Early hyperactivity and precocious maturation of corticostriatal circuits in Shank3B−/− mice

Journal name:
Nature Neuroscience
Volume:
19,
Pages:
716–724
Year published:
DOI:
doi:10.1038/nn.4260
Received
Accepted
Published online

Abstract

Some autistic individuals exhibit abnormal development of the caudate nucleus and associative cortical areas, suggesting potential dysfunction of cortico-basal ganglia (BG) circuits. Using optogenetic and electrophysiological approaches in mice, we identified a narrow postnatal period that is characterized by extensive glutamatergic synaptogenesis in striatal spiny projection neurons (SPNs) and a concomitant increase in corticostriatal circuit activity. SPNs during early development have high intrinsic excitability and respond strongly to cortical afferents despite sparse excitatory inputs. As a result, striatum and corticostriatal connectivity are highly sensitive to acute and chronic changes in cortical activity, suggesting that early imbalances in cortical function alter BG development. Indeed, a mouse model of autism with deletions in Shank3 (Shank3B−/−) shows early cortical hyperactivity, which triggers increased SPN excitatory synapse and corticostriatal hyperconnectivity. These results indicate that there is a tight functional coupling between cortex and striatum during early postnatal development and suggest a potential common circuit dysfunction that is caused by cortical hyperactivity.

At a glance

Figures

  1. Rapid development of striatal SPN excitatory input in mice after [sim]P10.
    Figure 1: Rapid development of striatal SPN excitatory input in mice after ~P10.

    (a) Whole-cell voltage-clamp recordings in SPNs of dorsomedial striatum in acute brain slices of Rbp4-Cre;ChR2-YFPf/f mice and optogenetic fiber stimulation using whole-field illumination. Scale bar represents 1 mm. (b) AMPAR oEPSCs recorded in SPNs under voltage clamp (Vh = −70 mV) at different postnatal days in response to brief pulses of 473-nm laser light (blue rectangle). (c,d) Mean oEPSC peak amplitude ± s.e.m. recorded in SPNs at Vh = −70 mV from P6–15 (c) and at Vh = −20 mV from P14–30 (d). (e) Developmental progression of oEPSC amplitude values normalized to P30. Error bars represent normalized s.e.m. (f) Representative traces of AMPAR (gray) and NMDAR (black) oEPSCs recorded in the same SPN of Rbp4-Cre;ChR2-YFPf/wt at Vh = −70 mV and Vh = +40 mV, respectively. Red circle indicates time of NMDAR current amplitude analysis at 50-ms post light stimulus (blue rectangle). (g,h) Mean AMPAR (g) and NMDAR (h) oEPSC peak amplitude ± s.e.m. in P10–11 and P14–15 SPNs. (i) Mean AMPAR to NMDAR ratio ± s.e.m. for each SPN represented in g and h. (j) Average (solid line) ± s.e.m. (shaded region) NMDAR oEPSCs from cells represented in g and h. (k) Mean NMDAR oEPSC decay time constant (τ) ± s.e.m. of P10–11 and P14–15 SPNs. (l) Coronal brain slice of P12 mouse infected with AAV8-CAG-EGFP. Ctx, cortex; Str, striatum. Scale bar represents 1 mm. (m) Representative images of EGFP-expressing SPN dendrites at different postnatal days. Scale bar represents 10 μm. (n) Average dendritic spine density ± s.e.m. from infected SPNs at P8–24. ***P <0.001; ****P <0.0001.

  2. Correlated increase in cortical and striatal activity in vivo from P10 to P16.
    Figure 2: Correlated increase in cortical and striatal activity in vivo from P10 to P16.

