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Nature, том 617, страницы 548–554 (2023 г.) Процитировать эту статью

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Изменения в моделях активности медиальной префронтальной коры позволяют грызунам, приматам и людям обновлять свое поведение, чтобы адаптироваться к изменениям в окружающей среде, например, при выполнении когнитивных задач1,2,3,4,5. Тормозные нейроны, экспрессирующие парвальбумин, в медиальной префронтальной коре важны для обучения новым стратегиям во время задачи по смене правил6,7,8, но взаимодействия цепей, которые переключают динамику префронтальной сети с поддержания на обновление паттернов активности, связанных с задачей, остаются неизвестными. Здесь мы описываем механизм, который связывает нейроны, экспрессирующие парвальбумин, новую мозолистую тормозную связь и изменения в представлении задач. В то время как неспецифическое ингибирование всех каллозальных проекций не препятствует обучению мышей изменениям правил или нарушению эволюции моделей активности, избирательное ингибирование только каллозальных проекций нейронов, экспрессирующих парвальбумин, ухудшает обучение сдвигу правил, десинхронизирует активность гамма-частоты, необходимую для обучения8 и подавляет реорганизацию моделей префронтальной активности, которая обычно сопровождает обучение смене правил. Эта диссоциация показывает, как мозолистые проекции, экспрессирующие парвальбумин, переключают режим работы префронтальных цепей с поддержания на обновление, передавая гамма-синхронность и ограничивая способность других мозолистых входов поддерживать ранее установленные нейронные репрезентации. Таким образом, мозолистые проекции, происходящие от нейронов, экспрессирующих парвальбумин, представляют собой ключевой локус цепи для понимания и коррекции дефицита поведенческой гибкости и гамма-синхронности, которые вовлечены в шизофрению и родственные состояния9,10.

Организмы должны постоянно обновлять свои поведенческие стратегии, чтобы адаптироваться к изменениям в окружающей среде. Неуместное упорство в использовании устаревших стратегий является отличительной чертой таких состояний, как шизофрения и биполярное расстройство, и классически проявляется в задаче по сортировке карточек штата Висконсин11 (WCST). Хорошо известно, что префронтальная кора отвечает за такой гибкий когнитивный контроль, обеспечивая активное поддержание представлений о правилах или целях12,13,14, адаптивное блокирование этих представлений15 и нисходящее смещение сенсорной обработки посредством обширной взаимосвязи с другими области мозга16,17. Исследования показали, что в медиальной префронтальной коре (mPFC) нормальная функция интернейронов, экспрессирующих парвальбумин (PV), необходима мышам для выполнения задач по «сдвигу правил», которые, как и WCST, включают в себя выявление неподсказанных изменений правил и изучение новых правила, в которых используются сигналы, которые ранее не имели отношения к результатам испытаний6,7. Более того, интернейроны PV играют ключевую роль в генерации синхронизированной ритмической активности в диапазоне гамма-частот (около 40 Гц)18,19. Действительно, во время задач по смене правил синхронность гамма-частотной активности между интернейронами PV в левой и правой mPFC увеличивается после попыток ошибок, то есть когда мыши получают обратную связь о том, что ранее усвоенное правило устарело, и оптогенетически нарушает эту синхронизацию. вызывает настойчивость8. Тем не менее, основные взаимоотношения между цепями (синаптическими связями), динамикой сети (межполушарная гамма-синхрония) и нейронными представлениями (зависимые от задачи изменения в моделях активности) остаются неизвестными.

