CB1 cannabinoid receptor-mediated changes of trabecular meshworkcellular properties

[摘要] Purpose: To evaluate the roles of CB1 cannabinoid receptors incellular functions of trabecular meshwork (TM) cells, including cellmigration, adhesion, morphology and cytoskeleton changes.Methods: Noladin ether, a selective CB1 receptor agonist, andSR141716A, a selective CB1 receptor antagonist, were used tocharacterize the cellular functions of cultured porcine TM cells.Fluorescence assisted transmigration invasion and motility assays(FATIMA) were conducted to study TM cell migration using solublefibronectin as a chemoattractant. Wound healing assays were used tofurther study TM cell migration. Standard cell adhesion assays of TMcells were performed on fibronectin-coated plates. In morphologicalstudies, Alexafluor 488-labeled phalloidin staining was used to examineactin filaments, and immunocytochemistry using anti-paxillin antibodieswas used to detect focal adhesions.Results: In cell migration assays, CB1 agonist noladin ether atnanomolar ranges led to a concentration-dependent inhibition ofmigration of TM cells toward soluble fibronectin. CB1 antagonistSR141716A antagonized noladin ether-induced inhibition of migration ofTM cells. In addition, noladin ether caused a delay in wound healing ofconfluent trabecular meshwork monolayers and this effect of noladinether was antagonized by SR141716A. In cell adhesion assays, noladinether treatment led to a moderate, but significant decrease of adhesionof TM cells to fibronectin-coated surface. This effect of noladin etherwas concentration-dependent, and was antagonized by SR141716A. Inmorphological studies, noladin ether treatment caused rounding of TMcells in contrast to well-spread control TM cells. In addition, therewas a reduction and fragmentation of actin stress fibers stained withAlexafluor 488-labeled phalloidin and a decrease of focal adhesionsdetected with an anti-paxillin antibody.Conclusions: Noladin ether modulates the migration, adhesion,morphology, and actin cytoskeleton of TM cells. These effects of noladinether are mediated through TM cell CB1 cannabinoid receptors.

[学科分类] 生物化学/生物物理