polyclonal goat anti calbindin  (Danaher Inc)

Journal: Molecular Neurobiology

Article Title: Calbindin Deficits May Underlie Dissociable Effects of 5-HT 6 and mGlu 7 Antagonists on Glutamate and Cognition in a Dual-Hit Neurodevelopmental Model for Schizophrenia

doi: 10.1007/s12035-020-01938-x

Figure Lengend Snippet: Impact of combined neonatal PCP and isolation rearing on hippocampal calbindin and 5-HT immunoreactivity in the dorsal hippocampus. Representative a , b calbindin immunoreactivity throughout a all subfields and b part of CA1 (indicated in a by a solid border), together with e higher magnification images of typical 5-HT immunoreactivity in CA1 strata oriens (s.o.) and radiatum (s.r.) locations (indicated in a by dashed borders). Red represents a , b calbindin or e 5-HT, and blue in a and the top portion of each image in e represents DAPI nuclear counterstain; the bottom portion of each image in e is a duplicate presented without the nuclear counterstain. Scale bars are equivalent to 100 μm a , b or 10 μm e . Mean ± SEM c number of calbindin-positive cells in s.o., s.r., and stratum lacunosum-moleculare (s.l-m) of the dorsal hippocampal subiculum/fasciola cinereum (sub/fc), CA1, and CA2/3, and f intensity of 5-HT immunofluorescence in s.o. and s.r. of dorsal hippocampal CA1 and CA3 and molecular (mol) and polymorphic (pol) layers of the dentate gyrus (DG), plus correlation analyses of d calbindin and g 5-HT immunofluorescence intensity in CA1 versus the NOD choice trial discrimination ratio (time exploring novel/total choice trial object exploration) following acute SB-399885. Male Lister hooded rats that received saline (1 ml/kg s.c.; Veh) or PCP (10 mg/kg) on PND 7, 9, and 11 were housed in groups (Gr; Veh only) or isolation (Iso; Veh and PCP) from weaning on PND 21, then underwent NOD on three occasions (PND 57–80) before tissue collection (PND 78–80), to receive vehicle (0.5% methylcellulose 1% Tween 80; 1 ml/kg i.p. 30 min before the familiarization trial), SB-399885 (10 mg/kg), or MMPIP (10 mg/kg) on separate test days in a pseudorandom order and serve as their own controls ( n = 13–14 per neurodevelopmental condition). Calbindin immunoreactivity a was strongest in the dentate gyrus and stratum pyramidale, and remaining cells matched the distribution of GABA interneurons. The b intensity of calbindin immunoreactivity in CA1 and c number of calbindin-positive cells throughout dorsal hippocampal subfields and cell layers were both influenced by neurodevelopmental condition ( P < 0.05 and P < 0.01, respectively), and although there were some reductions in single-hit Veh-Iso, these were much more extensive in PCP-Iso and correlated with d NOD performance following acute administration of SB-399885. Patterns of e 5-HT immunoreactivity were consistent with labeling of fine varicose axons (marked by arrowheads) but labeling intensity f was not influenced by neurodevelopmental condition and g did not correlate with NOD flowing SB-399885 administration. † P < 0.05 Veh-Iso and † P < 0.05; †† P < 0.01; †††† P < 0.0001 PCP-Iso versus Veh-Gr; # P < 0.05 PCP-Iso versus Veh-Iso (three-way repeated measures ANOVA with Tukey post hoc)

Article Snippet: Sections were washed (4 × 5 min) in phosphate-buffered saline (PBS), incubated (1 h) in 2% goat or donkey serum in buffer 1 (0.5% BSA, 0.3% Triton X-100), then (overnight, 4 °C) rabbit or goat polyclonal primary antibodies against calbindin or 5-HT (Abcam: 1:500 in buffer 1), or buffer alone for negative control.

Techniques: Isolation, Immunofluorescence, Labeling

Journal: Investigative Ophthalmology & Visual Science

Article Title: Critical Role for Phosphatidylinositol-3 Kinase Vps34/PIK3C3 in ON-Bipolar Cells

doi: 10.1167/iovs.19-26586

Figure Lengend Snippet: Maintenance of horizontal cells and rod synaptic terminals. (A, B) Immunostaining for ribeye (magenta), a component of the presynaptic ribbon, appears normal in Vps34 fl/fl ;Pcp2-Cre mice at 10 months, when PKCα (green, [A]) or TRPM1 (green, [B]) staining indicates near complete loss of ON-BCs. (C) Sections triple labeled for calbindin, a horizontal cell marker (green), ribeye (magenta), and PKCα (blue), indicate preservation of horizontal cell dendrites and OPL puncta. (D) Quantification of OPL synapses. OPL puncta containing closely-apposed ribeye and calbindin staining were counted in 50 × 50 μm fields from three to four images, each from a different animal.

