2-adrenergic receptor (2AR) agonists acutely relieve bonchoconstriction via cAMP-mediated relaxation of airway smooth muscle (ASM). Airway constrictor responsiveness may be significantly heightened, however, following protracted exposure to these agents, presumably reflecting the effects of 2AR-desensitization in ASM accompanying prolonged cAMP signaling. Since cAMP phosphodiesterase (PDE) activity can significantly modulate ASM contractility, we investigated the mechanism regulating PDE expression and its potential role in mediating changes in agonist-induced constrictor and relaxation responsiveness in ASM following heterologous desensitization of its 2AR by prolonged exposure to cAMP-elevating agents. Isolated rabbit ASM tissues and cultured human ASM cells treated for 24 hours with the receptor- or non-receptor-coupled cAMP-stimulating agent, PGE₂ or forskolin, respectively, exhibited constrictor hyperresponsiveness to acetylcholine and impaired 2AR-mediated relaxation and cAMP accumulation. These pro-asthmatic-like changes in ASM function were associated with upregulated PDE4 activity, reflective of increased transcription of the PDE4D5 isoform, and were prevented by pretreating the ASM with a PDE4 inhibitor. Extended studies using gene silencing and pharmacological approaches to inhibit specific intracellular signaling molecules demonstrated that the mechanism underlying PGE₂-induced transcriptional upregulation of PDE4D5 involves PKA-dependent activation of G_i protein signaling via the -subunit, the latter eliciting downstream activation of ERK1/2 and its consequent induction of PDE4D5 transcription. Collectively, these findings identify that 2AR desensitization in ASM following prolonged exposure to cAMP-elevating agents is associated with pro-asthmatic-like changes in ASM responsiveness that are mediated by upregulated PDE4 expression induced by an activated cross-talk between the PKA and ERK1/2 signaling pathways.