    (a) Experimental diagram of in vivo recordings in a sagittal view of a mouse brain showing cortex (CTX) and striatum (STR). (b) Representative recordings of multi-unit activity in cortex (left) and striatum (right) at P10 and P14. (c) Median ± interquartile range of average FR of cortical units from P10–11 to P14–16. (d,e) Median AP burst frequency (d) and intra-burst frequency ± interquartile range (e) of cortical units shown in c. (f) Median ± interquartile range of average FR of striatal units at different developmental time points. (g,h) Median AP burst frequency (g) and intra-burst frequency ± interquartile range (h) of striatal units shown in f. *P <0.05; **P <0.01; ***P <0.001; ****P <0.0001.

  3. Corticostriatal coupling during early development.
    Figure 3: Corticostriatal coupling during early development.

    (a) Experimental diagram of in vivo recordings and extracranial optogenetic stimulation using 473-nm laser (blue) in Rbp4-Cre;ChR2-YFPf/f mice showing cortex (CTX) and striatum (STR). (b) Example raster plot (top) and 20-ms bin peri-stimulus time histogram (PSTH, bottom) of action potentials of a P11 striatal unit during optogenetic stimulation of cortex with a 10-Hz light pulse train (blue). Note the robust response to the first pulse of the train. (c) Raster plot (top) and 5 ms bin PSTH (bottom) of the unit shown in b in response to individual optogenetic pulses (blue). (d) PSTH (20-ms bin) representing firing rate of cortical neurons during ChR2 stimulation (blue) at P10–11 (black) and P14–16 (red). Shaded regions represent ± s.e.m. (e) PSTH (5-ms bin) of units shown in d in response to the first pulse of the stimulation train. Shaded regions represent ± s.e.m. (f) PSTH (20-ms bin) of firing rate of striatal neurons during cortical stimulation (blue) at P10–11 (black) and P14–16 (red). Shaded regions represent ± s.e.m. (g) PSTH (5-ms bin) of striatal neurons in response to the first pulse of the stimulation train. Shaded regions represent ± s.e.m. Note the presence of secondary peaks indicative of burst firing in response to single light pulses.

  4. Hyperexcitability of SPNs during early development.
    Figure 4: Hyperexcitability of SPNs during early development.

    (a) Example membrane responses to discreet current injection steps in SPNs of dorsomedial striatum. (b,c) Mean resting membrane potential ± s.e.m. (b) and mean spike threshold potential ± s.e.m. (c) of SPNs recorded at different postnatal periods. (d) Mean ± s.e.m. current-voltage (I-V) relationship in SPNs across development. Dashed lines represent linear fits to voltage steps to 10, 25 and 50 pA, whose slopes were used to determine the input resistance. (e) Current-firing rate (I-F) plot of SPNs across development. Error bars represent ± s.e.m. (f) Mean SPN rheobase current ± s.e.m. from P10–11 to P16–17. **P <0.01; ***P <0.001; ****P <0.0001.

  5. Precocious maturation of striatal glutamatergic inputs in Shank3B-/- SPNs.
    Figure 5: Precocious maturation of striatal glutamatergic inputs in Shank3B−/− SPNs.

    (a) Representative mEPSC recordings in SPNs of dorsomedial striatum of WT and Shank3B−/− mice at P14. (b,c) Cumulative distribution of amplitude (b) and inter-event interval (c) of mEPSCs recorded from SPNs of WT and Shank3B−/− littermates at P14. (d) Representative mEPSC recordings of WT and Shank3B−/− SPNs at P60. (e,f) Cumulative distribution of amplitude (e) and inter-event interval (f) of mEPSCs recorded from WT and Shank3B−/− littermates at P60. (g,h) Mean mEPSC frequency (g) and amplitude (h) ± s.e.m. of SPNs from WT and Shank3B−/− animals at different developmental time points. WT maturation was characterized by a continuous increase in mEPSC frequency throughout development, whereas Shank3B−/− showed a precocious maturation followed by an arrest in later stages. (i) Experimental diagram depicting whole-cell voltage-clamp recordings in SPNs of dorsomedial striatum in acute brain slices of Shank3B−/−;Rbp4-Cre;ChR2-YFPf/wt mice and optogenetic fiber stimulation using whole-field illumination. Scale bar represents 1 mm. (j) Representative average AMPAR and NMDAR oEPSCs from WT (black) and Shank3B−/− (red) SPNs. Blue rectangle represents 5-ms 473-nm light stimulation. (k) Mean AMPAR oEPSC peak amplitude ± s.e.m. in P14 WT and KO SPNs. (l) Mean AMPAR to NMDAR ratio ± s.e.m. for SPNs represented in k. *P <0.05; ***P <0.001; ns, P ≥ 0.95.