Article Snippet: PKCα: mouse monoclonal H-7 (#sc-8393; Santa Cruz Biotechnology, Dallas, TX, USA), rabbit polyclonal (#2056; Cell Signaling Technology, Danvers, MA, USA); PCP2: mouse monoclonal F-3 (#sc-137064; Santa Cruz Biotechnology); calbindin: D28K goat polyclonal (#sc-7691; Santa Cruz Biotechnology), mouse monoclonal CB-955 (#C9848; Sigma-Aldrich Corp., St. Louis, MO, USA); calretinin: mouse monoclonal H-5 (#sc-365956; Santa Cruz Biotechnology); parvalbumin α: goat polyclonal (#sc-7449; Santa Cruz Biotechnology); ribeye: rabbit polyclonal anti-ribeye A-domain (#192-103; Synaptic Systems, Goettingen, Germany); LAMP1: mouse monoclonal LY1C6 (#428017 Calbiochem, Burlington, MA, USA); LC3: rabbit monoclonal D3U4C against LC3A/B (#12741; Cell Signaling Technology), mouse monoclonal (#MI52; MBL International, Woburn, MA, USA); Rab7: rabbit monoclonal D95F2 (#9367; Cell Signaling Technology); ubiquitin: mouse monoclonal P4D1 (#3936; Cell Signaling Technology), mouse monoclonal (#04-263 Millipore, Burlington, MA, USA); Cre: rabbit polyclonal (#69050; Millipore); p62: guinea pig polyclonal (#03-GP62-C; American Research Products, Waltham, MA, USA).

Techniques: Immunostaining, Staining, Labeling, Marker, Preserving

Journal: Scientific Reports

Article Title: Aberrant activation of the human sex-determining gene in early embryonic development results in postnatal growth retardation and lethality in mice

doi: 10.1038/s41598-017-04117-6

Figure Lengend Snippet: Impaired development of the lung and brain in hSRY-ON mice. ( A ) Immunostaining of hSRY (top), T1α for type I alveolar epithelial cells (middle), and surfactant protein C (SPC) for type II alveolar epithelial cells (bottom) on lung sections of newborn (P0) control (left, column) and hSRY-ON (right, column) mice. ( B ) Enlargements of boxed areas in T1α immunostaining in A. Arrowheads mark the thickness of the alveolar septa. ( C ) Average size of 432 septa thickness from the lung sections of 3 animals of hSRY-ON and control newborn pups. ( D ) H&E staining of lung sections for control (left) and hSRY-ON (right) mice at P0 and P9 age. ( E ) Average size of alveolar sacs from lung sections of 3 animals per group at P0 (350 total measurements) and P9 (550 total measurements) age of hSRY-ON and control mice. Control pups showed normal alveologenesis, starting with large alveolar sacs, which go through microvascular maturation and secondary septa development with reduced overall sac sizes. hSRY-ON pups showed minimal changes in morphology and alveolar sac sizes. ( F ) Gross morphology and green fluorescence expression in brain of an hSRY-ON mouse (right) and control littermate (left). ( G ) Double immunofluorescence of calbindin D28K (green, Purkinje-specific) and neuN (red, nuclear marker for neurons) on cerebella of control (left) and hSRY-ON (right) P9 mice. Yellow arrows indicate the transverse thickness of the molecular, Purkinje and granule cellular layers. Boxed areas on top represent areas of the enlargements in bottom figures. ( H ) Average thickness of cerebellar cortex, and ( I ) relative density of Purkinje cell dendrites in the cerebella between control and hSRY-ON mice.

Article Snippet: Antibodies specific to the following proteins were obtained from various vendors, as indicated: anti-FLAG mouse IgG (clone M2, Sigma-Aldrich), anti-Calbindin D28K goat polyclonal IgG (C-20, Santa Cruz Biotechnology, Dallas, TX), anti-NeuN mouse IgG (clone A60, EMD Millipore), anti-Prosurfactant Protein C rabbit antiserum (EMD Millipore), and anti-Podoplanin hamster IgG (clone 8.1.1, Developmental Studies Hybridoma Bank, Iowa, IA).

Techniques: Immunostaining, Control, Staining, Fluorescence, Expressing, Immunofluorescence, Marker