正常和急性高眼压兔眼视网膜的ACP组织化学研究

用光镜定量酶组织化学和电镜细胞化学方法,观察了兔眼视网膜酸性磷酸酶(acid phos-phatase,ACP)分布及急性高眼压对 ACP 分布及溶酶体的影响。ACP 活性强度在视网膜各层的顺序为(F 检验 P<0.05):由高到低依次①色素上皮层,②外网状层,③内网状层,④外核层和内核层,⑤杆锥体层和神经节细胞及神经纤维层,该结果与以往半定量判定有较大差异。高眼压后15天、30天组色素上皮层 ACP 活性增强,7天组杆锥体层 ACP 活性增强(P<0.01)。电镜下见高眼压后色素上皮层溶酶体数量增多,溶酶体外 ACP 活性增强,外段膜盘内溶酶体外 ACP 活性明显增强。提示 ACP(及其他溶酶体酶)可能在急性高眼压后的视网膜损伤过程中起重要作用。     

猫外层视网膜和脉络膜的年龄相关变化

取老年猫(12龄,2.5~3 kg)和青年猫(1~3龄,2~2.5 kg)各4只的视网膜,经4%多聚甲醛处理后用H.E染色以显示视网膜和脉络膜的结构。光学显微镜下观察感光细胞层、玻璃膜(Bruch’s membrane)结构的变化,计数色素上皮层(retinal pigment epithelium,RPE)细胞数、脉络膜毛细血管数,测量玻璃膜、脉络膜厚度,脉络膜毛细血管之间的距离。结果显示,与青年猫比较,老年猫视网膜感光细胞层结构杂乱;色素上皮细胞数显著下降;玻璃膜厚度无显著变化,出现较多碎片;脉络膜厚度明显变薄,脉络膜毛细血管数显著减小,脉络膜毛细血管之间的距离显著增大。推测老年猫脉络膜的退化可能是导致玻璃膜、色素上皮层的退化,进而导致感光细胞的功能衰退的重要原因。     

卵子皮层在受精中的作用

关于卵子皮层的概念,人们有两种不同的意见:一种意见指卵子质膜下的一簿层细胞质,厚度不到5μm,主要成分为皮层颗粒、色素颗粒以及肌动蛋白等,它在结构和功能上与其下的细胞质不同;另一种意见认为还应将质膜-细胞外被复合体包括在内。本文采纳后一种意见。 (一) 皮层在受精时的动态变化向海胆卵细胞质中注入一个微小的铁粒,并在已知强度的磁场内停留一定的时间,通过测铁粒子的移动速率可以测出皮层硬度。实验结果表明,铁粒子在细胞质中央区域比在皮层中运动快,说明皮层处在一种凝胶状态,皮层的     

封面说明:油松针叶的横切面

油松(Pinus tabulaeformis)叶两针一束,簇生于短技上,横切面呈半圆形.表皮和下皮层胞壁厚、胞腔小、排列紧密、外覆较厚的角质膜,气孔器的保卫细胞下陷到下皮层中,同叶肉细胞形成孔下室,在其外方田具喙的副卫细胞拱盖,可减少水分蒸腾.     

溴乙酰胺对钉螺超微结构的影响

本文应用电子显微镜技术观察了正常钉螺及溴乙酰胺作用后的钉螺头足部软体组织及肝脏组织的超微结构。头足部软体组织有上皮层、上皮下层及肌层。上皮层有三种细胞:无纤毛上皮细胞、纤毛上皮细胞及含有大量粘性分泌颗粒的腺体细胞。上皮层细胞内含有丰富的线粒体、内质网及张力细丝。基底膜将上皮细胞与上皮下层分开,上皮下层有含色素颗粒的细胞。肌层为梭形细胞,核居中央,无横纹。溴乙酰胺作用24小时后,钉螺头足部的上皮细胞与肝脏腺管细胞肿胀,核增大;染色质凝集,线粒体肿胀、嵴断裂呈空泡状,粗面内质网变粗呈短管状,排列混乱,或呈同心圆指纹状排列。这些病变与生化代谢结合进行了讨论。     

