(A) Synaptic circuit organization in the adult cerebellum. Granule cells (GCs) send excitatory inputs to Purkinje cells (PCs) and molecular layer interneurons (MLIs). MLIs form inhibitory synapses …
Adult cerebellar organization after cell-autonomous elimination of neurons.
Cerebellar granule cell (GC)-specific and Purkinje cell (PC)-specific conditional Stxbp1 homozygous mice (ΔGC mice and ΔPC mice, respectively) are generated, and cerebellar structures are examined …
(A) Vesicular glutamate transporter (VGluT1) signal from GC axons, that is, parallel fibers (PFs), is significantly decreased in the molecular layer (ML) of the ΔGC mice (n = 6 images from three …
Excitatory AMPARs remain at spiny synapses without presynaptic terminals.
(A) The number of postsynaptic density (PSD)-95 puncta is reduced (n = 6 images from three mice each) and individual puncta is enlarged in molecular layer (ML) of the ΔGC mice (n = 1421 puncta in …
Excitatory AMPARs remain at shaft synapses without presynaptic terminals.
(A) Immunofluorescence showing marked reductions of calbindin (Calb)-labeled Purkinje cells (PCs) in the cerebellar cortex and of vesicular inhibitory amino acid transporter (VIAAT)-labeled …
Inhibitory GABAARs require presynaptic terminals in the deep cerebellar nuclei (DCN).
(A) Wiring diagram between molecular layer interneurons (MLIs) and PCs. Lower MLIs, also known as basket cells, innervate PC somata and surround the axon initial segment of PCs with the pinceau …
Inhibitory postsynaptic sites at Purkinje cell (PC) somas require presynaptic terminals.
(A) Adenoassociated viruses (AAVs) carrying GFP or Nrxn3α fused with GFP (Nrxn3α GFP) are injected into the inferior olivary nucleus (IONs) that project excitatory CFs to Purkinje cells (PCs). (B) …
Ectopic Nrxn3 expression in excitatory climbing fibers (CFs) recruits inhibitory GABARs.
(A–C) Electron micrographs showing CA1 region of wild-type (A) and Stxbp1fl/fl mice injected with AAV-synapsin promoter-driven (pSynapsin) Cre (B) and wild-type injected with AAV-pSynapsin Cre and …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Strain, strain background (Escherichia coli) | Putative Nos1 promoter | BACPAC Resources Center (BPRC) | BAC: RP24-84E3 | |
Strain, strain background (Escherichia coli) | Putative Grin3a promoter | BACPAC Resources Center (BPRC) | BAC: RP23-104D24 | |
Strain, strain background (Escherichia coli) | Putative Kit promoter | BACPAC Resources Center (BPRC) | BAC: RP23-232H18 | |
Strain, strain background (Mus musculus) | Wild-type (C57BL/6 J) | The Jackson Laboratory | Stock# 00064 | |
Strain, strain background (Mus musculus) | Mouse: Stxbp1lox/lox | Verhage et al., 2000 | N/A | |
Strain, strain background (Mus musculus) | Mouse: Tg(Gabra6-Cre) | MMRRC | ID# 015966-UCD | |
Strain, strain background (Mus musculus) | Mouse:Tg(Pcp2-Cre) | The Jackson Laboratory | Stock# 004146 | |
Cell line (Homo sapiens) | 293AAV | Cell Biolab | Cat#: AAV-100 | |
Antibody | anti- GABAAR α1(Rabbit polyclonal) | Frontier Inst. | Cat# GABAARa1-Rb-Af660 RRID:AB_2571571 | IHC(1 μg/ml) |
Antibody | anti-GluD2(Rabbit polyclonal) | Frontier Inst. | Cat# GluD2C-Rb-Af1200 RRID:AB_2571601 | IHC (1 μg/ml) |
Antibody | anti-panAMPAR(Guinea pig polyclonal) | Frontier Inst. | Cat# panAMPAR-GP-Af580, RRID:AB_2571610 | IHC (1 μg/ml)IEM (5 μg/ml) |
Antibody | anti-VGluT1(Guinea pig polyclonal) | Frontier Inst. | Cat# VGluT1-GP-Af570, RRID:AB_2571534 | IHC (1 μg/ml)IEM (10 μg/ml) |
Antibody | anti-VGluT2(Guinea pig polyclonal) | Frontier Inst. | Cat# VGluT2-GP-Af810, RRID:AB_2341096 | IHC (1 μg/ml) |
Antibody | anti-VIAAT(Guinea pig polyclonal) | Frontier Inst. | Cat# VGAT-GP-Af1000, RRID:AB_2571624 | IHC (1 μg/ml) |
Antibody | anti-PSD95(Guinea pig polyclonal) | Frontier Inst. | Cat# PSD95-GP-Af660, RRID:AB_2571539 | IHC (1 μg/ml) |
