Journal: Cell reports

Article Title: Inappropriate Intrusion of an Axonal Mitochondrial Anchor into Dendrites Causes Neurodegeneration

doi: 10.1016/j.celrep.2019.09.012

Figure Lengend Snippet: (A–F) Immunohistochemistry technique for capturing SNPH intrusion in PC dendrites. Shown is SNPH (green), Syt2 (red), and Calbindin (blue) labeling in 3.5-month-old WT (A, C, and E) and Shi (B, D, and F) mice. Scale bar, 10 μm. (G and H) High magnification of the maximum intensity projection image from the z stack through dendritic regions of WT (G) and Shi (H). (I and J) Orthogonal (slice) view of SNPH punctum (indicated by arrows in panels G and H) in the dendritic region from WT (I) and Shi (J) in x-z and y-z orientations, respectively. (K) Quantification of percentage area occupied by SNPH within the dendritic volume from 3 mice of each group. Data are shown as mean ± SEM. *p < 0.05. (L–N) Capturing SNPH intrusion by pre-tagging dendritic mitochondria in vivo using viral transduction. (L) Technique to selectively transduce PCs with AAV-Mito-mCherry. (M) Demonstration of successful pre-tagging of mitochondria in dendrites of a single PC by Calbindin staining. Scale bar, 10 μm. (N) Demonstration of how pre-tagged dendritic mitochondria in PCs are used to capture SNPH intrusion by 3D rotation. (O and P) Single dendritic tree in WT (O) or Shi (P) pre-tagged with Mito-mCherry (red) and SNPH intrusion (green) captured by co-rotation with Mito-mCherry. Merged images show the fraction of dendritic mitochondria anchored by intruded SNPH (yellow). (Q and R) Percentage of PCs with SNPH intrusions (Q) and percentage of SNPH bound to mitochondria per dendritic tree (R) from WT (n = 73) and Shi (n = 106) PCs. Data are shown as mean ± SEM. ***p < 0.005.

Article Snippet: Polyclonal Calbindin , Boster , M03047–2.

Techniques: Immunohistochemistry, Labeling, In Vivo, Transduction, Staining

Journal: Cell reports

Article Title: Inappropriate Intrusion of an Axonal Mitochondrial Anchor into Dendrites Causes Neurodegeneration

doi: 10.1016/j.celrep.2019.09.012

Figure Lengend Snippet: (A and B) Representative images of lentivirally transduced GFP-SNPH (1–469) (A) and GFP-SNPH (B) in PCs of SNPH-KO mice injected with saline (no harmaline, vehicle only). (C-H) Effect of harmaline on GFP-SNPH (1–469)-transduced (C) and GFP-SNPH-transduced (F) PC dendrites. Degenerating dendrites in GFP-SNPH-transduced PCs can be seen in (F). Also shown is Calbindin labeling of GFP SNPH (1–469) (D) and GFP-SNPH (G) from (C) and (F). Merged images of GFP SNPH (1–469) and GFP-SNPH with Calbindin are shown in (E) and (H), respectively. (I–K) Representative image of a harmaline-induced degenerating PC (white arrow in I) transduced with GFP-SNPH. Calbindin staining from the same section is shown in (J), whereas a merged image is shown in (K). Scale bars, 20 μm. (L) Quantification of dendritic shrinkage in GFP-SNPH (1–469)- and GFP-SNPH-transduced PCs in the absence (n = 3 mice, vehicle only) or presence of harmaline (n = 5 mice). Data are shown as mean ± SEM. ***p < 0.001.

Article Snippet: Polyclonal Calbindin , Boster , M03047–2.

Techniques: Injection, Saline, Labeling, Transduction, Staining

Journal: Cell reports

Article Title: Inappropriate Intrusion of an Axonal Mitochondrial Anchor into Dendrites Causes Neurodegeneration

doi: 10.1016/j.celrep.2019.09.012

Figure Lengend Snippet:

Article Snippet: Polyclonal Calbindin , Boster , M03047–2.

Techniques: Virus, Plasmid Preparation, Recombinant, Software, Imaging

Journal: Scientific Reports

Article Title: Heatstroke-induced late-onset neurological deficits in mice caused by white matter demyelination, Purkinje cell degeneration, and synaptic impairment in the cerebellum

doi: 10.1038/s41598-022-14849-9

Figure Lengend Snippet: Purkinje cell number significantly decreased after heatstroke. To assess Purkinje cells, calbindin was immunostained in the brain and the calbindin-positive Purkinje cell numbers were counted. ( a ) Representative image of calbindin-immunoreactions in the cerebellum. Calbindin-immunoreactions stained the soma of Purkinje cells, the dendritic fibers that were widely stained in the molecular layer (ML), and the white matter. ( b ) Higher magnification images of the Purkinje cell layer in the control (CTL; left) group from 1 to 9 weeks after heatstroke. Purkinje cells in the CTL animals were aligned in an equidistant manner. However, the number of cells decreased after heatstroke. ( c ) The Purkinje cell numbers of the HS group were significantly decreased during the experimental periods after heatstroke (1, 3, and 9 weeks, n = 9, respectively) compared with the CTL group (n = 9). Data are expressed as means ± SD (* P < 0.05, Student’s t test). ML molecular layer, PL Purkinje cell layer, GL granular cell layer, WM white matter, SD standard deviation.

