Jumblatt Marcia M, McKenzie Richard W, Steele Pamela S, Emberts Celia G, Jumblatt James E
Department of Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, KY 40292, USA.
Cornea. 2003 Jan;22(1):41-5. doi: 10.1097/00003226-200301000-00010.
Several mucins including MUC1, MUC2, MUC4, and MUC5AC have been identified at the ocular surface and in tears. The lacrimal gland, however, is not generally considered a source of ocular mucin. Because the lacrimal glands are similar to the salivary glands, we hypothesized that the lacrimal gland would express MUC7, a distinctive salivary mucin. We report the presence of MUC7 RNA and protein in normal human lacrimal glands as determined by reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization, and Western blot analysis.
RNA from lacrimal glands and conjunctivae was isolated and subjected to RT-PCR with primers specific for MUC7. The identity of the PCR products was confirmed by sequencing. In situ hybridization with PCR product-based riboprobes was used to locate MUC7 transcripts in the lacrimal gland. MUC7 protein was detected by Western blot analysis.
Of six normal human lacrimal glands from which relatively intact mRNA could be extracted, four expressed MUC7. Hybridization with an antisense riboprobe for MUC7 indicates the presence of MUC7 transcripts in the cytoplasm of acinar cells. Western blot analysis confirms expression of the protein in the lacrimal gland. The presence of MUC1, MUC4, and MUC5B was also demonstrated by RT-PCR in lacrimal gland tissue. MUC7 transcripts and protein were also detected in normal human conjunctivae.
The mucin profile of the lacrimal gland resembles that of the salivary gland. Both RNA and protein corresponding to MUC7 are present in the normal human lacrimal gland. Reverse transcription-polymerase chain reaction indicates that other transcripts of MUC1, MUC4, and MUC5B are present as well. Ocular MUC7 is also produced by the conjunctival mucosa. The lacrimal gland, therefore, contributes not only to the aqueous component of tears but also, in concert with the conjunctiva, may contribute to the total pool of ocular surface mucins.
已在眼表和泪液中鉴定出包括MUC1、MUC2、MUC4和MUC5AC在内的多种黏蛋白。然而,泪腺通常不被认为是眼黏蛋白的来源。由于泪腺与唾液腺相似,我们推测泪腺会表达MUC7,一种独特的唾液黏蛋白。我们通过逆转录-聚合酶链反应(RT-PCR)、原位杂交和蛋白质印迹分析报告了正常人泪腺中MUC7 RNA和蛋白质的存在情况。
从泪腺和结膜中分离RNA,并使用针对MUC7的特异性引物进行RT-PCR。通过测序确认PCR产物的身份。使用基于PCR产物的核糖探针进行原位杂交,以定位泪腺中的MUC7转录本。通过蛋白质印迹分析检测MUC7蛋白。
在可提取相对完整mRNA的6个正常人泪腺中,有4个表达MUC7。用针对MUC7的反义核糖探针杂交表明,腺泡细胞的细胞质中存在MUC7转录本。蛋白质印迹分析证实泪腺中存在该蛋白。RT-PCR也在泪腺组织中证实了MUC1、MUC4和MUC5B的存在。在正常人结膜中也检测到了MUC7转录本和蛋白质。
泪腺的黏蛋白谱与唾液腺相似。正常人泪腺中存在与MUC7相对应的RNA和蛋白质。逆转录-聚合酶链反应表明也存在MUC1、MUC4和MUC5B的其他转录本。眼表的MUC7也由结膜黏膜产生。因此,泪腺不仅有助于泪液的水样成分,而且与结膜协同作用,可能有助于眼表黏蛋白的总量。