Wang J Y, Yaksh T L, Go V L
Brain Res. 1983 Nov 28;280(1):105-17. doi: 10.1016/0006-8993(83)91178-2.
In cat, radioimmunoassay revealed high concentrations of vasoactive intestinal polypeptide (VIP) and cholecystokinin (CCK) (approximately 10-90 ng/g wet weight tissue) in various regions of the cerebral cortex and in periventricular structures. Superfusion of cat cerebral cortex or perfusion from the lateral ventricle to the cisterna magna with artificial cerebrospinal fluid (CSF) was carried out in cats anesthetized with chloralose-urethane. The superfusate was assayed for VIP- and CCK-like immunoreactivity (VIP-LI and CCK-LI) simultaneously. Basal efflux of VIP-LI was 13.8 +/- 4.1 fmol/30 min/cm2 cortex and 31.6 +/- 5.0 fmol/30 min in ventriculo-cisternal superfusate. Basal efflux of CCK-LI, assayed in the same superfusate sample, was 10.7 +/- 2.5 fmol/30/min/cm2 cortex in cortical cup superfusate and 25.6 +/- 4.4 fmol/30 min in ventriculo-cisternal superfusate. The addition of potassium (50 mM) and veratridine (7.5 X 10(-5) M) to the superfusate produced significant increases in the levels of VIP-LI in both ventricular and cortical effluents. The potassium evoked release of VIP-LI and CCK-LI was inhibited by the substitution of cobalt (4 mM) for the calcium ion in the perfusion media; basal efflux was not affected. Separation of immunoreactivity by gel filtration chromatography demonstrated the CCK-LI in cat cortical tissue co-chromatographed with the COOH-terminal 4, 8, 33 and 39 amino acid peptide fragments of CCK and with a larger molecule. In contrast, the immunoreactivity in cortical and ventricular superfusate obtained during basal and evoked release co-migrated with authentic CCK octapeptide. For VIP, all immunoreactivity in tissue extract of cerebral cortex existed as a single peak which co-migrated with authentic VIP-28. The molecular pattern in cortical superfusate corresponded to that found in tissue extract.
在猫中,放射免疫分析显示,大脑皮层的各个区域以及脑室周围结构中,血管活性肠肽(VIP)和胆囊收缩素(CCK)的浓度很高(约为10 - 90纳克/克湿重组织)。在用氯醛糖 - 乌拉坦麻醉的猫中,对猫的大脑皮层进行灌流,或从侧脑室向大池灌注人工脑脊液(CSF)。同时对灌流液进行VIP样免疫反应性(VIP - LI)和CCK样免疫反应性(CCK - LI)检测。VIP - LI的基础流出量在皮层为13.8±4.1飞摩尔/30分钟/平方厘米,在脑室 - 脑池灌流液中为31.6±5.0飞摩尔/30分钟。在相同的灌流液样本中检测的CCK - LI基础流出量,在皮层杯灌流液中为10.7±2.5飞摩尔/30分钟/平方厘米,在脑室 - 脑池灌流液中为25.6±4.4飞摩尔/30分钟。向灌流液中加入钾(50毫摩尔)和藜芦碱(7.5×10⁻⁵摩尔)可使脑室和皮层流出液中的VIP - LI水平显著升高。钾诱导的VIP - LI和CCK - LI释放被灌注介质中用钴(4毫摩尔)替代钙离子所抑制;基础流出量不受影响。通过凝胶过滤色谱法分离免疫反应性表明,猫皮层组织中的CCK - LI与CCK的羧基末端4、8、33和39个氨基酸肽片段以及一个更大的分子共色谱。相比之下,在基础和诱导释放期间获得的皮层和脑室灌流液中的免疫反应性与真实的CCK八肽共迁移。对于VIP,大脑皮层组织提取物中的所有免疫反应性都以单峰形式存在,该单峰与真实的VIP - 28共迁移。皮层灌流液中的分子模式与组织提取物中的一致。