Обычно предполагается, что гамма-синхрония передается между регионами посредством возбуждающих синапсов20, которые являются преобладающей формой дальней связи в коре. Однако мы исследовали альтернативную гипотезу, предложенную недавними описаниями связей, высвобождающих γ-аминомасляную кислоту (ГАМКергические) дальнего действия, происходящих из нейронов PV в mPFC21. В частности, мы впервые продемонстрировали, что PV-экспрессирующие нейроны в mPFC дают начало мозолистым ГАМКергическим синапсам в контралатеральной mPFC (рис. 1). Мы идентифицировали эту анатомическую связь путем инъекции AAV-EF1α-DIO-ChR2-eYFP в одну mPFC мышей PV-cre и наблюдали меченные вирусом PV-концы в контралатеральной PFC, особенно в глубоких слоях 5 и 6 (рис. 1a). Чтобы охарактеризовать эту каллозальную проекцию ЛВ и ее нейроны-реципиенты, мы выполнили записи в контралатеральной мПФК (рис. 1б). Мы обнаружили, что каллозальные проекции PV иннервируют пирамидные нейроны (идентифицированные на основании их морфологии и физиологии небыстрых импульсов; 31 из 75 подключенных), но не быстроспайковые нейроны (0 из 18 подключенных) (рис. 1г). Ритмические серии PV-концевой оптогенетической стимуляции вызывали временные тормозные постсинаптические потенциалы (IPSP) в ChR2-негативных пирамидных нейронах (рис. 1д), которые не блокировались антагонистами глутаматергических рецепторов 6-циано-7-нитрохиноксалин-2,3. -диона динатриевая соль (CNQX) (10 мкМ) и d-2-амино-5-фосфонопентановая кислота (APV) (50 мкМ), но они были полностью отменены путем применения в ванне антагониста рецептора γ-аминомасляной кислоты (ГАМКА) типа А. габазин (10 мкМ; рис. 1д, е). Чтобы дополнительно охарактеризовать конкретные мишени каллозальных синапсов mPFC PV, мы ввели субъединицу B холерного токсина, конъюгированную с флуоресцентным красителем (CTb), в четыре нижестоящие мишени mPFC, а затем зарегистрировали данные от ретроградно меченных нейронов mPFC, которые проецируются на контралатеральную mPFC (т. туда, где были выполнены записи, и ипсилатерально от инъекции AAV-DIO-ChR2), дорсальное полосатое тело, медиодорсальный (MD) таламус или прилежащее ядро (NAc) (расширенные данные, рис. 1b–k). После одного светового импульса длительностью 5 мс для оптогенетической активации каллозальных терминалей PV+ в присутствии глутаматергических антагонистов (20 мкМ 6,7-динитрохиноксалин-2,3-диона (DNQX) и 50 мкМ APV) мы наблюдали заблокированные по времени IPSP у 22 из 22 пирамидальных нейронов, проецирующих MD, по сравнению с 0 из 18 пирамидальных нейронов, проецирующих мозолистое тело, 0 из 21 пирамидальных нейронов, проецирующих прилежащее тело, и 7 из 24 пирамидальных нейронов, проецирующих дорсальное полосатое тело.

50 Hz, fast after hyperpolarization (fAHP) amplitude > 14 mV, and spike-frequency accommodation (SFA) index < 2./p>1,000 times stronger than the measured scattered light power. Both 532 nm and 638 nm at this final light power ipsilateral to the viral injection site was used./p>

0.05 and were considered not significant. No statistical methods were used to pre-determine sample sizes but our sample size choice was based on previous studies6,8 and are consistent with those generally employed in the field. Data distribution was assumed to be normal, but this was not formally tested./p> 0.99). d,e,k,l, Two-way ANOVA (task day × virus) comparing rule-shift (RS) performance across groups (interaction: P < 0.0001), showed no group difference on day 1 but significant impairment on days 2 and 3 for DIO-eNpHR compared to DIO-mCherry and Syn-tdTomato. e, Optogenetic inhibition of mPFC callosal PV terminals impairs rule shifts in DIO-eNpHR mice compared to DIO-mCherry controls (day 1 to 2: P = 0.014; day 1 to 3: P = 0.012; day 2 to 3: P = 0.075). For Fig. 3f,g, full statistics are: 30–50 Hz synchronization is higher after RS errors than after RS correct decisions across task days in controls (n = 4 mice; two-way