LOCALIZATION OF TYPE-I, TYPE-III AND TYPE-IV COLLAGEN MESSENGER-RNAS IN RAT-HEART CELLS BY INSITU HYBRIDIZATION
LOCALIZATION OF TYPE-I, TYPE-III AND TYPE-IV COLLAGEN MESSENGER-RNAS IN RAT-HEART CELLS BY INSITU HYBRIDIZATION
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DOI：
10.1016/0022-2828(89)91498-3
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发表时间：
1989-01-01
影响因子：
5
通讯作者：
GIAMBRONE, MA
中科院分区：
文献类型：
--
作者：
EGHBALI, M;BLUMENFELD, OO;GIAMBRONE, MA
先前对幼年大鼠心脏中几种胶原的细胞起源的研究(Eghbali等人，1988)表明，I型和III型胶原的mRNAs存在于非心肌细胞中，主要是成纤维细胞，而IV型胶原的mRNAs存在于心肌细胞和非心肌细胞中。在本研究中，通过原位杂交在大鼠心脏组织和分离的心肌细胞中实现了胶原mRNAs的细胞定位。冰冻组织切片、分离的心肌细胞、培养的新生心肌细胞和成纤维细胞与型特异性胶原蛋白、肌动蛋白和肌球蛋白重链的DNA探针杂交。通过暗场成像观察到银粒。在心脏切片中，I型和III型mRNAs主要靠近心肌细胞和间质，在间质中已知存在成纤维细胞。相反，IV型胶原基因在心肌细胞和间质中均有表达。在新鲜分离的成人心肌细胞和培养的新生心肌细胞中，IV型胶原mRNA表达阳性，而I型胶原mRNA表达阴性。在培养的新生儿成纤维细胞中，IV型和I型胶原mRNAs含量均较高。
Previous studies investigating the cellular origins of several collagens in young adult rats hearts (Eghbali et al., 1988) demonstrated that the mRNAs for types I and III collagen occurred in non-myocyte cells, mostly fibroblasts, whereas the mRNA for type IV collagen was observed in both myocytes and non-myocyte cells. In the present study, cellular localization of collagen mRNAs has been achieved by in situ hybridization in rat heart tissue and in isolated heart cells. Frozen tissues sections, isolated cardiomycytes, cultured neonatal cardiomyocytes and fibroblasts were hybridized with DNA probes for type-specific collagens, actin, and myosin heavy chain. Silver grains were visualized by dark field imaging. In heart sections, types I and III mRNAs were observed predominantly adjacent to myocytes and in the interstitium, where fibroblasts are known to be present. In contrast, type IV collagen mRNA were identified both within the myocytes and the interstitium. In freshly isolated adult cardiomyocytes and in cultured neonatal cardiomyocytes, collagen type IV mRNA was observed but type I collagen mRNA was not. In cultured neonatal fibroblasts, both types IV and I collagen mRNAs were abundant.