DNA damage response induced by Etoposide promotes steroidogenesis via GADD45A in cultured adrenal cells.

Glucocorticoid production is regulated by adrenocorticotropic hormone (ACTH) via the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) pathway in the adrenal cortex, but the changes in steroidogenesis associated with aging are unknown. In this study, we show that cell-autonomous steroidogenesis is induced by non-ACTH- mediated genotoxic stress in human adrenocortical H295R cells. Low-dose etoposide (EP) was used to induce DNA damage as a genotoxic stress, leading to cellular senescence. We found that steroidogenesis was promoted in cells stained with γH2AX, a marker of DNA damaged cells. Among stress-associated and p53-inducible genes, the expression of GADD45A and steroidogenesis-related genes was significantly upregulated. Immunofluorescence analysis revealed that GADD45A accumulated in the nuclei. Metabolite assay using cultured media showed that EP-treated cells were induced to produce and secrete considerable amounts of glucocorticoid. Knockdown of GADD45A using small interfering RNA markedly inhibited the EP-induced upregulation of steroidogenesis-related gene expression, and glucocorticoid production. A p38MAPK inhibitor, but not a PKA inhibitor, suppressed EP-stimulated steroidogenesis. These results suggest that DNA damage itself promotes steroidogenesis via one or more unprecedented non-ACTH-mediated pathway. Specifically, GADD45A plays a crucial role in the steroidogenic processes triggered by EP-stimulated genotoxic stress. Our study sheds new light on an alternate mechanism of steroidogenesis in the adrenal cortex.

Seasonal variations of water column nutrients in the inner area of Ariake Bay, Japan: the role of muddy sediments.

To investigate seasonal variations of nutrient distribution in the mudflat-shallow water system, we conducted field surveys once a month from August 2007 to July 2008 in the inner area of Ariake Bay (IAB), Japan. The NH4 (+)-N concentration of the water column increased in autumn because of the high NH4 (+) release from the sediments, ranging from 850 to 3,001 µmol m(-2) day(-1). The NO3 (-)-N concentration was maximal in January, which was thought to be caused by NO3 (-) release from the oxic sediments and by NO3 (-) regeneration due to water column nitrification. The PO4 (3-)-P concentration of the water column was high in summer-autumn due to the high PO4 (3-) release from the reduced sediments, ranging from 22 to 164 µmol m(-2) day(-1). We estimated the total amounts of DIN and PO4 (3-)-P release (R DIN and [Formula: see text], respectively) from the muddy sediment area of the IAB. In summer-autumn, R DIN and [Formula: see text] corresponded to about 47.7 % of DIN input and about 116.6 % of PO4 (3-)-P input from the river, respectively. Thus, we concluded that the muddy sediments were an important source of nutrients for the water column of the IAB during summer-autumn. In addition, we found that phosphorus necessary for the growth of Porphyra (Porphyra yezoensis, Rhodophyceae) would be insufficient in the water column when phosphorus during the Porphyra aquaculture period is supplied only from the river. Therefore, the phosphorus release from the muddy sediments was thought to play an important role in the sustainable production of Porphyra in Ariake Bay.