a method for measuring electrical signals in a primary cilium方法测量电信号的初级纤毛.pdfVIP

Kleene and Kleene Cilia 2012, 1:17 /content/1/1/17 METHODOLOGY Open Access A method for measuring electrical signals in a primary cilium * Nancy K Kleene and Steven J Kleene Abstract Background: Most cells in the body possess a single primary cilium. These cilia are key transducers of sensory stimuli, and defects in cilia have been linked to several diseases. Evidence suggests that some transduction of sensory stimuli by the primary cilium depends on ion-conducting channels. However, the tiny size of the cilium has been a critical barrier to understanding its electrical properties. We report a novel method that allows sensitive, repeatable electrical recordings from primary cilia. Adherent cells were grown on small, spherical beads that could be easily moved within the recording chamber. In this configuration, an entire cilium could be pulled into a recording microelectrode. Results: In 47% of attempts, suction resulted in a seal with high input resistance. Single channels could be recorded while the cilium remained attached to the cell. When the pipette was raised into the air, the cell body was pulled off at the air-bath interface. The pipette retained the cilium and could then be immersed in various solutions that bathed the cytoplasmic face of the membrane. In excised cilia, ionic currents through ciliary channels were modulated by cytoplasmic Ca2+ and transmembrane voltage. Conclusions: Ciliary recording is a direct way to learn the effects of second messengers and voltage changes on ciliary transduction channels. Background signals in primary cilia are virtually non-existent. In- Most cells in the body possess a single primary cilium stead, hypotheses of