Rat cerebral mast cells undergo phenotypic changes during development

Abstract

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.