Learned spatiotemporal sequence recognition and prediction in primary visual cortex

Transcription

1 Supplementary Materials for Learned spatiotemporal sequene reognition and predition in primary visual ortex Jeffrey P. Gavornik and Mark F. Bear Howard Hughes Medial Institute Piower Institute for Learning and Memory Department of Brain and Cognitive Sienes Massahusetts Institute of Tehnology Nature Neurosiene: doi:.38/nn.3683

2 Baseline Day 5 V V 4 V 3 V 2 μv V +V 2 +V 3 +V 4 Sequene magnitude = 4 d 5 ms 7 6 Baseline Day Element magnitude (μv) Response lateny (ms) Element Element 2 Element 3 Element 4 Element Element 2 Element 3 Element 4 e f PSD (db) Experimental DCBA 3 Gray PSD (% ) Experimental PSD (db) Control Frequeny () Frequeny () Supplementary Figure. Sequene learning inreases response magnitudes, dereases response latenies and modulates spetral power. (a) Diagram illustrating sequene magnitude quantifiation. () Traes show the average response evoked y the sequene on day (Baseline, gray) and after training (Day 5, lue) in the experimental group from Fig.. () There is a signifiant effet of training (2 way RM ANOVA, P<.) and eah sequene element, plotted individually, inreases relative to aseline (P<. for A,B and C, P<.5 for D). (d) Training also has a signifiant effet on response lateny (defined as the interval etween stimulus onset and maximal negativity, 2 way RM ANOVA, P<.) and there is a derease in response lateny for eah element (P<.). (e) Plots showing the power spetral density (PSD, estimated using Welh s method, transparent shading shows 95% onfidene intervals around means indiated y solid lines) averaged aross all animals in eah group from Fig. d (Exp. n=6, Ctrl. n=4) during periods of ative stimulus viewing (, DCBA, and 3 ) or during inter stimulus gray sreen periods (Gray). (f) To highlight differene etween groups, spetral power is plotted as a perent of the average power at eah frequeny. In oth experimental and sramled ontrol groups visual stimuli drive higher spetral powers than gray sreen at frequenies up to approximately. In the experimental mie, spetral power is on average higher while viewing than DCBA at low frequenies ut this relationship reverses in the high gamma range (note, data was reorded with a 6 noth filter). In ontrol mie the familiar and novel sequene spetrums overlap at oth high and low frequenies. In oth groups, sequenes presented with familiar timing result in more spetral power than a sequene presented with novel timing. Although preise interpretation is diffiult, these results are roadly onsistent with the idea that familiar and novel sequene either eliit or are modulated y different attentional states. PSD (%) Control Nature Neurosiene: doi:.38/nn.3683

3 DCBA Normalized spike rate Normalized spike rate Supplementary Figure 2. Sequene reall is evident in multi-unit spiking ativity. (a) Eah panel shows the multi-unit spike rasters (top) and peri-stimulus time histogram (PSTH, ottom) reorded on a single wire in a multi-hannel undle implanted in a single mouse. Responses are evoked y the sequenes (trained, lue) and DCBA (novel, red). Dashed vertial lines indiate stimulus element onset times. () Average PSTH for the 5 hannels shown in a. Before averaging, the PSTHs were normalized to the maxium spike rate evoked y either or DCBA on a per hannel asis. Triangles indiate stimulus element onset times. () Average normalized PSTH from 6 mie (33 total reording hannels). Nature Neurosiene: doi:.38/nn.3683

4 8 7 Day Day 2 Day 3 Day 4 Day 5,DCBA (2 ms) (5 ms) (3 ms) Trained timing Novel timing Sequene magnitude (μv) Trained timing Novel timing Novel timing Trained timing μv μv μv 2ms 5ms (2 ms) DCBA (2 ms) (5 ms) (3 ms) 3ms Supplementary Figure 3. Sequene learning with 2 ms element hold time. (a) To further verify that sequene learning is not speifi to a 5 ms element hold time, a ohort of mie (n=4) was trained with the sequene using a 2 ms hold time. On the 5th day, the mie were tested on sequenes and DCBA with the trained time and with using novel hold times (5 and 3 ms). () The largest response ours when the trained sequene is presented with the trained timing. () There is a signifiant effet of sequene (-way RM ANOVA, P=.3) and with the trained timing is signifiantly larger (P<.5) than DCBA or presented with novel timing. Nature Neurosiene: doi:.38/nn.3683

