Jing's CC9&10 paper (2008)

They generated rat antibody for 5-HT.
Briefly, the antigen was prepared by coupling 2 mg serotonin oxalate to 10 mg BSA (Sigma-Aldrich) in 1 ml of 50 mM NaH2PO4, pH 7.2, using 100 microL 16% paraformaldehyde (EMS). Followingovernight incubation at 4°C, coupled antigen was purified from the reaction using a Microcon-30 spinning at 13,800 x g for 30 min at 4°C. After washing the retentate four times with 0.4 ml of 50 mM NaH2PO4, it was resuspended in 0.5 ml of the same buffer and transferred to a new tube...
Tissues were fixed in 4% paraformaldehyde, 0.2% picric acid, 25% sucrose, and 0.1MNaH2PO4, pH 7.6, for either 3 h at room temperature or overnight at 4°C.
Washing buffer (WB; 2% Triton X-100, 1% BSA, 154 mM NaCl, 50 mM EDTA, 0.01% thimerosal, and 10mM Na2HPO4, pH 7.4).

Both the dorsal and the ventral surfaces of the cerebral ganglion were desheathed. The cerebral ganglion was then twisted at the commissure to make it possible to access CC9/10 on the dorsal surface and contralateral CBIs on the ventral surface.

Frequent IPSPs are observed simultaneously in both CC9/10 cells (Fig. 2B). Activation of one CC9/10 can induce polysynaptic inhibition in the other CC9/10 (Fig. 2C1).

CC9/10 responds to noxious stimulus by tonic firing.
CC9/10 firing evokes locomotion.
The locomotion can be seen very well from PPCN (para-pedal commissural nerve).
The frequency of the locomotory rhythm is different depending on whether you stimulated P9 or AT4 nerve.

The cycle frequency of locomotor activity reflects the firing frequency of CC9/10 activity.
CC9 and CC10 may act as locomotion initiators.

One function of CC9/10 is to provide excitation to serotonergic modulatory neurons of the pedal ganglion.
It enhances the induction of the locomotory response.

CC9/10 weakly excites contralateral MCC and CBI-2. MCC>CBI-2

CC9/10 increase excitability (?) of MCC, but this could be just because of depolarization.

CC9–10 are broadly activated and their responses are not site-specific.

Aplysia locomotes, and then eats.


The Journal of Neuroscience, November 19, 2008 • 28(47):12349 –12361
Neural Analog of Arousal: Persistent Conditional Activation of a Feeding Modulator by Serotonergic Initiators of Locomotion
Jian Jing, Ferdinand S. Vilim, Elizabeth C. Cropper, and Klaudiusz R. Weiss