  6. Cortical hyperactivity in neonatal Shank3B-/- mice.
    Figure 6: Cortical hyperactivity in neonatal Shank3B−/− mice.

    (a) Experimental diagram of in vivo recordings in a sagittal view of a mouse brain showing cortex (CTX) and striatum (STR). (b,c) Representative recordings of multi-unit activity in cortex (b) and striatum (c) of WT and Shank3B−/− animals at P13–14. (d) Median ± interquartile range of average FR of cortical units from WT and Shank3B−/− mice. (e,f) Median frequency of AP bursts (e) and intra-burst frequency ± interquartile range (f) of cortical units shown in d. (g) Median ± interquartile range of average FR of striatal units from WT and Shank3B−/− mice. (h,i) Median frequency of AP bursts (h) and intra-burst firing rate ± interquartile range (i) of cortical units shown in e. *P <0.05; **P <0.01; ***P <0.001; ****P <0.0001.

  7. Elevated cortical activity during early development increases corticostriatal connectivity.
    Figure 7: Elevated cortical activity during early development increases corticostriatal connectivity.

    (a) Silencing of cortical interneuron output was achieved by injecting Cre-expressing adenovirus in the cortex of Slc32a1f/f animals at P4. (b) Local field potential (LFP) recordings from cortex of control and AAV-injected animals at P14 show epileptiform patterns of activity after VGAT deletion. (c) Spectrogram of LFPs shown in b. Scale bar represents 1 min. Color scale represents normalized power. (d) Example mEPSC recordings in SPNs of dorsomedial striatum of control and AAV-Cre–injected animals. (e,f) Cumulative distribution of mEPSC amplitude (e) and mEPSC inter-event interval values (f) for the total pool of mEPSCs recorded from control (black) and Cre-injected (red) littermates at P12–14. (g,h) Cell average mEPSC frequency (g) and amplitude ± s.e.m. (h) of SPNs from control and Cre-injected animals. (i) Schematic showing optogenetic cortical stimulation using extracranial implant of a low mass LED (blue) in Rbp4-Cre;ChR2f/f animals (top) and subsequent oEPSC measurements in SPNs in the ipsilateral (stimulated) and contralateral (control) hemispheres (bottom). (j) Example AMPAR oEPSCs recorded in SPNs located in dorsomedial striatum of the stimulated (ipsi, red) or opposite (contra, black) hemisphere in response to 5-ms pulses of 473-nm laser light (blue rectangle). (k) Mean oEPSC amplitude ± s.e.m. of control (contra) and stimulated (ipsi) SPNs. (l) Pair-wise comparison of average oEPSC amplitude in animals recorded in k. Error bars represent s.e.m. *P < 0.05; ***P <0.001.

  8. Early increase in corticostriatal drive in Shank3B-/- mice is a result of cortical hyperactivity.
    Figure 8: Early increase in corticostriatal drive in Shank3B−/− mice is a result of cortical hyperactivity.

    (a) Schematic representing bilateral injection of AAV8-DI-hM4Di into cortex of Shank3B−/−;Rbp4-Cre mice at P1–2 and bi-daily administration of CNO for 3 d before mEPSC recordings at P13–14. (b) Coronal brain slice of P13 Shank3B−/−;Rbp4-Cre mouse infected with AAV8-DI-hM4Di-mCherry. Ctx, cortex; Str, striatum. Scale bar represents 1 mm. (c) Example mEPSC recordings in SPNs of dorsomedial striatum of saline- or CNO-injected animals. (d,e) Cell average mEPSC frequency (d) and amplitude (e) ± s.e.m. of SPNs of saline- or CNO-injected animals ***P <0.001.