5 θ θ 2 2 θ Day 8 6 θ θ 2 Day 5 Day Day 2 Day 3 Day 4 Day 5 VEP mag (% Day ) SRP Training: CPP or Vehile θ SRP Testing: θ, θ CPP Vehile CPP Control ACBD d 6 5 ACBD 3 5ms 3 CPP Vehile Sequene magnitude (μv) μv 3ms CPP Vehile Supplementary Figure 4. CPP loks SRP ut not Sequene Learning. (a) Mie used in the CPP experiment (see Fig. 3) were reassigned after a 3 day washout period into CPP (n=6) and vehile ontrol (saline, n=9) groups and exposed to the SRP indution protool (desried previously in detail, see Frenkel et al. 26). Briefly, on eah of four training days the mie were injeted with CPP or saline (see methods) and exposed to a sinusoidal grating (.5 y/deg, % ontrast) oriented at θ and phase reversing at. On the 5th day, mie were shown phase reversing gratings oriented at the familiar and novel orientations (θ and θ 2 ). () Analysis of phase reversal loked VEP magnitudes shows a statistially signifiant interation etween treatment and reording day (days and 5, P<.). Post ho analysis reveals a signifiant differene etween CPP and ontrol treatments (P<.5) and a signifiant VEP inrease etween days and 5 for θ in the ontrol group (P<.), ut not in the CPP group. There is also a signifiant interation etween treatment and stimulus orientation on day 5 (P<.), a signifiant differene etween the CPP and saline groups (P<.5), and the response to the familiar orientation is signifiantly larger than to the novel orientation in the ontrol animals (P<.) ut not in the CPP animals. These findings are onsistent with previous reports that CPP loks SRP indution and provide a positive ontrol demonstrating the effiay of the CPP used in the sequene learning experiment. () Average voltage traes omparing the trained sequene,, with a novel sequene, ACBD, and novel timing, 3, on day 5 for the CPP (n=9) and vehile ontrol (n=6) groups from Figure 4. (d) There is not a signifiant differene etween CPP and ontrol groups on day 5 (2 way RM ANOVA, P=.752), and oth groups show the effet of sequene training ( way RM ANOVA, P<.5). Nature Neurosiene: doi:.38/nn.3683

6 Day Day 2 Day 3 Day 4 Day Day Day 2 Day 3 Sp.or Veh. Vehile, DCBA 3 Sopolamine Sequene magnitude (μv) Day 4 DCBA 5 Vehile Sopolamine 5ms 3 Sopolamine Vehile d Pre Sopolamine Post Sopolamine μv μv 3ms 5ms Supplementary Figure 5. Sequene learning requires musarini ACh reeptors. (a) Mie were trained on the sequene 25 minutes after daily injetion (i.p.) of either the musarini ACh reeptor loker sopolamine (3 mg/kg, n=5) or vehile (NaCl, n=5). On the fifth day (after drug washout) the mie were tested with, DCBA and 3. () Sequene magnitude quantifiation shows a signifiant interation etween treatment and reording day (P<.), a signifiant effet of treatment (P=.5). Signifiant within-treatment post-ho omparisons are indiated y * (P<.). Treatment effets on day are not signifiant (P=.429). () The effet of treatment and sequene on the 5th day of testing are shown. For quantifiation and statistis, see Fig. 3e. (d) To test the aute effets of sopolamine on evoked potententials, after testing on the 5th day the response to in mie from the vehile group (n=4) were reorded immediately efore and 25 minutes after sopolamine injetion. Evoked responses mathed those seen in d, indiating that musarini ACh reeptor lokade does not affet previously potentiated waveform morphology. Nature Neurosiene: doi:.38/nn.3683

