α1i  (Alomone Labs)

Journal: Brain Structure & Function

Article Title: Transmitter and ion channel profiles of neurons in the primate abducens and trochlear nuclei

doi: 10.1007/s00429-021-02315-7

Figure Lengend Snippet: Low-voltage activated calcium channel family (Cav3) subunits in the abducens and trochlear nucleus. a Consecutive coronal paraffin sections through the abducens nucleus (nVI) ( a ) depicting the immunolabeling for Cav3.1 (first panel), Cav3.2 (third panel) and Cav3.3 (fourth panel) subunits with combined immunostaining for ChAT (brown) and ACAN (black) (second panel) as reference. b, c Close-up of Cav subunit expression in nVI MIF (red arrowheads) and SIF motoneurons (MNs) (green arrows) and INTs (blue arrow). d Consecutive coronal paraffin sections through the trochlear nucleus (nVI) depicting the immunolabeling for Cav3.1 (first panel), Cav3.2 (second panel) and Cav3.3 (fourth panel) subunits with combined immunostaining for ChAT (brown) and ACAN (black) (third panel) as reference. Thin dashed lines in d indicate the border of nIV and thick dashed lines indicate the boundary between the MIF and SIF MNs. e Close-up of Cav subunit expression in MIF (red arrowheads) and SIF MNs (green arrow) on different sections as those illustrated in d . Note the weak Cav3.1 expression along the membrane of some SIF MNs (left column, black star). Red dashed lines indicate the tentative position of the border delineating the dorsal cap of nIV. f Cerebellar Purkinje cells located on the same consecutive sections as nVI as controls for immunopositivity of the different Cav subunits. Scale bar indicates 100 μm in a, d, f and 50 μm in b, c, e

Article Snippet: Voltage-dependent T-type calcium channel subunits Cav3.1 (CACNA1G, α1G; Cat #: ACC-021; RRID: AB_2039779), Cav3.2 (CACNA1H, α1H; Cat #: ACC-025; RRID: AB_2039781) and Cav3.3 (CACNA1I, α1H; Cat #: ACC-009; RRID: AB_2039783) were detected with polyclonal rabbit antibodies from (Alomone Labs, Jerusalem, ISRAEL).

Techniques: Immunolabeling, Immunostaining, Expressing

Journal: Science Advances

Article Title: Neuromodulatory control of inhibitory network arborization in the developing postnatal neocortex

doi: 10.1126/sciadv.abe7192

Figure Lengend Snippet: ChC axonal arborization requires Cav3.3-T-type VDCC genes in a dosage sensitive manner. ( A and B ) Confocal images of EP13 transplanted ZsGreen-labeled ChCs transfected with LacZ sgRNAs (A, left) or Cav3.3 sgRNAs (A, right) as well as EP13 transplanted GFP-labeled ChCs originating from Cav3.3 germline KO mice (B). Single optical sections indicating synaptic cartridges apposed to AnkG-labeled AISs are shown in the middle panels. The areas in white boxes with arrowheads are enlarged and overlaid with the AnkG channel. Bottom panels indicate reconstructed axonal arbors in the area defined in Fig. 4A . ( C ) The number of branch points in LacZ sgRNA –transfected ChCs ( n = 5 cells from three mice), Cav3.3 sgRNA– transfected homozygous KO ChCs ( n = 5 cells from three mice), transplanted ChCs originating from Cav3.3 homozygous KO mice ( n = 6 cells from three mice), and transplanted ChCs originating from Cav3.3 heterozygous KO mice ( n = 5 cells from three mice). One-way ANOVA, * P

Article Snippet: The following primary antibodies were used: goat anti-ChAT (1:200; Millipore, catalog no. AB144P), rabbit anti-Cav3.1 (1:100; Alomone Labs, catalog no.ACC-021), rabbit anti-Cav3.2 (1:100; Alomone Labs, catalog no. ACC-025), rabbit anti-Cav3.3 (1:100; Alomone Labs, catalog no. ACC-009), guinea pig polyclonal anti-PV (1:2000; Swant, PVG-213), rat anti-hemagglutinin (1:500; Roche, catalog no.11-867-423-001), chicken anti-GFP (1:800; Abcam, catalog no.ab13970), mouse anti-AnkG (1:500; UC Davis/NIH Neuromab, catalog no. clone N106/36 75-146), rabbit anti-ZsGreen (1:500; Clontech, catalog no. 632474), and rabbit anti-RFP (1:800; Rockland, catalog no. 600-401-379).

