Cyclic AMP-mediated cytoskeletal effects in adrenal cells are modified by serum, insulin, insulin-like growth factor-I, and an antibody against urokinase plasminogen activator

Peter J Hornsby, Sepehr Steve Maghsoudlou, Vicki Cheng, Charles Y. Cheng

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

In adrenocortical cells in culture, increased intracellular cyclic AMP resulting from exposure to agents such as ACTH and cholera toxin causes a change in cell morphoology termed 'retraction' or 'rounding'. The breakdown of actin-containing stress fibers in rounding suggested a role for microfilaments in steroidogenesis. Previously, we showed that cultured bovine adrenal cells under standard conditions (medium with 10% fetal bovine serum) do not round in response to intracellular cyclic AMP. Here, we show that these cells do round in defined, serum-free medium. Rounding was maximal within 1 h of addition of 1 nM cholera toxin and after 10 h most cells remained rounded. Cycloheximide at 100 μg/ml did not inhibit the response to cholera toxin. The rounding response was abolished when 10% fetal bovine serum, horse serum, or ether-extracted fetal bovine serum was included in the medium. The inhibitory effect of serum was not mimicked by growth factors with the exception that insulin and insulin-like growth factor-I (IGF-I), while not preventing rounding, accelerated the return of cells to a flattened morphology. A monoclonal antibody against urokinase plasminogen activator completely prevented rounding whereas a monoclonal antibody against tissue plasminogen activator had only a slight effect. Fluorescence visualization of F-actin with N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-phallacidin showed that rounding in cultured bovine adrenocortical cells resembles that defined earlier for human and rat adrenocortical cells and includes depolymerization of actin microfilaments. These cytoskeletal changes in adrenal cells are unlikely to play a role in steroidogenesis; however, they may be involved in tissue remodeling occurring as part of the indirect mitogenic effects of ACTH.

Original languageEnglish (US)
Pages (from-to)185-193
Number of pages9
JournalMolecular and Cellular Endocrinology
Volume67
Issue number2-3
DOIs
StatePublished - 1989
Externally publishedYes

Fingerprint

Plasminogen Activators
Cholera Toxin
Urokinase-Type Plasminogen Activator
Insulin-Like Growth Factor I
Cyclic AMP
Actins
Insulin
Adrenocorticotropic Hormone
Antibodies
Serum
Monoclonal Antibodies
Depolymerization
Serum-Free Culture Media
Tissue Plasminogen Activator
Cycloheximide
Ether
Rats
Intercellular Signaling Peptides and Proteins
Visualization
Fluorescence

Keywords

  • (Bovine adrenal cortex)
  • Cyclic AMP
  • Cytoskeleton
  • Insulin
  • Insulin-like growth factor-I
  • Serum
  • Urokinase plasminogen activator

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Cyclic AMP-mediated cytoskeletal effects in adrenal cells are modified by serum, insulin, insulin-like growth factor-I, and an antibody against urokinase plasminogen activator. / Hornsby, Peter J; Maghsoudlou, Sepehr Steve; Cheng, Vicki; Cheng, Charles Y.

In: Molecular and Cellular Endocrinology, Vol. 67, No. 2-3, 1989, p. 185-193.

Research output: Contribution to journalArticle

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abstract = "In adrenocortical cells in culture, increased intracellular cyclic AMP resulting from exposure to agents such as ACTH and cholera toxin causes a change in cell morphoology termed 'retraction' or 'rounding'. The breakdown of actin-containing stress fibers in rounding suggested a role for microfilaments in steroidogenesis. Previously, we showed that cultured bovine adrenal cells under standard conditions (medium with 10{\%} fetal bovine serum) do not round in response to intracellular cyclic AMP. Here, we show that these cells do round in defined, serum-free medium. Rounding was maximal within 1 h of addition of 1 nM cholera toxin and after 10 h most cells remained rounded. Cycloheximide at 100 μg/ml did not inhibit the response to cholera toxin. The rounding response was abolished when 10{\%} fetal bovine serum, horse serum, or ether-extracted fetal bovine serum was included in the medium. The inhibitory effect of serum was not mimicked by growth factors with the exception that insulin and insulin-like growth factor-I (IGF-I), while not preventing rounding, accelerated the return of cells to a flattened morphology. A monoclonal antibody against urokinase plasminogen activator completely prevented rounding whereas a monoclonal antibody against tissue plasminogen activator had only a slight effect. Fluorescence visualization of F-actin with N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-phallacidin showed that rounding in cultured bovine adrenocortical cells resembles that defined earlier for human and rat adrenocortical cells and includes depolymerization of actin microfilaments. These cytoskeletal changes in adrenal cells are unlikely to play a role in steroidogenesis; however, they may be involved in tissue remodeling occurring as part of the indirect mitogenic effects of ACTH.",
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