正常猕猴与人视网膜血管的比较

目的比较正常猕猴与人视网膜血管的异同,为进一步利用猕猴建立动物模型来研究视网膜血管打下基础。方法取健康成年猕猴眼球6只和人角膜移植供体剩余眼杯8只的视网膜,用ADP酶法进行血管染色,对两者视网膜血管的走行、血管分级、毛细血管分层以及黄斑区血管拱环等进行比较,测量结果进行统计学检验。结果猕猴与人的视网膜铺片经ADP酶法染色后见视网膜血管自穿出视盘后的一级血管逐渐分支变细,直至五级血管即毛细血管;在视盘旁、赤道部、周边部两者血管面积百分比没有差异;视盘旁血管分为多层,赤道部有两层,且深浅层间相互交通,周边部仅见一层毛细血管且较稀疏;两者黄斑区毛细血管均较密集,有形态完整呈不规则状的血管拱环,血管面积百分比以及血管拱环的面积、周长和直径没有差异。结论猕猴与人在视网膜血管走行、分级、毛细血管分层以及黄斑区血管拱环等多方面有良好的相似性,可用作人类视网膜血管、尤其是黄斑区视网膜血管研究的良好动物模型。     

正常猕猴与人的视网膜电图比较

目的比较正常猕猴与人视网膜电图异同,为进一步利用猕猴建立动物模型研究视网膜疾病打下基础。方法健康成年猕猴7只（14只眼）与8例（16只眼）正常人进行视网膜电图检测,对两者Rod-ERG中的b波,Max-ERG的a、b波,Cone-ERG的b波峰时值及波幅和OPs的O2值,Flicker-ERG的P值进行统计学检验。结果猕猴与人的视网膜电图波形结果较为相似,各指标与人的结果相比,潜伏期短,幅值低,但Cone-ERG和Flicker-ERG两者幅值差异不具有统计学意义。结论视网膜电图检测从功能上证明猕猴较其他常用实验动物更接近人,尤其表现在视锥细胞及黄斑区功能,可用作人类视网膜疾病尤其是黄斑区病变的良好动物模型。     

哺乳动物胰腺体部胰多肽（PP）免疫反应细胞的比较研究

采用SPA-GDN免疫组化染色技术,对人、大鼠、小鼠、豚鼠、猪、狗和猫等七种哺乳动物胰腺体部胰多肽(PP)免疫反应细胞的分布和形态进行比较研究,结果表明,上述七种动物PP细胞的分布和形态有明显的种间差异。人、大鼠和小鼠PP细胞主要位于胰岛周边部,形成环形结构,少量PP细胞散布在外分泌部的腺泡和导管;而豚鼠、猪和猫的PP细胞则主要分布在外分泌部腺泡和导管上皮间;狗的PP细胞在内、外分泌部均有分布。PP细胞的形态在上述动物间也有明显的差异,这可能与该细胞在不同动物的作用途径及功能不同有关。     

地塞米松对哮喘豚鼠肺内VIP分布的影响

为探讨哮喘豚鼠肺内血管活性肠肽（ＶＩＰ）的分布及地塞米松对其分布的影响,将豚鼠随机分成哮喘组、地塞米松组和对照组三组,采用兔抗ＶＩＰ多克隆血清,免疫组织化学ＡＢＣ法和葡萄糖氧化酶－ＤＡＢ－镍染色技术,发现对照豚鼠肺内各级气道壁均可见ＶＩＰ免疫反应（ＶＩＰ－ＩＲ）阳性纤维分布。在平滑肌层中,纤维成束走行,且有较多分枝;而在基底膜和气道上皮内,纤维成单根行走。随气道口径的变小,纤维分布密度也逐渐变小。哮喘豚鼠肺内ＶＩＰ－ＩＲ消失。地塞米松处理后使ＶＩＰ－ＩＲ部分恢复。上述结果提示,豚鼠经反复抗原攻击后,肺内ＶＩＰ－ＩＲ阳性纤维消失,这在实验性哮喘的发生过程中可能起一定作用。     