Techniques: Labeling, Transfection, Mouse Assay

Journal: Theranostics

Article Title: Neuromedin B receptor stimulation of Cav3.2 T-type Ca2+ channels in primary sensory neurons mediates peripheral pain hypersensitivity

doi: 10.7150/thno.62255

Figure Lengend Snippet: Activation of NmbR stimulates recombinant Cav3.2 channels heterologously expressed in HEK293 cells. A, western blot analysis of NmbR in HEK293 cells transiently transfected with NmbR cDNA. Blots depicted are representative of three independent experiments. B, membrane localization of NmbR in transfected HEK293 cells. Alphabets a through c in the diagram indicate the differential interference contrast (DIC, a ), the EGFP fluorescent signals of NmbR ( b ), and the merged image ( c ), respectively. C, exemplary current traces show the effect of Nmb (100 nM) on Cav3.1 (α1G), Cav3.2 (α1H) and Cav3.3 (α1I) channel currents. Currents were elicited by a 100 ms depolarizing step pulse from the holding potential of -110 mV to -30 mV. D, summary data show the effect of 100 nM Nmb on Cav3.1 ( n = 10), Cav3.2 ( n = 9) and Cav3.3 ( n = 7) channel currents. ** p

Article Snippet: The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween-20 (TBST) and probed with antibodies against Nmb (rabbit, 1:1000, Abcam, Cat. No. ab191499), Cav3.1 (rabbit, 1:1000, Alomone, Cat. No. ACC-021), Cav3.2 (mouse, 1:1000, Novus, Cat. No. NBP1-22444), Cav3.3 (rabbit, 1:1000, Alomone, Cat. No. ACC-009), NmbR (rabbit, 1:500, Sigma, Cat. No. SAB4502914), phospho-AMPKα1 (rabbit, 1:500, Abcam, Cat. No. ab92701), AMPKα1 (rabbit, 1:600, Abcam, Cat. No. ab110036), Gαq/11 (mouse, 1:600, Santa Cruz Biotechnology, Cat. No. SC-365906), and Gβ (mouse, 1:500, Santa Cruz Biotechnology, Cat. No. SC-515822).

Techniques: Activation Assay, Recombinant, Western Blot, Transfection

Journal: Molecular Psychiatry

Article Title: Developmental oxidative stress leads to T-type Ca2+ channel hypofunction in thalamic reticular nucleus of mouse models pertinent to schizophrenia

doi: 10.1038/s41380-021-01425-2

Figure Lengend Snippet: Hypofunction of T-Ca 2+ channels and decreased CaV3.3 expression in TRN neurons of adult Gclm -KO mice, resulting in alteration of their bursting profile. A Smaller proportion of TRN neurons generating burst firing at resting membrane potential (RMP) in KO as compared to WT mice (WT n = 13; KO n = 12; Fisher exact test, p = 0.02). B Weaker density of T-Ca 2+ currents activated at RMP in TRN neurons of KO ( n = 11) as compared to WT ( n = 10) mice ( p = 0.034, one-tailed t -test). C Representative recording of single bursting in a TRN neuron. D Threshold for the initial membrane potential required to induce single bursting upon depolarization. E Representative recording of repetitive bursting in a TRN neuron. F Threshold for the initial membrane potential required to induce repetitive bursting upon depolarization. Note that KO ( n = 6) TRN neurons require a more hyperpolarized membrane potential, particularly for exhibiting repetitive bursts, as compared to WT mice ( n = 8). G , H Top: Representative recordings of T-Ca 2+ and SK currents induced by a short constant depolarization step (from −110 to −40 mV), with their amplitudes increasing with greater hyperpolarizing initial membrane potential (going from −30 mV to −110 mV). Bottom: Density of T-Ca 2+ ( G ) and SK currents ( H ) activated from each of the initial membrane potentials. Compared to WT mice, TRN neurons in KO display overall smaller T-Ca 2+ ( F = 31.92 DF n = 1 DFd = 289; p

Article Snippet: We carried out CaV3.2/PV and CaV3.3/PV immunohistology as indicated in using the following primary [rabbit anti-CaV3.2 (Santa Cruz Biotechnology, USA); rabbit anti-CaV3.3 (Alomone labs, Israel), sheep anti-PV (R & D systems)] and secondary [goat anti-rabbit AF488 and donkey anti-sheep AF594 (Abcam, UK)] antibodies.

Techniques: Expressing, Mouse Assay, One-tailed Test