Fig 1.
1. The recording chamber; 2. inlet; 3. outlet; 4. Net (Filter); 5. Camera; 6. Spot light; 7. Reflectors. ASW in the recording chamber is shown in blue for illustration purpose only.
Fig 2.
(A) The raw image from a video frame; (B) The image after camera calibration and alignment; (C) The image after color detection and threshold value calculation showing animal outline in white; (D) object-recognition, the object (Aplysia) is circled in red line; (E) marking (yellow +) of the object center of gravity; (F) an example of movement path (shown in yellow) of Aplysia (1 hr).
Fig 3.
Plot of the paths (in mm) that animals traveled in six video segments determined both manually and automatically by the program. The thick line is the linear regression fit. The two sets of path data were highly correlated with a slope of 1.066.
Fig 4.
Absence of effects of GFFD in the isolated cerebral and pleural-pedal ganglia.
(A) The electrophysiological recordings of PPCN during control, after perfusion with 10−6 M or 10−5 M GFFD, and during wash, each recording lasting ~ 10 min. (B) Group data of the average PPCN frequency over 10 min under different conditions. GFFD had no significant effects on PPCN activity. Error bars indicate SEM.
Fig 5.
GdFFD induces activity in the isolated cerebral and pleural-pedal ganglia.
(A) The electrophysiological recordings of PPCN during control, after perfusion with 10−6 M or 10−5 M GdFFD, and during wash, each recording lasting ~ 10 min. (B) Group data. (B1) The average frequency of PPCN over 10 min; (B2) The average of frequency within bursts in PPCN over a period of 10 min; (B3) The average of frequency between bursts in PPCN over 10 min. Bonferroni post hoc tests: *p < 0.05; **p < 0.01; ***p < 0.001. Error bars indicate SEM.
Fig 6.
Comparison between network activity induced by GdFFD perfusion and by P9 stimulation.
(A) The electrophysiological recordings of PPCN during 10−5 M GdFFD perfusion or after P9 stimulation. The two traces in each condition are continuous. The black arrow marks P9 stimulation. Both GdFFD perfusion and P9 stimulation induced bursting activity, but activity between bursts was higher with GdFFD perfusion. Open bars indicate PPCN bursting activity. (B) Group data. (B1) The average of frequency within bursts of PPCN over 10 min; (B2) The average of frequency between bursts of PPCN over 10 min. (B3) The ratio of frequency within bursts to frequency between bursts. Paired two-tailed t-test: *p < 0.05; Error bars indicate SEM.
Fig 7.
GdFFD reduces Aplysia locomotion whereas GFFD has no effects based on video analysis.
(A) The locomotor path of Aplysia after injection with ASW or GdFFD. Left panel: ASW; right panel: 10−8 M GdFFD (The animal is present at top right corner as it typically stays in the area). Color of path: red: beginning of path; blue: end of the path. (B) Group data. The velocity (v) before (black bars) and after (gray bars) injections with ASW, 10−8 M GdFFD or 10−6 M GFFD. Error bars indicate SEM.