Krishna N S, Little S S, Getchell T V
Department of Surgery, University of Kentucky College of Medicine, Lexington 40536, USA.
J Comp Neurol. 1996 Sep 16;373(2):297-307. doi: 10.1002/(SICI)1096-9861(19960916)373:2<297::AID-CNE11>3.0.CO;2-I.
Olfactory receptor neurons are continuously replaced postnatally through the initiation of the division and terminal differentiation of progenitor cells located in the basal layer of the olfactory epithelium. Although the factors that regulate this process in vivo are not known, recent in vitro studies demonstrated that members of the epidermal growth factor (EGF) family including transforming growth factor-alpha (TGF alpha) and EGF are highly potent in promoting the proliferation of progenitor cells, suggesting a role for the EGF receptor (EGFR), which is the molecular receptor for both mitogens. We have examined the expression of EGFR mRNA and protein in the olfactory epithelium by using reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis and have examined their cellular localization with in situ RT-PCR and immunocytochemistry. RT-PCR and Southern blot analysis demonstrated that EGFR mRNA is expressed in the olfactory mucosa and also in the positive control tissues, kidney and tongue. The 170-kDa EGFR protein was identified with Western blot analysis in the olfactory epithelium and control tissues. Our results using in situ RT-PCR localized EGFR mRNA-expressing cells more extensively in the basal cell layer of the epithelium than did the immunocytochemical methods. These results suggest that EGFR mediates the mitogenic effect of TGF alpha and/or EGF on the quiescent basal cells to initiate the cell cycle.
嗅觉受体神经元在出生后通过位于嗅觉上皮基底层的祖细胞开始分裂和终末分化而不断更新。尽管体内调节这一过程的因素尚不清楚,但最近的体外研究表明,包括转化生长因子-α(TGFα)和表皮生长因子(EGF)在内的表皮生长因子(EGF)家族成员在促进祖细胞增殖方面具有高效能,提示作为这两种有丝分裂原分子受体的表皮生长因子受体(EGFR)发挥了作用。我们通过逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹分析检测了嗅觉上皮中EGFR mRNA和蛋白质的表达,并通过原位RT-PCR和免疫细胞化学检测了它们的细胞定位。RT-PCR和Southern印迹分析表明,EGFR mRNA在嗅觉黏膜以及阳性对照组织肾脏和舌中均有表达。通过蛋白质免疫印迹分析在嗅觉上皮和对照组织中鉴定出了170-kDa的EGFR蛋白。我们使用原位RT-PCR的结果显示,与免疫细胞化学方法相比,表达EGFR mRNA的细胞在上皮基底层中的定位范围更广。这些结果表明,EGFR介导TGFα和/或EGF对静止基细胞的促有丝分裂作用,从而启动细胞周期。