A novel biosensor to study cAMP dynamics in cilia and flagella

作者：Mukherjee Shatanik; Jansen Vera; Jikeli Jan F; Hamzeh Hussein; Alvarez Luis; Dombrowski Marco; Balbach Melanie; Struenker Timo; Seifert Reinhard; Kaupp U Benjamin; Wachten Dagmar

来源：eLife, 2016, 5: e14052.

DOI：10.7554/eLife.14052

<jats:p>The cellular messenger cAMP regulates multiple cellular functions, including signaling in cilia and flagella. The cAMP dynamics in these subcellular compartments are ill-defined. We introduce a novel FRET-based cAMP biosensor with nanomolar sensitivity that is out of reach for other sensors. To measure cAMP dynamics in the sperm flagellum, we generated transgenic mice and reveal that the hitherto methods determining total cAMP levels do not reflect changes in free cAMP levels. Moreover, cAMP dynamics in the midpiece and principal piece of the flagellum are distinctively different. The sole cAMP source in the flagellum is the soluble adenylate cyclase (SACY). Although bicarbonate-dependent SACY activity requires Ca2+, basal SACY activity is suppressed by Ca2+. Finally, we also applied the sensor to primary cilia. Our new cAMP biosensor features unique characteristics that allow gaining new insights into cAMP signaling and unravel the molecular mechanisms underlying ciliary function in vitro and in vivo.</jats:p>

• 出版日期2016-3-22