Comparative Neuroanatomy Laboratory, Biology, Ecology and Earth Science Department (DiBEST), University of Calabria, Ponte P. Bucci 4B, Arcavacata di Rende, Cosenza, 87036, Italy.
Institute on Membrane Technology, National Research Council of Italy, ITM-CNR, c/o University of Calabria, via P. Bucci cubo 17/C, I-87030, Rende (CS), Italy.
Mol Neurobiol. 2017 Nov;54(9):7369-7381. doi: 10.1007/s12035-016-0240-5. Epub 2016 Nov 5.
Depletion of oxygen and glucose even for brief periods is sufficient to cause cerebral ischemia, which is a predominant worldwide cause of motor deficits with the reduction of life quality and subsequently death. Hence, more insights regarding protective measures against ischemic events are becoming a major research goal. Among the many neuronal factors, N-methyl-D-aspartate receptors (NMDAR), orexinergic neuroreceptors (ORXR), and sympatho-inhibitory neuropeptide catestatin (CST) are widely involved with ischemic episodes. In this study, it was possible to induce in vitro ischemic conditions of the hamster (Mesocricetus auratus) hippocampal and hypothalamic neuronal cultures, grown on a newly compartmentalized membrane system, via oxygen and glucose deprivation (OGD). These cultures displayed notably differentiated NMDARergic and ORXergic receptor expression activities along with evident brain-derived neurotrophic factor (BDNF) plus orexin A (ORX-A) secretion, especially under co-cultured conditions. Interestingly, addition of CST in OGD-insulted hippocampal cells accounted for upregulated GluN1 and ORX1R transcripts that in the case of the latter neuroreceptor was very strongly (p < 0.001) increased when co-cultured with hypothalamic cells. Similarly, hypothalamic neurons supplied very evident upregulations of GluN1, ORX1R, and above all of GluN2A transcripts along with increased BDNF and ORX-A secretion in the presence of hippocampal cells. Overall, the preferential CST effects on BDNF plus ORX-A production together with altered NMDAR and ORXR levels, especially in co-cultured hypothalamic cells pointed to ORX-containing neurons as major protective constituents against ischemic damages thus opening new scenarios on the cross-talking roles of CST during ischemic disorders.
缺氧和葡萄糖的耗竭即使是短暂的,也足以导致脑缺血,这是全球范围内导致运动功能障碍的主要原因,降低了生活质量,随后导致死亡。因此,更多关于对抗缺血事件的保护措施的见解正在成为一个主要的研究目标。在许多神经元因素中,N-甲基-D-天冬氨酸受体(NMDAR)、食欲素能神经受体(ORXR)和交感神经抑制神经肽 catestatin(CST)广泛参与缺血发作。在这项研究中,有可能通过氧和葡萄糖剥夺(OGD)在体外诱导仓鼠(Mesocricetus auratus)海马和下丘脑神经元培养物的缺血条件,这些培养物在新的分隔膜系统上生长。这些培养物显示出明显分化的 NMDAR 能和 ORX 能受体表达活性,以及明显的脑源性神经营养因子(BDNF)加上食欲素 A(ORX-A)分泌,特别是在共培养条件下。有趣的是,在 OGD 损伤的海马细胞中添加 CST 导致 GluN1 和 ORX1R 转录物上调,在后一种神经受体中,当与下丘脑细胞共培养时,转录物上调非常强烈(p<0.001)。同样,下丘脑神经元在海马细胞存在的情况下,明显上调 GluN1、ORX1R,尤其是 GluN2A 转录物,同时增加 BDNF 和 ORX-A 分泌。总的来说,CST 对 BDNF 加 ORX-A 产生的优先影响以及 NMDAR 和 ORXR 水平的改变,特别是在共培养的下丘脑细胞中,表明含有食欲素的神经元是对抗缺血损伤的主要保护成分,从而为 CST 在缺血性疾病中的交叉对话作用开辟了新的场景。