Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P41181 (collecting duct)
 5,183 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Isolated human eccrine sweat glands have been microdissected into their secretory and reabsorptive components. Complete separation of these epithelia was confirmed by differential uptake of Neutral Red stain by an intermediate section of gland containing the junction between the secretory coil and the collecting duct. Primary cultures were obtained from explants of both tissues in medium RPMI-1640 or Williams E supplemented with foetal calf serum, insulin, transferrin, epidermal growth factor and hydrocortisone. The cells in the initial coil cultures had an elongated morphology while those of ductal origin were polyhedral. After 10 days both cultures were composed of polyhedral cells of varying diameter. All these morphological types were of epithelial lineage, as demonstrated by the binding of a monoclonal antibody to cytokeratin, the intermediate filament specific for epithelial cells. Outgrowth from both secretory and reabsorptive epithelia were multilayered, with plentiful desmosomal connections and an underlying basal lamina. Ultrastructural features typical of the epithelial cell types present in intact eccrine sweat glands were absent in a high proportion of the proliferating cells but domes, indicative of transepithelial active ion transport, were present in dense cultures from the reabsorptive duct. Outgrowth was also obtained from the secretory and reabsorptive epithelia of sweat glands from two cystic fibrotic patients. Since the most characteristic malfunction of cystic fibrosis is the impaired ion transport in the eccrine sweat gland, the availability of cultured epithelia should provide a useful model for study of the disease.
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PMID:The primary culture of epithelia from the secretory coil and collecting duct of normal human and cystic fibrotic eccrine sweat glands. 380 34

A new method of isolating human eccrine sweat glands by the repeated dissection of skin biopsies with scissors is described. The success of the technique is attributed to a potential line of weakness between the investing capsule and the surrounding connective tissue, which parts under shear forces. The yield is 20-50 glands per biopsy (5 cm X 0.5 cm). The glands are judged to be viable by: (i) light and electron microscopy; (ii) ATP, ADP and AMP contents of 81.0 +/- 12.7, 13.8 +/- 3.3 and 3.8 +/- 1.0 pmol/gland, respectively (mean +/- S.E.M.), which gave an energy charge of 0.90; (iii) the 28-fold rise in cyclic GMP content and the sevenfold rise in cyclic AMP content effected by treatment for 2 min with 10(-5) M-acetylcholine and for 10 min with 10(-5) M-isoprenaline, respectively; (iv) the rate of [3H]leucine uptake into protein; and (v) the concentration of Neutral Red by the collecting duct. Glands were maintained for 7 days on polycarbonate filters floating on RPMI 1640 tissue-culture medium. After this time the ATP, ADP and AMP contents were 63.2 +/- 7.3, 8.5 +/- 2.2 and 3.5 +/- 0.8 pmol/gland, respectively (mean +/- S.E.M.), which gave an energy charge of 0.90. During maintenance a dilatation of the intercellular spaces developed in both secretory coil and collecting duct. Following maintenance there was a significant rise in the rate of [3H]leucine uptake into protein. Maintained glands demonstrated a fivefold greater accumulation of cyclic AMP in response to isoprenaline than did freshly isolated glands, but there was no comparable maintenance hypersensitivity of cyclic GMP to acetylcholine. This pattern of adrenergic, but not cholinergic, maintenance hypersensitivity matches the known lack of denervation hypersensitivity of human eccrine sweat glands to acetylcholine in vivo.
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PMID:Biochemical and ultrastructural studies of human eccrine sweat glands isolated by shearing and maintained for seven days. 609 35

Cortical collecting duct fragments were manually dissected from 6-wk-old Sprague-Dawley rats. The fragments were enzymatically digested (collagenase A) into single cells, washed, and resuspended in serum-free RPMI 1640. Individual cells were examined electrophysiologically using the whole cell patch-clamp technique. Two morphologically distinct cell types were present in the cell suspension. Small round cells that had a capacitance of 7 pF and larger oval cells with a capacitance of 29 pF were consistently observed. Whole cell electrophysiological examination revealed that the small round cells had virtually no plasma membrane ionic conductance, whereas both inward and outward currents were observed in the larger oval-type cells. Also, superfusion of 250 pM arginine vasopressin specifically increased the inward conductance of only the larger cells. The effect could be completely inhibited by 2 microM amiloride or 100 mumol of the Rp diastereomer of 8-(4-chlorophenylthio)-adenosine 3',5'-cyclic monophosphate (a specific adenosine 3',5'-cyclic monophosphate inhibitor). These findings are consistent with the hypothesis that the larger cells are principal cells and the smaller cells are intercalated cells and directly demonstrate that an amiloride-sensitive whole cell conductance is readily observable in freshly isolated cortical collecting duct cells. Thus the whole cell configuration of the patch-clamp technique appears to be well suited for assessing cellular mechanisms that regulate the ionic conductances of cortical collecting duct cells.
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PMID:Whole cell sodium conductance of principal cells freshly isolated from rat cortical collecting duct. 757 11