不同方向撕除内界膜方式对特发性黄斑裂孔术后视网膜位移的影响

目的:比较玻璃体切割切术中不同方向撕除内界膜对特发性黄斑裂孔愈合后视网膜位移、视功能的影响。方法:纳入特发性黄斑裂孔患者25例(25眼),按照术中内界膜(ILM)撕除方向,以1:1随机分为NS-TI组(13眼)和TI-NS组(12眼)。NS-TI组患者接受内界膜撕除方向为鼻上起瓣,向颞下方向撕除ILM;TI-NS组患者接受内界膜撕除方向为颞下起瓣,向鼻上方向撕除ILM。术前、术后1月、术后3月采集患者自发荧光照相,通过影像学上血管标记点或交叉点的位置计算黄斑视盘距离(FMD)、颞侧血管至视盘距离(T-OD)、鼻侧血管至视盘距离(N-OD)、黄斑区垂直血管距离(VIAD)、黄斑区水平血管距离(HIAD)、黄斑区面积(PMA)。对比两种撕膜方式后术后1月、3月视网膜位移(包括FMD、T-OD、N-OD、VIAD、HIAD、PMA)、裂孔闭合率,术后最佳矫正视力,分析两种撕膜方式的异同。结果:术后1月及3月,两组患者的视网膜皆向视盘方向偏移,表现为FMD、T-OD、N-OD、VIAD、HIAD、PMA五项指标均较术前增大(p 0.05)。术后1月及3月,NS-TI组和TI-NS组FMD、T-OD、N-OD、VIAD、HIAD、PMA、黄斑裂孔愈合率(皆100%)和最佳矫正视力比较差异均无统计学意义(P0.05)。结论:不同方向撕除内界膜不是特发性黄斑裂孔术后视网膜位移的影响因素。     

玻璃体腔注射对裸小鼠视网膜组织形态学的影响

目的观察单纯的玻璃体腔注射对裸小鼠视网膜组织形态学的影响,为建立简单的制作视神经损伤动物模型奠定实验基础。方法在全身麻醉配合眼部局部麻醉情况下,利用微量注射器往裸小鼠玻璃体腔内迅速注入10μL生理盐水,然后在不同的时间点取注射眼进行固定、切片和HE染色,观察视网膜特别是视神经节细胞的变化。结果正常对照组视网膜层次清晰,各层排列整齐而致密,视网膜神经节细胞呈单层排列,大小不一,染色质分布均匀。实验组于注射后第1天、第3天和第5天视网膜神经节细胞减少的情况不明显,十层结构仍相对清晰。但于第7天,视网膜神经节细胞出现细胞明显缺失的现象,第14天为最严重,第30天和第60天与第14天相比无明显差别。结论玻璃体腔注射过量的生理盐水能够损伤视网膜组织,造成神经节细胞减少,有可能成为一种简单的制作视神经损伤动物模型的方法。     

屈光度和眼轴长度对豚鼠视网膜神经纤维层厚度和视乳头形态的影响

目的使用光学相干断层扫描(optical coherence tomography,OCT)观察豚鼠视网膜神经纤维层(retinal nerve fiber layer,RNFL)厚度及视乳头形态,并探讨豚鼠等效球镜和眼轴长度与这些参数的相关性。方法选用20只普通级豚鼠,进行等效球镜和眼轴长度测量,以及运用OCT观察豚鼠RNFL厚度及视乳头形态。结果豚鼠等效球镜与RNFL平均厚度、上方RNFL厚度、颞侧RNFL厚度、下方RNFL厚度、鼻侧RNFL厚度呈正相关;而眼轴长度与RNFL平均厚度、上方RNFL厚度、颞侧RNFL厚度、下方RNFL厚度、鼻侧RNFL厚度呈负相关。等效球镜和眼轴长度与盘沿面积、视盘面积、平均杯盘比、杯容积无相关性。等效球镜和垂直杯盘比无相关性,而眼轴长度与垂直杯盘比存在正相关。结论等效球镜和眼轴长度对豚鼠各方位RNFL厚度均有影响。在使用豚鼠作为高眼压动物模型时,需考虑其屈光状态和眼轴长度的影响。     