7 Veh. Sp. Cortial surfae 2/3 Day Day 2 Day 3 Day 4 Day 5 Layer White matter Sopolamine or vehile infused loally into V via indwelling annulae prior to stimulus presentation, no drug infusion Treatment: Sopolamine, Left hemisphere Vehile, Right hemisphere Sopolamine treated hemisphere Vehile treated hemisphere Day Day 2 Day 3 d Day Veh Day 3 Sp Day 2 Veh Day 4 Sp Day 3 Veh Day 5 Sp μv μv 5ms e Sequene magnitude (μv) 5ms Sp. trt. hemi. Veh. trt. hemi. ns ns ns f Day 2 BCD magnitude (μv) Untreated Loal Systemi Sopolamine g BCD magnitude (μv) Untreated day magnitudes < 25 μv Day Day 2 ns Veh: Sp: Day Day 3 Day 2 Day 4 Day 3 Day Day BCD magnitude (μv) Sopolamine Loal and Sys. Untreated Supplementary Figure 6. Sequene potentiation is prevented y a loal lokade of musarini ACh reeptors in V. (a) Shemati illustration of loal infusion. () μl of sopolamine (.6 mg/ml) or vehile (NaCL) was infused loally in V via ilateral indwelling annulae prior to sequene exposure on days and 2. No infusions were performed on days 3 through 5, n=7 mie. () While sequene responses in the vehile treated hemispheres potentiated normally, there was no potentiation in the sopolamine treated hemispheres. (d) After drug washout, responses in the drug treated hemispheres potentiated normally as seen y omparing the waveforms evoked on days 3 through 5 in the sopolamine treated hemisphere with those reorded in the vehile treated hemisphere on days through 3. (e) There is a signifiant effet of exposure day on response magnitude (2 Way RM ANOVA, P<.) ut no signifiant effet of treatment (P =.3) or interation etween treatment and reording day (P=.277) when omparing the first three days of sequene exposure asent sopolamine in the two hemispheres, demonstrating that sopolamine infusion does not damage the ortex. Although V is visually response under the aute influene of sopolamine, the day response magnitude of sequene elements B, C, and D is smaller on average than under vehile in oth the systemi and loal infusion experiments. This raises the possiility that potentiation is loked not y antagonism of the ACh reeptors ut rather eause there is not enough evoked ortial ativity to drive plastiity. Panels (f g) show day 2 magnitude driven y elements B C D as a funtion of day magnitude in 2 mie treated with sopolamine (7 loal, 5 systemi) and in 35 untreated mie. Untreated mie with small day response (< 25 μv, n=8) show more potentiation than do the sopolamine treated mie (points aove the dashed unity line indiate potentiation). There is a signifiant effet of treatment (2 Way RM ANOVA, P<.), interation etween treatment and reording day (P<.), and effet of reording day on response magnitude in the untreated mie (P<.). This demonstrates that small responses an drive sequene potentiation and suggests that musarini ACh reeptors are neessary for this to our. Nature Neurosiene: doi:.38/nn.3683

8 4 P69 35 Day Day 2 Day 3,DCBA 3 Trained timing (5ms) Novel timing (3ms) Sequene magnitude (μv) Day Day 3 μv 5ms μv Day DCBA Day 3 3 3ms Supplementary Figure 7. Sequene learning in adult mie. (a) Most experiments were onduted in adolesent mie, with training ommening around post-natal day 3. To verify that sequene learning ours in adult mie, a ohort of mie (n=5) were implanted with reording eletrodes on day P5 (inoular V, layer 4). Training with the sequene (5 ms hold time) ommened on P69, well after the developmental ritial period. After viewing the trained sequene on experiment day 3, the mie were also shown the novel sequenes DCBA and 3. () As in young mie, the average visually evoked response to the sequene inreased aross training days and was larger than the response driven y novel sequenes. () There is a signifiant effet of training and sequene (one way RM ANOVA, P<.), the response to on day 3 is signifiantly larger than on day (Holm Šidák post ho test, P<.) and is signifiantly larger than either DCBA (P=.2) or 3 (P=.). All other pairwise post ho omparisons are not statistially signifiant. Nature Neurosiene: doi:.38/nn.3683