title: Functional transplant of photoactivated adenylyl cyclase (PAC) into Aplysia sensory neurons.
abstract: In neural mechanisms of animal learning, intracellular cAMP has been known to play an important role. In the present experiments we attempted functional transplant of a photoactivated adenylyl cyclase (PAC) isolated from Euglena into Aplysia neurons, and explored whether PAC can produce cAMP in the neurons by light stimulation. Serotonergic modulation of mechanoafferent sensory neurons in Aplysia pleural ganglia has been reported to increase intracellular cAMP level and promotes synaptic transmission to motor neurons by increasing spike width of sensory neurons. When cAMP was directly injected into the sensory neurons, spike amplitude temporarily decreased while spike width temporarily increased. This effect was not substituted by injection of 5'AMP, and maintained longer in a bath solution containing IBMX, the phosphodiesterase inhibitor. We, therefore, explored these changes as indicators of appearance of the PAC function. PAC or the PAC expression vector (pNEX-PAC) was injected into cell bodies of sensory neurons. Spike amplitude decreased in both cases and spike width increased in the PAC injection when the neurons were stimulated with light, suggesting that the transplanted PAC works well in Aplysia neurons. These results indicate that we can control cAMP production in specific neurons with light by the functional transplant of PAC.
authors.affiliation: Department of Biophysics, Faculty of Pharmaceutical Science, Toho University, 2-2-1 Miyama, Funabashi 274-8510, Japan. nagahama@phar.toho-u.ac.jp
authors.forename: Tatsumi
authors.initials: T
authors.lastname: Nagahama
authors.forename: Takeshi
authors.initials: T
authors.lastname: Suzuki
authors.forename: Shinya
authors.initials: S
authors.lastname: Yoshikawa
authors.forename: Mineo
authors.initials: M
authors.lastname: Iseki
mesh_terms: D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001048:Aplysia; D001522:Behavior, Animal; D000242:Cyclic AMP / Q000494:pharmacology; D004347:Drug Interactions; D005055:Euglena / Q000737:chemistry; D017952:Ganglia, Invertebrate / Q000166:cytology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D008845:Microinjections / Q000379:methods; D009475:Neurons, Afferent / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects*; D018408:Patch-Clamp Techniques / Q000379:methods; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010775:Photic Stimulation; D012701:Serotonin / Q000494:pharmacology; D013997:Time Factors
chemical_list: D010726:Phosphodiesterase Inhibitors; D012701:Serotonin; D000242:Cyclic AMP; D000262:Adenylyl Cyclases; D015056:1-Methyl-3-isobutylxanthine