ANOVA; main effect of trial type: P = 0.0056; day 1: P = 0.004; day 2: P = 0.005; day 3: P = 0.022). g, Gamma synchrony is higher after RS errors than after RS correct decisions for DIO-eNpHR mice on day 1 (no light), but not days 2 and 3 (n = 5 mice; two-way ANOVA (trial type × task day); interaction: P = 0.0039; day 1: P = 0.0056; day 2: P = 0.16; day 3: P = 0.23). Differences in gamma synchrony between RS errors and correct trials are also significantly lower in DIO-eNpHR mice compared to controls on days 2 (two-way ANOVA (trial type × virus); interaction: P = 0.018) and 3 (two-way ANOVA (trial type × virus); interaction: P = 0.034). We then performed two-way ANOVA (task day × virus, type of error) comparing gamma synchrony across groups with appropriate post hoc tests corrected for multiple comparisons; gamma synchrony is lower for DIO-eNpHR mice compared to matched controls on error (interaction: P = 0.049), but not correct trials (interaction: P = 0.93) on both days 2 (P = 0.0089) and 3 (P = 0.016). For Fig. 3k–n, full statistics are: inhibiting callosal terminals does not affect rule shifts in Syn-eNpHR mice (n = 5) compared to controls (n = 4) across days (two-way ANOVA (task day × virus); P = 0.37). k, Performance of controls did not change across days (day 1 to 2: P = 0.48; day 1 to 3: P > 0.99; day 2 to 3: P > 0.99). l, Performance of Syn-eNpHR mice did not change across days (day 1 to 2: P > 0.99; day 1 to 3: P = 0.96; day 2 to 3: P = 0.85). m, Interhemispheric 30–50 Hz synchronization is higher after RS errors than RS correct decisions across days in controls (n = 4 mice; two-way ANOVA; main effect of trial type: P = 0.020; day 1: P = 0.015; day 2: P = 0.049; day 3: P = 0.025). n, Gamma synchrony is higher after RS errors than RS correct decisions for Syn-eNpHR mice on day 1 (no light). This was abolished with light on day 2, then restored in the absence of light on day 3 (n = 5 mice; two-way ANOVA; (trial type × task day); interaction: P = 0.011; day 1: P = 0.037; day 2: P = 0.46; day 3: P = 0.021). Gamma synchrony is lower for Syn-eNpHR mice compared to matched controls on error (two-way ANOVA; interaction: P = 0.045) but not correct trials (interaction: P = 0.74) on day 2 (P = 0.043), but not on day 3 (P = 0.58). Two-way ANOVA followed by Bonferroni post hoc comparisons was used./p> 0.99). f, Performance of Syn-eNpHR mice did not change (day 1 to 2: P = 0.58; day 1 to 3: P = 0.82; day 2 to 3: P > 0.99). For Fig. 4h,j, full statistics are: optogenetic inhibition changes the similarity of activity vectors specifically in DIO-eNpHR mice (two-way ANOVA, main effect of task day: P = 0.017; task day × group interaction: P = 0.32). h, For controls (n = 6 mice), there is no change across days in the similarity between activity vectors after RS errors and those after correct decisions (day 1 to 2: P = 0.57; day 1 to 3: P = 0.32; day 2 to 3: P = 0.90). i, In DIO-eNpHR mice (n = 6 mice), there is an increase in the similarity between activity vectors after RS errors and those after correct decisions from day 1 (before optogenetic inhibition) to days 2 and 3 (during and after inhibition, respectively) (day 1 to 2: P = 0.044; day 1 to 3: P = 0.0067; day 2 to 3: P = 0.71). j, For Syn-eNpHR mice (n = 6 mice), there is no change across days in the similarity between activity vectors after RS errors and those after correct decisions (day 1 to 2: P = 0.99; day 1 to 3: P = 0.93; day 2 to 3: P = 0.98). Two-way ANOVA to compare the change in activity vector similarity from day 1 to 2 in the DIO-eNpHR group to a group consisting of the eNpHR-negative and synapsin-eNpHR mice, using mouse, day, and day × group interaction as factors revealed a significant day × group interaction (P = 0.040). k–m, The similarity between activity vectors after decisions on error trials during the initial association (IA) and those during the RS. There is increased