Expression of the 67 kDa laminin receptor (67LR) during retinal development: Correlations with angiogenesis
Expression of the 67 kDa laminin receptor (67LR) during retinal development: Correlations with angiogenesis
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DOI：
10.1006/exer.2001.1013
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发表时间：
2001-07-01
影响因子：
3.4
通讯作者：
Stitt, AW
中科院分区：
文献类型：
--
作者：
McKenna, DJ;Simpson, DAC;Stitt, AW
血管细胞与基底膜层粘连蛋白组分的相互作用对正常细胞功能至关重要。同样。异常的相互作用可能在血管病理中起关键作用。先前已经证明，67kda层粘胶蛋白受体（67LR）在小鼠增生性视网膜病变期间高水平表达，在当前的研究中，我们在新生小鼠中检测了67LR，以确定该受体是否在发育中的小鼠视网膜的发育性血管生成方面发挥作用。各组C57/BL6小鼠分别于出生后第P1、P3、P5、P7、P9和P11天处死，观察视网膜血管状况。用fitc -葡聚糖灌注若干小鼠，取下眼睛，用4%多聚甲醛（PFA）固定，平装用于共聚焦扫描激光显微镜。其余小鼠的眼睛要么被置于4% PFA中并包埋在石蜡中，要么被剥离神经视网膜并提取总RNA或蛋白质。采用免疫荧光法、原位杂交法、定量逆转录酶聚合酶链反应法和Western blotting法定位和测定67LR的表达水平。67LR mRNA和蛋白的表达均表现出典型的双相表达模式，与小鼠视网膜血管发育的关键阶段相关。67LR在P1 （P < 0.05）和P7 （P < 0.05）高表达（与浅血管丛形成相关）。相反，当血管生成活性最低时，67LR表达降低。值得注意的是，视网膜平支架的光学切片显示，67LR在浅毛细血管丛和深毛细血管丛的发育段中都有高水平的表达，这种模式与层粘连蛋白强烈共定位。67LR在出生后视网膜血管发育过程中受到调节。在视网膜毛细血管丛形成的两个明确的阶段，高水平的67LR表明该受体可能在视网膜血管生成中起重要作用。(C) 2001学术出版社。
Interaction of vascular cells with the laminin component of basement membranes is important for normal cell function. Likewise. abnormal interactions may have a critical role in vascular pathology. It has been previously demonstrated that the 67 kDa laminin receptor (67LR) is expressed at high levels during proliferative retinopathy in a mouse model and in the current study we have examined 67LR in the neonatal mouse to determine if this receptor plays a role in aspects of developmental angiogenesis in the developing murine retina. Groups of C57/BL6 mice were killed at postnatal day P1, P3, P5, P7, P9 and P11 to assess the retinal vasculature. A number of mice were perfused with FITC-dextran and the eyes removed, fixed in 4 % paraformaldehyde (PFA) and flat-mounted for confocal scanning laser microscopy. The eyes from the remaining mice were either placed in 4 % PFA and embedded in paraffin-wax, or had the neural retina dissected off and total RNA or protein extracted. Immunofluorescence, in situ hybridization, quantitative reverse transcriptase polymerase chain reaction and Western blotting analysis were employed to locate and determine expression levels of 67LR. Both 67LR mRNA and protein expression showed a characteristic bi-phasic expression pattern which correlated with key stages of retinal vascular development in the murine retina. 67LR showed high expression levels at P1 (P < 0.05) (correlating with superficial vascular plexus formation) and at P7 (P < 0.05) (correlating with deep vascular plexus formation). Conversely, 67LR expression was decreased when active angiogenic activity was lowest. Significantly, optical sectioning of retinal flat-mounts revealed high levels of 67LR expression in developing segments of both superficial and deep capillary plexi, a pattern which co-localized strongly with laminin. 67LR is regulated during post-natal development of the retinal vasculature. High levels of 67LR during the two well-defined phases of retinal capillary plexus formation suggests that this receptor may play an important role in retinal angiogenesis. (C) 2001 Academic Press.