Article Snippet: After heat-induced antigen retrieval and blocking according to the protocol as mentioned above, the sections were incubated with either monoclonal rabbit anti-calbindin antibody (1:100, Cell Signaling, USA) or polyclonal rabbit anti-synaptophysin antibody (1:200, Proteintech, USA) with monoclonal mouse anti-postsynaptic density 95 (PSD95) antibody (1:400, BD, USA) overnight at 4 °C.

Techniques: Staining, Control, Standard Deviation

Journal: Scientific Reports

Article Title: Heatstroke-induced late-onset neurological deficits in mice caused by white matter demyelination, Purkinje cell degeneration, and synaptic impairment in the cerebellum

doi: 10.1038/s41598-022-14849-9

Figure Lengend Snippet: Immunostaining of postsynaptic density 95 in Purkinje cells after heatstroke. ( a ) Representative images of anti-postsynaptic density 95 (PSD95) (red), a postsynaptic marker, and calbindin (green) immunoreactions in the cerebellum in control mice. The PSD95 immunoreactions were observed in the Purkinje cells. Blue is due to 4′,6-diamidino-2-phenylindole nucleic staining. ( b ) The higher magnified images indicated that the PSD95-immunoreactions were located in the axon hillock of each Purkinje cell. The immunoreactions in the control group were widely recognized, such as plates, with relatively lower density in the Purkinje cells (arrowhead). The PSD95 immunoreactions 1 week after heatstroke (arrow) were gathered and concentrated in the axon hillock of the Purkinje cells like pen points. The immunoreactions appeared to have attenuated at 3 weeks and reappeared more heterogeneously at 9 weeks. ML molecular layer, PL Purkinje cell layer, GL granular cell layer, WM white matter, CTL control.

Article Snippet: After heat-induced antigen retrieval and blocking according to the protocol as mentioned above, the sections were incubated with either monoclonal rabbit anti-calbindin antibody (1:100, Cell Signaling, USA) or polyclonal rabbit anti-synaptophysin antibody (1:200, Proteintech, USA) with monoclonal mouse anti-postsynaptic density 95 (PSD95) antibody (1:400, BD, USA) overnight at 4 °C.

Techniques: Immunostaining, Marker, Control, Staining

Journal: International Journal of Molecular Sciences

Article Title: RNA-Seq Analysis Reveals an Essential Role of the Tyrosine Metabolic Pathway and Inflammation in Myopia-Induced Retinal Degeneration in Guinea Pigs

doi: 10.3390/ijms222212598

Figure Lengend Snippet: Primary and secondary antibodies used in immunofluorescence study.

Article Snippet: Calbindin , Rabbit , TA342845 , 1:50 , Origene , Horizontal cells.

Techniques: Immunofluorescence, Plasmid Preparation

Journal: Frontiers in Neuroscience

Article Title: Dystrophin Is Required for the Proper Timing in Retinal Histogenesis: A Thorough Investigation on the mdx Mouse Model of Duchenne Muscular Dystrophy

doi: 10.3389/fnins.2020.00760

Figure Lengend Snippet: Primary antibodies used for immunofluorescence.

Article Snippet: Rabbit anti-calbindin (HC) , 1:1000 , Merck Millipore (Billerica, MA, United States).

Techniques: Immunofluorescence

Journal: The Journal of Neuroscience

Article Title: Clustered Fine Compartmentalization of the Mouse Embryonic Cerebellar Cortex and Its Rearrangement into the Postnatal Striped Configuration

doi: 10.1523/JNEUROSCI.1710-12.2012

Figure Lengend Snippet: Antibody specification

Article Snippet: , Calbindin-D28k , Synthetic peptide derived from amino acids 185-199 of human Calbindin-D-28K , AnaSpec, rabbit polyclonal, Cat. # 53283, Lot # GL141 , 1:4000.

Techniques: Purification, Derivative Assay, Recombinant, Plasmid Preparation

Journal: Neurobiology of Stress

Article Title: Persistent pain intensifies recall of consolidated fear memories

doi: 10.1016/j.ynstr.2019.100163

Figure Lengend Snippet: Ongoing pain induced morphological changes in the BLA interneuronal population. Panels A and B show calbindin immunoreactivity (Calb-ir, green) and panels D and E show parvalbumin immunoreactivity (Parv-ir, red) expression in sham mice ( A and D ) and in cuffed mice ( B and E ). Panels G and H show the combined expression of the two markers in sham ( G ) and cuffed ( H ) mice, where the Calb/Parv-ir co-localization appears as yellow. The number of Calb-ir cells did not differ between the groups ( C) but the number of Parv immunopositive neurons increased in the cuffed mice ( F ) and most of these Parv-ir neurons were colocalized with Calb-ir, or the yellow labeled neurons in the G and H inserts; The graph in panel I compares the number of Calb/Parv-ir neurons of sham and cuffed mice. The cell counts were evaluated by T-test, * p < 0.05 and *** p < 0.001, bars represent mean ± SEM, n = 12 to 14 per group. BLA - basolateral amygdala, CeA - central amygdala, LA - lateral amygdala. Scale bar = 200 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: A similar immunostaining protocol was used for visualization of calbindin and parvalbumin in the BLA using rabbit anti-calbindin D28 antibody (Synaptic Systems, Goettingen, Germany) diluted 1:10 K and monoclonal mouse antibody against parvalbumin (Sawnt, Marly, Switzerland) diluted 1:5 K. The next step was incubation with secondary anti-rabbit Alexa 488 and anti-mouse Alexa 594 antibodies (Jackson ImmunoResearch Inc. West Grove, PA) for 4 h at room temperature.

Techniques: Expressing, Labeling