Lacey D L, Erdmann J M, Tan H L
Department of Pathology, Jewish Hospital at Washington University, St. Louis, Missouri 63110.
J Cell Biochem. 1994 Mar;54(3):365-71. doi: 10.1002/jcb.240540312.
Type 5 acid phosphatase is a lysosomal enzyme expressed in cells of monocyte/macrophage lineage frequently used as a marker of osteoclastic differentiation. Oligonucleotide primers for DNA amplification were designed following sequence alignment of rat bone and human macrophage type 5 acid phosphatases. DNA (330 bp in length) obtained using these primers and reverse transcribed total cell RNA from in vitro generated murine osteoclastic cells was cloned and sequenced. DNA sequence analysis of two clones demonstrates that the amplified material was 91% and 96% identical to rat bone type 5 acid phosphatase at the nucleotide and amino acid level, respectively. Northern blots of murine tissue RNA show the presence of 1.5-kb transcripts that are most highly expressed in the long bones. Total cell RNA from the osteoclastic cells contain a marked level of type 5 acid phosphatase mRNA when compared to the levels seen in the tissue samples. Additionally, osteoclastic cell RNA contains two additional transcripts of 2.5 and 5 kb. Bone marrow macrophages grown in the presence of M-CSF express low levels of the 1.5-kb transcript with no signal observed for either of the two larger transcripts that were seen in the osteoclastic RNA samples. Importantly, bone marrow macrophage 1.5-kb type 5 acid phosphatase transcript levels are increased by interleukin 4 treatment in both a time and concentration-dependent manner. These findings indicate that type 5 acid phosphatase, while a cytochemical marker for osteoclasts, can be induced in macrophages by agents that block in vitro osteoclastic differentiation. Increased type 5 acid phosphatase may play a role in interleukin 4-stimulated monocyte activities.
5型酸性磷酸酶是一种溶酶体酶,在单核细胞/巨噬细胞谱系的细胞中表达,常被用作破骨细胞分化的标志物。根据大鼠骨和人巨噬细胞5型酸性磷酸酶的序列比对设计了用于DNA扩增的寡核苷酸引物。使用这些引物获得的DNA(长度为330 bp)以及来自体外生成的小鼠破骨细胞的逆转录总细胞RNA被克隆并测序。对两个克隆的DNA序列分析表明,扩增产物在核苷酸和氨基酸水平上分别与大鼠骨5型酸性磷酸酶有91%和96%的同一性。小鼠组织RNA的Northern印迹显示存在1.5 kb的转录本,在长骨中表达最高。与组织样本中的水平相比,破骨细胞的总细胞RNA含有显著水平的5型酸性磷酸酶mRNA。此外,破骨细胞RNA还含有另外两个2.5 kb和5 kb的转录本。在M-CSF存在下生长的骨髓巨噬细胞表达低水平的1.5 kb转录本,在破骨细胞RNA样本中观察到的两个较大转录本均未检测到信号。重要的是,白细胞介素4处理可使骨髓巨噬细胞1.5 kb 5型酸性磷酸酶转录本水平呈时间和浓度依赖性增加。这些发现表明,5型酸性磷酸酶虽然是破骨细胞的细胞化学标志物,但可被阻断体外破骨细胞分化的因子在巨噬细胞中诱导产生。5型酸性磷酸酶的增加可能在白细胞介素4刺激的单核细胞活性中起作用。