糖尿病大鼠视网膜血管铺片技术的改进及形态学观察

近年来我国糖尿病的患病率在逐年增加 ,病例总数居世界第二。糖尿病视网膜病变 (DiabeticRetinopathy DR)是糖尿病最为常见和严重的微血管病的并发症之一 ,也是成人主要致盲的疾病之一 ,病程愈长致盲机率愈高。DR发病机理至今尚未完全阐明 ,认为与多种因素的协同作用有关。目前 ,临床尚无有效的防治手段。因此研究 DR的发病机理 ,研制出有效治疗药物的任务迫在眉睫。视网膜毛细血管铺片技术是一种研究 DR理想的方法 ,不仅可清楚地观察到毛细血管的结构、走行、分布。还可进行定量分析细胞之间的比例、血管的密度、管径的大小等。但是 ,…     

猫视网膜多巴胺能无长突细胞发育的中央—周边梯度

本文报道用酪氨酸羟化酶(TH)免疫细胞化学方法标记猫视网膜多巴胺(DA)能无长突细胞发育的中央—周边梯度。TH阳性反应的I型DA能无长突细胞在发育成熟过程中呈现时空顺序的中央—周边梯度:(1)P_1时期分化较高的细胞,即染色深,胞体大,具有2—4支树突,分枝分布于内网状层(IPL),最外缘的星状Ⅰ_1类细胞大都集中于视网膜的中央部位;而分化较低的细胞,即染色淡,胞体小,具有1—2支树突,分枝分布于IPL外层和中层的不规则形Ⅰ_3类细胞大都集中于视网膜的周边部位;介于两者之间的Ⅰ_2类细胞散在分布于整个视网膜。它们形成了空间分布上的中央—周边分化成熟梯度。(2)随着发育进程,Ⅰ_1类细胞数增多,分布区逐渐从中央向四周扩展,由占视网膜总面积的30%(P_1时)增至65%(P_6时),P_(13)时达97%。开眼后P_(13)时,由于Ⅰ_1类细胞分布已扩展至周边,中央区和周边区间细胞平均直径和树突发育成熟程度的差别逐渐缩小,Ⅰ型DA能无长突细胞发育成熟的中央—周边梯度明显减弱。至P_(23)时,周边区细胞对TH抗体免疫反应强度以及形态学上成熟程度均相似中央区者,上述中央—周边梯度特征则完全消失。I型DA能无长突细胞发育成熟过程中呈现时空顺序的中央—周边梯度特征是视网膜个体发育过程中的暂时现象,它与视网膜中一些神经发生过程存在平行关系。它在视网膜神经发生中的作用,文中进行了讨论。     

鼠兔子宫血管铸型的光镜和扫描电镜观察

用光镜和扫描电镜观察了ＡＢＳ丁酮溶液灌注的达乌尔鼠兔子宫血管与微血管构筑情况。子宫大部分血液来自子宫动脉,小部分来自生殖动脉。各弓状动脉进入宫壁后,即在宫壁内分支形成３个血管层：浆膜层、大血管层和粘肌层。研究发现鼠兔子宫内膜血管呈树杆状或有轻度弯曲向腔面垂直穿行,直至浅层分支形成毛细血管网和较大的窦状毛细血管;其内膜血管形态与有月经的人子宫内膜螺旋动脉明显差异。文中还对子宫微血管构筑与月经产生机制     

Divergent distribution in vascular and avascular mammalian retinae links neuroglobin to cellular respiration

The visual function of the vertebrate retina relies on sufficient supply with oxygen. Neuroglobin is a respiratory protein thought to play an essential role in oxygen homeostasis of neuronal cells. For further understanding of its function, we compared the distribution of neuroglobin and mitochondria in both vascular and avascular mammalian retinae. In the vascular retinae of mouse and rat, oxygen is supplied by the outer choroidal, deep retinal, and inner capillaries. We show that in this type of retina, mitochondria are concentrated in the inner segments of photoreceptor cells, the outer and the inner plexiform layers, and the ganglion cell layer. These are the same regions in which oxygen consumption takes place and in which neuroglobin is present at high levels. In the avascular retina of guinea pig the deep retinal and inner capillaries are absent. Therefore, only the inner segments of the photoreceptors adjacent to choroidal capillaries display an oxidative metabolism. We demonstrate that in the retina of guinea pigs both neuroglobin and mitochondria are restricted to this layer. Our results clearly demonstrate an association of neuroglobin and mitochondria, thus supporting the hypothesis that neuroglobin is a respiratory protein that supplies oxygen to the respiratory chain.     

