TISSUE-SPECIFIC AND DIFFERENTIATION-APPROPRIATE EXPRESSION OF THE HUMAN INVOLUCRIN GENE IN TRANSGENIC MICE - AN ABNORMAL EPIDERMAL PHENOTYPE
TISSUE-SPECIFIC AND DIFFERENTIATION-APPROPRIATE EXPRESSION OF THE HUMAN INVOLUCRIN GENE IN TRANSGENIC MICE - AN ABNORMAL EPIDERMAL PHENOTYPE
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DOI：
10.1111/j.1432-0436.1993.tb00708.x
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发表时间：
1993-07-01
期刊：
影响因子：
2.9
通讯作者：
ECKERT, RL
中科院分区：
文献类型：
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作者：
CRISH, JF;HOWARD, JM;ECKERT, RL
Involucrin是角质形成细胞角化包膜的前体，它特异地表达于表皮和其他复层鳞状上皮的超基底层。为了研究包衣蛋白基因的表达及其功能，我们在转基因小鼠中表达了一段6kb的人包衣蛋白基因DNA片段，其上游序列约为2.5kb，下游序列约为0.5kb。转基因产生68 kDa的蛋白，由人类包皮蛋白特异性抗体检测，并以组织特异性和分化适当的方式表达(即，表达限于表皮、宫颈外、气管、食道和结膜的超基底层)。与人包皮角质形成细胞相比，转基因表皮角质形成细胞中可溶的总蛋白水平高出两到四倍。新生的杂合子动物出生时体重正常，表皮外观正常，毛发在4至5天开始生长(即与非转基因动物的毛发生长时间相同)。在新生纯合子动物的一个亚群中，出生体重减轻，表皮有鳞，毛发开始生长较晚，大约在9至10天大。此外，杂合子和纯合子成年动物的毛发倾向于竖立，导致弥漫性脱发的外观。免疫荧光和电子显微镜分别定位于毛囊和角化的包膜中的总蛋白。这些结果提示，毛囊毛囊结构/功能和角化包膜结构的异常可能与毛囊蛋白的过度表达有关。这些动物为角化囊膜结构和功能的研究提供了新的模型。
Involucrin is a precursor of the keratinocyte cornified envelope that is specifically expressed in the suprabasal layers of the epidermis and other stratifying squamous epithelia. To study involucrin gene expression and the function of involucrin, we expressed a 6 kb DNA fragment of the human involucrin gene, containing approximately 2.5 kb of upstream sequence and 0.5 kb of downstream sequence, in transgenic mice. The transgene produces a 68 kDa protein that is detected by a human involucrin-specific antibody, and is expressed in a tissue-specific and differentiation-appropriate manner (i.e., expression is confined to the suprabasal layers of the epidermis, extocervix, trachea, esophagus and conjunctiva).Soluble involucrin levels are two to four times higher in transgenic epidermal keratinocytes compared to human foreskin keratinocytes. Newborn heterozygous animals have a normal birth weight and a normal appearing epidermis and hair growth begins at 4 to 5 days of age (i.e., the same time as hair growth in non-transgenic animals). In a subpopulation of the newborn homozygous animals birth weight is reduced, the epidermis is scaly and hair growth begins late, at around 9 to 10 days of age. In addition, the hair tends to stand erect on both heterozygous and homozygous adult animals giving the appearance of diffuse alopecia.Immunofluorescent and electron microscopy localize involucrin in the hair follicle and cornified envelope, respectively. These results suggest that overexpression of involucrin may cause abnormalities in hair follicle structure/function and cornified envelope structure. These animals provide a new model for the study of cornified envelope structure and function.