Antibody | anti-GFP(Goat polyclonal) | Frontier Inst. | Cat# GFP-Go-Af1480, RRID:AB_2571574 | IHC (1 μg/ml) |
Antibody | anti-calbindin(Goat polyclonal) | Frontier Inst. | Cat# Calbindin-Go-Af1040, RRID:AB_2532104 | IHC (1 μg/ml) |
Antibody | anti-MAP2(Goat polyclonal) | Frontier Inst. | Cat# MAP2-Go-Af860, RRID:AB_2571557 | IHC (1 μg/ml) |
Antibody | anti-Car8(Goat/Rabbit polyclonal) | Frontier Inst. | Cat# Car8-Go-Af780, RRID:AB_2571668Cat# Car8-Rb-Af330, RRID:AB_2571667 | IHC (1 μg/ml)IEM (10 μg/ml) |
Antibody | anti-Bassoon(Mouse monoclonal) | Enzo | Cat# SAP7F407, RRID:AB_2313990 | IHC (1 μg/ml)IEM (20 μg/ml) |
Antibody | anti-GABAAR γ2(Rabbit polyclonal) | Millipore | Cat#: AB5559, RRID: AB_11211236 | WB (1:2000) |
Antibody | anti-GARLH4(Rabbit polyclonal) | Yamasaki et al., 2017 | N/A | WB (0.1 μg/ml) |
Antibody | anti-GluA1(Rabbit polyclonal) | Millipore | Cat#: AB1504, RRID: AB_2113602 | WB (1:2000) |
Recombinant DNA reagent | pAAV-DJ(plasmid) | Cell Biolabs | VPK-410-DJ | |
Recombinant DNA reagent | pHelper(plasmid) | Cell Biolabs | VPK-410-DJ | |
Recombinant DNA reagent | pAAV-MCS(plasmid) | Cell Biolabs | VPK-410 | |
Recombinant DNA reagent | pAAV-Promoter less(plasmid) | This paper | N/A | pAAV-MCS (Cell Biolabs) |
Recombinant DNA reagent | Nrxn3alpha pXY-Asc(plasmid) | Horizon | Cat# MMM1013-202798372 | |
Sequence-based reagent | B1.Nrxn3a.For | This paper | PCR primers | GGGGACAAG TTTGTACAAAA AAGCAGGCTC CACCATGAG CTTTACCCTCCACTCAG TTTTCTTC |
Sequence-based reagent | Nrxn3a(-).B5.Rev | This paper | PCR primers | GGGGACAACT TTTGTATACAA AGTTGTCACAT AATACTCCTTG TCCTTGTTTTT CTGTTTC |
Sequence-based reagent | B5.(-M)AcGFP.Fo | This paper | PCR primers | GGGGACAACTTTGTATACAAAAGTTGTGAGCAAGGGCGCC GAGCTGTTC |
Sequence-based reagent | AcGFP*.B2.Rev | This paper | PCR primers | GGGGACCAC TTTGTACAAGA AAGCTGGGT TCACTTGTAC AGCTCATCCATGCC |
Sequence-based reagent | AsCI.DEST.for | This paper | PCR primers | TACATGGCGC GCCACAAGTT TGTACAAAAAAGC |
Sequence-based reagent | DEST.BsiWI.rev | This paper | PCR primers | ATGTACGTAC GACCACTTTG TACAAGAAAGC |
Sequence-based reagent | B5.cre.For | This paper | PCR primers | GGGGACAAC TTTGTATACAA AAGTTGCCACC ATGTCCAATTT ACTGACCG TACACC |
Sequence-based reagent | cre*.B2.Rev | This paper | PCR primers | GGGGACCACT TTGTACAAGAA AGCTGGGTTCA ATCGCCATCTT CCAGCAGGCG |
Sequence-based reagent | B1.pNOS.For | This paper | PCR primers | GGGGACAAGT TTGTACAAAAA AGCAGGCTC CCCTCACCCAT CCCCACCCAC CTCCATCCATAC |
Sequence-based reagent | pNOS.B5.Rev | This paper | PCR primers | GGGGACAACTTTTGTATACAAAGTTGTT GCCGTTCGGC CTTGGGTGG CATGATTTC |
Sequence-based reagent | B1.pGRIN3A.For | This paper | PCR primers | GGGGACAAGTTTGTACA AAAAAGCAG GCTCCCTGC CGTGCAAGGA CCACACATT CTACACTATAC |
Sequence-based reagent | pGRIN3A.B5.Rev | This paper | PCR primers | GGGGACAACTT TTGTATACAAA GTTGTCGGCCA CCTTACCGCGG GCTCCCCCA GCGCCTGG |
Sequence-based reagent | B1.pC-KIT.For | This paper | PCR primers | GGGGACAAGTTTGTACAAA AAAGCAGGCTG TCCACCCCCG GATAGCCACAG TGACTGTGAAATG |
Sequence-based reagent | pC-KIT.B5.Rev | This paper | PCR primers | GGGGACAACTTTTGTATACAAAGTTGT GTGCACCGAG CGCGGCAAA GCCGAGC |
Commercial assay or kit | Endofree QIAGEN Maxi kit | QIAGEN | QIAGEN Cat#12,362 | |
Chemical compound, drug | L-Glutamine | GIBCO | 25030–081 | |
Chemical compound, drug | Penicillin– streptomycin | GIBCO | 15140–122 | |
Chemical compound, drug | DMEM media | SIGMA | 11965092 | |
Chemical compound, drug | Iscove’s modified DM (IMDM) | Thermo | 12440053 | |
Chemical compound, drug | Benzonase nuclease | Sigma | E1014-25kU | |
Chemical compound, drug | Pfu Turbo DNA polymerase | Agilent | 600,250 | |
Software, algorithm | MetaMorph | Molecular Devices | RRID:SCR_002368 |