Journal: bioRxiv

Article Title: Behavioral state regulates the dynamics of memory consolidation

doi: 10.1101/2024.06.28.601231

Figure Lengend Snippet: (A) Diagram of cerebellar cortex (green), targeted here for reversible inactivations. (B) Schematic of guide cannula (grey) chronically implanted at cerebellar surface and internal cannula (white) acutely introduced after each training session to deliver muscimol in the eyelid area of cerebellar cortex. (C) Coronal brain slice. Purkinje cells stained for calbindin (red); fluorescent muscimol (GABAa receptor agonist), green. (D) Protocol for post-session reversible inactivations. Muscimol or vehicle (control) was infused after each session. Learning was assessed when training resumed 24h later with the subsequent session. (E) Left: Averaged acquisition curves (± s.e.m.) of mice trained on a self-paced running wheel, treated with vehicle-(black, N=10) or muscimol (green, N=15) after each training session. p =0.0002***; repeated-measures-ANOVA. Center: After training completion, muscimol was infused before test sessions (M), to verify cannula placement; Controls (black): p=0.00015***; Muscimol (green): p=0.0008***; paired-t-tests, before vs. after muscimol. Performance was not impaired with vehicle infusion (V) in a subsequent session; Controls (black): p=0.5n.s; Muscimol (green): p=0.2n.s.; paired-t-tests, before vs. after vehicle. Right: Controls then received muscimol infusions after additional training sessions to test the effect of reversible inactivations after acquisition (plateau sessions, N=5). (F) Quantification of learning onset session (see Methods). p=0.02*; Mann-Whitney-Wilcoxon test. Non-learners (NL) did not exhibit CRs within 14 training sessions. Each dot is one mouse. (G) Averaged eyelid closures (± s.e.m.) for CS-only trials from the last training sessions. Amplitudes: p=0.046*, controls (black, N=10) vs. muscimol (green, N=15), t-test. Blue: visual CS. Red dashed line and shading: timing of the (omitted) airpuff-US. (H) Same as (E) but for mice trained while locomoting at a constant speed (0.1 m/s) on a motorized running wheel (Vehicle controls, black, N=14; Muscimol, green, N=12). Left: Acquisition, p=0.7n.s.; repeated-measures-ANOVA. Middle: Before vs. after Muscimol test (M). Controls: p=1.7e-6***, Muscimol, p=2.9e-6***; paired-t-tests. Right: Before vs. after Vehicle test (V). Controls, p=0.4n.s; Muscimol, p=0.1n.s., paired t-tests. (I) Same as (F) but for mice in (H); Controls vs. Muscimol, p=0.5n.s., Mann-Whitney-Wilcoxon test. (J) Same as (G) but for mice in (H); Controls vs. Muscimol, p=0.9n.s., t-test.

Article Snippet: Brain sections (50um thick) were cut in a vibratome and stained for Purkinje cells (with chicken anti-calbindin primary antibody #214006 SYSY, and anti-chicken Alexa488 #703-545-155 or Alexa594 #703-545-155 secondary antibodies from Jackson Immunoresearch) and for cell viability (with DAPI marker).

Techniques: Slice Preparation, Staining, Control, MANN-WHITNEY

Journal: Cerebellum (London, England)

Article Title: The Primary Ciliary Deficits in Cerebellar Bergmann Glia of the Mouse Model of Fragile X Syndrome

doi: 10.1007/s12311-022-01382-8

Figure Lengend Snippet: (A) Immunostaining of Arl13b (red), NeuN (cyan, top), calbindin (green, middle), and BLBP (yellow, bottom) with DAPI nuclear staining (grey) in the posterior cerebellum of adult WT (left) and Fmr1 KO (right) mice. Blue arrows, Arl13b+ cells; Magenta arrowheads, Arl13b− cells. Scale bar, 10 μm. (B) Quantification of the percentage of Arl13b+ cells among NeuN+ cells (top), calbindin+ cells (middle), or BLBP+ cells (bottom) in the cerebellum from adult WT and Fmr1 KO mice. (C) Immunostaining of Arl13b (red) and BLBP (yellow) with DAPI nuclear staining (grey) in the PCL of P7, P10, P14, P28, and adult WT (left) and Fmr1 KO (right) mice. Blue arrows, Arl13b+ cells; Magenta arrowheads, Arl13b− cells. Scale bar, 10 μm. (D) Quantification of the percentage of Arl13b+ cells among BLBP+ cells in the cerebellum from WT and Fmr1 KO mice. n=4–8 for each group (A), n=8–24 for each group (C), mean ± SEM. Student’s unpaired t-test. *P<0.05; **P<0.01; ***P<0.001.

