Liang F, Li J Y, Lovisetti-Scamihorn P, Coen E, Depreitere J, Claeys M, Wechsung E, Dahlström A, Winkler H, De Potter W P
Laboratory of Neuropharmacology, Department of Medicine, University of Antwerp (UIA), Universiteitsplein 1, B-2610, Antwerp, Belgium.
Brain Res. 1999 Feb 13;818(2):459-67. doi: 10.1016/s0006-8993(98)01327-4.
The distribution of secretoneurin (SN), a peptide derived from secretogranin II (SgII), in the coeliac ganglion, the splenic nerve and the spleen was examined by immunohistochemistry. In the ganglion, SN immunoreactivity (IR) was unevenly distributed. Positive nerve terminals densely surrounded some postganglionic perikarya in which also intense SN-IR was present. In the crushed splenic nerves, intense immunoreactivities appeared proximal (but to a less extent also distal) to the crush of the nerve. Analysis by cytofluorimetric scanning (CFS) demonstrated that SN-IR and neuropeptide Y immunoreactivity (NPY-IR) were predominant in the axons proximal to the crush representing anterogradely transported components. Using radioimmunoassay (RIA) we demonstrated that upon electrical stimulation (10 Hz, 1 min) of the splenic nerve, significant amounts of SN-IR (64.2+/-2.3 fmol) were released together with NA (4. 1x106+/-0.2 fmol) and NPY (330.0+/-7.2 fmol) from the isolated perfused porcine spleen. To evaluate the processing of SgII in sympathetic neurons, boiled tissue extracts (coeliac ganglia and splenic nerve) and boiled spleen perfusate (used as a suitable source for vesicle derived peptides) were analysed by gel filtration chromatography followed by SN-RIA. In all cases immunoreactivity was present solely as SN, indicating that SgII was fully processed to the free peptide. The evidence that SN is transported to the nerve terminals and is released from the porcine spleen upon nerve stimulation, suggests that it may modulate adrenergic neurotransmission and may also play a role in the neuroimmune communication.
采用免疫组织化学方法,研究了源自分泌粒蛋白II(SgII)的一种肽——分泌神经元素(SN)在腹腔神经节、脾神经和脾脏中的分布情况。在神经节中,SN免疫反应性(IR)分布不均。阳性神经末梢密集地围绕着一些节后神经元胞体,这些胞体中也存在强烈的SN-IR。在挤压后的脾神经中,强烈的免疫反应出现在神经挤压部位的近端(但在远端也有一定程度的反应)。细胞荧光扫描(CFS)分析表明,SN-IR和神经肽Y免疫反应性(NPY-IR)在挤压部位近端的轴突中占主导地位,代表顺行运输成分。使用放射免疫测定法(RIA),我们证明在对脾神经进行电刺激(10Hz,1分钟)后,从离体灌注的猪脾脏中释放出大量的SN-IR(64.2±2.3fmol),同时还有去甲肾上腺素(NA,4.1×10⁶±0.2fmol)和NPY(330.0±7.2fmol)。为了评估交感神经元中SgII的加工过程,对煮沸的组织提取物(腹腔神经节和脾神经)和煮沸的脾脏灌流液(用作囊泡衍生肽的合适来源)进行凝胶过滤色谱分析,随后进行SN-RIA分析。在所有情况下,免疫反应性仅以SN形式存在,这表明SgII已完全加工成游离肽。SN被转运到神经末梢并在神经刺激后从猪脾脏中释放的证据表明,它可能调节肾上腺素能神经传递,也可能在神经免疫通讯中发挥作用。