Novel signaling of dynorphin at κ-opioid receptor/bradykinin B2 receptor heterodimers.

The κ-opioid receptor (KOR) and bradykinin B2 receptor (B2R) are involved in a variety of important physiological processes and share many similar characteristics in terms of their distribution and functions in the nervous system. We first demonstrated the endogenous expression of KOR and B2R in human SH-SY5Y cells and their co-localization on the membrane of human embryonic kidney 293 (HEK293) cells. Bioluminescence and fluorescence resonance energy transfer and the proximity ligation assay were exploited to demonstrate the formation of functional KOR and B2R heteromers in transfected cells. KOR/B2R heteromers triggered dynorphin A (1-13)-induced Gαs/protein kinase A signaling pathway activity, including upregulation of intracellular cAMP levels and cAMP-response element luciferase reporter activity, resulting in increased cAMP-response element-binding protein (CREB) phosphorylation, which could be dampened by the protein kinase A (PKA) inhibitor H89. This indicated that the co-existence of KOR and B2R is critical for CREB phosphorylation. In addition, dynorphin A (1-13) induced a significantly higher rate of proliferation in HEK293-KOR/B2R and human SH-SY5Y cells than in the control group. These results indicate that KOR can form a heterodimer with B2R and this leads to increased protein kinase A activity by the CREB signaling pathway, leading to a significant increase in cell proliferation. The nature of this signaling pathway has significant implications for the role of dynorphin in the regulation of neuroprotective effects.