Distinct regions control transcriptional activation of the alpha1(VI) collagen promoter in different tissues of transgenic mice.
Distinct regions control transcriptional activation of the alpha1(VI) collagen promoter in different tissues of transgenic mice.
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不同的区域控制转基因小鼠不同组织中α1（VI）胶原蛋白启动子的转录激活。
DOI：
10.1083/jcb.135.4.1163
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发表时间：
1996-11
期刊：
影响因子：
--
通讯作者：
Bressan GM
中科院分区：
文献类型：
--
作者：
Braghetta P;Fabbro C;Piccolo S;Marvulli D;Bonaldo P;Volpin D;Bressan GM
为了鉴定参与α 1（VI）胶原蛋白链转录的组织特异性调节的区域，产生了携带与E. coli β-半乳糖苷酶基因。通过胚胎的X-gal染色分析转基因表达模式显示：（a）近端0.6 kb的启动子序列激活了浅表肌肉腱膜插入皮肤的间充质细胞中的转录;肌腱也呈微弱阳性。(b)从转录起始位点开始的-4.0和-5.4kb之间的区域是神经中的转基因激活所必需的。它还驱动关节、椎间盘、表皮下和触须间充质中的表达。(c)在骨骼肌和脑膜中高水平转录所必需的片段包括在-6.2和-7.5kb之间。肌肉中的阳性细胞大多为单核细胞，可能包括结缔组织成分，但不排除成肌细胞染色。该片段还激活了关节、椎间盘以及表皮下和触须间充质中的表达。(d)由序列-4.0至-5.4和-6.2至-7.5诱导的触须中的β-半乳糖苷酶染色不一致：在后一种序列中，标记的核主要在腹侧和后象限中发现，并且在组织学上，在滤泡周围和之间的间充质外层中，而对于前者，其余象限为阳性，并且表达细胞主要位于真皮鞘的内层。(e)其他组织，特别是肺、肾上腺、消化道，产生大量的VI型胶原蛋白，未对β-半乳糖苷酶染色。(f)在内源基因失活的中枢神经系统和视网膜中，大多数品系表达lacZ转基因。这些数据表明，在不同的组织中的α 1（VI）的转录是由不同的序列元素在一个模块化的安排，一种机制，赋予高度的灵活性，在发展过程中的时空表达模式。
To identify regions involved in tissue specific regulation of transcription of the alpha1(VI) collagen chain, transgenic mice were generated carrying various portions of the gene's 5'-flanking sequence fused to the E. coli beta-galactosidase gene. Analysis of the transgene expression pattern by X-gal staining of embryos revealed that: (a) The proximal 0.6 kb of promoter sequence activated transcription in mesenchymal cells at sites of insertion of superficial muscular aponeurosis into the skin; tendons were also faintly positive. (b) The region between -4.0 and -5.4 kb from the transcription start site was required for activation of the transgene in nerves. It also drove expression in joints, in intervertebral disks, and in subepidermal and vibrissae mesenchyme. (c) The fragment comprised within -6.2 and -7.5 kb was necessary for high level transcription in skeletal muscle and meninges. Positive cells in muscle were mostly mononuclear and probably included connective tissue elements, although staining of myoblasts was not ruled out. This fragment also activated expression in joints, in intervertebral disks, and in subepidermal and vibrissae mesenchyme. (d) beta-Galactosidase staining in vibrissae induced by the sequences -4.0 to -5.4 and -6.2 to -7.5 was not coincident: with the latter sequence labeled nuclei were found mainly in the ventral and posterior quadrant, and, histologically, in the outer layers of mesenchyme surrounding and between the follicles, whereas with the former the remaining quadrants were positive and expressing cells were mostly in the inner layers of the dermal sheath. (e) Other tissues, notably lung, adrenal gland, digestive tract, which produce high amounts of collagen type VI, did not stain for beta-galactosidase. (f) Central nervous system and retina, in which the endogenous gene is inactive, expressed the lacZ transgene in most lines. The data suggest that transcription of alpha1(VI) in different tissues is regulated by distinct sequence elements in a modular arrangement, a mechanism which confers high flexibility in the temporal and spatial pattern of expression during development.