Figure 2. OFC^CamKII+, but not OFC^PV+, Ca²⁺ activity encodes prior action information.

(A) (top) Anatomical schematic and (bottom) representative histology of (A) OFC^CamKII+ in vivo Ca²⁺ fiber photometry experiments (C57BL/6J, n = 9 mice, 6 males, 3 females). Approximate bregma coordinate AP +2.68 mm.

(B) Representative data heat map of OFC^CamKII+ normalized fluorescence changes relative to lever press initiation, ordered by lever press duration.

(C–F) Ca²⁺ activity from OFC^CamKII+ populations z-scored normalized relative to a pre-lever press onset baseline period and aligned to (C) lever press onset, (D) lever press duration (presented as the relative percentage of total lever press duration), the (E) offset (i.e. termination) of a lever press, and the (F) first head entry made after a lever press.

(G) Representative traces indicating changes in OFC^CamKII+ Ca²⁺ fluorescence over time.

(H–J) β coefficients from LME models relating OFC^CamKII+ Ca²⁺ activity to current and prior durations for actual and order shuffled data (H) before press onset (Pre-LP Activity), (I) during the press (Ongoing LP Activity), and (J) after press offset (Post-LP Activity).

(K) (top) Anatomical schematic and (bottom) representative histology of OFC^PV+ in vivo Ca²⁺ fiber photometry experiments (PV^cre, n = 8, 5 males, 3 females). Approximate bregma coordinate AP +2.68 mm.

(L) Representative data heat map of OFC^PV+ normalized fluorescence changes relative to lever press initiation, ordered by lever press duration.

(M–P) Ca²⁺ activity from OFC^PV+ populations z-scored normalized relative to a pre-lever press onset baseline period and aligned to (M) lever press onset, (N) lever press duration (presented as the relative percentage of total lever press duration), the (O) offset (i.e. termination) of a lever press, and the (P) first head entry made after a lever press.

(Q) Representative traces indicating changes in OFC^PV+ Ca²⁺ fluorescence over time.

(R–T) β coefficients from LME models relating OFC^PV+ Ca²⁺ activity to current and prior durations for actual and order shuffled data (R) before press onset (Pre-LP Activity), (S) during the press (Ongoing LP Activity), and (T) after press offset (Post-LP Activity).

LP = lever press, HE = Head Entry. Black lines in C–F and M–P indicate significant differences between Rewarded and Unrewarded lever presses via permutation testing (p < 0.05). For B and L, dashed white lines indicate 1600 ms session criterion window. Orange markers indicate the termination of an unrewarded lever press. Blue markers indicate the termination of rewarded lever press. For G and Q, shaded regions indicate the duration of the current (n) or prior (n − 1) lever press. Ca²⁺ activity magnitude predicted by LME models includes −2s to 0s before (Pre), during (Ongoing), or 0 to 2s after (Post) the current lever press.

Shuffled data are the mean ± SEM of 1000 order shuffled β coefficients. ** p < 0.01, *** p < 0.001.

For a full report of permutation testing p-values related to Figures 2C–F and 2M–P, see publicly available data listed in the key resources table.

See also Figure S2, Table S2 and Table S3 for more data.