Li X, Zelenka P S, Piatigorsky J
Laboratory of Molecular and Developmental Biology, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892.
Dev Dyn. 1993 Feb;196(2):114-23. doi: 10.1002/aja.1001960205.
Chicken argininosuccinate lyase (ASL)/delta-crystallin, a lens enzyme-crystallin, is encoded in two linked genes (delta 1 and delta 2); only the delta 2 polypeptide contains ASL activity. Here we have quantified delta 1- and delta 2-crystallin mRNA in the lens, cornea, neural retina, heart, and brain at different stages of embryonic development and in 1-wk-old and 1-yr-old chickens by the polymerase chain reaction using internal delta 1 and delta 2 RNA standards. The delta 1/delta 2 mRNA ratio differed for every tissue and was regulated during development. In the embryo there was more delta 1 than delta 2 mRNA in the lens (50-100 times), cornea (3-4 times), and neural retina (2-20 times), about equal amounts of delta 1 and delta 2 mRNA in the heart, and more delta 2 mRNA in the brain (15 times). delta 1-Crystallin mRNA differentially decreased in every tissue after hatching; by contrast, the delta 2 mRNA remained about the same except for the lens, where it decreased 50-fold between 1 wk and 1 yr after hatching. In the 1-yr-old chicken, the delta 2/delta 1 mRNA ratios were 7 in the lens, 175 in the cornea, 22 in the neural retina, 107 in the heart, and 136 in the brain, indicating that delta 2-crystallin is strongly favored in all adult tissues of the chicken. The excess of delta 1 to delta 2 mRNA in the embryonic lens, cornea, and neural retina is intriguing, and suggests some connection with developing transparent eye tissues. Finally, we raise the possibility that expression of both delta-crystallin genes may create tetrameric ASL isoenzymes (perhaps with different specific activities). The unexpected predominance of delta 2 mRNA in the 1-yr-old lens suggests that both the enzymatic and refractive functions of ASL/delta-crystallin are operative and spatially separated, with the enzymatic role present in the cortical fibers and the refractive role in the center of the lens.
鸡精氨琥珀酸裂解酶(ASL)/δ-晶体蛋白,一种晶状体酶-晶体蛋白,由两个连锁基因(δ1和δ2)编码;只有δ2多肽具有ASL活性。在此,我们通过使用内部δ1和δ2 RNA标准的聚合酶链反应,对胚胎发育不同阶段以及1周龄和1岁鸡的晶状体、角膜、神经视网膜、心脏和大脑中的δ1-和δ2-晶体蛋白mRNA进行了定量。每个组织的δ1/δ2 mRNA比值不同,且在发育过程中受到调控。在胚胎期,晶状体(50 - 100倍)、角膜(3 - 4倍)和神经视网膜(2 - 20倍)中δ1 mRNA比δ2 mRNA多,心脏中δ1和δ2 mRNA量大致相等,大脑中δ2 mRNA更多(15倍)。孵化后,每个组织中δ1-晶体蛋白mRNA差异下降;相比之下,除晶状体外,δ2 mRNA保持大致相同,晶状体中δ2 mRNA在孵化后1周和1岁之间下降了50倍。在1岁鸡中,晶状体中δ2/δ1 mRNA比值为7,角膜中为175,神经视网膜中为22,心脏中为107,大脑中为136,表明δ2-晶体蛋白在鸡的所有成年组织中占优势。胚胎晶状体、角膜和神经视网膜中δ1 mRNA相对于δ2 mRNA的过量很有趣,提示与发育中的透明眼组织存在某种联系。最后,我们提出了δ-晶体蛋白两个基因的表达可能产生四聚体ASL同工酶(也许具有不同的比活性)的可能性。1岁晶状体中δ2 mRNA出人意料的优势表明,ASL/δ-晶体蛋白的酶促功能和屈光功能都是有效的且在空间上是分开的,酶促作用存在于皮质纤维中,屈光作用存在于晶状体中心。