Journal: PLoS ONE

Article Title: Endogenous Retinoic Acid Activity in Principal Cells and Intercalated Cells of Mouse Collecting Duct System

doi: 10.1371/journal.pone.0016770

Figure Lengend Snippet: A . Schematic representation of nephron and collecting ducts. Tamm-Horsfall protein (THP), calbindin D28K (CD28K), aquaporin 2 (AQP2) and vacuolar H + -ATPase B1 (V-ATPase) antibodies were used to identify thick ascending limbs, distal convoluted tubules, as well as principal cells and intercalated cells of collecting ducts, respectively. B . Left panel (merged image): CD28K signal (red) was observed only in the cortex, whereas THP signal (green) was detected in both cortex and outer medulla, but not in the deep inner medulla. Original magnification was 100×. Right panel (merged image): CD28K (red) and THP (green) signals were mutually exclusive. Original magnification was 400×. C . Left panel (merged image): CD28K (red) was detected only in the cortex whereas AQP2 (green) was observed in the outer medulla and inner medulla. Original magnification was 100×; right panel (merged image): distal convoluted tubules stained exclusively and intensely for CD28K (double arrow); tubules stained positive for both CD28K and AQP2 were the connecting tubules (arrowhead) whereas those showing weak or negative CD28K but a positive AQP2 signal were the cortical collecting ducts (arrow). Original magnification was 400×. D . Principal cells and intercalated cells that stained positive for AQP2 (green) and V-ATPase (red), respectively, co-localized to the same tubules in the cortex, outer medulla and inner medulla. Original magnification was 400×. No specific signal was noted on sections incubated with non-immune IgGs in place of primary antibodies (data not shown). Shown here are kidney paraffin sections of a 2-week-old mouse. Nuclei were visualized with DAPI (blue) counterstaining. C: cortex; OM: outer medulla; IM: inner medulla; G: glomeruli.

Article Snippet: The primary antibodies used were: chicken anti-β-gal (dilution 1∶200; Abcam Inc., Cambridge, UK), rabbit anti-aquaporin 2 (AQP2; dilution 1∶200; Milipore UK Ltd, Watford, UK), rabbit anti-Tamm-Horsfall protein (THP; dilution 1∶100; Insight Biotechnology Ltd, Wembley, UK), goat anti- vacuolar H + -ATPase B1 (V-ATPase; dilution 1∶500; Insight Biotechnology Ltd), goat anti-calbindin D28K (CD28K; dilution 1∶200; Insight Biotechnology Ltd) and rabbit anti-RARβ2 (dilution 1∶200; Insight Biotechnology Ltd).

Techniques: Staining, Incubation

Journal: PLoS ONE

Article Title: Endogenous Retinoic Acid Activity in Principal Cells and Intercalated Cells of Mouse Collecting Duct System

doi: 10.1371/journal.pone.0016770

Figure Lengend Snippet: In kidneys of 2-week-old ( A ) and 8-week-old ( B ) mice, tubules with intense CD28K signal (arrow) did not stain positive for β-gal protein. Some tubules with intermittent CD28K, likely connecting tubules or cortical collecting ducts, demonstrated positive β-gal signal (double arrow). Original magnification was 400×. No specific signal was detected on sections incubated with non-immune IgGs in place of primary antibodies (data not shown).

Article Snippet: The primary antibodies used were: chicken anti-β-gal (dilution 1∶200; Abcam Inc., Cambridge, UK), rabbit anti-aquaporin 2 (AQP2; dilution 1∶200; Milipore UK Ltd, Watford, UK), rabbit anti-Tamm-Horsfall protein (THP; dilution 1∶100; Insight Biotechnology Ltd, Wembley, UK), goat anti- vacuolar H + -ATPase B1 (V-ATPase; dilution 1∶500; Insight Biotechnology Ltd), goat anti-calbindin D28K (CD28K; dilution 1∶200; Insight Biotechnology Ltd) and rabbit anti-RARβ2 (dilution 1∶200; Insight Biotechnology Ltd).

Techniques: Staining, Incubation

Journal: The Journal of Neuroscience

Article Title: Combinations of AMPA Receptor Subunit Expression in Individual Cortical Neurons Correlate with Expression of Specific Calcium-Binding Proteins

doi: 10.1523/JNEUROSCI.17-05-01570.1997

Figure Lengend Snippet: Histograms of the number of CaBP-positive (parvalbumin, A; calbindin-D28k, B) neurons in the classified four types of neurons in each cortical layer (layers II/III, IV, V, and VI). The number of CaBP-positive neurons was counted in six 105 × 160 μm2 regions in three adult rats and summarized. The numbers of positive neurons are expressed as the percentage of the total number of counted parvalbumin- or calbindin-D28k-positive cells in all layers. A, The majority of parvalbumin-positive neurons was type 2A neurons. B, Calbindin-D28k immunoreactivity was observed mainly in type 1A neurons of the neocortical layer, with the exception of layer I, whereas type 1B neurons with calbindin-D28k signal were restricted to layerII/III. Dark gray bars, Type 1A neurons [GluR1(+)/GluR2(+)]; light gray bars, type 1B neurons [GluR1(−)/GluR2(+)]; black bars, type 2A neurons [GluR1(+)/GluR2(−)]; slashed bars, type 2B neurons [GluR1(−)/GluR2(−)].