Regulation of potassium levels by Müller cells in the vertebrate retina

The membrane properties of Müller cells, the principal glial cells of the vertebrate retina, have been characterized in a series of physiological experiments on freshly dissociated cells. In species lacking a retinal circulation (tiger salamander, rabbit, guinea pig), the end-foot of the Müller cell has a much higher K+ conductance than do other cell regions. In species with retinal circulation (mouse, cat, owl monkey) the K+ conductance of the end-foot is greater than the conductance of the proximal process of the cell. In these species, however, the K+ conductance of the soma and distal process is equal to, or greater than, the end-foot conductance. Müller cells also possess four voltage-dependent ion channels, including an inward rectifying K+ channel. These membrane specializations may aid in the regulation of extracellular K+ levels by Müller cells in the retina. High end-foot conductance shunts excess K+ out through the end-foot, where it diffuses into the vitreous humor. In vascularized retinae, excess K+ may also be transferred to the ablumenal wall of capillaries, where it could be transported into the blood.     

Opsin-like immunoreaction in the retinae and pineal organs of four mammalian species

Summary Opsin-like immunoreactivity was observed in the retinae and pineal organs of the mouse, rat and guinea pig, and the pineal organ of the cat. In the retina the immunoreaction was restricted to photoreceptor cells, which displayed immunostaining in their perikarya and outer and inner segments. Distinct pinealocytes endowed with characteristic processes were labelled in the pineal organs of the mouse and cat. However, in the cat the number of immunoreactive pinealocytes was very limited. In the pineal organs of the rat and guinea pig immunoreaction was very weak and diffuse. No immunoreaction was observed when the antibody was preabsorbed with purified bovine (rhod)opsin. These findings are in accord with the results of previous studies indicating molecular similarities between retinal photoreceptors and pinealocytes in mammals.Supported by grants from the Deutsche Forschungsgemeinschaft to HWK, the European Science Foundation to RGF, the Alexander-von-Humboldt Stiftung to PE, and the Dutch Foundation for the Advancement of Basic Research. The authors are greatly indebted to Dr. Willem de Grip, Nijmegen, and Profesor Andreas Oksche, Giessen, for their critical interest in this study     

Enzymes Related to Monoamine Transmitter Metabolism in Brain Microvessels

The activities of tyrosine hydroxylase, aromatic L-aminoacid decarboxylase, monoamine oxidase, and catechol-O-methyltransferase were measured in microvessel (capillaries and venules), parenchymal arterioles, and pial vessels from rat brains, and the decarboxylase activity was compared in brain microvessels from rabbit, cat, dog, pig, cow, baboon, and man. Cranial sympathectomy was performed to estimate the neuronal contribution to the enzyme activities. All vascular regions had substantial activities of the various enzymes studied. The activity of aromatic L-aminoacid decarboxylase in cerebral microvessels was high in rat, dog, pig, cow, and man; intermediate in rabbit and cat; and low in baboon. In addition to this enzyme, cerebral microvessels also contained tyrosine hydroxylase and monoamine oxidase. Aromatic aminoacid decarboxylase and monoamine oxidase serve an enzymatic barrier function at the microvascular level, whereas the main function of tyrosine hydroxylase is probably to synthesize monoamines within nerve terminals that remain in close association with microvessels under the conditions used for preparation of the microvascular fraction. In larger intracerebral and pial vessels monoamine oxidase was present both in the wall itself and in perivascular sympathetic nerves; the remaining two enzymes had a primarily neuronal localization. The latter types of vessels also contained catechol-O-methyltransferase in their walls.     

