Onoue H, Maeyama K, Nomura S, Kasugai T, Tei H, Kim H M, Watanabe T, Kitamura Y
Department of Pathology, Osaka University Medical School, Japan.
Am J Pathol. 1993 Apr;142(4):1001-7.
Ws/Ws rats have a small deletion at the tyrosine kinase domain of the c-kit gene, and practically no mast cells were detectable when the tissues were stained with alcian blue. Because alcian blue stains proteoglycans, there is a possibility that immature mast cells that do not contain a sufficient amount of proteoglycans are not detectable by this method. We examined this possibility by using other markers of mast cells. The histamine content in the skin of Ws/Ws rats was 0.3% that of control normal (+/+) rats. Because the number of alcian blue-positive mast cells in the skin of Ws/Ws rats was also 0.3% that of +/+ rats, histamine in the skin seemed to be concentrated to alcian blue-positive mast cells. Mast cells in the skin of +/+ rats express messenger RNA of Fc epsilon RI beta-subunit and c-kit protein. Because c-kit messenger RNA was normally expressed at least in the brain of Ws/Ws rats despite the small deletion, we examined the expression of Fc epsilon RI beta-subunit and c-kit messenger RNA in the skin and stomach of Ws/Ws rats by reverse transcriptase modification of polymerase chain reaction. Expression of either Fc epsilon RI beta-subunit or c-kit messenger RNA in the skin and stomach of Ws/Ws rats was estimated to be less than 1% that of +/+ rats. Moreover no Fc epsilon RI beta-subunit-expressing and no c-kit-expressing cells were detectable in the skin of Ws/Ws rats by in situ hybridization histochemistry. The present result suggests the absence of immature mast cells in tissues of Ws/Ws rats.
Ws/Ws大鼠的c-kit基因酪氨酸激酶结构域存在小的缺失,当用阿尔辛蓝对组织进行染色时,几乎检测不到肥大细胞。由于阿尔辛蓝可对蛋白聚糖进行染色,因此有可能通过这种方法检测不到含有不足量蛋白聚糖的未成熟肥大细胞。我们通过使用肥大细胞的其他标志物来研究这种可能性。Ws/Ws大鼠皮肤中的组胺含量为对照正常(+/+)大鼠的0.3%。由于Ws/Ws大鼠皮肤中阿尔辛蓝阳性肥大细胞的数量也是+/+大鼠的0.3%,皮肤中的组胺似乎集中在阿尔辛蓝阳性肥大细胞中。+/+大鼠皮肤中的肥大细胞表达FcεRIβ亚基的信使RNA和c-kit蛋白。尽管存在小的缺失,但c-kit信使RNA至少在Ws/Ws大鼠的脑中正常表达,因此我们通过聚合酶链反应的逆转录修饰来检测Ws/Ws大鼠皮肤和胃中FcεRIβ亚基和c-kit信使RNA的表达。据估计,Ws/Ws大鼠皮肤和胃中FcεRIβ亚基或c-kit信使RNA的表达量不到+/+大鼠的1%。此外,通过原位杂交组织化学在Ws/Ws大鼠的皮肤中未检测到表达FcεRIβ亚基和表达c-kit的细胞。目前的结果表明Ws/Ws大鼠组织中不存在未成熟肥大细胞。