similarity across days in DIO-eNpHR mice (n = 6; two-way ANOVA (task day × virus); interaction: P = 0.025). k, There is no change in the similarity of population activity vectors after IA errors and RS errors across days in controls (n = 6; day 1 to 2: P = 0.99; day 1 to 3: P = 0.52; day 2 to 3: P = 0.50). l, During and following optogenetic inhibition on days 2 and 3 in DIO-eNpHR mice, there is an increase in the similarity of population activity vectors following IA versus RS error trials (day 1 to 2: P = 0.0038; day 1 to 3: P = 0.0045; day 2 to 3: P = 0.95). m, In Syn-eNpHR mice, there is no change in the similarity of population activity vectors following IA versus RS errors trials across days (n = 6; day 1 to 2: P = 0.41; day 1 to 3: P = 0.90; day 2 to 3: P = 0.24). n, Controls have increases in average activity (fraction of frames in which a neuron is active, averaged across all neurons) during the 10 s following RS errors compared to the 10 s following RS correct decisions on all days (n = 6; two-way ANOVA, main effect of trial type: P = 0.010; day 1: Bonferroni P = 0.022; day 2: Bonferroni P = 0.015; day 3: Bonferroni P = 0.016). o, DIO-eNpHR mice have an increase in average activity after errors that depends on the day (n = 6; two-way ANOVA (task day × trial type); interaction: P = 0.0003). This difference occurs on day 1 (Bonferroni P < 0.0001), but is abolished with light delivery on day 2 (Bonferroni P > 0.99), and continues to be absent even without further light delivery on day 3 (Bonferroni P > 0.99). p, In Syn-eNpHR mice, there is an overall increase in average activity following error trials (n = 6; two-way ANOVA, main effect of trial type: P = 0.0088). This occurs on days 1 (Bonferroni P = 0.038) and 3 (Bonferroni P = 0.016), but not with light delivery on day 2 (Bonferroni P > 0.99). Two-way ANOVA followed by Tukey post hoc comparisons was used unless otherwise noted./p>

0.9999), Day 2 (post hoc t(28) = 0.6978, P > 0.9999), nor Day 3 (post hoc t(28) = 0.4652, P > 0.9999). o, Inhibition disrupts rule shift performance in DIO-eNpHR-expressing mice compared to controls when light is delivered continuously throughout the RS on Day 4 (post hoc t(28) = 9.886, P < 0.0001). Two-way ANOVA followed by Bonferroni post hoc comparisons was used. ****P < 0.0001./p>

0.9999; same and incorrect post hoc t(26) = 0.1486, P > 0.9999; different location two-way ANOVA (trial type × virus); interaction: F(1,13) = 13.87, P = 0.0026; different and correct post hoc t(26) = 3.724, P = 0.0019; different and incorrect post hoc t(26) = 3.724, P = 0.0019). i, Same as h but for the next 5 RS trials (n = 7 DIO-mCherry control mice; same location two-way ANOVA (trial type × virus); interaction: F(1,13) = 3.214, P = 0.0963; same and correct post hoc t(26) = 1.793, P = 0.1693; same and incorrect post hoc t(26) = 1.793, P = 0.1693; different location two-way ANOVA (trial type × virus); interaction: F(1,13) = 8.948, P = 0.0104; different and correct post hoc t(26) = 2.991, P = 0.0120; different and incorrect post hoc t(26) = 2.991, P = 0.0120). j–k, The overall speed (meters per second) of mice during the first 5 IA and RS trials across days in the cohort of mice used for microendoscopic Ca2+ imaging (n = 6 eNpHR-negative controls; n = 6 DIO-eNpHR mice; two-way ANOVA (IA vs. RS × task day) for control mice: interaction: F(1,10) = 0.00271, P = 0.9595; two way-ANOVA (IA vs. RS × task day) for DIO-eNpHR mice: interaction: F(1,10) = 1.378, P = 0.2677). l–m, There is no difference in the amount of time (seconds) mice spent exploring bowls before making a decision during the first 5 IA and RS trials across days in the microendoscope experimental dataset (n = 6 eNpHR-negative controls; n = 6 DIO-eNpHR mice; two-way ANOVA (IA vs. RS × task day) for control mice: interaction: F(1,10) = 0.5053, P = 0.4934; two way-ANOVA (IA vs. RS × task day) for DIO-eNpHR mice: interaction: