Rx同源异型盒基因与视觉神经系统发育的关系

Rx (retinal homeobox)家族是新发现的一类与视觉神经系统发育密切相关的同源异型盒基因,调控眼基质、视泡、视网膜、前脑以及中脑部分区域的发育.Rx基因的研究将对视觉神经系统发育的调控机制提供新的认识.     

视网膜发育与形成的“镶嵌模型”

视网膜上不同种类的神经细胞为秩序分布.在胚胎发育过程中,如何能形成这种有秩序的空间分布对于眼睛和视网膜的发育至关重要.研究表明,眼睛和视网膜的发育与形成受多种基因的调控,不同的基因决定了视觉系统发育的不同结构.目前有报道认为,动物的眼睛在空间和时间上是由不同组织和不同分化的细胞镶嵌而形成的.     

不同视觉经验中zif268基因的表达模式

zif268基因编码一转录因子ZIF268. 在发育的视皮层中,zif268基因的表达模式受发育的调节. 在具有正常视觉经验的视皮层中,zif268基因有较高水平的表达. 视觉废置后,视皮层内该基因的表达水平显著降低. 通过视觉刺激可显著增强该基因的表达. 有关zif268基因表达模式的研究对于阐明该基因在哺乳动物视觉系统中的生理功能起到借鉴的作用.     

河川沙塘鳢视觉器官的发育及其与摄食的关系

利用光学显微镜观察了河川沙塘鳢(Odontobutis potamophila)视觉器官的发育,并对其发育与摄食的关系进行了研究。河川沙塘鳢的眼囊起源于神经外胚层。当胚胎发育至心跳期时,眼囊内陷形成视杯;之后,视杯内表面的外胚层形成晶状体而与视杯分离,视杯进一步发育形成视网膜。随着胚胎的进一步发育,晶状体的直径增加,结构逐步发育完善。胚胎发育至眼黑色素出现期时,视网膜分化为6层,其中,外核层、内核层和神经节细胞层3个核层明显;胚胎发育至孵化前期时,视网膜已分化为10层。孵出后1d的仔鱼,其视网膜已能行使功能,仔鱼逐渐开口摄食。随着稚、幼鱼的发育,视网膜厚度进一步增加,结构发育完善。视网膜的结构和视觉特性显示河川沙塘鳢是要求光照条件好、白昼活动并具有较好视觉功能的鱼类。     

牛肝辅酶Ⅱ依赖性视黄醇脱氢酶cDNA的克隆及组织表达

迄今为止的研究证明 ,维生素A亦称视黄醇(retinol)的生理功能是通过其两步氧化代谢产物视黄醛与视黄酸 (亦称维甲酸 )来完成的 .视黄醛通过其光学异构体 1 1 顺式视黄醛与视觉细胞内的视蛋白 (opsin)结合组成视色素 .感光时 ,1 1 顺式视黄醛转变成全反式视黄醛从视蛋白脱落 ,这一过程同时传导到大脑产生视觉[1 ] .全反式维甲酸 (all transretinoicacid)则通过与其在核内受体 (RARα ,β ,γ)结合调节基因的转录来发挥其许多重要的生理功能 ,包括正常胚胎的发育 ,形态、神经系统的形成 ,成体动物的生长、发育、繁殖等 ,并通过调解组织及…     

NRSF与神经系统发育调控

神经系统发育全程中,基因表达模式处于不断变化。这一动态过程是受到机体的精密调控的,NRSF是参与调控的重要分子之一。NRSF是一种含有锌指结构的转录因子,它与神经限制性沉默元件（NRSE／RE-11结合后能募集一些协同作用蛋白,通过组蛋白去乙酰化等机制抑制NRSE下游基因的转录。很多神经特异性基因和一些神经发育调节相关分子的基因序列中载有NRSE,NRSF正是通过动态调控这些基因的表达来调节神经系统发育。由于NRSF靶基因表达模式的变化是神经系统发育进程的重要分子基础,近年来对NRSF与神经系统发育调控的研究备受关注,通过阻断NRSF的异常作用来治疗神经退行性疾病己成为新的研究热点。     

不同视觉经验中zif268基因的表达模式

zif268基因编码一转录因子ZIF268. 在发育的视皮层中,zif268基因的表达模式受发育的调节. 在具有正常视觉经验的视皮层中,zif268基因有较高水平的表达. 视觉废置后,视皮层内该基因的表达水平显著降低. 通过视觉刺激可显著增强该基因的表达. 有关zif268基因表达模式的研究对于阐明该基因在哺乳动物视觉系统中的生理功能起到借鉴的作用.     

