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Figures
- Fig. 1.
GABA receptor ligands of different action mechanisms modulate feeding behaviour in Prussian carp, Carassius gibelio. Dose–response relationships of feeding behaviour elements were registered 15 min after i.p. injection during 2 h time intervals. Bar pattern codes apply to A–D. FA, feeding act; PNZ, phenazepam; SR, SR-95531; Bic, bicuculline; FMZ, flumazenil.
- Fig. 2.
SR reverses Bic antagonism to PNZ effect on behaviour. (A) Time–response relationship for four GABAR ligands: 0.2 mg kg−1 PNZ, 2 mg kg−1 SR, 1.5 mg kg−1 Bic and 5 mg kg−1 FMZ. (B) Bic (1.5 mg kg−1) and SR were injected 30 min later than PNZ (0.2 mg kg−1). (C) Bic (1.5 mg kg−1) and SR were injected without PNZ. (D) Bic (1.5 mg kg−1) and SR were injected 30 min after PNZ (0.2 mg kg−1) and FMZ (5 mg kg−1). Doses of SR are symbol-coded. Asterisks denote significant difference from ‘no SR’ values (*P<0.05); data normalized to vehicle control; N=6–7, Bonferroni post hoc test.
- Fig. 3.
SR and Bic display different types of antagonism to GABA+PNZ effect on spontaneous IPSCs. (A) Example traces of GABAergic spontaneous activity. From top to bottom: control, PNZ, PNZ+SR, PNZ+Bic, PTX. Vertical dashed lines denote example IPSCs expanded on the right. (B) Sketch of C. gibelio brain. ‘T’ marks the telencephalon; vertical dashed line shows approximate direction of slice cutting. (C) Averaged spontaneous IPSCs normalized to peak amplitude of control event; note change of IPSC decay kinetics induced by Bic in contrast to SR. (D) Analysis paradigm for response decay kinetics with bi-exponential fitting. Lines through data points: best-fit bi-exponential functions; dashed lines of the same colour are plots of the fast and slow components alone. Ratio of fitting coefficients (RFC) of fast (τ=2.27) to slow (τ=20.41) component generated under PNZ obtained as 21.49/8.99=2.39; application of Bic augmented decay components ratio to 10.72/1.38=8.38. (E) Statistical summary of changes of ratio of fitting coefficients (RFC) induced by GABAR ligands. *P<0.05, N=6, Student's paired t-test.
- Fig. 4.
Pre-application of FMZ prevents effect of Bic on IPSC decay kinetics. (A) Example traces of GABAergic spontaneous activity. From top to bottom: control, PNZ, PNZ+FMZ, PNZ+FMZ+Bic, PNZ+FMZ+Bic+PTX. Vertical dashed lines denote example IPSCs expanded on the right. (B) Averaged spontaneous IPSCs normalized to peak amplitude of control event. (C) Statistical summary of changes of RFC induced by GABAR ligands. *P<0.05, ***P<0.001, N=6, Student's paired t-test.
- Fig. 5.
SR and Bic do not alter response kinetics generated by endogenous GABA. (A) Example traces of GABAergic spontaneous activity. From top to bottom: control, SR, Bic, PTX. Vertical dashed lines denote example IPSCs expanded on the right. (B) Averaged spontaneous IPSCs normalized to peak amplitude of control event; no change of response decay kinetics observed. (C) Statistical summary of changes of RFC induced by GABAR ligands.
- Fig. 6.
Bic, but not SR, antagonizes PNZ effect on single-channel GABAR openings. (A) Example traces, channel openings induced by GABA. From top to bottom: control (spontaneous openings), GABA, GABA+SR, GABA+Bic, GABA+PTX. (B) Example traces, channel openings induced by PNZ. From top to bottom: control, PNZ, PNZ+SR, PNZ+Bic, PNZ+ PTX. Scale bars apply to A and B. (C) Statistical summary of A. Charge transfer per second normalized to value generated by application of GABA; SR and Bic suppress GABA effect. (D) Statistical summary of B. Charge transfer per second normalized to value generated by application of PNZ. Bic, but not SR, suppresses effect of PNZ. Asterisks indicate significance of difference from unity: **P<0.1, ***P<0.01, N=6, Student's paired t-test.
- Fig. 7.
FMZ prevents conversion of Bic effect by SR on single-channel openings. (A) Traces from top to bottom: control (no ligands), GABA+PNZ, GABA+PNZ+Bic, GABA+PNZ+Bic+SR, GABA+PNZ+Bic+SR+PTX. (B) Traces from top to bottom: control, GABA+PNZ, GABA+PNZ+FMZ+Bic, GABA+PNZ+FMZ+Bic+SR, GABA+PNZ+FMZ+Bic+SR+PTX. Scale bars apply to A and B. (C) Statistical summary for A. (D) Statistical summary for B. In C and D, data on charge transfer are normalized to value generated by GABA+PNZ. **P<0.01, N=6, Student's paired t-test.
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