Kostetskii I, Linask K K, Zile M H
Department of Food Science & Human Nutrition, Michigan State University, 234C G.M. Trout Food Science Building, 48824-1224, East Lansing, MI, USA.
Division of Cardiology, The Children's Hospital of Philadelphia and University of Pennsylvania, 19104, Philadelphia, PA, USA.
Rouxs Arch Dev Biol. 1996 Feb;205(5-6):260-271. doi: 10.1007/BF00365804.
The vitamin A deficiency-associated lethal syndrome observed in avian embryos may be linked to dysfunction of vitamin A-dependent genes. We tested this hypothesis in a quail embryo model by examining the expression of retinoic acid receptors (RARs) and cytosolic retinoic acid binding protein (CRABP) in normal and vitamin A-deficient embryos during early development. RARα and RARγ mRNA were expressed at the same level in normal and vitamin A-deficient embryos during all stages studied. Expression of CRABP I was low in normal and vitamin A-deficient quail embryos during early development, but increased rapidly at later stages. Two transcripts of RARβ, 3.2 and 3.5 kb, were detected in quail embryos during developmental stages 6-12. In normal emryos the level of the 3.2-kb isoform increased as embryonic development advanced. The expression of the 3.5-kb transcript was significantly decreased in vitamin A-deficient embryos, while the 3.2-kb transcript was undetectable by northern analysis. In situ hybridization of stage 7-8 normal quail embryos using a chicken RARβ2 riboprobe revealed that RARβ2 expression was predominantly associated with the cell populations in heart-forming regions, somites, neural folds, notochord and the presumptive thyroid. In stark contrast, in the vitamin A-deficient quail embryo RARβ2 was not expressed in any of the above cell populations. We conclude that the expressions of RARβ and CRABP I are developmentally regulated. Additionally, the expression of RARβ2 during early embryogenesis is regulated by vitamin A status. We propose that RARβ2 plays an important role in the mechanism of action of retinoids in early avian development. The lack of expression of RARβ2 may be linked to abnormalities of the cardiovascular system.
在禽类胚胎中观察到的与维生素A缺乏相关的致死综合征可能与维生素A依赖基因的功能障碍有关。我们在鹌鹑胚胎模型中检验了这一假说,通过检测视黄酸受体(RARs)和胞质视黄酸结合蛋白(CRABP)在正常和维生素A缺乏胚胎早期发育过程中的表达情况。在所有研究阶段,正常和维生素A缺乏胚胎中RARα和RARγ mRNA的表达水平相同。早期发育阶段,正常和维生素A缺乏的鹌鹑胚胎中CRABP I的表达较低,但在后期迅速增加。在发育阶段6 - 12的鹌鹑胚胎中检测到RARβ的两种转录本,分别为3.2 kb和3.5 kb。在正常胚胎中,随着胚胎发育推进,3.2 kb异构体的水平升高。在维生素A缺乏的胚胎中,3.5 kb转录本的表达显著降低,而通过Northern分析未检测到3.2 kb转录本。使用鸡RARβ2核糖探针对7 - 8期正常鹌鹑胚胎进行原位杂交显示,RARβ2的表达主要与心脏形成区域、体节、神经褶、脊索和甲状腺原基中的细胞群体相关。与之形成鲜明对比的是,在维生素A缺乏的鹌鹑胚胎中,上述任何细胞群体均未表达RARβ2。我们得出结论,RARβ和CRABP I的表达受发育调控。此外,早期胚胎发生过程中RARβ2的表达受维生素A状态的调控。我们提出,RARβ2在禽类早期发育中类视黄醇的作用机制中起重要作用。RARβ2表达的缺失可能与心血管系统异常有关。