Cell Biology and Toxicology

, Volume 10, Issue 1, pp 35–43 | Cite as

Modulation of adrenal cell functions by cadmium salts: 3. Sites affected by CdCl2 during stimulated steroid synthesis

  • O. P. Mgbonyebi
  • C. T. Smothers
  • J. J. Mrotek
Article

Abstract

In previous studies cadmium chloride (CdCl2) nonlethally inhibited Y-1 mouse adrenal tumor cell 20-dihydroxyprogesterone (20DHP) secretion, affecting unstimulated and stimulated steroidogenic pathway sites differently. In addition, dibutyryl cAMP-stimulated 20DHP secretion was unaffected by CdCl2, while the site of the unstimulated effect was indirectly shown to involve steps between endogenous cholesterol utilization and 20-hydroxycholesterol association with mitochondrial cytochrome P450 side-chain cleavage enzyme. In the present study we determined CdCl2 effects on plasma membrane sites preceding pre-dbcAMP-stimulation of 20DHP secretion. Y-1 cells were incubated 0.5 h in medium with or without cadmium (using the concentration that inhibited adrenocorticotropin- (ACTH)-stimulated steroid secretion by 50%) together with exogenously added maximally stimulating concentrations of ACTH, cholera toxin, forskolin, or adenosine triphosphate Cholera toxin, forskolin and ATP bypass specific plasma membrane sites involved in the synthesis of intracellular cAMP and activate the steroid hormone biosynthetic pathway. Cadmium effects on ACTH-stimulated endogenous cAMP secretion were also examined. CdCl2 significantly reduced Y-1 cell 20DHP secretion following exposure to ACTH, cholera toxin, forskolin, and ATP; it also significantly decreased endogenous cAMP secretion into culture medium. These data may be interpreted to suggest that CdCl2 altered Y-1 cell regulation of adenyl cyclase activity, which reduced cAMP-activated cholesterol uptake by mitochondria as a consequence.

Keywords

cadmium chloride stimulated corticosteroid secretion Y-1 adrenal cells 

Abbreviations

ACTH

adrenocorticotropin

ATP

adenosine triphosphate

ANOVA

analysis of variance

CdCl2 or Cd2+

cadmium chloride

cAMP

cyclic 3′,5′-adenosine monophosphate

CTX

cholera toxin

dbcAMP

dibutyryl cAMP,N′,O′-dibutyryl-3′,5′-adenosine monophosphate

EGTA

ethylene glycol bis tetraacetic acid

FMEM

serum-free Eagle's Minimum Essential Medium with all other supplements

FSK

forskolin

Hepes

N-2-hydroxyethylpiperazine-N-1,2-ethanesulfonic acid

IC50'

concentration inhibiting stimulated steroid secretion by 50%

IU

international unit

MEM

Eagle's Minimum Essential Medium

P450scc

cytochrome P450 side-chain cleavage enzyme

PREG

pregnenolone

PROG

progesterone

SEM

standard error of the mean

SMEM

serum-containing Eagle's Minimum Essential Medium with supplements

20DHP

20-α-hydroxy-4-pregnen-3-one

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Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • O. P. Mgbonyebi
    • 1
  • C. T. Smothers
    • 1
  • J. J. Mrotek
    • 1
  1. 1.Physiology DepartmentMeharry Medical CollegeNashvilleUSA

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