International Journal of Physical Medicine & Rehabilitation
Open Access

Abstract

Inositol 1,4,5-Trisphosphate Receptor and Calcium Calmodulin-Dependent Protein Kinase are Involved in Endothelin Receptor Expression in Rat Cerebral Artery

Lars Edvinsson, Hilda Ahnstedt, Sajedeh Eftekhari and Roya Waldsee

Background: The present study was designed to examine the influence of inositol 1,4,5-trisphosphate receptor (IP3R) and calcium calmodulin-dependent protein kinase (CaMK) on endothelin receptor regulation during organ culture in rat middle cerebral arteries (MCA).

Methods: MCA segments were incubated with or without xestospongin C (XeC), an IP3R inhibitor, or KN93, a CaMKII blocker. The mRNA levels of the ETA and ETB receptors, nuclear factor of activated T cells activating protein (NFam1), CaMKII, IP3R and Downstream regulatory element antagonist modulator (DREAM) protein were determined by real-time PCR or immunohistochemistry. Contractile responses to endothelin-1 (ET-1) and sarafotoxin 6c (S6c) were studied by a sensitive myograph and the intracellular calcium levels [Ca2+]i were evaluated by the FURA-2AM.

Results: The contractile responses to ET-1 and S6c, the ETB receptors mRNA level and the baseline level of intracellular calcium [Ca2+]i were all increased after 24 h organ culture. Incubation with XeC or KN93 attenuated ETB receptor expression and increase in [Ca2+]i induced by ET-1 or S6c. XeC decreased the NFam1 mRNA levels while KN93 decreased DREAM protein levels.

Conclusions: The study suggests that CaMKII and calcium release via IP3R are involved in the mechanisms regulate ETB receptor putatively via DREAM and NFam1, respectively.