The 30 and 23S RNA components are likely to be precursors to 28 and 18S ribosomal RNA. The order of appearance of the monodisperse high molecular weight RNA fractions in the nucleus is in turn and order: (a) nucleolus, (b) chromosomes, and (c) nuclear sap. Since both 23 and 18S RNA are present in the chromosomes, the conversion to 18S RNA may take place there. On the other hand, 30S RNA is only found in the nucleus while 28S RNA can only be detected in the cytoplasm, suggesting that this conversion takes place in connection with the exit of the molecule from the nucleus.

ATP and UTP increase secretion of bronchial inhibitor by human tracheal gland cells in culture

1993 ◽  
Vol 265 (5) ◽  
pp. L479-L484 ◽  
Author(s):  
M. D. Merten ◽  
J. P. Breittmayer ◽  
C. Figarella ◽  
C. Frelin
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Nucleotide Receptor ◽  
Gland Cells ◽  
Cells In Culture ◽  
Alpha Beta ◽  
Gamma S

The effects of ATP and UTP on intracellular Ca2+ levels and on the secretion of the bronchial inhibitor and high-molecular-weight glycoproteins were studied in cultures of human bronchotracheal gland cells. ATP, adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S), and UTP increased intracellular Ca2+ levels in a manner that was partially dependent on the presence of extracellular Ca2+. Other nucleotides (ADP, alpha,beta-methylene ATP, beta,gamma-methylene ATP, and 2-methylthio ATP) and adenosine were ineffective, thus suggesting the presence of a “nucleotide” receptor specific for ATP and UTP. At concentrations similar to those that raised intracellular Ca2+ concentration, ATP, UTP, and ATP gamma S stimulate the secretion of the bronchial inhibitor. ATP and UTP also increase the production of sulfated high-molecular-weight glycoproteins. These results indicate the presence in human tracheal gland cells of a nucleotide receptor that mediates intracellular Ca2+ mobilization and controls the secretion of macromolecules.

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