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Nature volume 617、pages 548–554 (2023)この記事を引用

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内側前頭前皮質内の活動パターンの変化により、げっ歯類、非ヒト霊長類、およびヒトは、環境の変化に適応するために行動を更新することができます（たとえば、認知タスク中）。内側前頭前野のパルブアルブミン発現抑制性ニューロンは、ルールシフト課題中に新しい戦略を学習するのに重要である6,7,8が、前頭前野ネットワークのダイナミクスを課題に関連した活動パターンの維持から更新に切り替える回路相互作用は不明のままである。今回我々は、パルブアルブミン発現ニューロンを結び付ける機構、新しい脳梁抑制結合、および課題表現の変化について説明する。すべての脳梁投射を非特異的に阻害しても、マウスのルールシフトの学習は妨げられず、活動パターンの進化も阻害されませんが、パルブアルブミン発現ニューロンの脳梁投射のみを選択的に阻害すると、ルールシフト学習が損なわれ、学習に必要なガンマ周波数活動が非同期化します8。通常、ルールシフト学習に伴う前頭前野の活動パターンの再編成を抑制します。この解離は、脳梁のパルブアルブミン発現投影が、ガンマ同期を伝達し、以前に確立された神経表現を維持する他の脳梁入力の能力をゲートすることによって、前頭前回路の動作モードを維持から更新にどのように切り替えるかを明らかにする。したがって、パルブアルブミン発現ニューロンに由来する脳梁投射は、統合失調症および関連症状に関与していると考えられている行動の柔軟性とガンマ線同調性の欠損を理解し、修正するための重要な回路位置を表します9,10。

生物は環境の変化に適応するために行動戦略を継続的に更新しなければなりません。時代遅れの戦略に対する不適切な固執は、統合失調症や双極性障害などの症状の特徴であり、古典的にはウィスコンシンカードソーティングタスク 11 (WCST) に現れます。前頭前皮質は、規則または目標表現 12、13、14 の能動的な維持、これらの表現の適応的ゲート制御 15、および他のものとの広範な相互接続を介した感覚処理のトップダウンバイアスを提供することにより、このような柔軟な認知制御を担当していることが十分に文書化されています。脳領域16、17。研究によると、マウスが「ルールシフト」課題を実行するには、内側前頭前皮質（mPFC）内で正常なパルブアルブミン発現（PV）介在ニューロン機能が必要であることが示されている。この課題には、WCSTと同様に、手がかりのないルールの変更を特定し、新しいルールを学習することが含まれる以前は試験の結果とは無関係だった手がかりを使用するルール6,7。さらに、PV 介在ニューロンは、ガンマ周波数 (約 40 Hz) 範囲で同期したリズミカルな活動を生成する上で重要な役割を果たしています 18,19。実際、ルールシフト課題中、左右の mPFC の PV 介在ニューロン間のガンマ周波数活動の同期性は、エラー試行の後、つまり、以前に学習したルールが古くなったというフィードバックをマウスが受け取った後、光遺伝学的にこの同期を破壊することで増加します。忍耐力を引き起こす8. それにもかかわらず、回路（シナプス接続）、ネットワークダイナミクス（半球間のガンマ同期）、および神経表現（活動パターンのタスク依存の変化）の間の基本的な関係は不明のままです。

ガンマ同期は一般に、皮質における長距離通信の主な形式である興奮性シナプス 20 によって領域全体に伝達されると考えられています。しかし、我々は、mPFC21のPVニューロンに由来する長距離γ-アミノ酪酸放出（GABA作動性）結合に関する最近の記述によって示唆された代替仮説を調査した。具体的には、mPFCのPV発現ニューロンが対側のmPFCで脳梁のGABA作動性シナプスを生じさせることを初めて実証しました（図1）。 PV-creマウスの1つのmPFCにAAV-EF1α-DIO-ChR2-eYFPを注射することによってこの解剖学的リンクを特定し、対側PFC、特に深層5および6のウイルス標識されたPV末端を観察しました（図1a）。この脳梁のPV投影とそのレシピエントニューロンを特徴付けるために、対側のmPFCで記録を実行しました（図1b）。我々は、脳梁のPV投影が錐体ニューロンを神経支配する（形態と非高速スパイク生理学に基づいて特定、75個中31個が接続）が、高速スパイクニューロン（18個中0個が接続）を神経支配しないことを発見した（図1d）。 PV末端光遺伝学的刺激のリズミカルな列は、ChR2陰性錐体ニューロンのタイムロック抑制性シナプス後電位（IPSP）を誘発しました（図1e）。これは、グルタミン酸受容体拮抗薬6-シアノ-7-ニトロキノキサリン-2,3によってブロックされませんでした。 -ジオン二ナトリウム塩 (CNQX) (10 μM) および d-2-アミノ-5-ホスホペンタン酸 (APV) (50 μM) でしたが、A 型 γ-アミノ酪酸 (GABAA) 受容体アンタゴニストの浴適用により完全に消失しました。ガバジン（10μM;図1e、f）。脳梁 mPFC PV シナプスの特定の標的をさらに特徴付けるために、mPFC の 4 つの下流標的に蛍光色素結合コレラ毒素サブユニット B (CTb) を注入し、対側 mPFC (つまり、対側) に投射する逆行性標識 mPFC ニューロンから記録しました。記録が行われた場所およびAAV-DIO-ChR2注射の同側）、背側線条体、視床内側（MD）背側（MD）、または側坐核（NAc）（拡張データ図1b〜k）。グルタミン酸作動性アンタゴニスト（20 μM 6,7-ジニトロキノキサリン-2,3-ジオン（DNQX）および50 μM APV）の存在下で、単一の5ミリ秒の光パルスで脳梁PV+末端を光遺伝学的に活性化した後、タイムロックされたIPSPを観察しました。 MD投影錐体ニューロン22個中22個と比較して、脳梁投影錐体ニューロン18個中0個、側坐筋投影錐体ニューロン21個中0個、背側線条体投影錐体ニューロン24個中7個であった。

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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