| The correlation between vascular development and Reelin expression in the visual system of vertebrate mice was studied by means of frozen section technique,shaking technique,immunohistochemical staining,gelatin-ink infusion and HE staining.Reelin expression in adult carp,bullfrog,pigeon and rabbit were explored at the sametime.Part one: Study on the correlation between vascular development and Reelin expression in vertebrate mice's visual system.Methods: Immunofluorescence staining and gelatin-ink infusion were performed in wild type mouse of P0,P3,P7,P14,P20 and adult in retina,superior colliculus and visual cortex V1 area respectively to observe the development of blood vessels and Reelin expression.The whole-mounts and frozen sections of the retina were stained with Collagen ? antibody to observe the development of blood vessels.In the study of superior colliculus and the visual cortex V1 region,blood vessels were observed by gelatin-ink infusion to study the development of blood vessels in the region.Fluorescence staining was performed with GFAP antibody and Reelin antibody in the shock slices to show the expression of astrocytes associated with vascular development and Reelin expression.The retinas of Reeler mice at P0 and adult stages were immunostained with Collagen ? antibody to observe the development of blood vessels.Results:(1)The mouse retinal blood vessels were generated after birth.Surface blood vessels and deep blood vessels of retinas were out spread around with the optic disc was served as the center.At the P10 age,the surface blood vessels covered the entire retina.Deep blood vessels began to grow at P8,when to the P14 stage vessels covered the retina.At this time the mouse retinal vascular system matured.At the P0 stage,the blood vessels of Reeler mice have covered the entire retina,however the branches of the deep blood vessels of adult mice significantly reduced compared with that of the wild type.With the development of retina,Reelin first was expressed in the ganglion cells and the inner plexiform layer,vertical branches from the ganglion cell layer extended to the outer layer.In P14 to adult stage,Reelin mainly distributed in the ganglion cell layer,the inner nuclear layer and the outer plexiform layer.(2)At the P0 stage,the blood vessels of superior colliculus have been generated,which were vertical to the pia mater on the surface of the superior colliculus and connected with the blood vessels in the pia mater.With the development of mice,the vascular density of the superior colliculus gradually increased.No obvious Reelin staining was found in superior colliculus,but GFAP was expressed which mostly closed to the vascular of the pia mater in P0 to adult stage.(3)At the P0 stage,radial blood vessels were generated in visual cortex V1 area.which was vertical to the pia mater on the surface of V1 area and connected with the blood vessels in V1 area.With the development of the cortex,vascular branches were more and more,and the density gradually increased.Radial glial cells were associated with blood vessels.Reelin mainly was expressed in the molecular layer,however a small amount of positive staining could be seen in the pyramidal cell layer and the inner granular layer.Conclusion: In the development of mice's visual system,there was a certain correlation between the expression of Reelin and the development of visual system in time and space.Reelin may be involved in regulating the orderly development of blood vessels in the mice retina.Part two: Study on the expression of Reelin in retina of adult crucian carp,bullfrog,pigeon and rabbit.Methods: Immunofluorescence staining and HE staining were performed in retina of adult carp,bullfrog,pigeon and rabbit,which were sliced using frozen sections.Results:(1)Reelin could be expressed in the retina of adult wild carp,bullfrogs,pigeons and rabbits.(2)GFAP was expressed in the retina of adult wild carp,bullfrog and rabbit,but there was no obvious positive staining in the retina of domestic pigeon. |
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