Kondo J, Kusachi S, Ninomiya Y, Yoshioka H, Oohashi T, Doi M, Murakami T, Moritani H, Kumashiro H, Tsuji T
First Department of Internal Medicine, Okayama University Medical School, Japan.
Jpn Heart J. 1998 Mar;39(2):211-20. doi: 10.1536/ihj.39.211.
The type XVII collagen alpha 1 chain has been identified as a component of the type I hemidesmosome, and is thus thought to play a role in extracellular matrix (ECM) maintenance and signal transduction between the cell and the ECM. We examined the expression of type XVII collagen alpha 1 chain mRNA in the mouse heart by Northern blot analysis and determined the sequential changes of its expression in different developmental stages of the heart using the reverse transcriptase-polymerase chain reaction (RT-PCR) method. Northern blotting: Total RNA was extracted from 10 adult mouse hearts by the guanidine/cesium method. Hybridization was performed with mouse cDNA for alpha 1 (XVII) collagen. RT-PCR: Total RNA was extracted from 7 embryos, 4 neonates and 8 adult mice. Reverse transcription was performed using oligo-dT primer and MMLV. Amplification was carried out in alpha 1 (XVII) collagen and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). GAPDH served as an internal control. Northern blotting revealed a 5.6 kb signal that was identical to that of the alpha 1 (XVII) of skin and transformed keratinocyte reported previously. The sequences of the PCR products were also identical to those reported. The normalized expression ratios of alpha 1 (XVII) were 0.91 +/- 0.20 in the embryonic heart, 0.36 +/- 0.20 in the neonatal heart and 0.96 +/- 0.21 in the adult heart. In conclusion, we identified the expression of type XVII collagen alpha 1 chain mRNA in the mouse heart, suggesting that the type I hemidesmosome is located in the heart. The results of the RT-PCR at different developmental stages of the heart suggest that type XVII collagen contributes not only to cardiogenesis in the embryonic stage but also to maintenance of architecture and function in the adult heart.
XVII型胶原蛋白α1链已被确定为I型半桥粒的一个组成部分，因此被认为在细胞外基质（ECM）维持以及细胞与ECM之间的信号转导中发挥作用。我们通过Northern印迹分析检测了小鼠心脏中XVII型胶原蛋白α1链mRNA的表达，并使用逆转录聚合酶链反应（RT-PCR）方法确定了其在心脏不同发育阶段表达的顺序变化。Northern印迹法：采用胍/铯法从10只成年小鼠心脏中提取总RNA。用小鼠α1（XVII）胶原蛋白的cDNA进行杂交。RT-PCR：从7个胚胎、4只新生小鼠和8只成年小鼠中提取总RNA。使用寡聚dT引物和MMLV进行逆转录。在α1（XVII）胶原蛋白和甘油醛-3-磷酸脱氢酶（GAPDH）中进行扩增。GAPDH作为内对照。Northern印迹显示出一个5.6 kb的信号，与先前报道的皮肤和转化角质形成细胞的α1（XVII）信号相同。PCR产物的序列也与报道的序列相同。α1（XVII）的标准化表达率在胚胎心脏中为0.91±0.20，在新生心脏中为0.36±0.20，在成年心脏中为0.96±0.21。总之，我们确定了小鼠心脏中XVII型胶原蛋白α1链mRNA的表达，表明I型半桥粒存在于心脏中。心脏不同发育阶段的RT-PCR结果表明，XVII型胶原蛋白不仅在胚胎期对心脏发生有贡献，而且在成年心脏的结构和功能维持中也有作用。