Article Snippet: The following primary antibodies were used in this study: mouse anti-BLBP (1:200; Abcam; ab131137), mouse anti-Sox-2 (1:100; Santa Cruz; sc-365964), mouse anti-S-100β chain (1:100; Santa Cruz; sc-393919), mouse anti-NeuN (1:500; Millipore-Sigma; MAB-377), chicken anti-calbindin (1:400; Encor; CPCA-Calbindin), rabbit anti-Arl13b (1:700; Proteintech; 17711–1-AP), rabbit anti-BLBP (1:300; Abcam; ab32423), rat anti-BrdU (1:200; Novus; NB500–169), and rat anti-BrdU (1:200; Abcam; ab6326).

Techniques: Immunostaining, Staining

Journal: eLife

Article Title: LKB1 coordinates neurite remodeling to drive synapse layer emergence in the outer retina

doi: 10.7554/eLife.56931

Figure Lengend Snippet:

Article Snippet: Calbindin D-28k , Full-length recombinant human Calbindin D-28K , Horizontal cells; amacrine cells; retinal ganglion cells , Novus biologicals; chicken polyclonal; NBP2-50028; no RRID , 1:2000.

Techniques: RNAscope, Multiplex Assay, Software

Journal: Poultry science

Article Title: Effect of transient receptor potential vanilloid 6 gene silencing on the expression of calcium transport genes in chicken osteoblasts.

doi: 10.3382/ps/peu071

Figure Lengend Snippet: Figure 3. Expression levels of calcium transport genes in chicken osteoblasts 48 h after transfection. Levels were quantified by real-time PCR. (A) Expression levels of TRPV6. The data were expressed as the means ± SEM. ∗P < 0.05 and ∗∗P < 0.01 compared with the untreated group. (B) Expression levels of calbindin-D28K, PMCA1b and NCX1 after the transfection with pSIREN-TRPV6-3. The data were expressed as the means ± SEM. ∗∗P < 0.01 compared with the untreated group.

Article Snippet: The blots were incubated with the primary antibodies anti-calbindin-D28K (BM0203, Boshide, China), anti-TRPV6 (ACC-036, Alamone Labs, Israel) and anti-β-actin (Santa Cruz), which were diluted 1:500, 1:500, and 1:2000, respectively.

Techniques: Expressing, Transfection, Real-time Polymerase Chain Reaction

Journal: Poultry science

Article Title: Effect of transient receptor potential vanilloid 6 gene silencing on the expression of calcium transport genes in chicken osteoblasts.

doi: 10.3382/ps/peu071

Figure Lengend Snippet: Figure 4. Expression of TRPV6 and calbindin-D28K proteins in chicken osteoblasts 48 h after transfection. The protein expression levels of TRPV6 and calbindin-D28K were reduced by 40.2% (P < 0.01) and 29.8% (P < 0.01), respectively, 48 h after transfection with pSIREN-TRPV6-3 compared with the level obtained in the untreated group. There was no significant difference between the untreated and pSIREN-control groups. Representative blots for each group from trip- licate experiments were shown. The protein band density was quan- tified by densitometry using the Quantity One software. The protein level was expressed as a ratio relative to the expression of β-actin. The data were expressed as the means ± SEM. ∗∗P < 0.01 compared with the untreated group.

Article Snippet: The blots were incubated with the primary antibodies anti-calbindin-D28K (BM0203, Boshide, China), anti-TRPV6 (ACC-036, Alamone Labs, Israel) and anti-β-actin (Santa Cruz), which were diluted 1:500, 1:500, and 1:2000, respectively.

Techniques: Expressing, Transfection, Control, Software

Journal: PLOS One

Article Title: Impaired dynein function preserves spinal interneuron survival and positioning in an ALS-like mouse model

doi: 10.1371/journal.pone.0346246

Figure Lengend Snippet: Number of ChAT + (A-C), GAD-67 + (D), Calbindin + (E), Parvalbumin + (F), Parvalbumin + −Calbindin + (G), Parvalbumin + −GAD-67 + (H), and ChAT + −GAD-67 + (I) cells per biological replicate. The number of cells analysed is the sum of immunostained positive cells from eight spinal cord cross-sections (two sections per lumbar level L3–L6) per biological replicate. Data are shown as mean ± standard deviation and were analysed by an ordinary one-way ANOVA. N = 3 biological replicates per genotype.

Article Snippet: Following three 10-min PBS washes at 100 rpm, secondary probing was performed for 3 hours in the dark in PBS containing 0.05% Tween-20 and the relevant secondary antibodies for ChAT (1:250 dilution, donkey anti-goat IgG H + L Alexa Fluor ® 405, Abcam, ab175664), Calbindin (1:500 dilution, Rhodamine TRITC-AffiniPure donkey anti-guinea pig Ig G H + L, Jackson Immuno Research, 706-025-148-JIR), Parvalbumin (1:500 dilution, Alexa Fluor 647-AffiniPure donkey anti-rabbit IgG H + L, Jackson Immuno Research, 711-605-152), and GAD-67 (1:500 dilution, donkey anti-chicken IgY H + L Highly Cross Adsorbed Alexa Fluor ® 488, Life Technologies, A78948).

Techniques: Standard Deviation