降雪对荒漠地区藓类结皮中真藓生理生化的影响

生物土壤结皮的生存环境是地球上自养生物生存最为极端的生境之一,真藓是荒漠结皮中最为重要的组分之一。很少有研究涉及冬季降雪对结皮层生物体的影响。以宁夏沙坡头人工植被区内发育良好、长势均匀的真藓结皮为研究对象,系统研究降雪影响下荒漠地区藓类结皮层真藓的光合色素含量、可溶性糖含量、可溶性蛋白含量、丙二醛(MDA)含量以及脯氨酸含量的变化,并探讨其对降雪的生理生化响应。采用了4个降雪处理,分别是无降雪、0.5倍降雪、1倍降雪、2倍降雪。结果表明:随着降雪量的增加,其光合色素含量和可溶性蛋白含量显著增加;而可溶性糖含量、游离脯氨酸含量以及MDA含量均呈下降趋势。研究结果表明,作为生物土壤结皮重要水源之一的冬季降雪,能够为结皮层生物体提供适宜的水分条件以激发其生理生化活性,对维持荒漠生态系统的稳定性具有重要作用。     

碱蒿营养器官的扫描电镜观察研究

用扫描电子显微镜对碱蒿（Artemisia anethifolia）营养器官的解剖学特征进行了观察与研究。结果表明：碱蒿属于典型的泌盐盐生植物。叶肉质化,表皮上气孔为无规则型气孔,密集下陷,有较多的盐囊泡和分杈腺毛分布,表皮细胞外壁加厚,外壁的外层角质化,为等面叶,栅栏组织细胞排列疏松,叶中央为发达的储水组织,木质部有多束维管束;茎表皮上有少许气孔,并分布有腺毛和众多盐囊泡,表皮细胞外壁加厚形成角质层,皮层宽度较小,皮层薄壁组织细胞内可见淀粉粒,中柱后生木质部为口径大的导管,原生木质部为口径小的导管,管内充满盐晶体,中间有发达的贮水薄壁细胞;根的表皮细胞排列疏松,外皮层细胞排列紧密而整齐,中部皮层薄壁细胞层数较多,细胞中贮藏有许多淀粉粒,内皮层细胞排列紧密,初生木质部导管发达,内部周围存在大量盐晶体,根的次生木质部有通气组织。这些解剖结构均表现出碱蒿具有适应盐碱、干旱生境的结构特征。     

透明鱼及其在生物教学中的应用

透明金鱼,也叫“玻璃鱼”,它的透明特色主要是机体不含色素所致。根据其透明度可分为透明鱼、半透明鱼(有色透明鱼)、和镶嵌型透明鱼三种,它们主要存在于龙种和文种金鱼中,旦种金鱼中透明鱼较少。透明鱼的种类及其特征 1.透明鱼鳃盖和鳞片不含鸟粪素,皮层和肌肉不含色素,通过体侧可以看到它的鲜红色的鳃以及一切内脏。这种鱼在1—5月龄时全透明,长大后呈现肉红色。如常见的有玻璃龙金,玻璃文金和透明珍珠。有的透明度可以和银鱼相比。 2.半透明鱼又叫有色透明鱼,鳃盖、鳞片均不含鸟粪素,但皮层和肌肉色素呈可溶状态均匀分布,就象有色玻璃一样。如琥珀龙金、蓝色文金、乳白珍珠等。     

蒙古黄芪营养器官的解剖结构研究

通过对蒙古黄芪营养器官解剖结构研究表明,根为典型的双子叶植物根的结构,初生结构由表皮、皮层和维管柱组成,初生木质部和初生韧皮部在分化过程中呈外始式,初生木质部四原型,皮层内具凯氏带;茎由表皮、皮层和维管柱组成,维管束为外韧无限维管束;叶为异面叶,由表皮、叶内和叶脉组成,栅栏组织排列紧密且整齐,细胞呈长柱形,海绵组织排列疏松且不整齐,细胞呈海绵状,有大量气隙,叶脉维管束发达,为外韧无限维管束,叶上下表皮均具有气孔.     

