Department of Cell and Neurobiology, University of Southern California, Los Angeles, USA.
Curr Eye Res. 2011 Nov;36(11):971-8. doi: 10.3109/02713683.2011.602814.
To test the hypotheses that some epithelial cells in the rabbit lacrimal gland (LG) are mucin-secreting cells that are also particularly rich in aquaporin 5 (AQP5) and sodium potassium ATPase β(1) subunit (NKAβ(1)), LG-secreted mucins contribute to the total mucin pool in tear film, and that the rabbit LG is a heterogenic gland where proteins secreted in response to different agonists are varied.
LGs were obtained from adult female rabbits and processed for paraffin sections for hematoxylin and eosin (HE) staining, periodic acid-Schiff (PAS), mucicarmine, and Alcian blue (pH 0.4, 1.0, and 2.5) for the detection of mucins. Serial sections were used for immunohistochemistry (IHC) and PAS. LG lysates and fluids were assayed by dot blot for detection of mucins, and by SDS-PAGE to detect differences in protein profiles of LG fluids stimulated by different agonists.
HE staining demonstrated that the LG is a heterogeneous gland where most epithelial cells are serous, while all duct cells are mucous cells. Some acini and individual acinar cells within serous acini are also mucous or seromucous cells and these cells are particularly rich in AQP5 and NKAβ(1). Dot blot assay showed the presence of mucins in the LG fluids. The protein profiles of LG fluids from pilocarpine, phenylephrine, and isoproterenol varied significantly, particularly in the mid range.
Our data indicated that the rabbit LG is a heterogeneous gland that is composed of both serous and mucin-secreting cells, and mucins produced by the LG contribute to the mucin pool in the tear film. The heterogeneity of the rabbit LG supports the notion of differential secretion, i.e. the volume and composition of the LG fluids vary depending on various circumstances in the ocular surface and the body's needs.
验证以下假设,即在兔泪腺(LG)中,某些上皮细胞是分泌粘蛋白的细胞,且这些细胞富含水通道蛋白 5(AQP5)和钠钾 ATP 酶β(1)亚基(NKAβ(1));LG 分泌的粘蛋白有助于泪膜中总粘蛋白池的形成;并且兔 LG 是一种异质腺,其中对不同激动剂的反应分泌的蛋白质是不同的。
从成年雌性兔中获得 LG,并进行石蜡切片,用于苏木精和伊红(HE)染色、过碘酸希夫(PAS)染色、粘卡红和阿尔辛蓝(pH 值为 0.4、1.0 和 2.5)染色,以检测粘蛋白。使用连续切片进行免疫组织化学(IHC)和 PAS 染色。通过斑点印迹法检测 LG 裂解物和液体中的粘蛋白,通过 SDS-PAGE 检测不同激动剂刺激的 LG 液体中的蛋白质谱差异。
HE 染色显示 LG 是一种异质腺,其中大多数上皮细胞为浆液性,而所有导管细胞均为粘液性细胞。一些浆液性腺泡和浆液性腺泡内的单个腺泡细胞也是粘液性或浆液粘液性细胞,这些细胞富含 AQP5 和 NKAβ(1)。斑点印迹法检测到 LG 液体中存在粘蛋白。毛果芸香碱、苯肾上腺素和异丙肾上腺素刺激的 LG 液体的蛋白质谱差异显著,特别是在中间范围。
我们的数据表明,兔 LG 是一种异质腺,由浆液性和分泌粘蛋白的细胞组成,LG 产生的粘蛋白有助于泪膜中的粘蛋白池。兔 LG 的异质性支持差异分泌的概念,即 LG 液体的体积和组成取决于眼表和身体需要的各种情况。