were observed to gather around MNTB at P7, a phenomenon different from the wild type, and an increase in the number of GFAP-positive astrocytes that invaded inside MNTB was observed at P14. The result suggests that abnormal synaptic pruning may occur in calyx-MNTB synapses of Hapln4-KO (unpublished results).Further elucidation of the roles of HAPLN4 may highlight the developmental and physiological as well as pathological importance of PNN formation including psychiatric disorders. 1) Fawcett JW, Oohashi T, Pizzorusso T. The roles of perineuronal nets and the perinodal extracellular matrix in neuronal function. Nat. Rev. Neurosci. (20): 2) Oohashi T, Edamatsu M, Bekku Y, Carulli D. The hyaluronan and proteoglycan link proteins: Organizers of the brain extracellular matrix and key 3) Gandal MJ, Zhang P, Hadjimichael E, Walker RL, Chen C, Liu S, Won H, van Bakel H, Varghese M, Wang Y, Shieh AW, Haney J, Parhami S, Belmont J, Kim M, Moran Losada P, Khan Z, Mleczko J, Xia Y, Dai R, Wang D, Yang YT, Xu M, Fish K, Hof PR, Warrell J, Fitzgerald D, White K, Jaffe AE; PsychENCODE Consortium, Peters MA, Gerstein M, Liu C, Iakoucheva LM, Pinto D, Geschwind DH. Transcriptome-wide isoform-level dysregulation in ASD, schizophrenia, and bipolar disorder. Science. (362): eaat8127, 2018 4) Ji Y, Zhang X, Wang Z, Qin W, Liu H, Xue K, Tang J, Xu Q, Zhu D, Liu F, Yu C. Genes associated with gray matter volume alterations in schizophrenia. 5) Cicanic M, Edamatsu M, Bekku Y, Vorisek I, Oohashi T, Vargova L. A deficiency of the link protein Bral2 affects the size of the extracellular space in the 6) Sucha P, Chmelova M, Kamenicka M, Bochin M, Oohashi T, Vargova L. The effect of Hapln4 link protein deficiency on extracellular space diffusion 7) Pineda-Cirera L, Cabana-Domínguez J, Lee PH, Fernàndez-Castillo N, Cormand B. Identification of genetic variants influencing methylation in brain 8) Popelář J, Díaz Gómez M, Lindovský J, Rybalko N, Burianová J, Oohashi T, Syka J. The absence of brain-specific link protein Bral2 in perineuronal nets 9) Edamatsu M, Miyano R, Fujikawa A, Fujii F, Hori T, Sakaba T, Oohashi T. Hapln4/Bral2 is a selective regulator for formation and transmission of 451-465, 2019molecules for neuronal function and plasticity. Exp. Neurol. (274): 134-144, 2015Neuroimage. (225): 117526, 2021thalamus of aged mice. J. Neurosci. Res. (96): 313-327, 2018parameters and perineuronal nets in the auditory system during aging. Neurochem. Res. (45): 68-82, 2020with pleiotropic effects on psychiatric disorders. Prog. Neuropsychopharmacol. Biol. Psychiatry. (113): 110454, 2022hampers auditory temporal resolution and neural adaptation in mice. Physiol Res. (66): 867-880, 2017GABAergic synapses between Purkinje and deep cerebellar nuclei neurons. J. Neurochem. (147): 748-763, 201810) Nojima K, Miyazaki H, Hori T, Vargova L, Oohashi T. Assessment of possible contributions of Hyaluronan and Proteoglycan Binding Link Protein 4 to differential perineuronal net formation at the calyx of Held. Front. Cell. Dev. Biol. (9): 730550, 202111) Hori T, Takamori S. Physiological perspectives on molecular mechanisms and regulation of vesicular glutamate transport: Lessons from calyx of Held synapses. Front. Cell. Neurosci. (15): 811892, 2022pipette could visualize the morphology of presynaptic terminal (Figure 2B). Electrophysiological analysis showed that MNTB neurons receive multiple input in Hapln4-KO, whereas the neurons receive large single input in wild-type. The result suggests that Calyx-MNTB synapse formation during early postnatal development could be impaired in Hapln4-KO. To further investigate the underlying mechanism, we assessed the contributions of glial cells in synaptic pruning at postnatal days (P) 7 and 14, i.e., early and late stage of calyx-MNTB synapses and PNN formation. We found that GFAP-positive astrocytes References73
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