Journal: Developmental Neuroscience

Article Title: Developmental Activation of the Proteolipid Protein Promoter Transgene in Neuronal and Oligodendroglial Cells of Neostriatum in Mice

doi: 10.1159/000330321

Figure Lengend Snippet: PLP promoter transgene activation in neuronal and oligodendroglial phenotypes of neostriatal cells during the first postnatal week (P4). a EGFP+ neuronal group. Green fluorescent EGFP+ cells have medium spherical cell bodies with red fluorescent NeuN+ label in nuclei. Green and red labels colocalize in merged images. b EGFP+ oligodendroglial (OPC) group. EGFP+ cells have small bipolar cell bodies (prominent perinuclear label) with PDGFRα+ label in perikaryal cytoplasm. Labels colocalize in merged images. c EGFP+ oligodendroglial (OPC) group. EGFP+ cells have small bipolar and spherical cell bodies (prominent perinuclear label) with NG2+ label in perikaryal cytoplasm. Labels colocalize in merged images. Note lack of PDGFRα and NG2 labels in medium spherical EGFP+ cells adjacent to doubly labeled OPCs in b and c (arrows). d A significantly larger proportion of EGFP+/NeuN+ cells (30 of 30 cells) displays spherical cell bodies in comparison to EGFP+/PDGFRα+ (12 of 21 cells) or EGFP+/NG2+ (7 of 13 cells) profiles (NeuN+ vs. PDGFRα+: χ2 = 31.22, d.f. = 1; NeuN+ vs. NG2+: χ2 = 32.18, d.f. = 1). e A significantly larger proportion of EGFP+ cells contain NeuN (69 ± 11%) than PDGFRα+ (10 ± 4%) or NG2 [8 ± 3%; F (28.2) = 44.85; reliable NeuN vs. PDGFRα and NeuN vs. NG2 contrasts; ∗ pαs < 0.05]. a–c Scale bar for merged profiles = 10 μm.

Article Snippet: The following primary antibodies were tested: mouse monoclonal anti-NeuN (1:800, Millipore, Billerica, Mass., USA); mouse monoclonal anti-MAP2 AP-20 clone (1:800, Sigma-Aldrich); rabbit monoclonal anti-DARPP-32 (1:400, Cell Signaling Technology, Boston, Mass., USA); rabbit polyconal anti-NG2 (1:300, Millipore), rat monoclonal anti-PDGFRα (1:200, BD Biosciences, San Jose, Calif., USA); rat anti-PLP/DM20 clone AA3 (gift from Steven Pfeiffer, University of Connecticut Health Science Center, Conn., USA); rabbit polyconal anti-srPLP (1:400).

Techniques: Activation Assay, Labeling