Sensory perturbations in visual, auditory and tactile perception are core problems in fragile X syndrome (FXS). In the Fmr1 knockout mouse model of FXS, the maturation of synapses and circuits during critical period (CP) development in the somatosensory cortex is delayed, but it is unclear how this contributes to altered tactile sensory processing in the mature CNS. Here we demonstrate that inhibiting the juvenile chloride co-transporter NKCC1, which contributes to altered chloride homeostasis in developing cortical neurons of FXS mice, rectifies the chloride imbalance in layer IV somatosensory cortex neurons and corrects the development of thalamocortical excitatory synapses during the CP. Comparison of protein abundances demonstrated that NKCC1 inhibition during early development caused a broad remodeling of the proteome in the barrel cortex. In addition, the abnormally large size of whisker-evoked cortical maps in adult Fmr1 knockout mice was corrected by rectifying the chloride imbalance during the early CP. These data demonstrate that correcting the disrupted driving force through GABAA receptors during the CP in cortical neurons restores their synaptic development, has an unexpectedly large effect on differentially expressed proteins, and produces a long-lasting correction of somatosensory circuit function in FXS mice.
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Penagarikano O, Mulle JG, Warren ST. The pathophysiology of fragile x syndrome. Annu Rev Genome Hum Genet. 2007;8:109–29.
Darnell JC, Van Driesche SJ, Zhang C, Hung KY, Mele A, Fraser CE, et al. FMRP stalls ribosomal translocation on mRNAs linked to synaptic function and autism. Cell. 2011;146:247–61.
Contractor A, Klyachko VA, Portera-Cailliau C. Altered neuronal and circuit excitability in fragile X syndrome. Neuron. 2015;87:699–715.
Kau AS, Meyer WA, Kaufmann WE. Early development in males with fragile X syndrome: a review of the literature. Microsc Res Tech. 2002;57:174–8.
Baranek GT, Foster LG, Berkson G. Tactile defensiveness and stereotyped behaviors. Am J Occup Ther. 1997;51:91–95.
Cascio CJ. Somatosensory processing in neurodevelopmental disorders. J Neurodev Disord. 2010;2:62–69.
Rotschafer S, Razak K. Altered auditory processing in a mouse model of fragile X syndrome. Brain Res. 2013;1506:12–24.
Arnett MT, Herman DH, McGee AW. Deficits in tactile learning in a mouse model of fragile X syndrome. PLoS ONE. 2014;9:e109116.
Zhang Y, Bonnan A, Bony G, Ferezou I, Pietropaolo S, Ginger M, et al. Dendritic channelopathies contribute to neocortical and sensory hyperexcitability in Fmr1(-/y) mice. Nat Neurosci. 2014;17:1701–9.
He CX, Cantu DA, Mantri SS, Zeiger WA, Goel A, Portera-Cailliau C. Tactile defensiveness and impaired adaptation of neuronal activity in the Fmr1 knock-out mouse model of autism. J Neurosci. 2017;37:6475–87.
Goncalves JT, Anstey JE, Golshani P, Portera-Cailliau C. Circuit level defects in the developing neocortex of fragile X mice. Nat Neurosci. 2013;16:903–9.
Harlow EG, Till SM, Russell TA, Wijetunge LS, Kind P, Contractor A. Critical period plasticity is disrupted in the barrel cortex of FMR1 knockout mice. Neuron. 2010;65:385–98.
Cruz-Martin A, Crespo M, Portera-Cailliau C. Delayed stabilization of dendritic spines in fragile X mice. J Neurosci. 2010;30:7793–803.
Bureau I, Shepherd GM, Svoboda K. Circuit and plasticity defects in the developing somatosensory cortex of FMR1 knock-out mice. J Neurosci. 2008;28:5178–88.
Patel AB, Hays SA, Bureau I, Huber KM, Gibson JR. A target cell-specific role for presynaptic Fmr1 in regulating glutamate release onto neocortical fast-spiking inhibitory neurons. J Neurosci. 2013;33:2593–604.
Crair MC, Malenka RC. A critical period for long-term potentiation at thalamocortical synapses. Nature. 1995;375:325–8.
Inan M, Crair MC. Development of cortical maps: perspectives from the barrel cortex. Neuroscientist. 2007;13:49–61.
He Q, Nomura T, Xu J, Contractor A. The developmental switch in GABA polarity is delayed in fragile X mice. J Neurosci. 2014;34:446–50.
Wang DD, Kriegstein AR. Defining the role of GABA in cortical development. J Physiol. 2009;587(Pt 9):1873–9.
Ben-Ari Y, Gaiarsa JL, Tyzio R, Khazipov R. GABA: a pioneer transmitter that excites immature neurons and generates primitive oscillations. Physiol Rev. 2007;87:1215–84.
Kaila K, Price TJ, Payne JA, Puskarjov M, Voipio J. Cation-chloride cotransporters in neuronal development, plasticity and disease. Nat Rev Neurosci. 2014;15:637–54.
