Analysis of signaling pathways controlling flagellar movements in mammalian spermatozoa

The mammalian sperm flagellum is an example of organelles where sensory and signaling functions are integrated with motility. Structurally related to other ciliary appendages, it has unique cytoskeletal structures that serve to assemble signaling complexes as well as components of metabolic pathways. Flagellar motility is regulated by signaling events that control sperm ion milieu, energy production, and classical second messenger-dependent phosphorylation cascades. Here, we will concentrate on the cAMP signaling pathway associated with flagellar motility. We will describe methods to analyze the properties of a unique adenylyl cyclase termed sAC (gene name Sacy, Adcy10), which plays an essential function in cAMP accumulation in spermatozoa. This soluble adenylyl cyclase (sAC) is a sensor for bicarbonate, pH, and Ca(2+) and is likely involved in coupling energy homeostasis with signaling events. We will also describe methods to investigate the macromolecular complexes that bring together sAC and other signaling molecules.