PKA在盘基网柄菌(Dictyostelium discoideum)多细胞发育中的作用

在盘基网柄菌(Dictyosteliumdiscoideum)多细胞发育中,蛋白激酶A(proteinkinaseA,PKA)发挥多重作用.细胞聚集阶段,PKA调节腺苷酰环化酶的活性,中转cAMP,诱导dut、pdi等一些发育早期的基因表达;参与启动聚集后的细胞分化和形态构成,增强GBF活性,激活前孢子细胞特有基因的表达;它还精密调控前柄细胞特有基因ecmB的表达,准确启动拔顶发育,诱导孢柄和孢子的成熟.子实体形成后,PKA又是维持孢子休眠和保证孢子有效萌发的必需因子.在PKA调控下,盘基网柄菌有条不紊地完成整个发育过程.     

Organization and postembryonic development of glial cells in the visual system of the Chinese honeybee, Apis cerana cerana (Hymenoptera:Apidae )

神经胶质作为视觉系统的重要成分之一,对视觉系统的发育及功能起着重要的作用.本研究通过组织解剖观察、免疫组织化学等技术,对中华蜜蜂Apis cerana cerana幼虫和蛹的视觉系统中神经胶质的类型和发育过程进行了比较研究.研究表明:在中华蜜蜂视觉系统中,根据神经胶质的位置和形态主要分为表面神经胶质、皮层神经胶质和神经纤维网神经胶质3种类型;神经胶质主要来源于视柄和视叶中的神经胶质前体中心;神经胶质细胞数量的增加一方面来自于细胞的迁移,另一方面来自于神经胶质细胞自身的分裂增殖.本研究为昆虫神经胶质的发育以及功能研究提供理论基础.     

bcl-2在视网膜发育及病变治疗中的机制研究

在视网膜的正常发育与视网膜多种疾病中,细胞凋亡与视网膜细胞的死亡、疾病的发生发展有密切关系,是维持眼发育或视功能损伤的重要因素之一,而Bcl-2家族是主要的细胞凋亡调控蛋白,其中的抗凋亡蛋白又以Bcl-2基因最具有代表性,本文就视网膜发育及一些视网膜疾病中细胞凋亡与Bcl-2基因表达的变化与相关机制作一综述.     

Proper patterning of the optic fissure requires the sequential activity of BMP7 and SHH

The optic disc develops at the interface between optic stalk and retina, and enables both the exit of visual fibres and the entrance of mesenchymal cells that will form the hyaloid artery. In spite of the importance of the optic disc for eye function, little is known about the mechanisms that control its development. Here, we show that in mouse embryos, retinal fissure precursors can be recognised by the expression of netrin 1 and the overlapping distribution of both optic stalk (Pax2, Vax1) and ventral neural retina markers (Vax2, Raldh3). We also show that in the absence of Bmp7, fissure formation is not initiated. This absence is associated with a reduced cell proliferation and apoptosis in the proximoventral quadrant of the optic cup, lack of the hyaloid artery, optic nerve aplasia, and intra-retinal misrouting of RGC axons. BMP7 addition to organotypic cultures of optic vesicles from Bmp7-/- embryos rescues Pax2 expression in the ventral region, while follistatin, a BMP7 antagonist, prevents it in early, but not in late, optic vesicle cultures from wild-type embryos. The presence of Pax2-positive cells in late optic cup is instead abolished by interfering with Shh signalling. Furthermore, SHH addition re-establishes Pax2 expression in late optic cups derived from ocular retardation (or) embryos, where optic disc development is impaired owing to the near absence of SHH-producing RGC. Collectively, these data indicate that BMP7 is required for retinal fissure formation and that its activity is needed, before SHH signalling, for the generation of PAX2-positive cells at the optic disc.     