F(1,10) = 0.1147, P = 0.7419). n–o, The first move of the mouse toward the correct bowl (percent) during the first 5 IA and RS trials across days in the Ca2+ imaging experimental dataset (n = 6 eNpHR-negative controls; n = 6 DIO-eNpHR mice; two-way ANOVA (IA vs. RS × task day) for control mice: interaction: F(1,10) = 3.347, P = 0.0973; two way-ANOVA (IA vs. RS × task day) for DIO-eNpHR mice: interaction: F(1,10) = 0.1316, P = 0.7244). p–z, Effects of inhibiting callosal PV+ projections during a version of the RS task using a shorter (30 second) intertrial interval (ITI). p, Experimental design: Day 1, no light delivery; Day 2, continuous light during the RS for optogenetic inhibition of callosal PV terminals ; Day 3, no light was delivered. q, Representative image showing DIO-eYFP expression in one mPFC and a fiber-optic cannula implanted in the contralateral mPFC in a PV-Cre mouse. r, Representative image showing DIO-eNpHR-mCherry (DIO-eNpHR) expression in one mPFC and a fiber-optic cannula implanted in the contralateral mPFC in a PV-Cre mouse. s,w, IA performance with a 30 s ITI in eNpHR-negative mice (n = 6) and eNpHR-expressing mice (n = 8; two-way ANOVA (task day × virus); interaction: F(2,24) = 0.1585, P = 0.8543). t,x, Optogenetic inhibition of mPFC callosal PV terminals with a 30 s ITI impairs rule shift performance in DIO-eNpHR mice (n = 8) compared to controls (n = 6; two-way ANOVA (task day × virus); interaction: F(2,24) = 50.79, P < 0.0001). t, Performance of DIO-eYFP controls did not change from Day 1 to Day 2 (Tukey’s post hoc q(5) = 1.606, P = 0.5356), Day 1 to Day 3 (Tukey’s post hoc q(5) = 1.035, P = 0.7567), nor Day 2 to Day 3 (Tukey’s post hoc q(5) = 0.4344, P = 0.9498). x, Inhibition disrupts rule shift performance in DIO-eNpHR mice from Day 1 to Day 2 (Tukey’s post hoc q(7) = 15.34, P < 0.0001), Day 1 to Day 3 (Tukey’s post hoc q(7) = 21.75, P < 0.0001), but not Day 2 to Day 3 (Tukey’s post hoc q(7) = 2.679, P = 0.2101). u, y, Optogenetic inhibition of callosal PV terminals increases perseverative errors in DIO-eNpHR mice (n = 8 mice) compared to DIO-eYFP controls (n = 6 mice; two-way ANOVA (task day × virus) interaction: F(2,24) = 19.79, P < 0.0001). v, z, Optogenetic inhibition of callosal PV terminals has no effect on random errors in DIO-eNpHR mice (n = 8) compared to DIO-eYFP controls (n = 6; two-way ANOVA (task day × virus); interaction: F(2,24) = 1.079, P = 0.3559). u, v, Light delivery does not affect the number of perseverative (post hoc t(5) = 0.000 – 1.000, P > 0.9999) or random (post hoc t(5) = 0.4152 – 0.7906, P > 0.9999) errors in DIO-eYFP controls across days. y, z, Optogenetic inhibition of callosal PV terminals on Day 2 increased the number of perseverative (post hoc t(7) = 6.008, P = 0.0016 from Day 1 to Day 2; post hoc t(7) = 5.844, P = 0.0019 from Day 1 to Day 3; but not from Day 2 to Day 3: post hoc t(7) = 1.111, P = 0.9093) but not random (post hoc t(7) = 0.000 – 2.198, P = 0.1918 – > 0.9999) errors compared to no stimulation. Two-way ANOVA followed by Bonferroni post hoc comparisons was used unless otherwise noted. Data were expressed as mean ± s.e.m. **P < 0.01, ****P < 0.0001; scale bar, 100 μm./p>

0.9999) errors in DIO-eYFP controls. g, h, Optogenetic inhibition of callosal PV terminals on Day 2 increased the number of perseverative (post hoc t(13) = 10.75, P < 0.0001) and random (post hoc t(13) = 3.145, P = 0.0155) errors compared to no stimulation on Day 1. k, l, o, p, Optogenetic inhibition of all callosal projections has no effect on perseverative errors in Syn-eNpHR mice (n = 6) compared to controls (n = 4; two-way ANOVA (task day × virus); interaction: F(1,8) = 0.0, P > 0.9999) nor on random errors (two-way ANOVA (task day × virus); interaction: F(1,8) = 0.07805, P = 0.787). Two-way ANOVA followed by Bonferroni post hoc comparisons was used. *P < 0.05, ****P < 0.0001; scale bars, 250 μm and 100 μm, respectively./p>

