Regulation of crab Y-organ steroidogenesis in vitro

evidence that ecdysteroid production increases through activation of cAMP-phosphodiesterase by calcium-calmodulin

Mark P. Mattson, Eugene Spaziani

Research output: Contribution to journalArticle

Abstract

In decapod crustaceans steroidogenic glands (Y-organs) produce the molting hormone, ecdysone. A putative neuropeptide, molt-inhibiting hormone (MIH), released from eyestalk neurosecretory cells, directly regulates Y-organs by suppressing steroidogenesis; the effect is mediated by an increase in cAMP. We explored calcium-cAMP interactions in the regulation of Y-organs in vitro of the crab, Cancer antennarius. Basal ecdysteroid production was enhanced by extracellular calcium (EC). MIH suppression did not require EC but its action was blocked by high EC. The inhibitors of Ca2+ flux, lanthanum and ruthenium red, mimicked and enhanced MIH action. Calcium ionophore A23187 raised basal steroidogenesis dose-dependently (10-6 to 10-4 M) and with time course (effect evident after 2 h) similar to that of suppression by MIH. Low EC enhanced the suppressive effects on steroidogenesis of forskolin and dibutyryl cyclic AMP ((Bu)2cAMP) but not of MIH, lysine vasopressin (LVP), or 3-isobutyl-1-methylxanthine (IBMX); basal Y-organ cAMP levels were elevated by low EC and reduced by A23187. A23187 reduced the steroidogenic-suppressive effects of MIH, LVP, forskolin and (Bu)2cAMP but not of IBMX; rises in cAMP induced by MIH, LVP, and forskolin but not by IBMX were blunted by A23187. These findings suggested a stimulatory action of calcium on phosphodiesterase (PDE). The calmodulin (CM) inhibitor trifluoperazine (TFP; 10-5 to 10-4 M) reduced basal and A23187-stimulated steroidogenesis, enhanced the inhibitory effects of MIH and (Bu)21cAMP on ecdysteroid production, enhanced the stimulatory effects of MIH and forskolin on cAMP, and blocked the inhibition of cAMP by A23187. Y-organ PDE activity was enhanced by increasing free Ca2+ (10-7 to 10-5 M) and inhibited by TFP (10-5 to 10-4 M). Adenylate cyclase activity of Y-organ cell particulate fraction was unaffected by Ca2+ or TFP. Calcium stimulates steroidogenesis, apparently by activating a calcium-CM-dependent cAMP-PDE; the action is counter to the cAMP-mediated MIH-inhibitory system. Ca2+ fluxes were measured with dispersed Y-organ cells, in the presence and absence of agents that alter cAMP levels. The ionophore A23187, but not MIH or forskolin, increased 45Ca2+ entry by 45% over untreated control cells. Efflux from 45Ca2+-preloaded cells was increased 30% by MIH and forskolin, but not A23187. These data, together with those further above, suggest that MIH suppresses steroidogenesis in part by fostering Ca2+ depletion, and that the effect is mediated by cAMP.

Original languageEnglish (US)
Pages (from-to)135-151
Number of pages17
JournalMolecular and Cellular Endocrinology
Volume48
Issue number2-3
DOIs
StatePublished - 1986
Externally publishedYes

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Type 1 Cyclic Nucleotide Phosphodiesterases
Ecdysteroids
Phosphoric Diester Hydrolases
Calmodulin
Chemical activation
Calcimycin
Calcium
Colforsin
Lypressin
Ecdysone
molt-inhibiting hormone
In Vitro Techniques
Decapoda (Crustacea)
Fluxes
1-Methyl-3-isobutylxanthine
Ruthenium Red
Trifluoperazine
Lanthanum
Bucladesine
Foster Home Care

Keywords

  • calcium-calmodulin
  • cAMP-phosphodiesterase
  • ecdysteroid production
  • steroidogenesis
  • Y-organ, crab

