AKAP3 selectively binds PDE4A isoforms in bovine spermatozoa

Malini Bajpai, Sarah E. Fiedler, Zaohua Huang, Srinivasan Vijayaraghavan, Gary E. Olson, Gabriel Livera, Marco Conti, Daniel Carr

Research output: Contribution to journalArticle

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Abstract

Cyclic AMP plays an important role in regulating sperm motility and acrosome reaction through activation of cAMP-dependent protein kinase A (PKA). Phosphodiesterases (PDEs) modulate the levels of cyclic nucleotides by catalyzing their degradation. Although PDE inhibitors specific to PDE1 and PDE4 are known to alter sperm motility and capacitation in humans, little is known about the role or subcellular distribution of PDEs in spermatozoa. The localization of PKA is regulated by A-kinase anchoring proteins (AKAPs), which may also control the intracellular distribution of PDE. The present study was undertaken to investigate the role and localization of PDE4 during sperm capacitation. Addition of Rolipram or RS25344, PDE4-specific inhibitors significantly increased the progressive motility of bovine spermatozoa. Immunolocalization techniques detected both PDE4A and AKAP3 (formerly known as AKAP110) in the principal piece of bovine spermatozoa. The PDE4A5 isoform was detected primarily in the Triton X-100-soluble fraction of caudal epididymal spermatozoa. However, in ejaculated spermatozoa it was seen primarily in the SDS-soluble fraction, indicating a shift in PDE4A5 localization into insoluble organelles during sperm capacitation. AKAP3 was detected only in the SDS-soluble fraction of both caudal and ejaculated sperm. Immunoprecipitation experiments using COS cells cotransfected with AKAP3 and either Pde4a5 or Pde4d provide evidence that PDE4A5 but not PDE4D interacts with AKAP3. Pulldown assays using sperm cell lysates confirm this interaction in vitro. These data suggest that AKAP3 binds both PKA and PDE4A and functions as a scaffolding protein in spermatozoa to regulate local cAMP concentrations and modulate sperm functions.

Original languageEnglish (US)
Pages (from-to)109-118
Number of pages10
JournalBiology of Reproduction
Volume74
Issue number1
DOIs
StatePublished - Jan 2006

Fingerprint

Spermatozoa
Protein Isoforms
Sperm Capacitation
Cyclic AMP-Dependent Protein Kinases
Phosphoric Diester Hydrolases
Sperm Motility
Phosphodiesterase 4 Inhibitors
Rolipram
Acrosome Reaction
Phosphodiesterase Inhibitors
COS Cells
Cyclic Nucleotides
Octoxynol
Immunoprecipitation
Organelles
Cyclic AMP
Protein Kinases
Proteins

Keywords

  • Phosphodiesterases
  • Signal transduction
  • Sperm
  • Sperm motility
  • Transport

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Embryology

Cite this

Bajpai, M., Fiedler, S. E., Huang, Z., Vijayaraghavan, S., Olson, G. E., Livera, G., ... Carr, D. (2006). AKAP3 selectively binds PDE4A isoforms in bovine spermatozoa. Biology of Reproduction, 74(1), 109-118. https://doi.org/10.1095/biolreprod.105.043588

AKAP3 selectively binds PDE4A isoforms in bovine spermatozoa. / Bajpai, Malini; Fiedler, Sarah E.; Huang, Zaohua; Vijayaraghavan, Srinivasan; Olson, Gary E.; Livera, Gabriel; Conti, Marco; Carr, Daniel.

In: Biology of Reproduction, Vol. 74, No. 1, 01.2006, p. 109-118.

Research output: Contribution to journalArticle

Bajpai, M, Fiedler, SE, Huang, Z, Vijayaraghavan, S, Olson, GE, Livera, G, Conti, M & Carr, D 2006, 'AKAP3 selectively binds PDE4A isoforms in bovine spermatozoa', Biology of Reproduction, vol. 74, no. 1, pp. 109-118. https://doi.org/10.1095/biolreprod.105.043588
Bajpai M, Fiedler SE, Huang Z, Vijayaraghavan S, Olson GE, Livera G et al. AKAP3 selectively binds PDE4A isoforms in bovine spermatozoa. Biology of Reproduction. 2006 Jan;74(1):109-118. https://doi.org/10.1095/biolreprod.105.043588
Bajpai, Malini ; Fiedler, Sarah E. ; Huang, Zaohua ; Vijayaraghavan, Srinivasan ; Olson, Gary E. ; Livera, Gabriel ; Conti, Marco ; Carr, Daniel. / AKAP3 selectively binds PDE4A isoforms in bovine spermatozoa. In: Biology of Reproduction. 2006 ; Vol. 74, No. 1. pp. 109-118.
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