5-HT₆ receptor blockade regulates primary cilia morphology in striatal neurons

The 5-HT₆ receptor has been implicated in a variety of cognitive processes including habitual behaviors, learning, and memory. It is found almost exclusively in the brain, is expressed abundantly in striatum, and localizes to neuronal primary cilia. Primary cilia are antenna-like, sensory organelles found on most neurons that receive both chemical and mechanical signals from other cells and the surrounding environment; however, the effect of 5-HT₆ receptor function on cellular morphology has not been examined. We confirmed that 5-HT₆ receptors were localized to primary cilia in wild-type (WT) but not 5-HT₆ knockout (5-HT₆KO) in both native mouse brain tissue and primary cultured striatal neurons then used primary neurons cultured from WT or 5-HT₆KO mice to study the function of these receptors. Selective 5-HT₆ antagonists reduced cilia length in neurons cultured from wild-type mice in a concentration and time-dependent manner without altering dendrites, but had no effect on cilia length in 5-HT₆KO cultured neurons. Varying the expression levels of heterologously expressed 5-HT₆ receptors affected the fidelity of ciliary localization in both WT and 5-HT₆KO neurons; overexpression lead to increasing amounts of 5-HT₆ localization outside of the cilia but did not alter cilia morphology. Introducing discrete mutations into the third cytoplasmic loop of the 5-HT₆ receptor greatly reduced, but did not entirely eliminate, trafficking of the 5-HT₆ receptor to primary cilia. These data suggest that blocking 5-HT₆ receptor activity reduces the length of primary cilia and that mechanisms that regulate trafficking of 5-HT₆ receptors to cilia are more complex than previously thought.