  9. Correlated oEPSC peak amplitude recorded at Vh=-70 and -20 mV.
    Supplementary Fig. 1: Correlated oEPSC peak amplitude recorded at Vh=−70 and −20 mV.

    Average oEPSC peak amplitude recorded in P14 SPNs of Rbp4-Cre;ChR2f/f mice at Vh=−70 and −20 mV. Note the linear relationship between oEPSCs recorded under the two conditions across a wide range of oEPSC amplitudes.

  10. Average cortical and striatal multi-unit responses to optogenetic stimulation of Rbp4-Cre;ChR2f/f mice.
    Supplementary Fig. 2: Average cortical and striatal multi-unit responses to optogenetic stimulation of Rbp4-Cre;ChR2f/f mice.

    (a) Peri-stimulus time histogram (5 ms bin) of APs fired by P10−11 (black) and P14−16 (red) cortical and (b) striatal units in response to extracranial optogenetic stimulation of Rbp4-Cre;ChR2f/f mice with 100 pulses of 473 nm light (blue rectangle). Shaded regions represent ± SEM. Arrows point to time bin with peak firing rate. Note the 5 ms shift in response latency of striatal units from P10−11 to P14−16.

  11. Similar presynaptic release properties in WT and Shank3B-/- SPNs across development.
    Supplementary Fig. 3: Similar presynaptic release properties in WT and Shank3B−/ SPNs across development.

    (a) Example traces of eEPSCs evoked in SPNs of P14 WT and Shank3B−/ mice in response to paired electrical pulses (P1 and P2) with 50 ms ISI. (b) Mean ± SEM ratio of eEPSC amplitude in response to the paired stimulation pulses (P2/P1) in P14 and (c) P9 WT and Shank3B−/ SPNs.

  12. Increased Rbp4+ excitatory input onto Shank3B-/- SPNs during early development.
    Supplementary Fig. 4: Increased Rbp4+ excitatory input onto Shank3B−/− SPNs during early development.

    (a) Experimental diagram depicting whole cell recordings in P14 SPNs of dorsomedial striatum in acute brain slices of Shank3B−/;Rbp4-Cre;ChR2-YFPf/wt mice and optogenetic fiber stimulation using whole field illumination (blue cone). Scale bar, 1 mm. (b) Representative traces of AMPAR oEPSCs recorded in SPNs of Shank3B+/ or Shank3B−/ mice under voltage clamp (Vh= -70 mV) in response to brief pulses of 473 nm laser light (blue rectangle). (c) Mean oEPSC peak amplitude ± SEM recorded in Shank3B+/ or Shank3B−/ SPNs. (d) Pair-wise comparison of average oEPSC amplitude in animals recorded in (c). Note that SPNs from Shank3B−/ animals have consistently larger oEPSC amplitude compared to SPNs from Shank3B+/ heterozygous littermates.

  13. Similar intrinsic excitability of WT and Shank3B-/- SPNs at P13-14.
    Supplementary Fig. 5: Similar intrinsic excitability of WT and Shank3B−/ SPNs at P13−14.

    (a) Mean ± SEM current-voltage (I-V) relationship in WT and KO SPNs. (b) Mean spike threshold potential (c) rheobase current (d) Vrest and (e) Current-firing rate (I-F) plot of WT and KO SPNs recorded at P13−14. Error bars represent ± SEM.

  14. Decreased locomotion of AAV-Cre injected vGATf/f mice.
    Supplementary Fig. 6: Decreased locomotion of AAV-Cre injected vGATf/f mice.