Chancey JH, Adlaf EW, Sapp MC, Pugh PC, Wadiche JI, Overstreet-Wadiche LS. GABA depolarization is required for experience-dependent synapse unsilencing in adult-born neurons. J Neurosci. 2013;33:6614–22.
Oh WC, Lutzu S, Castillo PE, Kwon HB. De novo synaptogenesis induced by GABA in the developing mouse cortex. Science (New York). 2016;353:1037–40.
Deidda G, Allegra M, Cerri C, Naskar S, Bony G, Zunino G, et al. Early depolarizing GABA controls critical-period plasticity in the rat visual cortex. Nat Neurosci. 2015;18:87–96.
Cleary RT, Sun H, Huynh T, Manning SM, Li Y, Rotenberg A, et al. Bumetanide enhances phenobarbital efficacy in a rat model of hypoxic neonatal seizures. PLoS ONE. 2013;8:e57148.
Loscher W, Puskarjov M, Kaila K. Cation-chloride cotransporters NKCC1 and KCC2 as potential targets for novel antiepileptic and antiepileptogenic treatments. Neuropharmacology. 2013;69:62–74.
Isaac JT, Crair MC, Nicoll RA, Malenka RC. Silent synapses during development of thalamocortical inputs. Neuron. 1997;18:269–80.
Tang B, Wang T, Wan H, Han L, Qin X, Zhang Y, et al. Fmr1 deficiency promotes age-dependent alterations in the cortical synaptic proteome. Proc Natl Acad Sci USA. 2015;112:E4697–4706.
Li Z, Adams RM, Chourey K, Hurst GB, Hettich RL, Pan C. Systematic comparison of label-free, metabolic labeling, and isobaric chemical labeling for quantitative proteomics on LTQ Orbitrap Velos. J Proteome Res. 2012;11:1582–90.
Butko MT, Savas JN, Friedman B, Delahunty C, Ebner F, Yates JR 3rd, et al. In vivo quantitative proteomics of somatosensory cortical synapses shows which protein levels are modulated by sensory deprivation. Proc Natl Acad Sci USA. 2013;110:E726–735.
Mi H, Poudel S, Muruganujan A, Casagrande JT, Thomas PD. PANTHER version 10: expanded protein families and functions, and analysis tools. Nucleic Acids Res. 2016;44(D1):D336–342.
Deidda G, Parrini M, Naskar S, Bozarth IF, Contestabile A, Cancedda L. Reversing excitatory GABAAR signaling restores synaptic plasticity and memory in a mouse model of Down syndrome. Nat Med. 2015;21:318–26.
Tang X, Kim J, Zhou L, Wengert E, Zhang L, Wu Z, et al. KCC2 rescues functional deficits in human neurons derived from patients with Rett syndrome. Proc Natl Acad Sci USA. 2016;113:751–6.
Banerjee A, Rikhye RV, Breton-Provencher V, Tang X, Li C, Li K, et al. Jointly reduced inhibition and excitation underlies circuit-wide changes in cortical processing in Rett syndrome. Proc Natl Acad Sci USA. 2016;113:E7287–E7296.
Li Y, Cleary R, Kellogg M, Soul JS, Berry GT, Jensen FE. Sensitive isotope dilution liquid chromatography/tandem mass spectrometry method for quantitative analysis of bumetanide in serum and brain tissue. J Chromatogr B. 2011;879:998–1002.
Topfer M, Tollner K, Brandt C, Twele F, Broer S, Loscher W. Consequences of inhibition of bumetanide metabolism in rodents on brain penetration and effects of bumetanide in chronic models of epilepsy. Eur J Neurosci. 2014;39:673–87.
Owens DF, Kriegstein AR. Is there more to GABA than synaptic inhibition? Nat Rev Neurosci. 2002;3:715–27.
Lombardi LM, Baker SA, Zoghbi HY. MECP2 disorders: from the clinic to mice and back. J Clin Invest. 2015;125:2914–23.
Benowitz LI, Routtenberg A. GAP-43: an intrinsic determinant of neuronal development and plasticity. Trends Neurosci. 1997;20:84–91.
Itami C, Kimura F, Nakamura S. Brain-derived neurotrophic factor regulates the maturation of layer 4 fast-spiking cells after the second postnatal week in the developing barrel cortex. J Neurosci. 2007;27:2241–52.
Wen TH, Afroz S, Reinhard SM, Palacios AR, Tapia K, Binder DK et al. Genetic Reduction of Matrix Metalloproteinase-9 Promotes Formation of Perineuronal Nets Around Parvalbumin-Expressing Interneurons and Normalizes Auditory Cortex Responses in Developing Fmr1 Knock-Out Mice. Cereb Cortex. 2017;13:1–14.
