Chegini N, Zhao Y, McLean F W
Department of Obstetrics and Gynecology, University of Florida, Gainesville 32610.
Biol Reprod. 1994 May;50(5):1049-58. doi: 10.1095/biolreprod50.5.1049.
Reverse transcription polymerase chain reaction (RT-PCR) revealed that the Fallopian tubes express epidermal growth factor (EGF), transforming growth factor (TGF alpha), and EGF receptor (EGF-R) mRNA. The RT-PCR product was verified by restriction enzyme digestion analysis. Immunohistochemically, EGF, TGF alpha, and EGF-R were localized in Fallopian tubes by use of specific antibodies to human EGF, mature fragments of human TGF alpha, and monoclonal antibodies to the extracellular binding domain of EGF-R. The tubal epithelial cells were the primary site of immunoreactive EGF, TGF alpha, and EGF-R, which were present to a lesser extent in the stromal cells, smooth muscle cell layers, fibroblasts of serosal tissue, and arterial endothelial and smooth muscle cells. Using antibodies generated against the amino and carboxy termini of TGF alpha precursor produced a similar cellular distribution to that observed for mature TGF alpha. The intensity of immunoreactive TGF alpha with these antibodies was similar to that seen with EGF. The ciliated and nonciliated epithelial cells in the ampullary and isthmus regions immunostained with similar intensity for EGF, TGF alpha, and EGF-R. The immunostaining for EGF, TGF alpha, and EGF-R was cycle-dependent, was considerably higher during late proliferative and early-to-mid-secretory phases than during early proliferative and late secretory phases of the menstrual cycle, and was reduced during the postmenopausal period. Specimens obtained 5-12 yr after tubal ligation immunostained for EGF, TGF alpha, and EGF-R similarly to sections from unligated tubes taken during the same phase of the cycle. Quantitative autoradiography of 125I-EGF binding generated a pattern similar to that of immunostaining for EGF-R binding. Net grain density/100 microns 2 calculated for different cell types indicated that the epithelial cells had a significantly higher grain density than did other tubal cell types (p < 0.05) without the cycle dependency seen in the immunohistochemical study. In summary, the results demonstrate that the human Fallopian tube expresses mRNA and contains immunoreactive proteins for EGF, TGF alpha, and EGF-R as well as binding sites for 125I-EGF. The cycle dependency and lower immunostaining in postmenopausal tubes suggest a potential regulation of their expression by ovarian steroids. The results imply the importance of EGF/TGF alpha in a variety of tubal biochemical and physiological functions and possibly early embryonic development.
逆转录聚合酶链反应（RT-PCR）显示，输卵管表达表皮生长因子（EGF）、转化生长因子（TGFα）和EGF受体（EGF-R）的mRNA。RT-PCR产物经限制性内切酶消化分析验证。免疫组织化学方法中，利用针对人EGF、人TGFα成熟片段的特异性抗体以及针对EGF-R细胞外结合域的单克隆抗体，将EGF、TGFα和EGF-R定位在输卵管中。输卵管上皮细胞是免疫反应性EGF、TGFα和EGF-R的主要部位，在基质细胞、平滑肌细胞层、浆膜组织成纤维细胞以及动脉内皮和平滑肌细胞中含量较少。使用针对TGFα前体氨基和羧基末端产生的抗体，其细胞分布与成熟TGFα观察到的相似。这些抗体免疫反应性TGFα的强度与EGF相似。壶腹部和峡部区域的纤毛和非纤毛上皮细胞对EGF、TGFα和EGF-R的免疫染色强度相似。EGF、TGFα和EGF-R的免疫染色呈周期依赖性，在月经周期的晚增殖期和早至中分泌期显著高于早增殖期和晚分泌期，绝经后降低。输卵管结扎术后5 - 12年获得的标本，其EGF、TGFα和EGF-R的免疫染色与同一周期未结扎输卵管的切片相似。125I-EGF结合的定量放射自显影产生的模式与EGF-R结合的免疫染色相似。针对不同细胞类型计算的每100平方微米净颗粒密度表明，上皮细胞的颗粒密度显著高于其他输卵管细胞类型（p < 0.05），且无免疫组织化学研究中所见的周期依赖性。总之，结果表明人输卵管表达mRNA，并含有EGF、TGFα和EGF-R的免疫反应性蛋白以及125I-EGF的结合位点。绝经后输卵管的周期依赖性和较低的免疫染色表明其表达可能受卵巢甾体激素调节。结果表明EGF/TGFα在多种输卵管生化和生理功能以及可能的早期胚胎发育中具有重要作用。