    (a) Heat map representing locomotion of vGATf/f mice (control) and vGATf/f littermates injected with AAV-Cre-EGFP (Cre) in an open chamber for 15 min. Color scale represents normalized time spent at each location. Scale bar, 10 cm. (b) Mean average velocity ± SEM of control and Cre injected vGATf/f animals. (c) Mean average time moving ± SEM of control and Cre injected vGATf/f animals. (d) Mean average total distance moved ± SEM of control and Cre injected vGATf/f animals.

  15. Similar PPR of eEPSC in SPNs of vGATf/f (control) and vGATf/f mice injected with AAV-Cre-EGFP.
    Supplementary Fig. 7: Similar PPR of eEPSC in SPNs of vGATf/f (control) and vGATf/f mice injected with AAV-Cre-EGFP.

    (a) Example traces of eEPSCs evoked in SPNs of control and Cre-injected littermates in response to paired electrical pulses (P1 and P2) with 50 ms ISI. (b) Mean ± SEM ratio of eEPSC amplitude in response to the paired stimulation pulses (P2/P1).

References

  1. Shepherd, G.M.G. Corticostriatal connectivity and its role in disease. Nat. Rev. Neurosci. 14, 278291 (2013).
  2. Oldenburg, I.A. & Sabatini, B.L. Antagonistic but not symmetric regulation of primary motor cortex by basal ganglia direct and indirect pathways. Neuron 86, 11741181 (2015).
  3. Difiglia, M., Pasik, P. & Pasik, T. Early postnatal development of the monkey neostriatum: a Golgi and ultrastructural study. J. Comp. Neurol. 190, 303331 (1980).
  4. Levine, M.S., Fisher, R.S., Hull, C.D. & Buchwald, N.A. Postnatal development of identified medium-sized caudate spiny neurons in the cat. Brain Res. 389, 4762 (1986).
  5. Tepper, J.M., Sharpe, N.A., Koós, T.Z. & Trent, F. Postnatal development of the rat neostriatum: electrophysiological, light- and electron-microscopic studies. Dev. Neurosci. 20, 125145 (1998).
  6. Kozorovitskiy, Y., Saunders, A., Johnson, C.a., Lowell, B.B. & Sabatini, B.L. Corrigendum: Recurrent network activity drives striatal synaptogenesis. Nature 489, 326326 (2012).
  7. Langen, M., Durston, S., Staal, W.G., Palmen, S.J.M.C. & van Engeland, H. Caudate nucleus is enlarged in high-functioning medication-naive subjects with autism. Biol. Psychiatry 62, 262266 (2007).
  8. Wolff, J.J., Hazlett, H.C., Lightbody, A.A., Reiss, A.L. & Piven, J. Repetitive and self-injurious behaviors: associations with caudate volume in autism and fragile X syndrome. J. Neurodev. Disord. 5, 12 (2013).
  9. Langen, M. et al. Changes in the developmental trajectories of striatum in autism. Biol. Psychiatry 66, 327333 (2009).
  10. Langen, M. et al. Changes in the development of striatum are involved in repetitive behavior in autism. Biol. Psychiatry 76, 405411 (2014).
  11. Lord, C., Cook, E.H., Leventhal, B.L. & Amaral, D.G. Autism spectrum disorders. Neuron 28, 355363 (2000).
  12. DeLong, M. & Wichmann, T. Changing views of basal ganglia circuits and circuit disorders. Clin. EEG Neurosci. 41, 6167 (2010).
  13. Abrahams, B.S. & Geschwind, D.H. Advances in autism genetics: on the threshold of a new neurobiology. Nat. Rev. Genet. 9, 341355 (2008).
