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. 2023 May 26;26(6):106985. doi: 10.1016/j.isci.2023.106985

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© 2023 The Author(s)

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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Lack of emotional valence induced by optogenetic activation of L5 subpopulations

(A) An illustration depicting a freely moving mouse in a two-spout choice test with one water spout paired with LED illumination and the other unpaired with LED illumination. Note that the two spouts deliver the same amount of water per lick.

(B and D) Left: the number of bouts in which the mouse chose the spout with versus without illumination for mice expressing ChR2 (black circles) or tdTomato (white squares) in the L5a population (ChR: 42.3 ± 5.1 for [LED+] versus 43.7 ± 7.7 for [LED-], Wilcoxon signed rank test, p = 0.56; tdTom: 41.0 ± 6.3 for [LED+] versus 41.8 ± 5.4 for [LED-], Paired t test, p = 0.78, B) and L5b population (ChR: 39.9 ± 4.7 for [LED+] versus 31.3 ± 3.8 for [LED-], Paired t test, p = 0.024; tdTom: 45.9 ± 5.9 for [LED+] versus 46.1 ± 5.8 for [LED-], Paired t test, p = 0.91, D). Inset bar graphs represent the percentage of bouts paired with illumination for (L5a: 50.3 ± 3.7% for ChR versus 48.8 ± 1.7% for tdTom, Wilcoxon rank-sum test, p = 0.30, B; L5b: 56.2 ± 1.8% for ChR versus 49.9 ± 1.3% for tdTom, Student’s t test, p = 0.013, D). Right: the number of bouts in which the mouse switched versus unswitched choice next to the bouts with illumination (ChR: 17.8 ± 2.5 for switched choice versus 23.2 ± 3.1 for unswitched choice, Paired t test, p = 0.071; tdTom: 20.3 ± 3.5 for switched choice versus 19.8 ± 3.2 for unswitched choice, Paired t test, p = 0.84 for the L5a population, B; ChR: 14.8 ± 1.5 for switched choice versus 24.5 ± 4.4 for unswitched choice, Paired t test, p = 0.047; tdTom: 23.4 ± 1.9 for switched choice versus 21.8 ± 4.0 for unswitched choice, Paired t test, p = 0.56 for the L5b population, D). Inset bar graphs represent the percentage of bouts paired with illumination for (L5a: 43.6 ± 2.9% for ChR versus 51.1 ± 2.6% for tdTom, Wilcoxon rank-sum test, p = 0.24, B; L5b: 40.0 ± 4.0% for ChR versus 53.4 ± 2.7% for tdTom, Student’s t test, p = 0.015, D).

(C and E) Left: the ratio of the mean licks per bout with illumination relative to those without illumination for mice expressing ChR2 (black) or tdTomato (white) in the L5a population (0.47 ± 0.04 for ChR versus 1.02 ± 0.14 for tdTom, Welch’s t test, p < 0.01, C) and L5b population (1.81 ± 0.17 for ChR versus 0.97 ± 0.12 for tdTom, Student’s t test, p < 0.01, E). Right: the percentage of bouts with illumination in the top 10 longest bouts in the sessions (L5a: 33.0 ± 5.2% for ChR versus 49.3 ± 2.4% for tdTom, Student’s t test, p < 0.01, C; L5b: 68.0 ± 1.0% for ChR versus 47.5 ± 4.6% for tdTom, Welch’s t test, p < 0.01, E). Gray dots represent individual mice (n = 6 for ChR, n = 8 for tdTom in B and C; n = 8 for ChR, n = 8 for tdTom in D and E).

(F) An illustration depicting a freely moving mouse in a two-spout choice test with one spout paired with and the other unpaired with LED illumination. Note that both spouts were devoid of water.

(G) Left: the number of bouts in which the mouse chose the spout with versus without illumination for mice whose L5b population expressed ChR2 (black circles) or tdTomato (white squares) (ChR: 15.1 ± 6.4 for [LED+] versus 10.6 ± 2.2 for [LED-], Wilcoxon signed rank test, p = 0.64; tdTom: 10.1 ± 3.0 for [LED+] versus 10.4 ± 2.9 for [LED-], Wilcoxon signed rank test, p = 0.42). Inset bar graphs represent the percentage of bouts paired with illumination for (51.9 ± 6.6% for ChR versus 47.5 ± 3.6% for tdTom, Student’s t test, p = 0.57). Right: the number of bouts in which the mouse switched versus unswitched choice next to the bouts with illumination (ChR: 5.6 ± 1.6 for switched choice versus 8.9 ± 4.8 for unswitched choice, Wilcoxon signed rank test, p = 0.53; tdTom: 4.6 ± 1.6 for switched choice versus 5.0 ± 1.7 for unswitched choice, Wilcoxon signed rank test, p = 0.88). Inset bar graphs represent the percentage of bouts paired with illumination for (54.1 ± 9.3% for ChR versus 49.9 ± 6.4% for tdTom, Student’s t test, p = 0.71).

(H) Left: the ratio of the mean licks per bout with illumination relative to those without illumination for mice whose L5b population expressed ChR2 or tdTomato (1.43 ± 0.35 for ChR versus 1.31 ± 0.22 for tdTom, Wilcoxon rank-sum test, p = 0.88). Right: the ratio of the total number of licks in the session (2.70 ± 1.20 for ChR versus 1.21 ± 0.25 for tdTom, Wilcoxon rank-sum test, p = 0.88). Gray dots represent individual mice (n = 8 for ChR, n = 8 for tdTom). Dotted lines in B-E, G, and H indicate the chance level. See also Figure S7.