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

@article{7217f833dc1a497da85eb17cbf73b50d,
title = "Regulation of crab Y-organ steroidogenesis in vitro: evidence that ecdysteroid production increases through activation of cAMP-phosphodiesterase by calcium-calmodulin",
abstract = "In decapod crustaceans steroidogenic glands (Y-organs) produce the molting hormone, ecdysone. A putative neuropeptide, molt-inhibiting hormone (MIH), released from eyestalk neurosecretory cells, directly regulates Y-organs by suppressing steroidogenesis; the effect is mediated by an increase in cAMP. We explored calcium-cAMP interactions in the regulation of Y-organs in vitro of the crab, Cancer antennarius. Basal ecdysteroid production was enhanced by extracellular calcium (EC). MIH suppression did not require EC but its action was blocked by high EC. The inhibitors of Ca2+ flux, lanthanum and ruthenium red, mimicked and enhanced MIH action. Calcium ionophore A23187 raised basal steroidogenesis dose-dependently (10-6 to 10-4 M) and with time course (effect evident after 2 h) similar to that of suppression by MIH. Low EC enhanced the suppressive effects on steroidogenesis of forskolin and dibutyryl cyclic AMP ((Bu)2cAMP) but not of MIH, lysine vasopressin (LVP), or 3-isobutyl-1-methylxanthine (IBMX); basal Y-organ cAMP levels were elevated by low EC and reduced by A23187. A23187 reduced the steroidogenic-suppressive effects of MIH, LVP, forskolin and (Bu)2cAMP but not of IBMX; rises in cAMP induced by MIH, LVP, and forskolin but not by IBMX were blunted by A23187. These findings suggested a stimulatory action of calcium on phosphodiesterase (PDE). The calmodulin (CM) inhibitor trifluoperazine (TFP; 10-5 to 10-4 M) reduced basal and A23187-stimulated steroidogenesis, enhanced the inhibitory effects of MIH and (Bu)21cAMP on ecdysteroid production, enhanced the stimulatory effects of MIH and forskolin on cAMP, and blocked the inhibition of cAMP by A23187. Y-organ PDE activity was enhanced by increasing free Ca2+ (10-7 to 10-5 M) and inhibited by TFP (10-5 to 10-4 M). Adenylate cyclase activity of Y-organ cell particulate fraction was unaffected by Ca2+ or TFP. Calcium stimulates steroidogenesis, apparently by activating a calcium-CM-dependent cAMP-PDE; the action is counter to the cAMP-mediated MIH-inhibitory system. Ca2+ fluxes were measured with dispersed Y-organ cells, in the presence and absence of agents that alter cAMP levels. The ionophore A23187, but not MIH or forskolin, increased 45Ca2+ entry by 45{\%} over untreated control cells. Efflux from 45Ca2+-preloaded cells was increased 30{\%} by MIH and forskolin, but not A23187. These data, together with those further above, suggest that MIH suppresses steroidogenesis in part by fostering Ca2+ depletion, and that the effect is mediated by cAMP.",
keywords = "calcium-calmodulin, cAMP-phosphodiesterase, ecdysteroid production, steroidogenesis, Y-organ, crab",
author = "Mattson, {Mark P.} and Eugene Spaziani",
year = "1986",
doi = "10.1016/0303-7207(86)90036-5",
language = "English (US)",
volume = "48",
pages = "135--151",
journal = "Molecular and Cellular Endocrinology",
issn = "0303-7207",
publisher = "Elsevier Ireland Ltd",
number = "2-3",

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TY - JOUR

T1 - Regulation of crab Y-organ steroidogenesis in vitro

T2 - evidence that ecdysteroid production increases through activation of cAMP-phosphodiesterase by calcium-calmodulin

AU - Mattson, Mark P.