  14. Jiang, Y.H. & Ehlers, M.D. Modeling autism by SHANK gene mutations in mice. Neuron 78, 827 (2013).
  15. Durand, C.M. et al. Mutations in the gene encoding the synaptic scaffolding protein SHANK3 are associated with autism spectrum disorders. Nat. Genet. 39, 2527 (2007).
  16. Gauthier, J. et al. Novel de novo SHANK3 mutation in autistic patients. Am. J. Med. Genet. B. Neuropsychiatr. Genet. 150B, 421424 (2009).
  17. Roussignol, G. et al. Shank expression is sufficient to induce functional dendritic spine synapses in aspiny neurons. J. Neurosci. 25, 35603570 (2005).
  18. Arons, M.H. et al. Autism-associated mutations in ProSAP2/Shank3 impair synaptic transmission and neurexin-neuroligin-mediated transsynaptic signaling. J. Neurosci. 32, 1496614978 (2012).
  19. Han, K. et al. SHANK3 overexpression causes manic-like behavior with unique pharmacogenetic properties. Nature 503, 7277 (2013).
  20. Verpelli, C. et al. Importance of Shank3 protein in regulating metabotropic glutamate receptor 5 (mGluR5) expression and signaling at synapses. J. Biol. Chem. 286, 3483934850 (2011).
  21. Bozdagi, O. et al. Haploinsufficiency of the autism-associated Shank3 gene leads to deficits in synaptic function, social interaction and social communication. Mol. Autism 1, 15 (2010).
  22. Yang, M. et al. Reduced excitatory neurotransmission and mild autism-relevant phenotypes in adolescent Shank3 null mutant mice. J. Neurosci. 32, 65256541 (2012).
  23. Peça, J. et al. Shank3 mutant mice display autistic-like behaviours and striatal dysfunction. Nature 472, 437442 (2011).
  24. Wang, X. et al. Synaptic dysfunction and abnormal behaviors in mice lacking major isoforms of Shank3. Hum. Mol. Genet. 20, 30933108 (2011).
  25. Harris, J.A. et al. Anatomical characterization of Cre driver mice for neural circuit mapping and manipulation. Front. Neural Circuits 8, 76 (2014).
  26. Petralia, R.S. et al. Selective acquisition of AMPA receptors over postnatal development suggests a molecular basis for silent synapses. Nat. Neurosci. 2, 3136 (1999).
  27. Busetto, G., Higley, M.J. & Sabatini, B.L. Developmental presence and disappearance of postsynaptically silent synapses on dendritic spines of rat layer 2/3 pyramidal neurons. J. Physiol. (Lond.) 586, 15191527 (2008).
  28. Khazipov, R. et al. Early motor activity drives spindle bursts in the developing somatosensory cortex. Nature 432, 758761 (2004).
  29. Carter, A.G., Soler-Llavina, G.J. & Sabatini, B.L. Timing and location of synaptic inputs determine modes of subthreshold integration in striatal medium spiny neurons. J. Neurosci. 27, 89678977 (2007).
  30. Choi, S. & Lovinger, D.M. Decreased probability of neurotransmitter release underlies striatal long-term depression and postnatal development of corticostriatal synapses. Proc. Natl. Acad. Sci. USA 94, 26652670 (1997).
  31. Moody, W.J. & Bosma, M.M. Ion channel development, spontaneous activity, and activity-dependent development in nerve and muscle cells. Physiol. Rev. 85, 883941 (2005).
  32. Kozorovitskiy, Y. et al. Neuromodulation of excitatory synaptogenesis in striatal development. eLife 4, e10111 (2015).
