Chang W Y, Kulp S K, Sugimoto Y, Canatan H, Shidaifat F, Inpanbutr N, Lin Y C
College of Veterinary Medicine, The Ohio State University, 309 Sisson Hall, 1900 Coffey Rd., 43210-1092, Columbus, OH.
Endocrine. 1996 Dec;5(3):247-55. doi: 10.1007/BF02739057.
Keratinocyte growth factor (KGF) was originally discovered in human embryonic lung fibroblasts and is a member of the fibroblast growth factor (FGF) family. Members of the FGF family have been shown to regulate testicular function. However, the recently discovered KGF has not been studied in the testis. KGF has been detected in many other tissues, including the prostate, an organ whose development and function have been associated with presence of the testis. In this study, KGF mRNA was detected in the whole testis using reverse transcription polymerase chain reaction (RT-PCR). The 575-bp KGF-specific product was detected along with a 594-bp β-actin-specific product. To identify the cell types in which KGF mRNA was predominantly expressed, interstitial cells were physically separated from seminiferous tubules. The interstitial cells were then sorted on a discontinuous Percoll gradient and total cellular mRNAs isolated. Using RT-PCR and Southern hybridization with specific cDNA probes, the KGF mRNA was detected in interstitial cells. KGF expression levels were then evaluated semiquantitatively with a competitive RT-PCR assay. KGF expression levels were highest in interstitial cells that equilibrated between 20 and 30% Percoll. Enriched Leydig cells and seminiferous tubules expressed low levels of KGF. Finally, immunohistochemical analysis was performed on canine testes using a rabbit anti-KGF polyclonal antibody. The KGF protein was localized predominantly to peritubular cells of the canine testis. These results suggest that KGF is synthesized in the canine testis.
角质形成细胞生长因子（KGF）最初是在人胚胎肺成纤维细胞中发现的，它是成纤维细胞生长因子（FGF）家族的成员。FGF家族的成员已被证明可调节睾丸功能。然而，最近发现的KGF尚未在睾丸中进行研究。KGF已在许多其他组织中被检测到，包括前列腺，该器官的发育和功能与睾丸的存在有关。在本研究中，使用逆转录聚合酶链反应（RT-PCR）在整个睾丸中检测到了KGF mRNA。检测到了575 bp的KGF特异性产物以及594 bp的β-肌动蛋白特异性产物。为了确定KGF mRNA主要表达的细胞类型，将间质细胞与曲细精管进行了物理分离。然后将间质细胞在不连续的Percoll梯度上进行分选，并分离出总细胞mRNA。使用RT-PCR和与特异性cDNA探针的Southern杂交，在间质细胞中检测到了KGF mRNA。然后用竞争性RT-PCR测定法对KGF表达水平进行半定量评估。在平衡于20%至30% Percoll之间的间质细胞中，KGF表达水平最高。富集的睾丸间质细胞和曲细精管表达低水平的KGF。最后，使用兔抗KGF多克隆抗体对犬睾丸进行了免疫组织化学分析。KGF蛋白主要定位于犬睾丸的管周细胞。这些结果表明KGF是在犬睾丸中合成的。