Article Snippet: On the next day, the sections were washed three times for 5 min each time in PBS, treated in biotinylated anti-mouse IgG (diluted 1:200, second antibody for parvalbumin and calbindin-D28k; Vector Laboratories, Burlingame, CA) for 1 hr at RT, and washed three times for 10 min each again.

Techniques:

Journal: The Journal of Neuroscience

Article Title: Combinations of AMPA Receptor Subunit Expression in Individual Cortical Neurons Correlate with Expression of Specific Calcium-Binding Proteins

doi: 10.1523/JNEUROSCI.17-05-01570.1997

Figure Lengend Snippet: Detection of calbindin-D28k in classified neurons using the triple-staining technique. A–C, The type 1A (arrowheads) and type 1B (arrows) neurons in layer II/III had calbindin-D28k immunoreactivity.D–F, The nonpyramidal type 1A neurons in layer VI (arrowhead) were stained with calbindin-D28k antibody.CB, Calbindin-D28k. Scale bar, 50 μm.

Article Snippet: On the next day, the sections were washed three times for 5 min each time in PBS, treated in biotinylated anti-mouse IgG (diluted 1:200, second antibody for parvalbumin and calbindin-D28k; Vector Laboratories, Burlingame, CA) for 1 hr at RT, and washed three times for 10 min each again.

Techniques: Staining

Journal: The Journal of Neuroscience

Article Title: Combinations of AMPA Receptor Subunit Expression in Individual Cortical Neurons Correlate with Expression of Specific Calcium-Binding Proteins

doi: 10.1523/JNEUROSCI.17-05-01570.1997

Figure Lengend Snippet: Parvalbumin and calbindin-D28k expression in 100 neurons classified by GluR subunit combinations in each layer of somatosensory cortex

Article Snippet: On the next day, the sections were washed three times for 5 min each time in PBS, treated in biotinylated anti-mouse IgG (diluted 1:200, second antibody for parvalbumin and calbindin-D28k; Vector Laboratories, Burlingame, CA) for 1 hr at RT, and washed three times for 10 min each again.

Techniques: Expressing

Journal: Frontiers in Molecular Neuroscience

Article Title: Dopamine Modulates Homeostatic Excitatory Synaptic Plasticity of Immature Dentate Granule Cells in Entorhino-Hippocampal Slice Cultures

doi: 10.3389/fnmol.2018.00303

Figure Lengend Snippet: Immature dentate granule cells (GCs) of 1-week-old slice cultures express homeostatic excitatory synaptic plasticity. (A) Example of a 1-week-old entorhino-hippocampal slice culture stained for calbindin (Calb, magenta) and doublecortin (DCX, green). The GC layer (GCL) is shown at higher magnification. Scale bars, 50 μm and 10 μm. (B) Quantification of DCX and Calb expression in 1-week-old slice cultures. Some cells expressed both or neither of the markers (n.c., not classified; n = 17 cultures per group). (C) Sample traces of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor-mediated miniature excitatory postsynaptic currents (mEPSCs) recorded from immature GCs in tetrodotoxin TTX (2 μM, 3 days) and vehicle-only (control)-treated cultures. (D) Example of a recorded (biocytin-filled), post hoc identified (streptavidin-Alexa568, white) immature GC in a DCX-stained slice culture. Scale bars, 10 μm and 5 μm (inset). (E) Group data of mEPSC recordings (amplitudes and frequencies) from immature GCs of vehicle-only-(Ctrl, control) and TTX-treated cultures ( n control = 9 cells, n TTX = 9 cells; Mann-Whitney test; U-value = 6). Individual data points are indicated by gray dots. Values represent mean ± SEM (** p < 0.01; NS, non-significant difference).

Article Snippet: Slice cultures were fixed, re-sliced and immunostained as described previously (Lenz et al., ) with antibodies against doublecortin (DCX; C-18; goat; Santa Cruz; 1:500) and calbindin (Calb; D28-K, mouse; Swant; 1:1,000).

Techniques: Staining, Expressing, Control, MANN-WHITNEY