Merriman-Smith R, Tunstall M, Kistler J, Donaldson P, Housley G, Eckert R
School of Biological Sciences, University of Auckland, New Zealand.
Invest Ophthalmol Vis Sci. 1998 Dec;39(13):2791-6.
To verify at the molecular level that P2 receptors are expressed in the lens and to determine their expression profiles.
The reverse transcription-polymerase chain reaction (RT-PCR) was used to screen rat lens epithelial and fiber cells for the expression of the ionotropic P2X2 receptor and the G-protein-linked receptor isoforms P2Y1 and P2Y2. Northern blot analysis was used to confirm the level of expression of P2Y1 and P2Y2. The profile of P2-receptor isoform expression in the lens was identified using an indirect RT-PCR in situ hybridization procedure on paraffin sections of whole rat eyes.
P2X2-receptor transcripts could not be detected in lens epithelial or fiber cells. P2Y1- and P2Y2-receptor transcripts were detected in lens fiber cells but not in the epithelial cells. Their expression profiles were maximal in the lens cortex and mostly overlapped each other, except that transcripts for the P2Y2-receptor isoform appeared earlier in the not yet fully elongated fiber cells in the lens bow region.
The molecular data support physiological evidence previously reported by others that P2Y receptors are expressed in the lens and identifies cortical fiber cells as the principal site of expression.
在分子水平验证P2受体在晶状体中表达，并确定其表达谱。
采用逆转录聚合酶链反应（RT-PCR）筛选大鼠晶状体上皮细胞和纤维细胞中离子型P2X2受体以及G蛋白偶联受体亚型P2Y1和P2Y2的表达。用Northern印迹分析确认P2Y1和P2Y2的表达水平。采用间接RT-PCR原位杂交法对大鼠全眼球石蜡切片进行检测，以确定晶状体中P2受体亚型的表达谱。
在晶状体上皮细胞或纤维细胞中未检测到P2X2受体转录本。在晶状体纤维细胞中检测到P2Y1和P2Y2受体转录本，但在上皮细胞中未检测到。它们的表达谱在晶状体皮质中最高，且大部分相互重叠，不过P2Y2受体亚型的转录本在晶状体弓区尚未完全伸长的纤维细胞中出现得更早。
分子数据支持了其他人之前报道的生理学证据，即P2Y受体在晶状体中表达，并确定皮质纤维细胞是主要的表达部位。
