The distinction of astrocytomas and oligodendrogliomas, mainly pilocytic astrocytomas (PILOs) from infiltrating astrocytomas and oligodendrogliomas (ODs), and high-grade oligodendrogliomas from glioblastomas (GBMs), poses a serious clinical problem. There is no useful immunohistochemical (IHC) marker to differentiate these gliomas, and sometimes the differential diagnosis between them is arbitrary. We identified galectin-3 (Gal-3) as a possible tool to differentiate them based on gene expression profiles of GBMs. We confirmed the differential expression in 45 surgical samples (thirteen GBMs; seven PILOs; 5 grade II ODs; 5 anaplastic oligodendrogliomas [AODs], including 2 Oligo-astrocytomas; 8 diffuse astrocytomas [ASTs], and 7 non-neoplastic samples) by quantification of Gal-3 gene expression by real-time quantitative PCR (rt-PCR). Higher expression of Gal-3 was observed in GBMs and PILOs than in OD, AODs and ASTs. The IHC expression of Gal-3 was evaluated in 90 specimens (fifteen PlLOs, fourteen ASTs, 10 anaplastic astrocytomas, fifteen GBMs, eleven ODs, fifteen AODs, and 10 dysembryoplastic neuroepithelial tumors). The mean labeling score for Gal-3 determined according to the percentage of labeled cells in the tumor bulk was significantly different in GBMs versus AODs and in PILOs versus ASTs. Hence, Gal-3 is differentially expressed in central nervous system tumors, making IHC detection of Gal-3 a useful tool in distinguishing between these gliomas.