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Related Experiment Videos

Clonal sublines that are morphologically and functionally distinct from parental OK cells.

J A Cole1, L R Forte, W J Krause

  • 1Department of Pharmacology, School of Medicine, University of Missouri, Columbia.

The American Journal of Physiology
|April 1, 1989
PubMed
Summary
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Opossum kidney cells were cloned to study parathyroid hormone (PTH) effects on phosphate transport. Clonal lines revealed specific responses to PTH, forskolin, and prostaglandin E1, aiding research into kidney cell signaling.

Area of Science:

  • Cell Biology
  • Renal Physiology
  • Molecular Endocrinology

Background:

  • The opossum kidney (OK) cell line, a common model for renal studies, exhibits heterogeneous apical microvilli distribution.
  • Heterogeneity suggests potential variations in cellular function and signaling pathways within the parental OK cell line.

Purpose of the Study:

  • To isolate and characterize clonal subpopulations of OK cells to investigate the regulation of sodium-dependent phosphate transport.
  • To determine the specific roles of parathyroid hormone (PTH), forskolin (FSK), and prostaglandin E1 (PGE1) in modulating phosphate transport in distinct OK cell clones.
  • To explore the potential existence of PTH receptor subclasses coupled to different effector systems (cAMP and/or calcium) in kidney cells.

Main Methods:

  • Derivation of three clonal subpopulations (OK/B, OK/P, OK/H) from a parental OK cell line.

Related Experiment Videos

  • Assessment of apical microvilli distribution to confirm homogeneity in clonal sublines.
  • Measurement of adenosine 3',5'-cyclic monophosphate (cAMP) formation in response to PTH, FSK, and PGE1.
  • Quantification of sodium-dependent phosphate transport inhibition by PTH, FSK, PGE1, and phorbol 12-myristate 13-acetate in parental and clonal cells.

    • Main Results:

    • All clonal sublines exhibited uniform microvilli distribution, indicating homogeneity.
    • PTH, FSK, and PGE1 increased cAMP formation in all cell types.
    • PTH inhibited phosphate transport in parental, OK/B, and OK/P cells, but not in OK/H cells.
    • FSK inhibited phosphate transport in parental, OK/B, and OK/P cells, with less effect on OK/H cells.
    • PGE1 inhibited phosphate transport in OK/B and OK/P cells but was ineffective in parental and OK/H cells.
    • Phorbol 12-myristate 13-acetate, an inhibitor in parental cells, had minimal effect on clonal sublines.

    Conclusions:

    • Clonal OK cell lines are phenotypically stable and useful for studying kidney phosphate transport regulation.
    • Different clonal subpopulations display distinct sensitivities to PTH, FSK, and PGE1, suggesting cell-specific signaling pathways.
    • The differential response to these agents supports the hypothesis of PTH receptor subclasses mediating distinct cellular responses in kidney cells.