Postnatal development and the differential expression of presynaptic terminal-associated proteins in the developing retina of the Brazilian opossum, Monodelphis domestica.

M H Greenlee, J J Swanson, J J Simon, J K Elmquist, C D Jacobson, D S Sakaguchi
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Abstract

In the present study we have characterized the postnatal (PN) development of the retina in the Brazilian opossum, Monodelphis domestica. Monodelphis, a small, pouchless marsupial, undergoes a protracted period of postnatal development. Using bromodeoxyuridine immunohistochemistry, we have investigated postnatal neurogenesis of the retina. In addition, we have examined the differentiation of the retina by using antibodies directed against the presynaptic terminal-associated proteins synaptotagmin, Rab3A, synaptophysin and synaptosomal-associated protein-25 (SNAP-25), and have characterized their spatial and temporal distribution during postnatal development. This study is the first systematic comparison of the developmental expression of multiple presynaptic terminal-associated proteins in relation to retinal neurogenesis and differentiation. At birth (1PN), the Monodelphis retina was relatively undifferentiated morphologically and birthdating analysis revealed mitotically active cells throughout the retina. The 8PN retina was organized into two cellular layers: an outer region of mitotically active neuroepithelial cells and an inner region of postmitotic cells. The inner plexiform layer formed between 5PN and 10PN, and exhibited unique patterns of immunoreactivity with the antibodies used in this analysis. By 25PN the retina was well laminated, and synaptotagmin-, Rab3A-, synaptophysin- and SNAP-25-like immunoreactivities exhibited distinct and specific patterns within the plexiform layers, although they had not yet achieved their mature, adult patterns. These results indicate that each of these proteins exhibits developmentally regulated changes in its cellular localization, and therefore may play important roles during morphogenesis and synaptogenesis of the vertebrate retina.

出生后发育和突触前终末相关蛋白在巴西负鼠视网膜发育中的差异表达。
在目前的研究中,我们描述了巴西负鼠(Monodelphis domestica)视网膜的产后(PN)发育。一种小的,无袋的有袋动物,在出生后经历了漫长的发育时期。利用溴脱氧尿苷免疫组化技术,我们研究了视网膜出生后的神经发生。此外,我们还利用针对突触前终末相关蛋白synaptotagmin、Rab3A、synaptophysin和synaptosomal-associated protein-25 (SNAP-25)的抗体检测了视网膜的分化,并表征了它们在出生后发育过程中的时空分布。本研究首次系统比较了多种突触前终末相关蛋白在视网膜神经发生和分化中的发育表达。出生时(1PN), Monodelphis视网膜在形态学上相对未分化,出生年代分析显示整个视网膜有丝分裂活跃细胞。8PN视网膜被组织成两个细胞层:有丝分裂活跃的神经上皮细胞的外部区域和有丝分裂后细胞的内部区域。内部丛状层形成于5PN和10PN之间,与本分析中使用的抗体表现出独特的免疫反应性模式。到25PN时,视网膜层压良好,突触tagmin-, Rab3A-,突触体素-和snap -25样免疫反应在网状层内表现出独特的特异性模式,尽管它们尚未达到成熟的成体模式。这些结果表明,这些蛋白在其细胞定位中表现出发育调节的变化,因此可能在脊椎动物视网膜的形态发生和突触发生中发挥重要作用。
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