Alam K Y, Frostholm A, Hackshaw K V, Evans J E, Rotter A, Chiu I M
Department of Internal Medicine, College of Medicine, The Ohio State University, Columbus, Ohio 43210, USA.
J Biol Chem. 1996 Nov 22;271(47):30263-71. doi: 10.1074/jbc.271.47.30263.
The present study elucidates the molecular structure of a murine fibroblast growth factor 1 (FGF-1) promoter and describes its distribution in the adult and developing mouse brain. A cDNA clone coding for FGF-1 was isolated from a mouse brain cDNA library. Nucleotide sequence analysis revealed that the clone contained, in addition to the protein coding region, an untranslated exon (FGF-1B) 34 base pairs upstream of the translation start codon ATG. The mouse cDNA clone corresponded to the sole FGF-1 transcript in the brain. An RNase protection assay was used to map the transcription start site of the 1B promoter. The sequences upstream from the major transcription initiation site lacked consensus TATA or CAAT boxes. In situ hybridization with cRNA probes specific for the 1B transcript showed the message to be restricted largely to sensory and motor nuclei in the brainstem, and to the ventral spinal cord and cerebellum. Although occasional brainstem nuclei were labeled at low levels by embryonic day 18, the majority of nuclei became detectable autoradiographically during postnatal weeks 1 and 2, and adult levels of grain density were reached during the 3rd and 4th postnatal weeks. FGF-1B mRNA was expressed in phylogenetically older brain regions, which are involved primarily in processing information from exteroceptive sensory mechanoreceptors and in motor control. The relatively late developmental expression suggests a role for FGF-1 in neuronal maturation, rather than in neurogenesis.
本研究阐明了小鼠成纤维细胞生长因子1(FGF-1)启动子的分子结构,并描述了其在成年和发育中小鼠大脑中的分布情况。从鼠脑cDNA文库中分离出编码FGF-1的cDNA克隆。核苷酸序列分析表明,该克隆除了蛋白质编码区外,在翻译起始密码子ATG上游34个碱基对处含有一个非翻译外显子(FGF-1B)。小鼠cDNA克隆与脑中唯一的FGF-1转录本相对应。采用核糖核酸酶保护试验来定位1B启动子的转录起始位点。主要转录起始位点上游的序列缺乏共有TATA或CAAT框。用针对1B转录本的cRNA探针进行原位杂交显示,该信息主要局限于脑干中的感觉和运动核,以及脊髓腹侧和小脑。虽然在胚胎第18天偶尔有低水平的脑干核被标记,但大多数核在出生后第1周和第2周通过放射自显影可检测到,并且在出生后第3周和第4周达到成年时的颗粒密度水平。FGF-1B mRNA在系统发育较古老的脑区表达,这些脑区主要参与处理来自外部感受性感觉机械感受器的信息以及运动控制。相对较晚的发育表达表明FGF-1在神经元成熟而非神经发生中起作用。