Department of Oral and Maxillofacial Surgery, National University of Singapore, Singapore.
J Chem Neuroanat. 2010 Jul;39(4):242-7. doi: 10.1016/j.jchemneu.2010.02.002. Epub 2010 Feb 11.
Phospholipases A(2) (PLA(2)) are enzymes which cleave the sn-2 ester bond in membrane phospholipids to release free fatty acids and lysophospholipids. The present study aimed to elucidate the expression profile of multiple secretory phospholipase A(2) (sPLA(2)) isoforms in the normal rat CNS with focus on sPLA(2)-IIA in the brainstem and spinal cord. Quantitative RT-PCR analysis showed that sPLA(2)-IB expression was low throughout the CNS, sPLA(2)-IIA expression was high in the brainstem and spinal cord, sPLA(2)-IIC expression was high in the cerebral neocortex, hippocampus and thalamus/hypothalamus, sPLA(2)-V expression was high in the olfactory bulb and cerebellum, and sPLA(2)-X was expressed at very low levels in the normal CNS. Of the isoforms, sPLA(2)-IIA mRNA expression was highest in the brainstem and spinal cord suggesting that this could be the most relevant isoform in the ascending pain pathway. Western blot analysis showed high level of sPLA(2)-IIA expression in the brainstem and cervical, thoracic and lumbar spinal segments but low level of expression in other parts of the brain. sPLA(2)-IIA was localized by immunohistochemistry to the spinal trigeminal and facial motor nuclei and dorsal- and ventral-horns of the spinal cord. The enzyme was found on the endoplasmic reticulum of neuronal cell bodies and small diameter dendrites or dendritic spines at electron microscopy. The expression of sPLA(2)-IIA in the dorsal horn and spinal trigeminal nucleus is consistent with previous results which showed an important role of CNS sPLA(2) in nociceptive transmission.
磷脂酶 A(2) (PLA(2)) 是一种能够在膜磷脂的 sn-2 酯键处进行切割的酶,从而释放出游离脂肪酸和溶血磷脂。本研究旨在阐明多种分泌型磷脂酶 A(2) (sPLA(2)) 同工型在正常大鼠中枢神经系统中的表达谱,重点研究脑干和脊髓中的 sPLA(2)-IIA。定量 RT-PCR 分析显示,sPLA(2)-IB 在整个中枢神经系统中的表达水平较低,sPLA(2)-IIA 在脑干和脊髓中的表达水平较高,sPLA(2)-IIC 在大脑新皮质、海马体和丘脑/下丘脑中的表达水平较高,sPLA(2)-V 在嗅球和小脑中的表达水平较高,sPLA(2)-X 在正常中枢神经系统中的表达水平非常低。在这些同工型中,sPLA(2)-IIA mRNA 的表达在脑干和脊髓中最高,这表明它可能是上行疼痛通路中最相关的同工型。Western blot 分析显示,sPLA(2)-IIA 在脑干和颈段、胸段和腰段脊髓中表达水平较高,而在大脑的其他部位表达水平较低。免疫组织化学显示,sPLA(2)-IIA 定位于脊髓三叉神经和面运动核以及脊髓背角和腹角。在电子显微镜下,发现该酶位于神经元细胞体和小直径树突或树突棘的内质网上。sPLA(2)-IIA 在背角和三叉神经核中的表达与先前的研究结果一致,该研究表明中枢神经系统 sPLA(2) 在伤害性传递中起着重要作用。