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I型前胶原α1 mRNA在冈上肌腱撕裂处的定位。使用地高辛标记的寡核苷酸探针进行原位杂交。

Localization of mRNA of procollagen alpha 1 type I in torn supraspinatus tendons. In situ hybridization using digoxigenin labeled oligonucleotide probe.

作者信息

Hamada K, Okawara Y, Fryer J N, Tomonaga A, Fukuda H

机构信息

Department of Orthopaedic Surgery, Tokai University Oiso Hospital, Kanagawa, Japan.

出版信息

Clin Orthop Relat Res. 1994 Jul(304):18-21.

PMID:8020212
Abstract

A tendon is predominantly composed of collagen Type I. To determine the synthesis of collagen Type I after a rotator cuff tear, an in situ hybridization technique was applied to localize cells containing procollagen alpha 1 Type I in the proximal stump of five torn supraspinatus tendons obtained during surgery. Specimens were fixed in 10% buffered formalin, embedded in paraffin, and sectioned at 6 microns. A 22 mer oligonucleotide corresponding to a sequence coding a part of human procollagen alpha 1 Type I messenger RNA (mRNA) was used as a hybridization probe. The probe was 3'-end labeled with digoxigenin-11-dUTP, and the probe-mRNA hybrids were enzymatically visualized using conventional chromogens for alkaline phosphatase. The procollagen alpha 1 type I mRNA was clearly observed in the cells near the margin of the tear. However, they were not consistently found in the vicinity of the intratendinous extension of the tear and in cells of the subacromial bursa. It is concluded that this method should be used to study the characteristics of collagen synthesis in a torn rotator cuff tendon.

摘要

肌腱主要由I型胶原蛋白组成。为了确定肩袖撕裂后I型胶原蛋白的合成情况,采用原位杂交技术对手术中获取的5条撕裂的冈上肌腱近端残端中含I型前胶原α1的细胞进行定位。标本用10%缓冲福尔马林固定,石蜡包埋,切成6微米厚的切片。一条与编码人I型前胶原α1信使核糖核酸(mRNA)一部分的序列对应的22聚体寡核苷酸用作杂交探针。该探针用洋地黄毒苷-11-dUTP进行3′端标记,探针-mRNA杂交体用碱性磷酸酶的常规显色剂进行酶促显色。在撕裂边缘附近的细胞中清晰观察到I型前胶原α1 mRNA。然而,在撕裂的腱内延伸附近和肩峰下滑囊的细胞中并未始终发现它们。得出的结论是,该方法应用于研究撕裂的肩袖肌腱中胶原蛋白合成的特征。

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