Alonso G, Ridet J L, Oestreicher A B, Gispen W H, Privat A
INSERM U 336, University of Montpellier II, France.
Exp Neurol. 1995 Jul;134(1):35-48. doi: 10.1006/exnr.1995.1034.
The persistence of high levels of B-50 (GAP-43) in fibers innervating various regions of the adult central nervous system is generally thought to characterize neuronal systems capable of undergoing morphological plasticity. In a recent series of in situ hybridization studies, it has been shown that most catecholaminergic and serotonergic neurons of the adult rat brain express high levels of B-50 mRNA. The present study addresses the question whether high expression of B-50 mRNA in the catecholaminergic and serotonergic perikarya corresponds with detectable high levels of the B-50 protein in the efferent axonal fibers that innervate various regions of the adult rat brain and spinal cord. For this purpose, vibratome sections were doubly immunostained for B-50 and for tyrosine hydroxylase or serotonin and were analyzed by laser scanning confocal microscope. Colocalizations were investigated either (1) in regions of intact rat brain and spinal cord in which particular concentrations of B-50 immunoreactive fibers appeared codistributed with catecholaminergic or serotonergic fibers or (2) in intrahypothalamic portions of the medial forebrain bundle in which a surgical lesion was made. In the intact brain, frequent colocalizations of B-50 and tyrosine hydroxylase were detected in fibers innervating both the mediobasal hypothalamus and the neurointermediate hypophysial lobe. In all the other regions examined, the analysis of thin optical sections demonstrated that immunoreactivity to B-50 was only rarely associated with axonal profiles immunoreactive to tyrosine hydroxylase or to serotonin. By contrast, in the lesioned medial forebrain bundle B-50 immunoreactivity was found to be associated with numerous catecholaminergic and serotonergic axonal sprouts that regenerate around the surgical lesion. These data indicate that the majority of intact catecholaminergic and serotonergic axons innervating the adult rat brain and spinal cord contains low levels of B-50. However, following axotomy, B-50 is immunocytochemically detectable in the regenerating sprouts produced by both types of axonal fibers. This suggests that under basal conditions the relatively high content of B-50 mRNA in monoaminergic perikarya does not lead to appreciable accumulation of B-50 within corresponding axonal fibers and terminals, whereas conditions of morphological reorganization induce increased production of B-50 that accumulates within monoaminergic axonal sprouts.
一般认为,支配成年中枢神经系统不同区域的纤维中高水平的B - 50(GAP - 43)持续存在,是能够发生形态可塑性的神经元系统的特征。在最近一系列原位杂交研究中,已表明成年大鼠脑中大多数儿茶酚胺能和5 - 羟色胺能神经元表达高水平的B - 50 mRNA。本研究探讨了儿茶酚胺能和5 - 羟色胺能神经元胞体中B - 50 mRNA的高表达是否与支配成年大鼠脑和脊髓不同区域的传出轴突纤维中可检测到的高水平B - 50蛋白相对应。为此,用振动切片机切片,对B - 50以及酪氨酸羟化酶或5 - 羟色胺进行双重免疫染色,并通过激光扫描共聚焦显微镜进行分析。共定位研究在以下两种情况中进行:(1)在成年大鼠完整的脑和脊髓区域,其中特定浓度的B - 50免疫反应性纤维似乎与儿茶酚胺能或5 - 羟色胺能纤维共分布;(2)在进行了手术损伤的松果体束下丘脑内侧部分。在完整大脑中,在支配下丘脑中间基底部和神经垂体中间叶的纤维中,检测到B - 50和酪氨酸羟化酶频繁共定位。在所有其他检查区域,对薄光学切片的分析表明,对B - 50的免疫反应性仅很少与对酪氨酸羟化酶或5 - 羟色胺免疫反应的轴突轮廓相关。相比之下,在损伤的松果体束中,发现B - 50免疫反应性与在手术损伤周围再生的许多儿茶酚胺能和5 - 羟色胺能轴突发芽相关。这些数据表明,支配成年大鼠脑和脊髓的大多数完整儿茶酚胺能和5 - 羟色胺能轴突含有低水平的B - 50。然而,轴突切断后,在两种类型轴突纤维产生的再生芽中可通过免疫细胞化学检测到B - 50。这表明在基础条件下,单胺能神经元胞体中相对较高的B - 50 mRNA含量不会导致相应轴突纤维和终末内B - 50的明显积累,而形态重组条件会诱导B - 50产生增加并在单胺能轴突发芽中积累。