Tong B J, Das S K, Threadgill D, Magnuson T, Dey S K
Department of Physiology, Ralph L. Smith Research Center, University of Kansas Medical Center, Kansas City, 66160, USA.
Endocrinology. 1996 Apr;137(4):1492-6. doi: 10.1210/endo.137.4.8625928.
The present investigation examined the differential expression of the full-length (fl) and the truncated (tr) forms of the epidermal growth factor receptor (EGFr) in the preimplantation mouse uterus and blastocyst. Northern blot hybridization using a complementary RNA probe specific to the full-length form (EGFr-fl) detected a 6.5-kb transcript, whereas that of the truncated form (EGFr-tr) detected a 2.7-kb transcript in the preimplantation mouse uterus on days 1 and 4 of pregnancy (day 1 = vaginal plug). In situ hybridization using these probes detected the EGFr-fl transcripts only in the stroma and myometrium, but not in the epithelium, whereas EGFr-tr transcripts were detected in all major uterine cell-types. To confirm the results of in situ hybridization, RT-PCR was performed on RNA isolated from separated uterine cell-types on day 4 of pregnancy using sequence specific primers for the two forms of the receptor. The results concur that the EGFr-tr transcript is expressed in the epithelium, stroma and myometrium, whereas that of the EGFr-fl transcript is not expressed in the epithelium. In the preimplantation blastocyst, RT-PCR detected the EGFr-fl messenger RNA, but not the EGFr-tr messenger RNA. These results suggest that the blastocyst, not the uterine epithelium, is the target for EGF family of growth factors in embryo-uterine interaction during implantation.
本研究检测了表皮生长因子受体(EGFr)全长(fl)和截短(tr)形式在植入前小鼠子宫和囊胚中的差异表达。使用针对全长形式(EGFr-fl)的互补RNA探针进行的Northern印迹杂交在妊娠第1天和第4天(第1天 = 阴道栓)的植入前小鼠子宫中检测到一个6.5 kb的转录本,而针对截短形式(EGFr-tr)的杂交检测到一个2.7 kb的转录本。使用这些探针进行的原位杂交仅在基质和肌层中检测到EGFr-fl转录本,而在上皮中未检测到,而在所有主要的子宫细胞类型中均检测到EGFr-tr转录本。为了证实原位杂交的结果,在妊娠第4天使用针对受体两种形式的序列特异性引物对从分离的子宫细胞类型中分离的RNA进行了RT-PCR。结果一致表明,EGFr-tr转录本在上皮、基质和肌层中表达,而EGFr-fl转录本在上皮中不表达。在植入前的囊胚中,RT-PCR检测到了EGFr-fl信使RNA,但未检测到EGFr-tr信使RNA。这些结果表明,在植入过程中胚胎-子宫相互作用中,囊胚而非子宫上皮是表皮生长因子家族生长因子的作用靶点。