Wright D J, Ritchie T C, Coulter J D
Department of Anatomy, College of Medicine, University of Iowa, Iowa City 52242.
J Comp Neurol. 1991 Feb 22;304(4):530-43. doi: 10.1002/cne.903040403.
Previous studies of the nerve terminal protein NT75 in the developing spinal cord have suggested an association between the appearance of NT75 immunoreactivity and the process of synaptogenesis. To examine the time course of NT75 expression further, the current study compared the localization of NT75 and the synaptic vesicle protein synaptophysin in the adult and developing rat cerebellum and in cerebellar tissue cultures. In the adult cerebellum, dense NT75 staining is confined to the molecular layer, where it is associated with parallel fiber endings of cerebellar granule cells. During development, NT75 immunoreactivity is first detectable in the cerebellar cortex as a dense band of staining in the deepest portion of the molecular layer at postnatal day 10. The stained zone expands to occupy a progressively greater portion of the molecular layer until about postnatal day 20. Synaptophysin staining occurs in granule cell processes earlier than NT75 and is found throughout the molecular layer by postnatal day 7. Quantitatively, rapid increases in both NT75 and synaptophysin occur in the first three postnatal weeks, with NT75 activity reaching levels exceeding the adult value by 50% over postnatal days 20 through 30, whereas synaptophysin plateaus at near adult levels by postnatal day 20. In cerebellar cultures, NT75 staining in neurites develops over several days, increasing coincidentally with development of synaptic contacts, whereas synaptophysin staining is already present in most neurites after only 1 day in vitro. The results indicate that NT75 expression in developing cerebellar granule cell nerve terminals is closely associated with the appearance of mature nerve terminals, suggesting that the protein may have a role in the formation/stabilization of the synaptic ending or in the mechanisms of synaptic transmission.
以往对发育中的脊髓中神经末梢蛋白NT75的研究表明,NT75免疫反应性的出现与突触发生过程之间存在关联。为了进一步研究NT75表达的时间进程,本研究比较了NT75和突触囊泡蛋白突触素在成年和发育中的大鼠小脑以及小脑组织培养物中的定位。在成年小脑中,密集的NT75染色局限于分子层,与小脑颗粒细胞的平行纤维末梢相关。在发育过程中,NT75免疫反应性在出生后第10天首先在小脑皮质中作为分子层最深处的一条密集染色带被检测到。染色区域逐渐扩大,占据分子层中越来越大的部分,直到出生后约第20天。突触素染色比NT75更早出现在颗粒细胞突起中,到出生后第7天在整个分子层中都能发现。定量分析表明,NT75和突触素在出生后的前三周都迅速增加,NT75活性在出生后第20天至30天超过成年值50%,而突触素在出生后第20天达到接近成年水平并趋于平稳。在小脑培养物中,神经突中的NT75染色在几天内逐渐形成,与突触接触的发育同时增加,而突触素染色在体外培养仅1天后就在大多数神经突中已经存在。结果表明,发育中的小脑颗粒细胞神经末梢中NT75的表达与成熟神经末梢的出现密切相关,提示该蛋白可能在突触末梢的形成/稳定或突触传递机制中发挥作用。