A novel biosensor to study cAMP dynamics in cilia and flagella

Autor: Shatanik Mukherjee, Vera Jansen, Jan F Jikeli, Hussein Hamzeh, Luis Alvarez, Marco Dombrowski, Melanie Balbach, Timo Strünker, Reinhard Seifert, U Benjamin Kaupp, Dagmar Wachten
Jazyk: angličtina
Rok vydání: 2016
Předmět:
Zdroj: eLife, Vol 5 (2016)
Druh dokumentu: article
ISSN: 2050-084X
DOI: 10.7554/eLife.14052
Popis: 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.
Databáze: Directory of Open Access Journals