Shah Dhara, Ali Marwan, Pasha Zeeshan, Jaboori Assraa Jassim, Jassim Sarmad H, Jain Sandeep, Aakalu Vinay K
Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, Illinois, United States of America.
PLoS One. 2016 Jan 29;11(1):e0148018. doi: 10.1371/journal.pone.0148018. eCollection 2016.
Study of human lacrimal cell biology is limited by poor access to tissue samples, heterogeneous cell composition of tissue and a lack of established lacrimal epithelial markers. In order to further our understanding of lacrimal cell biology, we sought to find a better marker for human lacrimal epithelial cells, compared to what has been reported in the literature.
We utilized human Muller's muscle conjunctival resection (MMCR) specimens containing accessory lacrimal gland (ALG) and cadaveric main lacrimal gland (MLG) as sources of lacrimal tissue. Candidate markers were sought using human ALG tissue from MMCR specimens, isolated by laser capture microdissection (LCM). Affymetrix® analysis was performed on total RNA isolated from FFPE samples to profile transcription in ALG. MMCR tissue sections were assessed by immunofluorescence using antibodies for histatin-1, lactoferrin, E-cadherin (E-cad) and alpha-smooth muscle actin (ASMA). Reverse transcriptase polymerase chain reaction (RT-PCR) analysis was performed to analyze the expression of histatin-1, E-cad and lactoferrin from cadaveric MLG.
Histatin-1 is expressed in ALG and MLG, localizes to lacrimal epithelium, and to a greater degree than do other putative lacrimal epithelial markers.
Histatin-1 is a good marker for human lacrimal epithelium in ALG and MLG and can be used to identify lacrimal cells in future studies.
人类泪腺细胞生物学研究受到组织样本获取困难、组织细胞组成异质性以及缺乏既定泪腺上皮标志物的限制。为了进一步了解泪腺细胞生物学,我们试图找到一种比文献报道更好的人类泪腺上皮细胞标志物。
我们利用含有副泪腺(ALG)的人类米勒肌结膜切除术(MMCR)标本和尸体主泪腺(MLG)作为泪腺组织来源。使用通过激光捕获显微切割(LCM)分离的MMCR标本中的人类ALG组织寻找候选标志物。对从福尔马林固定石蜡包埋(FFPE)样本中分离的总RNA进行Affymetrix®分析,以分析ALG中的转录情况。使用针对组蛋白-1、乳铁蛋白(lactoferrin)、E-钙黏蛋白(E-cad)和α-平滑肌肌动蛋白(ASMA)的抗体,通过免疫荧光评估MMCR组织切片。进行逆转录聚合酶链反应(RT-PCR)分析,以分析尸体MLG中组蛋白-1、E-cad和乳铁蛋白的表达。
组蛋白-1在ALG和MLG中表达,定位于泪腺上皮,并且比其他假定的泪腺上皮标志物的定位程度更高。
组蛋白-1是ALG和MLG中人类泪腺上皮的良好标志物,可用于未来研究中鉴定泪腺细胞。