Soluble adenylyl cyclase is required for netrin-1 signaling in nerve growth cones.
Growth cones at the tips of nascent and regenerating axons direct axon elongation. Netrin-1, a secreted molecule that promotes axon outgrowth and regulates axon pathfinding, elevates cyclic AMP (cAMP) levels in growth cones and regulates growth cone morphology and axonal outgrowth. These morphological effects depend on the intracellular levels of cAMP. However, the specific pathways that regulate cAMP levels in response to netrin-1 signaling are unclear. Here we show that 'soluble' adenylyl cyclase (sAC), an atypical calcium-regulated cAMP-generating enzyme previously implicated in sperm maturation, is expressed in developing rat axons and generates cAMP in response to netrin-1. Overexpression of sAC results in axonal outgrowth and growth cone elaboration, whereas inhibition of sAC blocks netrin-1-induced axon outgrowth and growth cone elaboration. Taken together, these results indicate that netrin-1 signals through sAC-generated cAMP, and identify a fundamental role for sAC in axonal development.
Wu
Karen Y
KY
Department of Pharmacology, Weill Medical College, Cornell University, New York, New York 10021, USA.
Zippin
Jonathan H
JH
Huron
David R
DR
Kamenetsky
Margarita
M
Hengst
Ulrich
U
Buck
Jochen
J
Levin
Lonny R
LR
Jaffrey
Samie R
SR
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D000242:Cyclic AMP / Q000378:metabolism; D004347:Drug Interactions; D004622:Embryo, Mammalian; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005455:Fluorescent Antibody Technique / Q000379:methods; D005727:Ganglia, Spinal / Q000166:cytology; D020439:Growth Cones / Q000187:drug effects* / Q000502:physiology*; D016086:Lentivirus / Q000302:isolation & purification; D009414:Nerve Growth Factors / Q000494:pharmacology*; D000075388:Netrin-1; D009475:Neurons, Afferent / Q000166:cytology*; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D046509:Tissue Culture Techniques; D014162:Transfection / Q000379:methods; D025521:Tumor Suppressor Proteins / Q000494:pharmacology*
D004791:Enzyme Inhibitors; D009414:Nerve Growth Factors; C000617354:Ntn1 protein, rat; D034741:RNA, Small Interfering; D025521:Tumor Suppressor Proteins; D000075388:Netrin-1; D000242:Cyclic AMP; D000262:Adenylyl Cyclases