视网膜脱离围手术期的护理

视网膜脱离是视网膜的神经上皮层和它本身的色素上皮层分离。它是一种常见的严重性、致盲性眼底疾病,手术是唯一有效的治疗方法[1]。特别是原发性视网膜脱离应争取早日手术治疗。2003年1月~2005年1月,我院采用巩膜外冷凝外加压环扎术治疗视网膜脱离58例,效果满意。现将护理体会总结如下。1临床资料本组58例患者中,男38例,女20例,年龄最小12岁,最大68岁。全部患者均行冷凝外加压环扎术。术后全部患者视网膜均达到解剖复位,且无感染发生。2护理2·1术前护理(1)术前心理护理。患者因视网膜脱离而视力减退,心理负担很重,既希望尽快手术,又对手术…     

Effect of pigmented epithelium from different developmental stages of chick embryo on cell growth of retinal explants

The model system to investigate the effect of retinal pigmented epithelium (PE) on the retinal development in vitro has been established in this laboratory. Chick retina separated from 5-day-old embryo (E 5) were cut into strips and explanted on the collagen substratum either in close contact with retinal PE (RPE), or without PE (R). The rates of cell proliferation of retinal strips cultured for 48 hr were measured by the uptake of radioactive thymidine and DNA contents. Both parameters in RPE were increased to values ranging from 137 to 167% when PE was taken from E5 and E6. However PE taken from E7, E8 and E9 had no effect on cell proliferation. The rate of cell proliferation of retina were increased both when separated retina and PE of E5 either from same or from an other eye closely contact again and when retina and PE of E5 were explanted together without separation. However the rates of cell proliferation were remained without much change when a millipore filter existed between retina and PE of E5 as well as the retina was inverted, the ganglion cell layer contacted with PE. The neurite outgrowth from retina explant with and without PE of E5 or E6 were also different. After culture for 24 hr the fiber length of neurite growing in RPE was only 36-39% of that in R. After 48 hr it was about 70% of that in R. This results suggested that the developmental stage of PE and the direct cell-to-cell contact of PE from E5 with photoreceptor layer of retina was important for the retinal cell proliferation. But PE had negative influence on neurite growth of retina in culture.     

Oriented axon outgrowth from avian embryonic retinae in culture

The neural retina of avian embryos was spread on a membrane filter and cut in any desired orientation. Strips cut across the retina of 4- to 7-day chick or 3- to 6-day quail embryos were explanted onto collagen gels. Vigorous neurite outgrowth was seen for about 3 days, by which time many neurites were 3 mm long. Horseradish peroxidase (HRP) labeling showed that the cells producing the neurites were large and formed a layer near the inner limiting membrane, indicating that the neurites in vitro were axons of retinal ganglion cells. The size of the neurite population and the regions from which neurites emerged vaired with the donor age, while most neurites sprouted from the side of the explant formerly closest to the optic fissure. This pattern closely resembled that of axon growth in the normal retina, as revealed by SEM, silver staining, and HRP labeling. Mitotic inhibitors (Ara-C and FUdR) did not alter the neurite outgrowth. Pretreatment of retinae with trypsin or collagenase did not disorganize axons at the time of explantation, but tended to equalize neurite emergence on each side of the retinal strips. We suggest that microenvironmental factors, especially the enzyme-labile inner limiting membrane, are important for axon guidance in the retina.     

Pigmented epithelium to retinal transdifferentiation and Pax6 expression in larval Xenopus laevis

This study examines the retinal transdifferentiation (TD) of retinal pigmented epithelium (RPE) fragments dissected from Xenopus laevis larvae and implanted into the vitreous chamber of non-lentectomized host eyes. In these experimental conditions, most RPE implants transformed into polarized vesicles in which the side adjacent to the lens maintained the RPE phenotype, while the side adjacent to the host retina transformed into a laminar retina with the photoreceptor layer facing the cavity of the vesicle and with the ganglionar cell layer facing the host retina. The formation of a new retina with a laminar organization is the result of depigmentation, proliferation and differentiation of progenitor cells under the influence of inductive factors from the host retina. The phases of the TD process were followed using BrdU labelling as a marker of the proliferation phase and using a monoclonal antibody (mAbHP1) as a definitive indicator of retina formation. Pigmented RPE cells do not express Pax6. In the early phase of RPE to retinal TD, all depigmented and proliferating progenitor cells expressed Pax6. Changes in the Pax6 expression pattern became apparent in the early phase of differentiation, when Pax6 expression decreased in the presumptive outer nuclear layer (ONL) of the new-forming retina. Finally, during the late differentiation phase, the ONL, which contains photoreceptors, no longer expressed Pax6, Pax6 expression being confined to the ganglion cell layer and the inner nuclear layer. These results indicate that Pax6 may have different roles during the different phases of RPE to retinal TD, acting as an early retinal determinant and later directing progenitor cell fate.     