Sonic hedgehog is involved in formation of the ventral optic cup by limiting Bmp4 expression to the dorsal domain

Accumulating evidence suggests that Sonic hedgehog (Shh) signaling plays a crucial role in eye vesicle patterning in vertebrates. Shh promotes expression of Pax2 in the optic stalk and represses expression of Pax6 in the optic cup. Shh signaling contributes to establishment of both proximal–distal and dorsal–ventral axes by activating Vax1, Vax2, and Pax2. In the dorsal part of the developing retina, Bmp4 is expressed and antagonizes the ventralizing effects of Shh signaling through the activation of Tbx5 expression in chick and Xenopus. To examine the roles of Shh signaling in optic cup formation and optic stalk development, we utilized the Smoothened (Smo) conditional knockout (CKO) mouse line. Smo is a membrane protein which mediates Shh signaling into inside of cells. Cre expression was driven by Fgf15 enhancer. The ventral evagination of the optic cup deteriorated from E10 in the Smo-CKO, whereas the dorsal optic cup and optic stalk develop normally until E11. We analyzed expression of various genes such as Pax family (Pax2/Pax6), Vax family (Vax1/Vax2) and Bmp4. Bmp4 expression was greatly upregulated in the optic vesicle by the 21-somite stage. Then Vax1/2 expression was decreased at the 20- to 24-somite stages. Pax2/6 expression was affected at the 27- to 32-somite stages. Our data suggest that the effects of the absence of Shh signaling on Vax1/Vax2 are mediated through increased Bmp4 expression throughout the optic cup. Also unchanged patterns of Raldh2 and Raldh3 suggest that retinoic acid is not the downstream to Shh signaling to control the ventral optic cup morphology.     

Vax2 inactivation in mouse determines alteration of the eye dorsal-ventral axis, misrouting of the optic fibres and eye coloboma

Vax2 is a homeobox gene whose expression is confined to the ventral region of the prospective neural retina. Overexpression of this gene at early stages of development in Xenopus and in chicken embryos determines a ventralisation of the retina, thus suggesting its role in the molecular pathway that underlies eye development. We describe the generation and characterisation of a mouse with a targeted null mutation of the Vax2 gene. Vax2 homozygous mutant mice display incomplete closure of the optic fissure that leads to eye coloboma. This phenotype is not fully penetrant, suggesting that additional factors contribute to its generation. Vax2 inactivation determines dorsalisation of the expression of mid-late (Ephb2 and Efnb2) but not early (Pax2 and Tbx5) markers of dorsal-ventral polarity in the developing retina. Finally, Vax2 mutant mice exhibit abnormal projections of ventral retinal ganglion cells. In particular, we observed the almost complete absence of ipsilaterally projecting retinal ganglion cells axons in the optic chiasm and alteration of the retinocollicular projections. All these findings indicate that Vax2 is required for the proper closure of the optic fissure, for the establishment of a physiological asymmetry on the dorsal-ventral axis of the eye and for the formation of appropriate retinocollicular connections.     

Retinoic acid is required for specification of the ventral eye field and for Rathke's pouch in the avian embryo

We have investigated the role of retinoic acid (RA) in eye development using the vitamin A deficient quail model system, which overcomes problems of retinoic acid synthesising enzyme redundancy in the embryo. In the absence of retinoic acid, the ventral optic stalk and ventral retina are missing, whereas the dorsal optic stalk and dorsal retina develop appropriately. Other ocular abnormalities observed were a thinner retina and the lack of differentiation of the lens. In an attempt to explain this, we studied the expression of various dorsally and ventrally expressed genes such as Pax2, Pax6, Tbx6, Vax2, Raldh1 and Raldh3 and noted that they were unchanged in their expression patterns. In contrast, the RA catabolising enzymes Cyp26A1 and Cyp26B1 which are known to be RA-responsive were not expressed at all in the developing eye. At much earlier stages, the expression domain of Shh in the prechordal plate was reduced, as was Nkx2.1 and we suggest a model whereby the eye field is specified according to the concentration of SHH protein that is present. We also describe another organ, Rathke's pouch which fails to develop in the absence of retinoic acid. We attribute this to the down-regulation of Bmp2, Shh and Fgf8 which are known to be involved in the induction of this structure.     

Hedgehog signalling maintains the optic stalk-retinal interface through the regulation of Vax gene activity

During early formation of the eye, the optic vesicle becomes partitioned into a proximal domain that forms the optic nerve and a distal domain that forms the retina. In this study, we investigate the activity of Nodal, Hedgehog (Hh) and Fgf signals and Vax family homeodomain proteins in this patterning event. We show that zebrafish vax1 and vax2 are expressed in overlapping domains encompassing the ventral retina, optic stalks and preoptic area. Abrogation of Vax1 and Vax2 activity leads to a failure to close the choroid fissure and progressive expansion of retinal tissue into the optic nerve, finally resulting in a fusion of retinal neurons and pigment epithelium with forebrain tissue. We show that Hh signals acting through Smoothened act downstream of the Nodal pathway to promote Vax gene expression. However, in the absence of both Nodal and Hh signals, Vax genes are expressed revealing that other signals, which we show include Fgfs, contribute to Vax gene regulation. Finally, we show that Pax2.1 and Vax1/Vax2 are likely to act in parallel downstream of Hh activity and that the bel locus (yet to be cloned) mediates the ability of Hh-, and perhaps Fgf-, signals to induce Vax expression in the preoptic area. Taking all these results together, we present a model of the partitioning of the optic vesicle along its proximo-distal axis.     

