Perera A D, Lagenaur C F, Plant T M
Department of Cell Biology and Physiology, Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261.
Endocrinology. 1993 Dec;133(6):2729-35. doi: 10.1210/endo.133.6.7694845.
Puberty in primates is triggered by a gonad-independent reinitiation of a pulsatile mode of GnRH release. The purpose of the present study was to begin to examine the hypothesis that this neuroendocrine event is the result of structural or plastic changes within the neural network governing the activity of GnRH neurons. Specifically, we sought to determine whether polysialic acid neural cell adhesion molecule (PSA-NCAM), a plasma membrane-associated glycoprotein that has previously been proposed to be a marker for postnatal neuronal plasticity, was expressed within GnRH neuron containing areas of the rhesus monkey hypothalamus. The study employed male monkeys that were castrated prepubertally. Immunocytochemistry of hypothalamic tissue from four animals of pubertal age employing a monoclonal antibody (12F8) specific for PSA-NCAM revealed the presence of PSA-NCAM immunoreactivity within the region of the arcuate nucleus and median eminence of the medial basal hypothalamus (MBH) and in the region of the organum vasculosum of the lamina terminalis of the rostral hypothalamus, two areas in the monkey brain where GnRH neurons are concentrated. As expected, immunostaining for total NCAM using a polyclonal rabbit antibody to mouse total NCAM was uniformly distributed throughout hypothalamic sections containing the MBH. Double staining showed that some, though not all, GnRH cell bodies of the MBH were located within the PSA-NCAM-immunopositive region of the arcuate nucleus and the median eminence. The pattern of PSA-NCAM immunoreactivity in the MBH of three prepubertal monkeys was similar to that seen for the older animals. Western analysis of a membrane extract from the MBH of a monkey of pubertal age, employing antibody 12F8, identified a broad band of staining at the expected molecular weight for this adhesion molecule. A similar, but less intense, immunoreactive band was observed for the preoptic area. In contrast, an immunoblot of a membrane extract of cerebral cortex was only faintly positive for PSA-NCAM. Taken together, the foregoing findings are consistent with the notion that structural changes within the MBH may underlie the pubertal reinitiation of pulsatile GnRH release. Moreover, the presence of PSA-NCAM in the MBH of prepubertal monkeys suggests that the role, if any, of this molecule in the onset of sexual maturation in primates is permissive in nature.
灵长类动物的青春期是由促性腺激素释放激素(GnRH)释放的脉冲模式在性腺非依赖性的情况下重新启动所触发的。本研究的目的是开始检验这样一种假说,即这种神经内分泌事件是控制GnRH神经元活动的神经网络内结构或可塑性变化的结果。具体而言,我们试图确定多唾液酸神经细胞黏附分子(PSA-NCAM),一种先前被认为是出生后神经元可塑性标志物的质膜相关糖蛋白,是否在恒河猴下丘脑含有GnRH神经元的区域中表达。该研究使用了青春期前阉割的雄性猴子。使用针对PSA-NCAM的单克隆抗体(12F8)对四只青春期年龄动物的下丘脑组织进行免疫细胞化学分析,结果显示在下丘脑内侧基底部(MBH)的弓状核和正中隆起区域以及下丘脑前部终板血管器区域存在PSA-NCAM免疫反应性,这是猴子大脑中GnRH神经元集中的两个区域。正如预期的那样,使用针对小鼠总NCAM的多克隆兔抗体对总NCAM进行免疫染色,在包含MBH的整个下丘脑切片中均匀分布。双重染色显示,MBH中部分(而非全部)GnRH细胞体位于弓状核和正中隆起的PSA-NCAM免疫阳性区域内。三只青春期前猴子的MBH中PSA-NCAM免疫反应性模式与年龄较大动物的相似。使用抗体12F8对一只青春期年龄猴子的MBH膜提取物进行蛋白质免疫印迹分析,在该黏附分子预期分子量处鉴定出一条宽条带染色。视前区观察到类似但强度较弱的免疫反应条带。相比之下,大脑皮质膜提取物的免疫印迹对PSA-NCAM仅呈弱阳性。综上所述,上述发现与MBH内的结构变化可能是青春期GnRH脉冲释放重新启动的基础这一观点一致。此外,青春期前猴子的MBH中存在PSA-NCAM表明该分子在灵长类动物性成熟开始过程中(如果有作用的话),其作用本质上是允许性的。