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甘丙肽信使核糖核酸在大鼠脑出生后发育过程中的表达:浦肯野细胞中的表达模式区分小脑的前叶和后叶。

Galanin messenger RNA during postnatal development of the rat brain: expression patterns in Purkinje cells differentiate anterior and posterior lobes of cerebellum.

作者信息

Ryan M C, Loiacono R E, Gundlach A L

机构信息

The University of Melbourne, Department of Medicine, Austin and Repatriation Medical Centre, Heidelberg, Australia.

出版信息

Neuroscience. 1997 Jun;78(4):1113-27. doi: 10.1016/s0306-4522(96)00652-5.

Abstract

Following our initial mapping of preprogalanin messenger RNA in adult brain and its presence in a subpopulation of cerebellar Purkinje neurons [Ryan M. C. and Gundlach A. C. (1996) Neuroscience 70, 709-728], the present study examined the ontogenic expression of preprogalanin messenger RNA in the postnatal rat brain focussing on the Purkinje cells of the cerebellar cortex. Using in situ hybridization histochemistry, preprogalanin messenger RNA was detected in the developing forebrain and hindbrain from postnatal day 4 to day 60 (adult). On postnatal day 4 very light hybridization signal (labelling) was observed in cells of a number of nuclei including the central amygdaloid nucleus, the medial preoptic area, paraventricular nucleus and dorsomedial hypothalamic nucleus of the forebrain while lightly-labelled cells were detected in neurons of the nucleus of the solitary tract and locus coeruleus of the hindbrain. Hybridization signal was not apparent in other nuclei until later, with positively-labelled neurons first apparent in the dorsal cochlear nucleus at postnatal day 21. The abundance of preprogalanin messenger RNA-positive neurons and the intensity of the hybridization signal increased, in most regions, until postnatal day 28 when labelling resembled that of the mature rat. Preprogalanin messenger RNA was first detected in the cerebellum on postnatal day 10 only in Purkinje cells of lobule 10 of the posterior vermis and increased in distribution throughout Purkinje cell layers of the entire cerebellar cortex by postnatal day 13. The intensity of hybridization signal in Purkinje cells varied between lobules, with Purkinje cells in lobule 10 displaying a moderate to heavy degree of labelling, while lobules 6-9 and the more posterior lobules of the hemisphere including crus 2 of the ansiform lobule, the paramedian lobule and the copula pyramis, displayed only light labelling. The intensity of labelling in the anterior vermis and the remaining lobules of the hemisphere including crus 1 of the ansiform lobule, the simple lobule, the paraflocculus and the flocculus, was homogeneously weak. By postnatal day 21, Purkinje cell labelling reached maximum intensity in all lobules. Regional differences were still apparent, however, with labelling in the posterior vermis and hemisphere ranging from moderate to heavy, with only light to moderate labelling detected in the anterior vermis. The intensity of labelling in the posterior vermis and most lobules of the hemisphere was similar from postnatal day 21 to adulthood, while, in the anterior vermis, crus 1 of the ansiform lobule and the simple lobule, the intensity of hybridization decreased slightly by postnatal day 28 and was completely absent in Purkinje cells of the adult rat. Differential expression of preprogalanin messenger RNA in Purkinje cells of the developing rat cerebellum and transient expression in certain lobules suggests that galanin gene products may have a role in both the developing and mature rat brain and that galanin gene expression may represent a useful marker for differentiating the anterior and posterior cerebellar lobes.

摘要

在我们首次绘制成年大脑中甘丙肽原信使核糖核酸(preprogalanin messenger RNA)图谱并发现其存在于小脑浦肯野神经元亚群之后[瑞安·M.C.和冈德拉克·A.C.(1996年)《神经科学》70卷,709 - 728页],本研究检测了出生后大鼠大脑中甘丙肽原信使核糖核酸的个体发育表达,重点关注小脑皮质的浦肯野细胞。采用原位杂交组织化学方法,在出生后第4天至第60天(成年)的发育中的前脑和后脑检测到了甘丙肽原信使核糖核酸。在出生后第4天,在前脑的一些核团细胞中观察到非常微弱的杂交信号(标记),包括中央杏仁核、内侧视前区、室旁核和背内侧下丘脑核,而后脑的孤束核和蓝斑核的神经元中检测到轻度标记的细胞。直到后来,其他核团中才出现明显的杂交信号,出生后第21天,在背侧耳蜗核首次出现阳性标记的神经元。在大多数区域,甘丙肽原信使核糖核酸阳性神经元的数量和杂交信号的强度增加,直到出生后第28天,此时标记与成年大鼠相似。出生后第10天,仅在后蚓部第10小叶的浦肯野细胞中首次检测到甘丙肽原信使核糖核酸,到出生后第13天,其分布扩展到整个小脑皮质的浦肯野细胞层。浦肯野细胞中杂交信号的强度在不同小叶之间有所不同,第10小叶的浦肯野细胞显示中度至重度标记,而第6 - 9小叶以及半球更靠后的小叶,包括半月小叶的脚2、旁正中小叶和锥体连合,仅显示轻度标记。前蚓部和半球其余小叶,包括半月小叶的脚1、简单小叶、旁绒球和绒球的标记强度均匀较弱。到出生后第21天,所有小叶的浦肯野细胞标记达到最大强度。然而,区域差异仍然明显,后蚓部和半球的标记从中度到重度不等,前蚓部仅检测到轻度至中度标记。从出生后第21天到成年,后蚓部和半球大多数小叶的标记强度相似,而在前蚓部、半月小叶的脚1和简单小叶,杂交强度在出生后第28天略有下降,成年大鼠的浦肯野细胞中完全没有标记。发育中大鼠小脑浦肯野细胞中甘丙肽原信使核糖核酸的差异表达以及某些小叶中的短暂表达表明,甘丙肽基因产物可能在发育中和成熟的大鼠大脑中都起作用,并且甘丙肽基因表达可能是区分小脑前后叶的一个有用标记。

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