Biomedical Sciences Program, College of Health Sciences, Midwestern University, Glendale, AZ 85308, USA.
Cell Biol Int. 2011 Nov;35(11):1131-9. doi: 10.1042/CBI20110063.
Fish keratocytes are used as a model system for the study of the mechanics of cell motility because of their characteristic rapid, smooth gliding motion, but little work has been done on the regulation of fish keratocyte movement. As TGFβ (transforming growth factor β) plays multiple roles in primary human keratinocyte cell migration, we investigated the possible involvement of TGFβ in fish keratocyte migration. Studying the involvement of TGFβ1 in 24 h keratocyte explant allows the examination of the cells before alterations in cellular physiology occur due to extended culture times. During this initial period, TGFβ levels increase 6.2-fold in SFM (serum-free medium) and 2.4-fold in SFM+2% FBS (fetal bovine serum), while TGFβ1 and TGFβRII (TGFβ receptor II) mRNA levels increase ∼3- and ∼5-fold respectively in each culture condition. Two measures of motility, cell sheet area and migration distance, vary with the amount of exogenous TGFβ1 and culture media. The addition of 100 ng/ml exogenous TGFβ1 in SFM increases both measures [3.3-fold (P = 4.5×10-5) and 26% (P = 2.1×10-2) respectively]. In contrast, 100 ng/ml of exogenous TGFβ1 in medium containing 2% FBS decreases migration distance by 2.1-fold (P = 1.7×10-7), but does not affect sheet area. TGFβ1 (10 ng/ml) has little effect on cell sheet area in SFM cultures, but leads to a 1.8-fold increase (P = 1.5×10-2) with 2% FBS. The variable response to TGFβ1 may be, at least in part, explained by the effect of 2% FBS on cell morphology, mode of motility and expression of endogenous TGFβ1 and TGFβRII. Together, these results suggest that expression of TGFβ and its receptor are up-regulated during zebrafish keratocyte explant culture and that TGFβ promotes fish keratocyte migration.
鱼类角膜细胞因其特征性的快速、平滑滑行运动而被用作研究细胞运动力学的模型系统，但对鱼类角膜细胞运动的调节研究甚少。由于 TGFβ（转化生长因子β）在人角质形成细胞迁移中发挥多种作用，我们研究了 TGFβ 可能参与鱼类角膜细胞迁移。研究 TGFβ1 在 24 小时角膜细胞外植体中的作用，可以在细胞因培养时间延长而发生细胞生理变化之前检查细胞。在这个初始阶段，TGFβ 水平在 SFM（无血清培养基）中增加 6.2 倍，在 SFM+2%FBS（胎牛血清）中增加 2.4 倍，而 TGFβ1 和 TGFβRII（TGFβ 受体 II）mRNA 水平在每种培养条件下分别增加约 3 倍和 5 倍。两种运动性测量指标，细胞片面积和迁移距离，随外源性 TGFβ1 和培养基的数量而变化。在 SFM 中添加 100ng/ml 外源性 TGFβ1 会增加这两个测量值[分别增加 3.3 倍（P=4.5×10-5）和 26%（P=2.1×10-2）]。相比之下，100ng/ml 外源性 TGFβ1 在含有 2%FBS 的培养基中使迁移距离减少 2.1 倍（P=1.7×10-7），但不影响片面积。TGFβ1（10ng/ml）对 SFM 培养物中的细胞片面积影响不大，但在含 2%FBS 时会导致 1.8 倍的增加（P=1.5×10-2）。对 TGFβ1 的可变反应至少部分可以解释为 2%FBS 对细胞形态、运动方式以及内源性 TGFβ1 和 TGFβRII 表达的影响。综上所述，这些结果表明，TGFβ 及其受体的表达在斑马鱼角膜细胞外植体培养过程中上调，并且 TGFβ 促进鱼类角膜细胞迁移。