Spatial specification of mammalian eye territories by reciprocal transcriptional repression of Pax2 and Pax6

We have studied the molecular basis of the Pax2 and Pax6 function in the establishment of visual system territories. Loss-of-function mutants have revealed crucial roles for Pax2 in the generation of the optic stalk and for Pax6 in the development of the optic cup. Ectopic expression of Pax6 in the optic stalk under control of Pax2 promoter elements resulted in a shift of the optic cup/optic stalk boundary indicated by the presence of retinal pigmented cells on the optic stalk. By studying mouse embryos at early developmental stages we detected an expansion of Pax2 expression domain in the Pax6(-/-) mutant and of Pax6 expression domain in the Pax2(-/-) embryo. These results suggest that the position of the optic cup/optic stalk boundary depends on Pax2 and Pax6 expression, hinting at a possible molecular interaction. Using gel shift experiments, we confirmed the presence of Pax2- and Pax6-binding sites on the retina enhancer of the Pax6 gene and on the Pax2 upstream control region, respectively. Co-transfection experiments revealed a reciprocal inhibition of Pax2 promoter/enhancer activity by Pax6 protein and vice versa. Based on our findings, we propose a model for Pax gene regulation that establishes the proper spatial regionalization of the mammalian visual system.     

中华蜜蜂视觉系统中神经胶质的组成和胚后发育

神经胶质作为视觉系统的重要成分之一, 对视觉系统的发育及功能起着重要的作用。本研究通过组织解剖观察、 免疫组织化学等技术, 对中华蜜蜂Apis cerana cerana幼虫和蛹的视觉系统中神经胶质的类型和发育过程进行了比较研究。研究表明： 在中华蜜蜂视觉系统中, 根据神经胶质的位置和形态主要分为表面神经胶质、 皮层神经胶质和神经纤维网神经胶质3种类型; 神经胶质主要来源于视柄和视叶中的神经胶质前体中心; 神经胶质细胞数量的增加一方面来自于细胞的迁移, 另一方面来自于神经胶质细胞自身的分裂增殖。本研究为昆虫神经胶质的发育以及功能研究提供理论基础。     

Pre-existing neuronal pathways in the developing optic lobes of Drosophila

We have identified a set of larval neurones in the developing adult optic lobes of Drosophila by selectively labelling cells that have undergone only a few mitoses. A cluster of three cells is located in each of the optic lobes near the insertion site of the optic stalk. Their axons fasciculate with fibres of the larval optic nerve, the Bolwig's nerve, and then form part of the posterior optic tract. These cells are likely to be first order interneurones of the larval visual system. Unlike the Bolwig's nerve, they persist into the adult stage. The possibility of a pioneering function of the larval visual system during formation of the adult optic lobe neuropil is discussed.     

Retinal ganglion cell-derived sonic hedgehog signaling is required for optic disc and stalk neuroepithelial cell development

The development of optic stalk neuroepithelial cells depends on Hedgehog (Hh) signaling, yet the source(s) of Hh protein in the optic stalk is unknown. We provide genetic evidence that sonic hedgehog (Shh) from retinal ganglion cells (RGCs) promotes the development of optic disc and stalk neuroepithelial cells. We demonstrate that RGCs express Shh soon after differentiation, and cells at the optic disc in close proximity to the Shh-expressing RGCs upregulate Hh target genes, which suggests they are responding to RGC-derived Shh signaling. Conditional ablation of Shh in RGCs caused a complete loss of optic disc astrocyte precursor cells, resulting in defective axon guidance in the retina, as well as conversion of the neuroepithelial cells in the optic stalk to pigmented cells. We further show that Shh signaling modulates the size of the Pax2(+) astrocyte precursor cell population at the optic disc in vitro. Together, these data provide a novel insight into the source of Hh that promotes neuroepithelial cell development in the mammalian optic disc and stalk.