Journal: Toxicologic pathology

Article Title: Loss of Immunohistochemical Reactivity in Association With Handling-induced Dark Neurons in Mouse Brains

doi: 10.1177/0192623319896263

Figure Lengend Snippet: Representative images of the left dorsal hippocampal formation from two middle-aged (14–15 months) male C57BL/6N mice (in columns of adjacent tissue sections, original scan 20x). On hematoxylin and eosin staining, (A) dark neurons having contracted hyperbasophilic cell bodies and dendrites are shown in the stratum pyramidale (between arrows), in comparison with (B) well-preserved neurons. On diaminobenzidine immunohistochemistry for (C, D) caveolin-3 (CAV3, neuronal perikarya/neurites, with hematoxylin counterstaining), (E, F) retinoblastoma binding protein-4 chromatin remodeling factor (RBBP4, neuronal nuclei), (G, H) glial fibrillary acidic protein (GFAP, astroglia), and (I, J) allograft inflammatory factor-1 (AIF1, microglia), focal loss of immunoreactivity for CAV3, RBBP4, and GFAP is observed in a pattern that spatially matches the presence of dark neurons, whereas the variation of AIF1 immunoreactivity is not conspicuous. Insets = high magnification (original scan 40x).

Article Snippet: Primary antibodies used were directed against CAV3 (mouse monoclonal clone A-3 specific for an epitope mapping amino acid residues 3–40 at the N-terminus of mouse CAV3, sc-5310, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:10,000 dilution), RBBP4 (mouse monoclonal clone 11G10, GTX70232, GeneTex, Irvine, CA, USA; 1:40,000 dilution), 7 GFAP (rabbit polyclonal, Z0334, Dako, Carpinteria, CA, USA; 1:1,000 dilution), and AIF1 (rabbit polyclonal, 019-19741, Wako, Richmond, VA, USA; 1:1,000 dilution).

Techniques: Staining, Comparison, Immunohistochemistry, Binding Assay

Journal: Toxicologic pathology

Article Title: Loss of Immunohistochemical Reactivity in Association With Handling-induced Dark Neurons in Mouse Brains

doi: 10.1177/0192623319896263

Figure Lengend Snippet: Scatter plots showing the relationship between the hyperbasophilia density of dark neurons and the immunoreactivity density of each protein marker in the left dorsal hippocampal formation. The negative correlation is observed for (A) caveolin-3 (CAV3; r = −0.444, p = 0.0032), (B) retinoblastoma binding protein-4 chromatin remodeling factor (RBBP4; r = −0.614, p < 0.0001), and (C) glial fibrillary acidic protein (GFAP; r = −0.527, p = 0.0003) but not statistically significant for (D) allograft inflammatory factor-1 (AIF1; r = −0.246, p = 0.116; n = 42, Spearman’s rho).

Article Snippet: Primary antibodies used were directed against CAV3 (mouse monoclonal clone A-3 specific for an epitope mapping amino acid residues 3–40 at the N-terminus of mouse CAV3, sc-5310, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:10,000 dilution), RBBP4 (mouse monoclonal clone 11G10, GTX70232, GeneTex, Irvine, CA, USA; 1:40,000 dilution), 7 GFAP (rabbit polyclonal, Z0334, Dako, Carpinteria, CA, USA; 1:1,000 dilution), and AIF1 (rabbit polyclonal, 019-19741, Wako, Richmond, VA, USA; 1:1,000 dilution).

Techniques: Marker, Binding Assay

Journal: Toxicologic pathology

Article Title: Loss of Immunohistochemical Reactivity in Association With Handling-induced Dark Neurons in Mouse Brains

doi: 10.1177/0192623319896263

Figure Lengend Snippet: The hyperbasophilia density of dark neurons and the immunoreactivity density of protein markers in the hippocampal formation by age/diet mouse groups a .

Article Snippet: Primary antibodies used were directed against CAV3 (mouse monoclonal clone A-3 specific for an epitope mapping amino acid residues 3–40 at the N-terminus of mouse CAV3, sc-5310, Santa Cruz Biotechnology, Santa Cruz, CA, USA; 1:10,000 dilution), RBBP4 (mouse monoclonal clone 11G10, GTX70232, GeneTex, Irvine, CA, USA; 1:40,000 dilution), 7 GFAP (rabbit polyclonal, Z0334, Dako, Carpinteria, CA, USA; 1:1,000 dilution), and AIF1 (rabbit polyclonal, 019-19741, Wako, Richmond, VA, USA; 1:1,000 dilution).

Techniques: