cAMP-Specific Phosphodiesterases 8A and 8B, Essential Regulators of Leydig Cell Steroidogenesis

Shimizu‐Albergine, Masami; Tsai, Li Chun Lisa; Patrucco, Enrico; Beavo, Joseph A.

doi:10.1124/mol.111.076125

Cited by 65 publications

(87 citation statements)

References 38 publications

(58 reference statements)

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“…Both PDE7 and PDE8 are most active at low cAMP concentrations (K m !100 nM) and PDE8 has been suggested to regulate the resting levels of cAMP (Bender & Beavo 2006, Tsai et al 2011, Shimizu-Albergine et al 2012 and it can speculated that PDE7 and PDE8 share the same function in the ovary. In contrast, PDE4 has a lower affinity for cAMP (K m Z1-10 mM) meaning that this enzyme is mainly active at higher cAMP concentrations observed during hormonal stimulation of cAMP production as seen for instance with hCG stimulation (Park et al 2003, Bender & Beavo 2006.…”

Section: Phosphodiesterases In the Rat Ovarymentioning

confidence: 99%

Phosphodiesterases in the rat ovary: effect of cAMP in primordial follicles

2015

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Phosphodiesterases (PDEs) are important regulators of the intracellular cAMP concentration, which is a central second messenger that affects a multitude of intracellular functions. In the ovaries, cAMP exerts diverse functions, including regulation of ovulation and it has been suggested that augmented cAMP levels stimulate primordial follicle growth. The present study examined the gene expression, enzyme activity and immunolocalization of the different cAMP hydrolysing PDEs families in the rat ovary. Further, the effect of PDE4 inhibition on primordial follicle activation in cultured neonatal rat ovaries was also evaluated. We found varied expression of all eight families in the ovary with Pde7b and Pde8a having the highest expression each accounting for more than 20% of the total PDE mRNA. PDE4 accounted for 15-26% of the total PDE activity. Immunoreactive PDE11A was found in the oocytes and PDE2A in the corpora lutea. Incubating neonatal rat ovaries with PDE4 inhibitors did not increase primordial follicle activation or change the expression of the developing follicle markers Gdf9, Amh, Inha, the proliferation marker Mki67 or the primordial follicle marker Tmeff2. In addition, the cAMP analogue 8-bromo-cAMP did not increase AKT1 or FOXO3A phosphorylation associated with follicle activation or increase the expression of Kitlg known to be associated with follicle differentiation but did increase the Tmeff2, Mki67 and Inha expression in a dose-dependent manner. In conclusion, this study shows that both Pde7b and Pde8a are highly expressed in the rodent ovary and that PDE4 inhibition does not cause an increase in primordial follicle activation. Reproduction (2015) 150 11-20

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Section: Phosphodiesterases In the Rat Ovarymentioning

confidence: 99%

Phosphodiesterases in the rat ovary: effect of cAMP in primordial follicles

2015

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“…Among the different PDE enzymes, inhibition of PDE4 and PDE8 is known to stimulate steroidogenesis (3)(4)(5)(6). Within these gene families, PDE8A and PDE8B both regulate steroidogenesis (5), whereas the role for specific PDE4 family gene products is less well understood. PDE4A, PDE4B, and PDE4C, but not PDE4D, have been shown to be expressed in Leydig cells (5,7,8), but the role for specific PDE4s in regulating steroidogenesis is unknown.…”

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confidence: 99%

“…Within these gene families, PDE8A and PDE8B both regulate steroidogenesis (5), whereas the role for specific PDE4 family gene products is less well understood. PDE4A, PDE4B, and PDE4C, but not PDE4D, have been shown to be expressed in Leydig cells (5,7,8), but the role for specific PDE4s in regulating steroidogenesis is unknown. We have recently demonstrated that simultaneous inhibition of PDE4 and PDE8 in MA10 Leydig cells results in an increase of a large number of phosphorylated PKA consensus sites in multiple proteins (8) concurrent with a very large increase in steroid production (5).…”

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confidence: 99%

“…A large number of cyclic nucleotide phosphodiesterases (PDEs) serve to limit the intracellular temporal and spatial effects of cAMP by hydrolyzing cAMP to 5′-AMP (1,2). Among the different PDE enzymes, inhibition of PDE4 and PDE8 is known to stimulate steroidogenesis (3)(4)(5)(6). Within these gene families, PDE8A and PDE8B both regulate steroidogenesis (5), whereas the role for specific PDE4 family gene products is less well understood.…”

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confidence: 99%

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SCAP/SREBP pathway is required for the full steroidogenic response to cyclic AMP