0.9999). By contrast, rule shift performance in mice expressing both NpHR and ChR2 (h; n = 8) is significantly different across days than that of mice which express NpHR-only (two-way ANOVA (task day × virus); interaction: F(3,27) = 6.747, P = 0.0015). Optogenetic inhibition of mPFC callosal PV terminals causes NpHR+ChR2-expressing mice (n = 8) to take a large number of trials to learn rule shifts on Day 1 and this does not change on Day 2 (no light) (post hoc t(7) = 1.446, P > 0.9999). However, RS learning is rescued by 40 Hz optogenetic stimulation on Day 3 (post hoc t(7) = 10.91, P < 0.0001) and this improvement does not change on ‘Day 4’ of testing which occurs one week later (post hoc t(7) = 0.6394, P > 0.9999). f, i: Optogenetic inhibition followed by stimulation of callosal PV terminals changes the number of perseverative errors in mice expressing both NpHR and ChR2 (n = 8 mice) compared to controls expressing only NpHR (n = 3 mice; two-way ANOVA; main effect of task day: F(2.015,18.13) = 7.167, P = 0.0050; main effect of virus: F(1,9) = 14.31, P = 0.0043; interaction: F(3,27) = 5.324, P = 0.0052). By contrast, there is no difference in numbers of random errors (two-way ANOVA; main effect of task day: F(1.491,13.42) = 1.706, P = 0.2189; main effect of virus: F(1,9) = 1.523, P = 0.2483; interaction: F(3,27) = 0.2901, P = 0.8322). f, Once mice expressing NpHR only receive optogenetic inhibition on Day 1, the number of perseverative errors is stable across days (n = 3 mice; Day 1 to Day 2: post hoc t(2) = 1.222, P > 0.9999; Day 1 to Day 3: post hoc t(2) = 1.299, P > 0.9999; Day 1 to Day 4: post hoc t(2) = 0.3111, P > 0.9999; Day 2 to Day 3: post hoc t(2) = 0.3780, P > 0.9999; Day 2 to Day 4: post hoc t(2) = 1.606, P > 0.9999; Day 3 to Day 4: post hoc t(2) = 2.000, P > 0.9999). g, In mice that express NpHR only (n = 3 mice), numbers of random errors are also stable across days (Day 1 to Day 2: post hoc t(2) = 1.387, P > 0.9999; Day 1 to Day 3: post hoc t(2) = 1.732, P > 0.9999; Day 1 to Day 4: post hoc t(2) = 1.000, P > 0.9999; Day 2 to Day 3: post hoc t(2) = 1.000, P > 0.9999; Day 2 to Day 4: post hoc t(2) = 1.000, P > 0.9999; Day 3 to Day 4: post hoc t(2) = 0.3780, P > 0.9999). i, 40 Hz stimulation of callosal PV terminals on Day 3 reduces the number of perseverative errors in mice expressing both NpHR and ChR2 (n = 8; Day 1 to Day 2: post hoc t(7) = 0.6494, P > 0.9999; Day 1 to Day 3: post hoc t(7) = 5.218, P = 0.0074; Day 1 to Day 4: post hoc t(7) = 6.416, P = 0.0022; Day 2 to Day 3: post hoc t(7) = 4.822, P = 0.0115; Day 2 to Day 4: post hoc t(7) = 12.33, P < 0.0001; Day 3 to Day 4: post hoc t(7) = 0.4971, P > 0.9999). j, 40 Hz stimulation on Day 3 does not affect the number of random errors in mice expressing both NpHR and ChR2 (n = 8 mice; Day 1 to Day 2: post hoc t(7) = 0.7061, P > 0.9999; Day 1 to Day 3: post hoc t(7) = 0.6417, P > 0.9999; Day 1 to Day 4: post hoc t(7) = 1.210, P > 0.9999; Day 2 to Day 3: post hoc t(7) = 0.3140, P > 0.9999; Day 2 to Day 4: post hoc t(7) = 0.4237, P > 0.9999; Day 3 to Day 4: post hoc t(7) = 0.7977, P > 0.9999). Two-way ANOVA followed by Bonferroni post hoc comparisons was used. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; scale bars, 100 μm and 50 μm, respectively./p>

0.9999) or random (Day 1 to Day 2: post hoc t(14) = 0.284, P > 0.9999; Day 1 to Day 3: post hoc t(14) = 0.284, P > 0.9999; Day 2 to Day 3: post hoc t(14) = 0.0, P > 0.9999) errors in controls across days. g, h, Optogenetic inhibition of callosal PV terminals induces perseveration on Day 2 and Day 3 compared to no stimulation on Day 1 (Day 1 to Day 2: post hoc t(14) = 4.092, P = 0.0033; Day 1 to Day 3: post hoc t(14) = 6.405, P < 0.0001; Day 2 to Day 3: post hoc t(14) = 2.313, P = 0.1094), but has no effect on random errors (Day 1 to Day 2: post hoc t(14) = 1.524, P = 0.4493; Day 1 to Day 3: post