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, 164 (5), 1717-25

Anti-human Olig2 Antibody as a Useful Immunohistochemical Marker of Normal Oligodendrocytes and Gliomas

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Anti-human Olig2 Antibody as a Useful Immunohistochemical Marker of Normal Oligodendrocytes and Gliomas

Hideaki Yokoo et al. Am J Pathol.

Abstract

Olig2 is a recently identified transcription factor involved in the phenotype definition of cells in the oligodendroglial lineage. The expression of Olig2 transcript has been demonstrated in human oligodendroglial tumors, although the protein expression has not been studied extensively. We developed a polyclonal antibody to human Olig2 and analyzed it immunohistochemically. The antibody depicted a single distinct band of predicted molecular weight by Western blotting, and did not cross-react with human Olig1. In normal human brain tissue, the nuclei of oligodendrocytes of interfascicular, perivascular, and perineuronal disposition were clearly labeled by the antibody. Similarly, the nuclei of oligodendroglial tumors were labeled. There was no apparent correlation between the staining intensity and histological grade. Astrocytic components within the tumors were generally less or not stained. Astrocytic tumors were also positive with the Olig2 antiserum to a lesser extent, and the difference between oligodendroglial and astrocytic tumors was demonstrated by a statistical analysis. Olig2 and glial fibrillary acidic protein were expressed in a mutually exclusive manner, and Olig2 expression was cell-cycle related. Neither central neurocytoma nor schwannoma cases were stained. Our antibody was demonstrated to be useful in recognizing normal oligodendrocytes on paraffin sections, and applicable in diagnosis of some brain tumors.

Figures

Figure 1
Figure 1
Western blotting of Olig2-C. A distinct band of predicted molecular size (32 kd) is identified in the left lane. In the right lane, the band is not illustrated when the primary antibody is replaced with preabsorbed Olig2-C.
Figure 2
Figure 2
NIH3T3 cells co-transfected with human Olig1 and EGFP are immunostained (red) either by anti-Olig1 antiserum (a) or Olig2-C (b). Nuclei are counterstained with 4′,6-diamidino-2-phenylindole (blue), and EGFP emits green fluorescence. Positive red signals are recognized in a, while negative in b, demonstrating that transfection is successful, and Olig2-C does not react with Olig1. Similarly, cytoplasmic Olig1 signal was noted at the murine Olig1 transfection (data not shown). Scale bars, 25 μm.
Figure 3
Figure 3
a to c: Immunohistochemistry with Olig2-C labels nuclei of the cells of perineuronal (a), perivascular (b), and interfascicular (c) dispositions. The size of immunolabeled nuclei is smaller than that of neurons (asterisk) and astrocytes. These characteristics are compatible with oligodendrocytes. d: Double immunostaining of Olig2-C (brown) and anti-glial fibrillary acidic protein antiserum (deep purple) demonstrates a nonoverlapping positive reaction. e: The nuclear staining is remarkably reduced when the primary antibody is replaced with a preabsorbed fraction of Olig2-C. f: Immunohistochemistry with Olig2-N displays weaker nuclear staining with diffuse background noise. Original magnifications, ×500 (a–f).
Figure 4
Figure 4
a and b: Oligodendroglioma cells with perinuclear haloes (a) and those showing perineuronal satellitosis (b) are stained by Olig2-C. Note that not all nuclei are immunostained. c: Olig2-C reaction is noted in anaplastic oligodendroglioma. This specimen is highly mitotic and accompanied by abundant apoptotic bodies. The apoptotic nuclei are Olig2-C-negative. d: Double staining of Olig2-C and H&E on anaplastic oligodendroglioma illustrates that cells with astrocytic differentiation are weakly or not stained by Olig2-C. e: A nucleus under mitotic phase is Olig2-C-negative (anaplastic oligodendroglioma). f: Dual-immunofluorescent staining with MIB-1 (red) and Olig2-C (green) illustrates MIB-1-positive/Olig2-C-negative cells in addition to some double-positive cells (anaplastic oligodendroglioma). g and h: Representative cases of astrocytic tumors (g, fibrillary astrocytoma; h, glioblastoma). Scattered Olig2-C-positive nuclei are noted. The cell bodies possessing labeled nuclei appear vacant or inconspicuous, whereas truly astrocytic cells with rich cytoplasm tend to be Olig2-C-negative. i and j: Central neurocytoma (i) and schwannoma (j) are negative with Olig2-C. Original magnifications: ×250 (a, f, g, i, j); ×500 (b–e, h).

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