An interspecies comparison of gangliosides and neutral glycolipids in adrenal glands

Gangliosides and neutral glycolipids of adrenal glands of mouse, rat, guinea pig, rabbit, cat, pig, cow, monkey, and chicken were analyzed by thin layer chromatography (TLC). The major gangliosides common to all species had lactosylceramide in their core structure. GM3 containing N-acetylneuraminic acid (NeuAc) was the major ganglioside in rat, guinea pig, rabbit, and cat, whereas GM3 containing N-glycolylneuraminic acid (NeuGc) was the major one in mouse, cow, and monkey. GD3 was also detected in all species except mouse and GD3(NeuAc)2 was the major in pig adrenal gland. GM4(NeuAc) was detected in the adrenal glands of guinea pig and chicken but not in those of the other species. In the neutral glycolipid fractions, galactosylceramide, glucosylceramide, lactosylceramide, globotriaosylceramide and globotetraosylceramide were detected and the proportions of these glycolipids varied among the species. Guinea pig and chicken adrenal glands contained large amounts of galactosylceramide, this being consistent with the presence of GM4 in these two species. Globopentaosylceramide was detected in mouse, guinea pig, cat, and chicken by the TLC-immunostaining procedure.     

Astrocyte invasion and vasculogenesis in the developing ferret retina

We studied the time course of astrocyte invasion and blood vessel formation in the developing ferret retina using glial fibrillary acidic protein (GFAP)-immunohistochemistry for astrocytes and isolectin B4 histochemistry for blood vessels. As in other mammals, strongly GFAP positive astrocytes invade the ferret retina from the optic nerve. At birth, strongly GFAP positive astrocytes have reached about 22% of the distance between optic disc and outer retinal edge whereas weakly GFAP positive processes already extend to the edge of the retina. At postnatal days P30–P37 about 82% of the distance between optic disc and outer retinal edge and in the adult 88% of this distance is covered with strongly labelled astrocytes. Superficial blood vessels form from the optic disc. They reach up to about 24% of the retinal radius at birth and grow radially across the retina during further development. At P30–P37, the whole retina is covered with superficial blood vessels. The deep vascular layer forms later (around P30) through sprouting from superficial vessels. The radial pattern of astrocyte and vessel growth from the optic disc is not affected by the formation of the area centralis and visual streak.     

Abnormal findings in the ocular fundi of colony-born cynomolgus monkeys

The ocular fundi of 1,151 apparently healthy colony-born cynomolgus monkeys (Macaca fascicularis) ranging in age from newborn to 19 years were examined using an ophthalmoscope. Two hundred and thirty-eight abnormal findings were recorded in 219 of the 1,151 monkeys. Of these, 23 were related to the optic disc and 115 to the retinal vessels. Of the remaining 100 abnormal findings, 91 were retinal degenerations and 9 were retinal hemorrhages. The 23 optic disc abnormalities consisted of 3 cases of micropapilla, 4 of ectasia and 16 of myelination of the retinal nerve fibers. Of the 115 retinal vascular abnormalities, 87 were arterial tortuosity, one was venous tortuosity, 2 were tortuosity of both artery and vein, 2 were artery-vein crossing, 20 were copper-wire artery, one was inosculation of the artery, one was vascularization of the vein and one was persistent hyaloid artery. Of the 91 retinal degenerations, one was degeneration of the periphery of the macular and the optic disc, 8 were macular degeneration and 82 were peripheral degeneration. Nine cases of retinal hemorrhages appeared under 6 years of age.     

Fluorescence microscopical and chemical characterization of the adrenergic innervation in mammalian liver tissue

Liver tissue from 12 different mammalian species was studied with a fluorescence histochemical technique for the cellular localization of amines (Falck-Hillarp technique) and with a chemical method for the determination of norepinephrine (HPLC-technique). Adrenergic nerve plexus were found in interlobular blood vessels derived from the portal vein and hepatic artery. Varicose adrenergic nerve fibres were, generally, seen to branch from the fibres around the blood vessels and to enter the liver parenchyma, where they formed a randomly distributed intralobular network. The density of these intralobular fibres showed marked species variation. Human liver and liver from the rhesus monkey, baboon, cynomolgus monkey and guinea pig showed a high density of parenchymal adrenergic nerves. Rabbit, cat, pig, cow and horse liver formed an intermediate group, having fewer varicose adrenergic nerve fibres but an unequivocal distribution of these nerves to the liver parenchyma. In rat and mouse liver no parenchymal innervation could be demonstrated. The density of the parenchymal innervation generally correlated with the concentration of norepinephrine in the liver tissue.