Shimizu‐Albergine

Yserloo

Golkowski

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Luteinizing hormone (LH) stimulates steroidogenesis largely through a surge in cyclic AMP (cAMP). Steroidogenic rates are also critically dependent on the availability of cholesterol at mitochondrial sites of synthesis. This cholesterol is provided by cellular uptake of lipoproteins, mobilization of intracellular lipid, and de novo synthesis. Whether and how these pathways are coordinated by cAMP are poorly understood. Recent phosphoproteomic analyses of cAMP-dependent phosphorylation sites in MA10 Leydig cells suggested that cAMP regulates multiple steps in these processes, including activation of the SCAP/SREBP pathway. SCAP [sterol-regulatory element-binding protein (SREBP) cleavage-activating protein] acts as a cholesterol sensor responsible for regulating intracellular cholesterol balance. Its role in cAMP-mediated control of steroidogenesis has not been explored. We used two CRISPR (clustered regularly interspaced short palindromic repeat)-Cas9 (CRISPR associated protein 9) knockout approaches to test the role of SCAP in steroidogenesis. Our results demonstrate that SCAP is required for progesterone production induced by concurrent inhibition of the cAMP phosphodiesterases PDE4 and PDE8. These inhibitors increased SCAP phosphorylation, SREBP2 activation, and subsequent expression of cholesterol biosynthetic genes, whereas SCAP deficiency largely prevented these effects. Reexpression of SCAP in SCAP-deficient cells restored SREBP2 protein expression and partially restored steroidogenic responses, confirming the requirement of SCAP-SREBP2 in steroidogenesis. Inhibitors of 3-hydroxy-3-methylglutaryl-Coenzyme A reductase and isoprenylation attenuated, whereas exogenously provided cholesterol augmented, PDE inhibitor-induced steroidogenesis, suggesting that the cholesterol substrate needed for steroidogenesis is provided by both de novo synthesis and isoprenylation-dependent mechanisms.Overall, these results demonstrate a novel role for LH/cAMP in SCAP/ SREBP activation and subsequent regulation of steroidogenesis.

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“…For example, both PDE3 and PDE4 inhibitors will need to be tested in all combinations with PDE1/7/8 inhibitors. It is known, for example, that a combination of PDE4 and PDE8 inhibitors is particularly effective in MA-10 cells (35,36), so it seems quite likely that additional PDE-regulated functional compartments will be identified as different combinations of PDE inhibitors are investigated.…”

Section: Discussionmentioning

confidence: 99%

Analyses of PDE-regulated phosphoproteomes reveal unique and specific cAMP-signaling modules in T cells

Beltejar

Lau

Golkowski

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Specific functions for different cyclic nucleotide phosphodiesterases (PDEs) have not yet been identified in most cell types. Conventional approaches to study PDE function typically rely on measurements of global cAMP, general increases in cAMP-dependent protein kinase (PKA), or the activity of exchange protein activated by cAMP (EPAC). Although newer approaches using subcellularly targeted FRET reporter sensors have helped define more compartmentalized regulation of cAMP, PKA, and EPAC, they have limited ability to link this regulation to downstream effector molecules and biological functions. To address this problem, we have begun to use an unbiased mass spectrometry-based approach coupled with treatment using PDE isozyme-selective inhibitors to characterize the phosphoproteomes of the functional pools of cAMP/PKA/EPAC that are regulated by specific cAMP-PDEs (the PDE-regulated phosphoproteomes). In Jurkat cells we find multiple, distinct PDE-regulated phosphoproteomes that can be defined by their responses to different PDE inhibitors. We also find that little phosphorylation occurs unless at least two different PDEs are concurrently inhibited in these cells. Moreover, bioinformatics analyses of these phosphoproteomes provide insight into the unique functional roles, mechanisms of action, and synergistic relationships among the different PDEs that coordinate cAMP-signaling cascades in these cells. The data strongly suggest that the phosphorylation of many different substrates contributes to cAMP-dependent regulation of these cells. The findings further suggest that the approach of using selective, inhibitor-dependent phosphoproteome analysis can provide a generalized methodology for understanding the roles of different PDEs in the regulation of cyclic nucleotide signaling.

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cAMP-Specific Phosphodiesterases 8A and 8B, Essential Regulators of Leydig Cell Steroidogenesis

Cited by 65 publications

References 38 publications

Phosphodiesterases in the rat ovary: effect of cAMP in primordial follicles

Phosphodiesterases in the rat ovary: effect of cAMP in primordial follicles

SCAP/SREBP pathway is required for the full steroidogenic response to cyclic AMP

Analyses of PDE-regulated phosphoproteomes reveal unique and specific cAMP-signaling modules in T cells

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