  33. Sohur, U.S., Padmanabhan, H.K., Kotchetkov, I.S., Menezes, J.R.L. & Macklis, J.D. Anatomic and molecular development of corticostriatal projection neurons in mice. Cereb. Cortex 24, 293303 (2014).
  34. Colonnese, M.T. et al. A conserved switch in sensory processing prepares developing neocortex for vision. Neuron 67, 480498 (2010).
  35. Kilb, W., Kirischuk, S. & Luhmann, H.J. Electrical activity patterns and the functional maturation of the neocortex. Eur. J. Neurosci. 34, 16771686 (2011).
  36. Etherington, S.J. & Williams, S.R. Postnatal development of intrinsic and synaptic properties transforms signaling in the layer 5 excitatory neural network of the visual cortex. J. Neurosci. 31, 95269537 (2011).
  37. Gertler, T.S., Chan, C.S. & Surmeier, D.J. Dichotomous anatomical properties of adult striatal medium spiny neurons. J. Neurosci. 28, 1081410824 (2008).
  38. Kirkby, L.A., Sack, G.S., Firl, A. & Feller, M.B. A role for correlated spontaneous activity in the assembly of neural circuits. Neuron 80, 11291144 (2013).
  39. Grabrucker, A.M. et al. Concerted action of zinc and ProSAP/Shank in synaptogenesis and synapse maturation. EMBO J. 30, 569581 (2011).
  40. Gogolla, N., Takesian, A.E., Feng, G., Fagiolini, M. & Hensch, T.K. Sensory integration in mouse insular cortex reflects GABA circuit maturation. Neuron 83, 894905 (2014).
  41. Martin, L.J. & Cork, L.C. The non-human primate striatum undergoes marked prolonged remodeling during postnatal development. Front. Cell. Neurosci. 8, 294 (2014).
  42. Lewine, J.D. et al. Magnetoencephalographic patterns of epileptiform activity in children with regressive autism spectrum disorders. Pediatrics 104, 405418 (1999).
  43. Gonçalves, J.T., Anstey, J.E., Golshani, P. & Portera-Cailliau, C. Circuit level defects in the developing neocortex of Fragile X mice. Nat. Neurosci. 16, 903909 (2013).
  44. Bateup, H.S. et al. Excitatory/inhibitory synaptic imbalance leads to hippocampal hyperexcitability in mouse models of tuberous sclerosis. Neuron 78, 510522 (2013).
  45. Peñagarikano, O. et al. Absence of CNTNAP2 leads to epilepsy, neuronal migration abnormalities, and core autism-related deficits. Cell 147, 235246 (2011).
  46. Han, S. et al. Autistic-like behaviour in Scn1a+/− mice and rescue by enhanced GABA-mediated neurotransmission. Nature 489, 385390 (2012).
  47. Wolff, J.J. et al. Longitudinal patterns of repetitive behavior in toddlers with autism. J. Child Psychol. Psychiatry 55, 945953 (2014).
  48. Goldman, S. et al. Motor stereotypies in children with autism and other developmental disorders. Dev. Med. Child Neurol. 51, 3038 (2009).
  49. Goldman, S. & Greene, P.E. Stereotypies in autism: a video demonstration of their clinical variability. Front. Integr. Neurosci. 6, 121 (2012).
  50. Harris, K.M., Mahone, E.M. & Singer, H.S. Nonautistic motor stereotypies: clinical features and longitudinal follow-up. Pediatr. Neurol. 38, 267272 (2008).
  51. Legéndy, C.R. & Salcman, M. Bursts and recurrences of bursts in the spike trains of spontaneously active striate cortex neurons. J. Neurophysiol. 53, 926939 (1985).

