While many neurons are known to contain multiple neurotransmitters, the specific roles played by each co-transmitter within a neuron are often poorly understood. Here, we investigated the roles of the co-transmitters of the pyloric suppressor (PS) neurons, which are located in the stomatogastric nervous system (STNS) of the lobster Homarus americanus. The PS neurons are known to contain histamine; using RT-PCR, we identified a second co-transmitter as the FMRFamide-like peptide crustacean myosuppressin (Crust-MS). The modulatory effects of Crust-MS application on the gastric mill and pyloric patterns, generated in the stomatogastric ganglion (STG), closely resembled those recorded following extracellular PS neuron stimulation. To determine whether histamine plays a role in mediating the effects of the PS neurons in the STG, we bath-applied histamine receptor antagonists to the ganglion. In the presence of the antagonists, the histamine response was blocked, but Crust-MS application and PS stimulation continued to modulate the gastric and pyloric patterns, suggesting that PS effects in the STG are mediated largely by Crust-MS. PS neuron stimulation also excited the oesophageal rhythm, produced in the commissural ganglia (CoGs) of the STNS. Application of histamine, but not Crust-MS, to the CoGs mimicked this effect. Histamine receptor antagonists blocked the ability of both histamine and PS stimulation to excite the oesophageal rhythm, providing strong evidence that the PS neurons use histamine in the CoGs to exert their effects. Overall, our data suggest that the PS neurons differentially utilize their co-transmitters in spatially distinct locations to coordinate the activity of three independent networks.
M.A.K., E.R.G., K.E.H., M.C.C., A.E.C. and P.S.D. were involved in the conception, design and execution of the study and the interpretation of findings. M.A.K., A.E.C. and P.S.D. drafted and revised the article.
No competing interests declared.
This project was supported by a grant from the Arnold and Mabel Beckman Foundation, by National Science Foundation award 1121973 from the division of Integrative Organismal Systems, by the National Center for Research Resources (award 5P20RR016463-12) and the National Institute of General Medical Sciences (award 8 P20 GM103423-12) from the National Institutes of Health, and by the Cades Foundation of Honolulu, Hawaii. Deposited in PMC for release after 12 months.
LIST OF ABBREVIATIONS
- cardiac ganglion
- commissural ganglion
- central pattern generator
- dorsal lateral ventricular nerve
- gastric mill neuron
- inferior oesophageal nerve
- inferior ventricular nerve
- lateral gastric neuron
- lateral pyloric neuron
- modulatory proctolin neuron
- medial ventricular nerve
- oesophageal dilator 1 neuron
- oesophageal ganglion
- pyloric dilator neuron
- pyloric suppressor neuron
- pyloric neuron
- pyloric nerve
- reverse transcriptase-polymerase chain reaction
- superior oesophageal nerve
- stomatogastric ganglion
- stomatogastric nerve
- stomatogastric nervous system
- ventral dilator neuron
- ventral lateral ventricular nerve
- ventral pyloric dilator nerve
- ventral posterior oesophageal nerve
- © 2013. Published by The Company of Biologists Ltd