Go to JCI Insight
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Alerts
  • Advertising/recruitment
  • Subscribe
  • Contact
  • Current Issue
  • Past Issues
  • By specialty
    • COVID-19
    • Cardiology
    • Gastroenterology
    • Immunology
    • Metabolism
    • Nephrology
    • Neuroscience
    • Oncology
    • Pulmonology
    • Vascular biology
    • All ...
  • Videos
    • Conversations with Giants in Medicine
    • Author's Takes
  • Reviews
    • View all reviews ...
    • 100th Anniversary of Insulin's Discovery (Jan 2021)
    • Hypoxia-inducible factors in disease pathophysiology and therapeutics (Oct 2020)
    • Latency in Infectious Disease (Jul 2020)
    • Immunotherapy in Hematological Cancers (Apr 2020)
    • Big Data's Future in Medicine (Feb 2020)
    • Mechanisms Underlying the Metabolic Syndrome (Oct 2019)
    • Reparative Immunology (Jul 2019)
    • View all review series ...
  • Viewpoint
  • Collections
    • Recently published
    • In-Press Preview
    • Commentaries
    • Concise Communication
    • Editorials
    • Viewpoint
    • Top read articles
  • Clinical Medicine
  • JCI This Month
    • Current issue
    • Past issues

  • Current issue
  • Past issues
  • Specialties
  • Reviews
  • Review series
  • Conversations with Giants in Medicine
  • Author's Takes
  • Recently published
  • In-Press Preview
  • Commentaries
  • Concise Communication
  • Editorials
  • Viewpoint
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Alerts
  • Advertising/recruitment
  • Subscribe
  • Contact
Cyclic nucleotide phosphodiesterase 3A–deficient mice as a model of female infertility
Silvia Masciarelli, … , Marco Conti, Vincent Manganiello
Silvia Masciarelli, … , Marco Conti, Vincent Manganiello
Published July 15, 2004
Citation Information: J Clin Invest. 2004;114(2):196-205. https://doi.org/10.1172/JCI21804.
View: Text | PDF
Article Reproductive biology

Cyclic nucleotide phosphodiesterase 3A–deficient mice as a model of female infertility

  • Text
  • PDF
Abstract

Since cAMP blocks meiotic maturation of mammalian and amphibian oocytes in vitro and cyclic nucleotide phosphodiesterase 3A (PDE3A) is primarily responsible for oocyte cAMP hydrolysis, we generated PDE3A-deficient mice by homologous recombination. The Pde3a–/– females were viable and ovulated a normal number of oocytes but were completely infertile, because ovulated oocytes were arrested at the germinal vesicle stage and, therefore, could not be fertilized. Pde3a–/– oocytes lacked cAMP-specific PDE activity, contained increased cAMP levels, and failed to undergo spontaneous maturation in vitro (up to 48 hours). Meiotic maturation in Pde3a–/– oocytes was restored by inhibiting protein kinase A (PKA) with adenosine-3′,5′-cyclic monophosphorothioate, Rp-isomer (Rp-cAMPS) or by injection of protein kinase inhibitor peptide (PKI) or mRNA coding for phosphatase CDC25, which confirms that increased cAMP-PKA signaling is responsible for the meiotic blockade. Pde3a–/– oocytes that underwent germinal vesicle breakdown showed activation of MPF and MAPK, completed the first meiotic division extruding a polar body, and became competent for fertilization by spermatozoa. We believe that these findings provide the first genetic evidence indicating that resumption of meiosis in vivo and in vitro requires PDE3A activity. Pde3a–/– mice represent an in vivo model where meiotic maturation and ovulation are dissociated, which underscores inhibition of oocyte maturation as a potential strategy for contraception.

Authors

Silvia Masciarelli, Kathleen Horner, Chengyu Liu, Sun Hee Park, Mary Hinckley, Steven Hockman, Taku Nedachi, Catherine Jin, Marco Conti, Vincent Manganiello

×

Figure 2

Options: View larger image (or click on image) Download as PowerPoint
Oocyte cAMP content and PDE3 activities in tissues from Pde3a+/+ and Pde...
Oocyte cAMP content and PDE3 activities in tissues from Pde3a+/+ and Pde3a–/– mice. (A) PDE activities in Pde3a+/+ and Pde3a–/– oocytes. PDE activity was assayed in oocytes from 6 Pde3A–/– and 6 Pde3a+/+ mice with [3H]-cAMP as substrate as described (22, 23) and is reported as fmol cAMP hydrolyzed/min/oocyte (mean ± SEM; n = 6). (B) cAMP content in denuded Pde3a–/– and Pde3a+/+ oocytes. As described in Methods, oocytes from 11 Pde3a+/+ and 15 Pde3a–/– mice were assayed for cAMP content, reported as fmol cAMP/oocyte. (C) PDE3 activities in heart, lung, liver, and adipose tissues from Pde3a+/+ and Pde3a–/– mice. Homogenates were prepared and PDE3 activities assayed as described in Methods. Data are means ± SEM of values (pmol cAMP hydrolyzed/min/mg protein), n = 6 mice. (D) Gel filtration chromatography. Solubilized proteins (∼4 mg), prepared from lung tissues from Pde3a+/+ or Pde3a–/– mice in buffer containing 1% NP-40 as described in Methods, were subjected to gel filtration chromatography. Left panel: protein (AU; 280 nm) (open circles, open triangles) and PDE3 activity (PDE3 cpm/10 μl) (filled circles, filled triangles) in indicated fractions from Pde3a+/+ (WT, open circles, filled circles) and Pde3a–/– (KO, open triangles, filled triangles) mice; >90% of the applied PDE3 activity was recovered in indicated fractions from +/+ mice (filled circles). Molecular weight standards: I, thyroglobulin; II, g-globulin; III, ovalbumin; IV myoglobin; V, Vit B12. Right panel: Western blots of material applied (input 10 μl) and indicated fractions (40 μl) from +/+ and –/– mice with rabbit anti-PDE3A (upper panel) and anti-PDE3B (lower panel) IgG. Recombinant rat PDE3A (r3A) was used as positive control in PDE3A Western blots.
Follow JCI:
Copyright © 2021 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

Sign up for email alerts