Dimitriadou V, Rouleau A, Tuong M D, Ligneau X, Newlands G F, Miller H R, Schwartz J C, Garbarg M
Laboratoire de Physiologie, Faculté de Pharmacie, Université René Descartes, Paris, France.
Brain Res Dev Brain Res. 1996 Nov 22;97(1):29-41. doi: 10.1016/s0165-3806(96)00127-7.
The evolution of rat cerebral mast cell phenotype during development was studied using antibodies against the granule chymases, rat mast cell protease I (RMCP-I) and rat mast cell protease II (RMCP-II) and their gene transcripts, as markers for serosal and mucosal mast cells, respectively. In situ hybridization using specific oligoprobes for RMCP-II permitted visualization of RMCP-II mRNA-containing cells as early as day 15 of embryonic development (E15). From E19 to day 4 postpartum (D4) their number increased whilst they migrated from the pia mater to the choroid fissure; at D8 cells expressing RMCP-II gene transcripts were no longer observed. The 3'-end untranslated nucleotide sequence of the RMCP-I cDNA was established in order to design selective cDNA probes for Northern blot analysis of both enzymes. Northern blot analysis revealed a strong expression of RMCP-I and RMCP-II mRNAs at D2. At D4, RMCP-I mRNA expression was still high, whereas that of RMCP-II was decreased. In adult brain, mRNA expression for both proteases was low, but detectable. Quantification of both proteases by ELISA showed that, from E19 to D4, levels of RMCP-II were maximal at E19 and remained constant until D4, whereas RMCP-I increased as a function of age. Thereafter, levels of both proteases decreased progressively, but were still present in the adult brain, with RMCP-II being uniformly distributed and RMCP-I concentrated in the thalamus. Immunohistochemical staining showed RMCP-II-immunoreactive cells within the pia mater at E19; on D2 and D4, cells with both RMCP-I and RMCP-II immunoreactivities were found within the choroid fissure and from D8, only RMCP-I-immunoreactive mast cells were observed. In the thalamus of adult rats, the latter had a perivascular localization. This study shows that in the adult, both types of mast cells are present, although in small numbers, except for RMCP-I-immunoreactive mast cells which are abundant in the thalamus. The changes in the number and phenotype of cerebral mast cells may result from the influence of a number of growth factors during development.
利用抗颗粒糜蛋白酶、大鼠肥大细胞蛋白酶I（RMCP-I）和大鼠肥大细胞蛋白酶II（RMCP-II）的抗体及其基因转录本，分别作为浆膜肥大细胞和黏膜肥大细胞的标志物，研究了大鼠大脑发育过程中肥大细胞表型的演变。使用针对RMCP-II的特异性寡核苷酸探针进行原位杂交，最早在胚胎发育第15天（E15）就能观察到含有RMCP-II mRNA的细胞。从E19到产后第4天（D4），它们的数量增加，同时从软脑膜迁移到脉络膜裂；在D8时，不再观察到表达RMCP-II基因转录本的细胞。确定了RMCP-I cDNA的3'端非翻译核苷酸序列，以便设计用于这两种酶的Northern印迹分析的选择性cDNA探针。Northern印迹分析显示，在D2时RMCP-I和RMCP-II mRNA有强烈表达。在D4时，RMCP-I mRNA表达仍然很高，而RMCP-II的表达则下降。在成年大脑中，两种蛋白酶的mRNA表达都很低，但仍可检测到。通过ELISA对两种蛋白酶进行定量分析表明，从E19到D4，RMCP-II的水平在E19时最高，并一直保持到D4，而RMCP-I则随年龄增加。此后，两种蛋白酶的水平逐渐下降，但在成年大脑中仍然存在，RMCP-II均匀分布，RMCP-I集中在丘脑。免疫组织化学染色显示，在E19时软脑膜内有RMCP-II免疫反应性细胞；在D2和D4时，在脉络膜裂内发现了同时具有RMCP-I和RMCP-II免疫反应性的细胞，从D8开始，只观察到RMCP-I免疫反应性肥大细胞。在成年大鼠的丘脑中，后者具有血管周围定位。这项研究表明，在成年大鼠中，两种类型的肥大细胞都存在，尽管数量很少，但丘脑中有大量的RMCP-I免疫反应性肥大细胞除外。大脑肥大细胞数量和表型的变化可能是发育过程中多种生长因子影响的结果。
