Involvement of Ca-dependent PKCs in the adaptive changes of μ opioid pathways to sympathetic denervation in the guinea pig colon

C. Giaroni , E. Zanetti , A. Pascale , R. Oldrini , L. Canciani , D. Giuliani , M. Amadio , A.M. Chiaravalli , S. Lecchini , G.M. Frigo

In the guinea pig colon, chronic sympathetic denervation entails supersensitivity to inhibitory μ-opioid agents modulating cholinergic neurons. The mechanism underlying such adaptive change has not yet been unravelled, although protein kinase C (PKC) may be involved. A previous study indirectly demonstrated that activation of μ-opioid receptors on myenteric neurons facilitates PKC activity. Such coupling may counteract the inhibitory action of μ-opioid agents on acetylcholine overflow, since PKC, per sè, increases this parameter. After chronic sympathetic denervation such restraint abates, representing a possible mechanism for development of supersensitivity to μ-opioid agents. In the present study, this hypothesis was further investigated. After chronic sympathetic denervation, Ca-dependent PKC activity was reduced in colonic myenteric plexus synaptosomes. The μ-opioid agent, DAMGO, increased Ca-dependent PKC activity in synaptosomes obtained from normal, but not from denervated animals. In myenteric synaptosomes obtained from this experimental group, protein levels of Ca-dependent PKC isoforms βI, βII and γ decreased, whereas α levels increased. In whole-mount preparations, the four Ca-dependent PKC isoforms co-localized with μ-opioid receptors on subpopulations of colonic myenteric neurons. The percentage of neurons staining for PKCβII, as well as the number of μ-opioid receptor-positive neurons staining for PKCβII, decreased in denervated preparations. The same parameters related to PKCα, βI or γ remained unchanged. Overall, the present data strengthen the concept that μ-opioid receptors located on myenteric neurons are coupled to Ca-dependent PKCs. After chronic sympathetic denervation, a reduced efficiency of this coupling may predominantly involve PKCβII, although also PKCβI and γ, but not PKCα, may be implicated.

Mots clés: Ca++-dependent PKC; μ opioid receptor; Sympathetic denervation; Changes in sensitivity; Myenteric plexus

C. Giaroni, E. Zanetti, A. Pascale, R. Oldrini, L. Canciani, et al.. Involvement of Ca-dependent PKCs in the adaptive changes of μ opioid pathways to sympathetic denervation in the guinea pig colon. Biochemical Pharmacology, 2009, 78 (9), pp.1233. ⟨10.1016/j.bcp.2009.06.107⟩. ⟨hal-00519083⟩