Combs J W, Lagunoff D, Benditt E P
J Cell Biol. 1965 Jun;25(3):577-92. doi: 10.1083/jcb.25.3.577.
Histochemical reactions and radioautography were used to investigate the sequence of mast cell development in rat embryos. Mast cells arise ubiquitously in and are confined to the loose connective tissue in the embryo. The alcian blue-safranin reaction distinguishes between weakly sulfated and strongly sulfated mucopolysaccharides by a shift from alcian blue to safranin staining. Based on this reaction and morphologic characteristics, four stages were identified. Stage I mast cells are lymphocyte-like cells with cytoplasmic granules which invariably stain blue with the alcian blue-safranin reaction. In Stage II cells the majority of granules are alcian blue-positive, but some safranin-positive granules have appeared. Stage III mast cells are distinguished by a majority of safranin-positive cytoplasmic granules; some alcian blue-positive granules still remain. Stage IV cells contain only safranin-positive granules. Thymidine-H(3) uptake and identification of mitotic figures indicates that mast cells in Stages I and II comprise a mitotic pool while those in Stages III and IV are mitotically inactive. The pattern of S(35)O(4) incorporation and the sequence of appearance of histochemically identifiable mast cell constituents corroborates division of the proliferation and differentiation of embryonic mast cells into the four stages described above. The process of formation of mast cell granules is interpreted as reflecting the synthesis and accumulation of a heparin precursor in alcian blue positive granules followed by the synthesis and accumulation of highly N-sulfated heparin along with mast cell chymase and finally histamine in safranin-positive granules.
采用组织化学反应和放射自显影术研究大鼠胚胎中肥大细胞的发育顺序。肥大细胞在胚胎中普遍产生,且局限于疏松结缔组织。阿尔辛蓝-番红反应通过从阿尔辛蓝染色转变为番红染色来区分弱硫酸化和强硫酸化的黏多糖。基于此反应和形态学特征,确定了四个阶段。I期肥大细胞是淋巴细胞样细胞,其胞质颗粒在阿尔辛蓝-番红反应中总是染成蓝色。在II期细胞中,大多数颗粒为阿尔辛蓝阳性,但出现了一些番红阳性颗粒。III期肥大细胞的特征是大多数胞质颗粒为番红阳性;仍有一些阿尔辛蓝阳性颗粒。IV期细胞仅含有番红阳性颗粒。胸腺嘧啶-H(3)摄取和有丝分裂图像的鉴定表明,I期和II期的肥大细胞构成一个有丝分裂池,而III期和IV期的肥大细胞无有丝分裂活性。S(35)O(4)掺入模式和组织化学可识别的肥大细胞成分的出现顺序证实了胚胎肥大细胞的增殖和分化分为上述四个阶段。肥大细胞颗粒的形成过程被解释为反映了阿尔辛蓝阳性颗粒中肝素前体的合成和积累,随后是高度N-硫酸化肝素以及肥大细胞糜蛋白酶的合成和积累,最后是番红阳性颗粒中组胺的合成和积累。