Söderpalm A, Szél A, Caffé A R, van Veen T
Department of Zoology, University of Göteborg, Sweden.
Invest Ophthalmol Vis Sci. 1994 Oct;35(11):3910-21.
The authors have established an organ culture method in which the the postnatal development and the structural integrity of the mouse retina can be maintained for at least 6 weeks. Additionally, they have examined the emergence and in vitro morphogenesis of the photoreceptors and the development of insoluble components of the interphotoreceptor matrix.
Neural retinas and retinal pigment epithelia from 48-hour-old C3H ++/++ mice were cultured. At various ages, the tissues were fixed and cryosectioned or wholemounted. Photoreceptor development was studied by immunocytochemistry with visual pigment antibodies and by lectin cytochemistry. The ultrastructure of the photoreceptors was studied by electron microscopy.
Immunopositive rods and short-wave sensitive cones were detectable as early as 3 days after explantation. From this time on, matrix domains around cones were also identifiable and labelled with peanut agglutinin lectin. However, the antibody specific to the middle-wave sensitive cone pigment failed to recognize any cones throughout the 6-week culture period.
Both basic photoreceptor types appeared and developed in this organ culture system according to a timetable comparable to normal in vivo development. Surprisingly, under these circumstances, one of the two cone pigments was not expressed by any photoreceptors.
作者建立了一种器官培养方法,在此方法中,小鼠视网膜的出生后发育和结构完整性可维持至少6周。此外,他们研究了光感受器的出现及体外形态发生,以及光感受器间基质不溶性成分的发育。
培养来自48小时龄C3H ++/++小鼠的神经视网膜和视网膜色素上皮。在不同年龄,将组织固定、冷冻切片或整装。通过用视色素抗体进行免疫细胞化学和凝集素细胞化学研究光感受器发育。通过电子显微镜研究光感受器的超微结构。
早在植入后3天就能检测到免疫阳性的视杆细胞和短波敏感视锥细胞。从此时起,视锥细胞周围的基质区域也可识别并用花生凝集素进行标记。然而,在整个6周培养期内,中波敏感视锥色素特异性抗体未能识别任何视锥细胞。
在这个器官培养系统中,两种基本光感受器类型按照与正常体内发育相当的时间表出现并发育。令人惊讶的是,在这些情况下,两种视锥色素之一未被任何光感受器表达。