hoc t(14) = 0.254, P > 0.9999; Day 2 to Day 3: post hoc t(14) = 1.27, P = 0.6744). i, PV-Cre Ai14 mice had bilateral AAV-DIO-Ace2N-4AA-mNeon (Ace-mNeon) injections, an ipsilateral AAV-Synapsin-eNpHR-BFP (Syn-eNpHR) or AAV-Synapsin-mCherry (Syn-mCherry) injection and multimode fiber-optic implants in both prefrontal cortices. j, Representative images from mice injected with a control virus (Syn-mCherry), showing mCherry, Ace-mNeon, and tdTomato expression in the mPFC ipsi to the virus injection, and Ace-mNeon and tdTomato in the contra hemisphere. k, n, Experimental design: Day 1, no light delivery; Day 2, continuous light for inhibition during the R; Day 3, no light delivery. l, m, o, p, Optogenetic inhibition of callosal terminals does not change the number of perseverative errors in Syn-eNpHR mice (n = 5 mice, l-m) compared to Syn-mCherry controls (n = 4 mice, l-m; two-way ANOVA (task day × virus); interaction: F(2,14) = 1.933, P = 0.1814), and has no effect on random errors (two-way ANOVA (task day × virus); interaction: F(2,14) = 0.3789, P = 0.6914). l, m, Light delivery does not affect the number of perseverative (Day 1 to Day 2: post hoc t(3) = 1.464, P = 0.7183; Day 1 to Day 3: post hoc t(3) = 0.8783, P > 0.9999; Day 2 to Day 3: post hoc t(3) = 0.4804, P > 0.9999) or random (Day 1 to Day 2: post hoc t(3) = 1.732, P = 0.5451; Day 2 to Day 3: post hoc t(3) = 1.732, P = 0.5451) errors in controls across days. o, p, Nonspecific optogenetic inhibition of all callosal projections does not affect the number of perseverative (n = 5 mice; Day 1 to Day 2: post hoc t(4) = 0.6882, P > 0.9999; Day 1 to Day 3: post hoc t(4) = 2.108, P = 0.3081; Day 2 to Day 3: post hoc t(4) = 1.121, P = 0.9752) or random (Day 1 to Day 2: post hoc t(4) = 0.4082, P > 0.9999; Day 1 to Day 3: post hoc t(4) = 1.000, P > 0.9999; Day 2 to Day 3: post hoc t(4) = 0.000, P > 0.9999) errors in Syn-eNpHR mice across days. Two-way ANOVA followed by Bonferroni post hoc comparisons was used. **P < 0.01, ****P < 0.0001; scale bars, 50 μm./p>

0.9999; Day 3: post hoc t(24) = 1.763, P = 0.2717). g–i, We also re-analyzed the TEMPO data collected from PV-Cre Ai14 mice injected in one mPFC with DIO-eNpHR to identify specific times when optogenetic inhibition of callosal PV+ projections disrupts gamma synchrony. We measured the change in gamma synchrony (calculated in 1 s windows for various time points relative to behavioral events) between Day 1 (control) and Day 2 (optogenetic inhibition) for both errors (filled circles) and correct choices (open circles) during the first 5 RS trials in PV-Cre mice expressing DIO-eNpHR. g, This change in gamma synchrony (change = Day 2 – Day 1) is not significantly different for error vs. correct trials around the time of trial start (n = 5 mice; two-way ANOVA (time point × error vs. correct); interaction: F(15,64) = 1.642, P = 0.0874). h, This change in gamma synchrony is more negative for error vs. correct trials around the start of digging (n = 5 mice; two-way ANOVA (time point × error vs. correct); interaction: F(15,64) = 1.931, P = 0.0360). i, This change in gamma synchrony is more negative for error vs. correct trials around the end of digging (n = 5 mice; two-way ANOVA (time point × error vs. correct); interaction: F(10,44) = 2.096, P = 0.0454). j, Example traces of left (L) and right (R) Ace2n-4AA-mNeon (Ace-mNeon) traces (green and red, respectively), from 0–10 s after dig start on RS correct and error trials in a DIO-eNpHR-expressing mouse and k, zoomed in on the period 6–7 s after dig start. l, The time course of gamma synchrony (correlation values calculated from filtered and cleaned Ace-mNeon signals) from 0–10 s after dig start for these two example trials. Two-way ANOVA followed by Bonferroni post hoc comparisons was used. Data were expressed as mean ± s.e.m. *P < 0.05, **P < 0.01./p>

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