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Author information

Affiliations

  1. Department of Neurobiology, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA.

    • Rui T Peixoto,
    • Wengang Wang,
    • Donyell M Croney,
    • Yevgenia Kozorovitskiy &
    • Bernardo L Sabatini

Contributions

R.T.P. and B.L.S. conceived the study and wrote the manuscript. R.T.P. carried out in vivo recordings and analyzed the data. R.T.P., W.W. and Y.K. carried out in vitro slice recordings and R.T.P. analyzed the data. D.M.C. performed the behavioral experiments and dendritic spine imaging and analysis.

Competing financial interests

The authors declare no competing financial interests.

Corresponding author

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Supplementary information

Supplementary Figures

  1. Supplementary Figure 1: Correlated oEPSC peak amplitude recorded at Vh=−70 and −20 mV. (39 KB)

    Average oEPSC peak amplitude recorded in P14 SPNs of Rbp4-Cre;ChR2f/f mice at Vh=−70 and −20 mV. Note the linear relationship between oEPSCs recorded under the two conditions across a wide range of oEPSC amplitudes.

  2. Supplementary Figure 2: Average cortical and striatal multi-unit responses to optogenetic stimulation of Rbp4-Cre;ChR2f/f mice. (81 KB)

    (a) Peri-stimulus time histogram (5 ms bin) of APs fired by P10−11 (black) and P14−16 (red) cortical and (b) striatal units in response to extracranial optogenetic stimulation of Rbp4-Cre;ChR2f/f mice with 100 pulses of 473 nm light (blue rectangle). Shaded regions represent ± SEM. Arrows point to time bin with peak firing rate. Note the 5 ms shift in response latency of striatal units from P10−11 to P14−16.

  3. Supplementary Figure 3: Similar presynaptic release properties in WT and Shank3B−/ SPNs across development. (44 KB)

    (a) Example traces of eEPSCs evoked in SPNs of P14 WT and Shank3B−/ mice in response to paired electrical pulses (P1 and P2) with 50 ms ISI. (b) Mean ± SEM ratio of eEPSC amplitude in response to the paired stimulation pulses (P2/P1) in P14 and (c) P9 WT and Shank3B−/ SPNs.

  4. Supplementary Figure 4: Increased Rbp4+ excitatory input onto Shank3B−/− SPNs during early development. (73 KB)

    (a) Experimental diagram depicting whole cell recordings in P14 SPNs of dorsomedial striatum in acute brain slices of Shank3B−/;Rbp4-Cre;ChR2-YFPf/wt mice and optogenetic fiber stimulation using whole field illumination (blue cone). Scale bar, 1 mm. (b) Representative traces of AMPAR oEPSCs recorded in SPNs of Shank3B+/ or Shank3B−/ mice under voltage clamp (Vh= -70 mV) in response to brief pulses of 473 nm laser light (blue rectangle). (c) Mean oEPSC peak amplitude ± SEM recorded in Shank3B+/ or Shank3B−/ SPNs. (d) Pair-wise comparison of average oEPSC amplitude in animals recorded in (c). Note that SPNs from Shank3B−/ animals have consistently larger oEPSC amplitude compared to SPNs from Shank3B+/ heterozygous littermates.

  5. Supplementary Figure 5: Similar intrinsic excitability of WT and Shank3B−/ SPNs at P13−14. (80 KB)

    (a) Mean ± SEM current-voltage (I-V) relationship in WT and KO SPNs. (b) Mean spike threshold potential (c) rheobase current (d) Vrest and (e) Current-firing rate (I-F) plot of WT and KO SPNs recorded at P13−14. Error bars represent ± SEM.

  6. Supplementary Figure 6: Decreased locomotion of AAV-Cre injected vGATf/f mice. (63 KB)

    (a) Heat map representing locomotion of vGATf/f mice (control) and vGATf/f littermates injected with AAV-Cre-EGFP (Cre) in an open chamber for 15 min. Color scale represents normalized time spent at each location. Scale bar, 10 cm. (b) Mean average velocity ± SEM of control and Cre injected vGATf/f animals. (c) Mean average time moving ± SEM of control and Cre injected vGATf/f animals. (d) Mean average total distance moved ± SEM of control and Cre injected vGATf/f animals.

  7. Supplementary Figure 7: Similar PPR of eEPSC in SPNs of vGATf/f (control) and vGATf/f mice injected with AAV-Cre-EGFP. (53 KB)

    (a) Example traces of eEPSCs evoked in SPNs of control and Cre-injected littermates in response to paired electrical pulses (P1 and P2) with 50 ms ISI. (b) Mean ± SEM ratio of eEPSC amplitude in response to the paired stimulation pulses (P2/P1).

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