Transdifferentiation of Pigmented Epithelium Induced by the Influence of Lens Epithelium in Frogs

Abstract. The influence of lens epithelium (LE) of adult frogs on the character of transdifferentiation of retinal pigmented epithelium (RPE) of adult frogs and tadpoles of Rana temporaria has been studied. After a period of intense proliferation RPE cultured in vivo in contact with LE in the tadpole orbit almost exclusively transforms into retina. RPE precultivated in vitro in contact with LE for three days in protein-free medium does not manifest cell divisions and mostly transdifferentiates into lentoids. The problem of the relative significance of inducing determinants and the role of activation or inhibition of proliferation in transdifferentiation is discussed.     

ATP released via gap junction hemichannels from the pigment epithelium regulates neural retinal progenitor proliferation

The retinal pigment epithelium (RPE) plays an essential role in the normal development of the underlying neural retina, but the mechanisms by which this regulation occurs are largely unknown. Ca2+ transients, induced by the neurotransmitter ATP acting on purinergic receptors, both increase proliferation and stimulate DNA synthesis in neural retinal progenitor cells. Here, we show that the RPE regulates proliferation in the underlying neural retina by the release of a soluble factor and identify that factor as ATP. Further, we show that this ATP is released by efflux through gap junction connexin 43 hemichannels, the opening of which is evoked by spontaneous elevations of Ca2+ in trigger cells in the RPE. This release mechanism is localized within the RPE cells to the membranes facing the neural retina, a location ideally positioned to influence neural retinal development. ATP released from RPE hemichannels speeds both cell division and proliferation in the neural retina.     

Study on RNA synthesis in the retina and retinal pigment epithelium of mice by light microscopic radioautography.

With the aim of determining the distribution of the incorporation of 3H-uridine in both retina and retinal pigment epithelium (RPE), the mouse eyes at embryonic day 9.5 (E 9.5), E 12.5, E 14.5, E 16.5, E 18.5 of gestational ages, and postnatal day 1 (P 1), P 3, P 7, P 14 were analyzed by light microscopic radioautography. Small pieces of the ocular tissues were labelled with 3H-uridine in vitro and light microscopic radioautographs were prepared. The average grain numbers per cell of the respective regions of tissues were calculated. In the retina, the grain numbers increased gradually from E 9.5 to P 1 and reached the maximal value at P 1, and then decreased until P 14. However, the grain numbers were more in the vitreal portion than those in the scleral portion at E 16.5 and then became more in the scleral portion from E 18.5 to P 14. It is considered that the ganglion and bipolar cells finish the RNA synthesis earlier, while the photoreceptor cells do it later during the fetal and postnatal development. In the RPE, the grain numbers gradually increased from E 12.5 to P 7 and then decreased until P 14. Considering the same ages, the grain numbers increased in the following order, anterior, equatorial and posterior regions during embryonic stages, but decreased in the same order after birth. Therefore, it is suggested that the activity of RNA synthesis in PE cells is higher in the posterior region than in the anterior region during embryonic stages. But the activity ascends generally and becomes relatively higher in the anterior region, after birth. Comparing the retina and RPE, it was noted that the grain numbers in the RPE were more important than in the retina and that the maximal value was at P 1 in the retina, while it was at P 7 in the RPE. From these results, it can be concluded that the RNA synthesis ceases earlier in the retina than in the RPE.     