AU - Spaziani, Eugene

PY - 1986

Y1 - 1986

N2 - In decapod crustaceans steroidogenic glands (Y-organs) produce the molting hormone, ecdysone. A putative neuropeptide, molt-inhibiting hormone (MIH), released from eyestalk neurosecretory cells, directly regulates Y-organs by suppressing steroidogenesis; the effect is mediated by an increase in cAMP. We explored calcium-cAMP interactions in the regulation of Y-organs in vitro of the crab, Cancer antennarius. Basal ecdysteroid production was enhanced by extracellular calcium (EC). MIH suppression did not require EC but its action was blocked by high EC. The inhibitors of Ca2+ flux, lanthanum and ruthenium red, mimicked and enhanced MIH action. Calcium ionophore A23187 raised basal steroidogenesis dose-dependently (10-6 to 10-4 M) and with time course (effect evident after 2 h) similar to that of suppression by MIH. Low EC enhanced the suppressive effects on steroidogenesis of forskolin and dibutyryl cyclic AMP ((Bu)2cAMP) but not of MIH, lysine vasopressin (LVP), or 3-isobutyl-1-methylxanthine (IBMX); basal Y-organ cAMP levels were elevated by low EC and reduced by A23187. A23187 reduced the steroidogenic-suppressive effects of MIH, LVP, forskolin and (Bu)2cAMP but not of IBMX; rises in cAMP induced by MIH, LVP, and forskolin but not by IBMX were blunted by A23187. These findings suggested a stimulatory action of calcium on phosphodiesterase (PDE). The calmodulin (CM) inhibitor trifluoperazine (TFP; 10-5 to 10-4 M) reduced basal and A23187-stimulated steroidogenesis, enhanced the inhibitory effects of MIH and (Bu)21cAMP on ecdysteroid production, enhanced the stimulatory effects of MIH and forskolin on cAMP, and blocked the inhibition of cAMP by A23187. Y-organ PDE activity was enhanced by increasing free Ca2+ (10-7 to 10-5 M) and inhibited by TFP (10-5 to 10-4 M). Adenylate cyclase activity of Y-organ cell particulate fraction was unaffected by Ca2+ or TFP. Calcium stimulates steroidogenesis, apparently by activating a calcium-CM-dependent cAMP-PDE; the action is counter to the cAMP-mediated MIH-inhibitory system. Ca2+ fluxes were measured with dispersed Y-organ cells, in the presence and absence of agents that alter cAMP levels. The ionophore A23187, but not MIH or forskolin, increased 45Ca2+ entry by 45% over untreated control cells. Efflux from 45Ca2+-preloaded cells was increased 30% by MIH and forskolin, but not A23187. These data, together with those further above, suggest that MIH suppresses steroidogenesis in part by fostering Ca2+ depletion, and that the effect is mediated by cAMP.

AB - In decapod crustaceans steroidogenic glands (Y-organs) produce the molting hormone, ecdysone. A putative neuropeptide, molt-inhibiting hormone (MIH), released from eyestalk neurosecretory cells, directly regulates Y-organs by suppressing steroidogenesis; the effect is mediated by an increase in cAMP. We explored calcium-cAMP interactions in the regulation of Y-organs in vitro of the crab, Cancer antennarius. Basal ecdysteroid production was enhanced by extracellular calcium (EC). MIH suppression did not require EC but its action was blocked by high EC. The inhibitors of Ca2+ flux, lanthanum and ruthenium red, mimicked and enhanced MIH action. Calcium ionophore A23187 raised basal steroidogenesis dose-dependently (10-6 to 10-4 M) and with time course (effect evident after 2 h) similar to that of suppression by MIH. Low EC enhanced the suppressive effects on steroidogenesis of forskolin and dibutyryl cyclic AMP ((Bu)2cAMP) but not of MIH, lysine vasopressin (LVP), or 3-isobutyl-1-methylxanthine (IBMX); basal Y-organ cAMP levels were elevated by low EC and reduced by A23187. A23187 reduced the steroidogenic-suppressive effects of MIH, LVP, forskolin and (Bu)2cAMP but not of IBMX; rises in cAMP induced by MIH, LVP, and forskolin but not by IBMX were blunted by A23187. These findings suggested a stimulatory action of calcium on phosphodiesterase (PDE). The calmodulin (CM) inhibitor trifluoperazine (TFP; 10-5 to 10-4 M) reduced basal and A23187-stimulated steroidogenesis, enhanced the inhibitory effects of MIH and (Bu)21cAMP on ecdysteroid production, enhanced the stimulatory effects of MIH and forskolin on cAMP, and blocked the inhibition of cAMP by A23187. Y-organ PDE activity was enhanced by increasing free Ca2+ (10-7 to 10-5 M) and inhibited by TFP (10-5 to 10-4 M). Adenylate cyclase activity of Y-organ cell particulate fraction was unaffected by Ca2+ or TFP. Calcium stimulates steroidogenesis, apparently by activating a calcium-CM-dependent cAMP-PDE; the action is counter to the cAMP-mediated MIH-inhibitory system. Ca2+ fluxes were measured with dispersed Y-organ cells, in the presence and absence of agents that alter cAMP levels. The ionophore A23187, but not MIH or forskolin, increased 45Ca2+ entry by 45% over untreated control cells. Efflux from 45Ca2+-preloaded cells was increased 30% by MIH and forskolin, but not A23187. These data, together with those further above, suggest that MIH suppresses steroidogenesis in part by fostering Ca2+ depletion, and that the effect is mediated by cAMP.

KW - calcium-calmodulin

KW - cAMP-phosphodiesterase

KW - ecdysteroid production

KW - steroidogenesis

KW - Y-organ, crab

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U2 - 10.1016/0303-7207(86)90036-5

DO - 10.1016/0303-7207(86)90036-5

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