Reiner A, Eldred W D, Beinfeld M C, Krause J E
J Neurosci. 1985 Jun;5(6):1527-44. doi: 10.1523/JNEUROSCI.05-06-01527.1985.
Single-label and double-label immunohistochemical techniques were used to demonstrate the coexistence of substance P-like immunoreactivity (SPLI) and cholecystokinin-8-like immunoreactivity (CCK-8-LI) in an extensive fiber system within the telencephalic cortex of turtle. All SPLI-containing fibers and terminals of this system contain CCK-8-LI and vice versa. The fibers of this system course from more medial cortical regions to more lateral ones, originating either from neurons in the more medial cortices or from extracortical neurons, the axons of which ascend the medial wall of the cortex. The precise location of the neurons that give rise to this cortical projection system is uncertain, but a hypothalamic location seems most likely at present. The fibers and terminals of this system are found throughout the entire mediolateral and rostrocaudal extent of the telencephalic cortex of turtle and are largely confined to the cell body layer of the cortex. Fewer SPLI/CCK-8-LI-containing fibers are found in pyriform (olfactory) cortex than in the other cortices. Ultrastructural studies indicate that SPLI/CCK-8-LI-containing terminals make asymmetric synapses on cell bodies or their proximal dendrites. Both SPLI and CCK-8-LI are found in large dense core vesicles in these labeled terminals. Labeled terminals also contained numerous small, round, unlabeled vesicles clustered near synaptic release sites and a number of unlabeled large dense core vesicles. Quantification of the percentage of the large dense core vesicles that were labeled in SP-labeled terminals, in CCK-8-labeled terminals, and in terminals labeled for both SP and CCK-8 provided suggestive evidence that SPLI and CCK-8-LI must be contained within the same large dense core vesicles. Radioimmunoassay indicated that the SP/CCK-8-containing system of turtle cortex contains 0.93 +/- 0.090 pg of SP/microgram of cortical tissue protein and 0.31 +/- 0.11 pg of CCK-8/micrograms of cortical tissue protein. The CCK-8-like material in turtle cortex coelutes with CCK-8-sulfate, using gradient elution high pressure liquid chromatography (HPLC). The SP-like material, although immunologically highly similar to undecapeptide SP (Reiner, A., J. E. Krause, K. T. Keyser, W. D. Eldred, and J. F. McKelvy (1984) J. Comp. Neurol. 226: 50-75), does not coelute with undecapeptide SP using gradient elution HPLC.(ABSTRACT TRUNCATED AT 400 WORDS)
采用单标记和双标记免疫组织化学技术,以证明在龟端脑皮质广泛的纤维系统中,P物质样免疫反应性(SPLI)和胆囊收缩素-8样免疫反应性(CCK-8-LI)共存。该系统中所有含SPLI的纤维和终末均含有CCK-8-LI,反之亦然。该系统的纤维从内侧皮质区域延伸至外侧皮质区域,起源于内侧皮质中更多的神经元或皮质外神经元,其轴突沿皮质内侧壁上行。产生该皮质投射系统的神经元的确切位置尚不确定,但目前看来最有可能位于下丘脑。该系统的纤维和终末见于龟端脑皮质整个内外侧和前后范围,且主要局限于皮质的细胞体层。梨状(嗅觉)皮质中含SPLI/CCK-8-LI的纤维比其他皮质中少。超微结构研究表明,含SPLI/CCK-8-LI的终末在细胞体或其近端树突上形成不对称突触。在这些标记终末的大致密核心囊泡中发现了SPLI和CCK-8-LI。标记终末还含有许多小的、圆形的、未标记的囊泡聚集在突触释放部位附近,以及一些未标记的大致密核心囊泡。对SP标记终末、CCK-8标记终末以及同时标记SP和CCK-8的终末中被标记的大致密核心囊泡百分比进行定量,提供了提示性证据,表明SPLI和CCK-8-LI必定包含在同一大致密核心囊泡内。放射免疫分析表明,龟皮质含SP/CCK-8的系统每微克皮质组织蛋白含0.93±0.090 pg的SP和0.31±0.11 pg的CCK-8。采用梯度洗脱高压液相色谱(HPLC)法,龟皮质中的CCK-8样物质与硫酸化CCK-8共洗脱。尽管该SP样物质在免疫上与十一肽SP高度相似(雷纳,A.,J.E.克劳斯,K.T.凯泽,W.D.埃尔德雷德,和J.F.麦凯尔维(1984年)《比较神经学杂志》226:50 - 75),但采用梯度洗脱HPLC法时,它与十一肽SP不共洗脱。(摘要截短于400字)