Nomura T, Musial TF, Marshall JJ, Zhu Y, Remmers CL, Xu J, et al. Delayed Maturation of Fast-Spiking Interneurons Is Rectified by Activation of the TrkB Receptor in the Mouse Model of Fragile X Syndrome. J Neurosci. 2017;37:11298–310.
Liu T, Wan RP, Tang LJ, Liu SJ, Li HJ, Zhao QH, et al. A MicroRNA Profile in Fmr1 Knockout Mice Reveals MicroRNA Expression Alterations with Possible Roles in Fragile X Syndrome. Mol Neurobiol. 2015;51:1053–63.
Lippi G, Fernandes CC, Ewell LA, John D, Romoli B, Curia G et al. MicroRNA-101 regulates multiple developmental programs to constrain excitation in adult neural networks. Neuron. 2016;92:1337–1351.
Nimchinsky EA, Oberlander AM, Svoboda K. Abnormal development of dendritic spines in FMR1 knock-out mice. J Neurosci. 2001;21:5139–46.
Meredith RM. Sensitive and critical periods during neurotypical and aberrant neurodevelopment: a framework for neurodevelopmental disorders. Neurosci Biobehav Rev. 2015;50:180–8.
Dansie LE, Phommahaxay K, Okusanya AG, Uwadia J, Huang M, Rotschafer SE, et al. Long-lasting effects of minocycline on behavior in young but not adult Fragile X mice. Neuroscience. 2013;246:186–98.
Tyzio R, Nardou R, Ferrari DC, Tsintsadze T, Shahrokhi A, Eftekhari S, et al. Oxytocin-mediated GABA inhibition during delivery attenuates autism pathogenesis in rodent offspring. Science. 2014;343:675–9.
Marguet SL, Le-Schulte VTQ, Merseburg A, Neu A, Eichler R, Jakovcevski I, et al. Treatment during a vulnerable developmental period rescues a genetic epilepsy. Nat Med. 2015;21:1436–44.
Feldmeyer D, Egger V, Lubke J, Sakmann B. Reliable synaptic connections between pairs of excitatory layer 4 neurones within a single ‘barrel’ of developing rat somatosensory cortex. J Physiol. 1999;521:169–90.
Daw MI, Ashby MC, Isaac JT. Coordinated developmental recruitment of latent fast spiking interneurons in layer IV barrel cortex. Nat Neurosci. 2007;10:453–61.
Feldman DE, Nicoll RA, Malenka RC, Isaac JT. Long-term depression at thalamocortical synapses in developing rat somatosensory cortex. Neuron. 1998;21:347–57.
Mostany R, Portera-Cailliau C. A method for 2-photon imaging of blood flow in the neocortex through a cranial window. J Vis Exp. 2008; pii 678.
Holtmaat A, de Paola V, Wilbrecht L, Trachtenberg JT, Svoboda K, Portera-Cailliau C. Imaging neocortical neurons through a chronic cranial window. Cold Spring Harb Protoc. 2012;2012:694–701.
McAlister GC, Nusinow DP, Jedrychowski MP, Wuhr M, Huttlin EL, Erickson BK, et al. MultiNotch MS3 enables accurate, sensitive, and multiplexed detection of differential expression across cancer cell line proteomes. Anal Chem. 2014;86:7150–8.
He L, Diedrich J, Chu YY, Yates JR 3rd. Extracting accurate precursor information for tandem mass spectra by RawConverter. Anal Chem. 2015;87:11361–7.
Park SK, Aslanian A, McClatchy DB, Han X, Shah H, Singh M, et al. Census 2: isobaric labeling data analysis. Bioinformatics. 2014;30:2208–9.
We thank Dr. Máté Marosi for help with the intrinsic signal imaging experiments. This work was funded by grants from NIH/NIMH (1R21MH104808) to AC, DOD USAMRMC W81XWH-14-1-0433 and a John Merck Fund award to AC and CP-C, the Simons Foundation (SFARI Award 295438) and NIH/NICHD (5R01HD054453) to CP-C. JNS was supported by NIH/NIDCD (R00DC-013805) and The Hartwell Foundation. QH was supported by a fellowship from the FRAXA Research Foundation. Data from LC-MS experiments were uploaded to a public repository ftp://MSV000081526@massive.ucsd.edu
QH, EA, SNS, CP performed experiments, analyzed data, and contributed to writing of the manuscript. JNS, CP-C and AC provided direction for the study, analyzed data, and wrote the manuscript. CP-C and AC generated funding to support the experiments. All authors contributed to the conception and design of the experiments.
Conflict of interest
The authors declare that they have no conflict of interest.
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He, Q., Arroyo, E.D., Smukowski, S.N. et al. Critical period inhibition of NKCC1 rectifies synapse plasticity in the somatosensory cortex and restores adult tactile response maps in fragile X mice. Mol Psychiatry 24, 1732–1747 (2019). https://doi.org/10.1038/s41380-018-0048-y
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