Perkovich C L, Ubels J L, Lee S Y, Soprano D R
Medical College of Wisconsin, Milwaukee.
Exp Eye Res. 1993 May;56(5):513-9. doi: 10.1006/exer.1993.1065.
Cells that require or metabolize vitamin A contain cellular retinol-binding proteins (CRBP) and cellular retinoic acid-binding proteins (CRABP) which apparently participate in metabolism and transport of retinoids within the cell. Since the lacrimal gland secretes and metabolizes vitamin A, the cellular retinoid-binding proteins and their mRNAs should also be present in this gland. Total RNA from rat and rabbit lacrimal glands was analysed by RNase protection using 32P-labeled antisense cRNA probes. CRBP-I mRNA is present in rat lacrimal gland at 2% of the level in liver but CRBP-I mRNA could not be identified conclusively in rabbit lacrimal gland using the available rat cRNA probe. CRABP-I mRNA is present in rat lacrimal gland at 5% of the level in the 12-day gestation rat fetus, but was not detectable in the rabbit lacrimal gland. Rabbit or rat lacrimal gland cytosol was incubated with [3H]retinol or [3H] retinoic acid followed by ion-exchange chromatography using a Tris buffer and NaCl gradient. CRBP is present in rabbit and rat lacrimal gland at 78.5 +/- 14.5 and 71.1 +/- 11 pmol g-1 protein, respectively. CRABP is present in rat lacrimal gland at 190.6 +/- 13.8 pmol g-1 protein but was not detectable in rabbit lacrimal gland. The presence of CRBP in the lacrimal gland is consistent with its role in secretion of retinol into the lacrimal gland fluid. The apparent lack of CRABP in rabbit lacrimal gland as compared with rat suggests species differences in the role of retinoids in differentiation and maintenance of this organ.
需要或代谢维生素A的细胞含有细胞视黄醇结合蛋白(CRBP)和细胞视黄酸结合蛋白(CRABP),它们显然参与细胞内类视黄醇的代谢和转运。由于泪腺分泌和代谢维生素A,细胞类视黄醇结合蛋白及其mRNA也应存在于该腺体中。使用32P标记的反义cRNA探针通过RNA酶保护分析法分析了大鼠和兔泪腺的总RNA。CRBP-I mRNA在大鼠泪腺中的含量为肝脏中的2%,但使用现有的大鼠cRNA探针在兔泪腺中未能明确鉴定出CRBP-I mRNA。CRABP-I mRNA在大鼠泪腺中的含量为妊娠12天大鼠胎儿的5%,但在兔泪腺中未检测到。将兔或大鼠泪腺细胞溶质与[3H]视黄醇或[3H]视黄酸一起孵育,然后使用Tris缓冲液和NaCl梯度进行离子交换色谱分析。CRBP分别以78.5±14.5和71.1±11 pmol g-1蛋白的含量存在于兔和大鼠泪腺中。CRABP以190.6±13.8 pmol g-1蛋白的含量存在于大鼠泪腺中,但在兔泪腺中未检测到。泪腺中CRBP的存在与其在将视黄醇分泌到泪腺液中的作用一致。与大鼠相比,兔泪腺中明显缺乏CRABP,这表明类视黄醇在该器官的分化和维持中的作用存在物种差异。