Blanchette-Mackie E J, Scow R O
Anat Rec. 1976 Apr;184(4):599-609. doi: 10.1002/ar.1091840402.
The effects of lipolysis on the structure of chylomicrons were studied with the scanning electron microscope using rat chylomicrons incubated with purified bovine milk lipoprotein lipase for 20 minutes at pH 8.1. Since the amount of albumin added to the medium was limited, some of the free fatty acids and partial glycerides formed by lipolysis accumulated in the chylomicrons. Lipolyzed chylomicrons fixed with OSO4 at pH 7.4 appeared in scanning electron micrographs as spheres with multiply idented irregular surfaces, while those fixed at pH 5.5, AS WELL AS CONTROL CHYLOMICRONS FIXED AT BOTH PHs, appeared as spheres with smooth surfaces. Sections of OSO4-fixed specimens, viewed with the transmission electron microscope, showed that the core of lipolyzed chylomicrons fixed at pH 7.4 contained numerous circular electron-lucent areas at the periphery, thus accounting for the indented surfaces observed above, while the core surfaces of the other specimens were circular and smooth. These findings confirm an earlier report that aqueous spaces form in chylomicrons during lipolysis when albumin in the medium is limited, and that the aqueous spaces disappear when specimens are prepared at pH 5.5 for microscopy. Thin sections of specimens that had been prepared for scanning electron microscopy showed that the gold-palladium coating was desposited directly on the indented surface of the lipid core of lipolyzed chylomicrons fixed at pH 7.4. It is concluded that vacuum dehydration during specimen preparation ruptures the outer wall of the aqueous spaces in lipolyzed chylomicrons and thereby exposes the interior of the spaces to gold-palladium coating and viewing with the scanning electron microscope.
利用扫描电子显微镜,对脂解作用对乳糜微粒结构的影响进行了研究。研究中使用大鼠乳糜微粒,在pH 8.1条件下与纯化的牛乳脂蛋白脂肪酶一起孵育20分钟。由于添加到培养基中的白蛋白量有限,脂解作用产生的一些游离脂肪酸和部分甘油酯在乳糜微粒中积累。在pH 7.4条件下用四氧化锇固定的脂解乳糜微粒,在扫描电子显微镜照片中呈现为表面有多处凹陷的不规则球体,而在pH 5.5条件下固定的脂解乳糜微粒以及在这两个pH值下固定的对照乳糜微粒,均呈现为表面光滑的球体。用透射电子显微镜观察四氧化锇固定标本的切片,结果显示,在pH 7.4条件下固定的脂解乳糜微粒的核心在外围含有许多圆形的电子透明区域,这就解释了上述观察到的凹陷表面,而其他标本的核心表面则是圆形且光滑的。这些发现证实了早期的一份报告,即在培养基中白蛋白有限时,脂解过程中乳糜微粒内会形成水相空间,而在pH 5.5条件下制备用于显微镜观察的标本时,这些水相空间会消失。为扫描电子显微镜制备的标本的薄片显示,金钯涂层直接沉积在pH 7.4条件下固定的脂解乳糜微粒脂质核心的凹陷表面上。得出的结论是,标本制备过程中的真空脱水会使脂解乳糜微粒中水相空间的外壁破裂,从而使这些空间的内部暴露于金钯涂层并能用扫描电子显微镜观察。