Ultrastructural Changes and Expression of PCNA and RPE65 in Sodium Iodate-Induced Acute Retinal Pigment Epithelium Degeneration Model

Alteration in retinal pigment epithelium (RPE) results in the visual dysfunction and blindness of retinal degenerative diseases. Injection of sodium iodate (NaIO₃) generates degeneration of RPE. We analyzed the sequential ultrastructure and expression of proliferating cell nuclear antigen (PCNA) and retina-specific RPE65 in NaIO₃-induced retinal degeneration model. Adult male rats were injected 1% NaIO₃ (50 mg/kg) and eyes were enucleated at 1, 3, 5, 7 and 14 days post-injection (DPI), fixed, and processed for histological analysis. NaIO₃-induced retinal degeneration was successfully established. At 1 DPI, most RPE cells were degenerated and replaced by a few proliferating RPE cells in the peripheral area. At 3 DPI, the RPE and photoreceptor out segments (POS) underwent a marked morphological change, including POS disruption, accumulation of residual bodies in RPE and POS, and hyperplasia of the RPE cell. At 5 DPI, POS showed a maximum increase in the outer segment debris and the retina showed partial detachment. These abnormal morphological changes gradually decreased by day 7. At 14 DPI, the damaged RPE and POS were partially regenerated from the peripheral to the central region. Expression of PCNA and RPE65 increased from day 3 onward. The damaged RPE showed earlier expression of PCNA and RPE65 than POS. The RPE damaged by NaIO₃ rapidly proliferated to put down roots on Bruch’s membrane from the peripheral retina and proliferation and hyperplasia of the RPE had a regular direction of progress. Therefore, NaIO₃-induced acute changes in retina mimic the patho-morphologic features of RPE-related diseases.     

Axonal pathfinding in organ-cultured embryonic avian retinae

Eye cups from stage 14-28 (E2 to E5) chick and quail embryos consisting of neural retina, lens, and vitreous body were cultured for 1 or 2 days. These eyes expanded by proliferation of the retinal cells and the surface areas of the retinae increased several-fold. The area covered by ganglion cells and axons also expanded in vitro. [3H]Thymidine labeling showed extensive proliferation of the neuroepithelial cells including the formation of new ganglion cells. Culturing eyes from embryos before stage 17 results, as in vivo, in the generation of the first ganglion cells of the retina, but unlike in the in vivo situation, the outgrowing axons always formed a random fiber net in the central portion of the retina. A defined axonal pattern identical to the in vivo developed only in specimens from embryos of stage 17 and older. Some aberrant axons, however, were also observed at the retinal periphery in specimens from embryos of more advanced stages (20-24), but only during the second day of culturing. Axons in retinae from embryos of stages 23 to 26 heading toward the optic fissure often crossed the fissure and, in contrast to the situation in vivo, invaded the opposite retinal side. These axons of wrong polarity followed the pathways of axons growing centripetally but in reverse direction. This suggests that the polarity of growing nerve fibers and their course are determined by different factors. Culturing the eyes of embryos from stages 20 to 25 in the presence of antibodies showed that the antibodies penetrated the entire retina with 6 hr. Neither anti-N-CAM nor the T-61 antibody--both recognizing membrane proteins of retinal cells--affected the growth of the eyes in vitro. The development of the axonal pattern in vitro was not affected by incubation with N-CAM-antibodies at concentrations up to 500 micron/ml, whereas the T-61 antibody which is known to block neurite extention in vitro (S. Henke-Fahle, W. Reckhaus, and R. Babiel (l984). "Developmental Neuroscience: Physiological, Pharmacological, and Clinical Aspects," pp. 393-398. Elsevier, Amsterdam/New York) showed inhibition of axonal growth in retina cultures at 50 micron/ml. These results indicate that the eye cultures can be used as a test system for antibodies against antigens which could be involved in axon extension and neurite pathfinding in situ.     

The possibility of lens formation in explants of the retina and pigment epithelium of the eye in chick embryos

Undissociated tissue explants of the retina and retinal pigment epithelium (RPE) of 3,5-, 4-, 5- and 8-day-old chick embryos were cultured in vitro. After 7 days in culture, lentoids were observed in explants of either retina or RPE from 3,5-, 4- and 5-day-old embryos. As demonstrated by immunohistochemistry, these lentoids contained specific chick lens proteins (alpha-, beta- and delta-crystallins). No crystallin-containing cells were found in eye tissue explants from 8-day-old embryos. However, when 5-bromo-deoxyuridine (25 microM) was introduced into the medium at the beginning of culturing (for 12 h), large eosinophilic cells containing alpha-, beta- and delta-crystallins were detected in retinal explants of the 8-day old embryos. Thus, retina and RPE of 3,5-5-day-old chick embryos are capable of lens differentiation after explantation in vitro without dissociation into individual cells. This capacity is lost during development.