LHRH CELLS MIGRATE ON PERIPHERIN FIBERS IN EMBRYONIC OLFACTORY EXPLANT CULTURES - AN IN-VITRO MODEL FOR NEUROPHILIC NEURONAL MIGRATION
LHRH CELLS MIGRATE ON PERIPHERIN FIBERS IN EMBRYONIC OLFACTORY EXPLANT CULTURES - AN IN-VITRO MODEL FOR NEUROPHILIC NEURONAL MIGRATION
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DOI：
10.1006/dbio.1994.1319
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发表时间：
1994-11-01
影响因子：
2.7
通讯作者：
WRAY, S
中科院分区：
文献类型：
--
作者：
FUESHKO, S;WRAY, S
促黄体生成激素释放激素（LHRH）神经元起源于嗅基板的祖细胞。在产前发育期间，这些细胞通过亲神经性相互作用迁移，在轨道样安排沿着轴突的嗅觉复合体。这些细胞获得成人样分布的机制尚不清楚。在这项研究中，我们建立了一个在体外，胚胎小鼠嗅外植体模型，以检查的因素，指导LHRH细胞迁移。培养物由E11.5胚胎产生，并维持长达3周。通常，总LHRH神经元群体的20-50%（160-400个LHRH细胞）存活并在这些外植体中维持基因表达。纤连蛋白和层粘连蛋白染色描绘了从第1-7天从整个染色组织的外周同心扩散的基质产生细胞。与此相反，LHRH细胞迁移专门从内嗅窝区的表面培养3天后。从双侧嗅窝中出现的LHRH细胞组之间的关系不是随机的，而是高度组织化的;在93%的培养物中，这些组之间的角度关系为180-270度。在体外培养5天或更长时间后，发现LHRH细胞的定向轨迹与体内观察到的相似，形成从嗅窝到基底层的连续体，其中许多双极LHRH细胞离散地位于基底层。检测到的LHRH细胞距嗅窝的最大距离为0.9 mm，这与LHRH细胞在体内穿过鼻区域的距离一致。LHRH细胞的可重复时空外观与外植体的纤维连接蛋白和层粘连蛋白产生细胞的同心扩散无关。两者合计，这些数据表明，LHRH细胞在这些外植体中定向迁移，并且在没有脑组织的情况下，控制这些神经元的初始迁移的分子线索在该系统中保持完整。双标记免疫细胞化学表明，至少有三个群体的神经元纤维存在于外植体：N-CAM阳性，外周蛋白阳性，和N-CAM/外周蛋白阳性。虽然所有三个纤维组表现出高度组织化的时空分布模式，只有外周蛋白阳性纤维与LHRH细胞的位置。LHRH细胞之前总是，并在密切相关，外周蛋白阳性纤维。我们假设，信号所产生的外周阳性轴突提供适当的指导线索，LHRH细胞，因为他们从嗅窝移民。(C)1994年出版社出版。
Luteinizing hormone releasing hormone (LHRH) neurons arise from progenitor cells in the olfactory placode. During prenatal development, these cells migrate via neurophilic interactions, in track-like arrangements along axons of the olfactory complex. The mechanisms by which these cells attain an adult-like distribution are unknown. In this study, we established an in vitro, embryonic mouse olfactory explant model to examine the factors directing LHRH cell migration. Cultures were generated from E11.5 embryos and maintained for up to 3 weeks. Typically 20-50% (160-400 LHRH cells) of the total LHRH neuronal population survived and maintained gene expression in these explants. Fibronectin and laminin staining delineated substratum producing cells which concentrically spread, from Days 1-7, from the periphery of the entire explanted tissue. In contrast, LHRH cells emigrated exclusively from inside olfactory pit areas to the surface of the culture after 3 days. The relationship between groups of LHRH cells emerging from bilateral olfactory pits was not random, but highly organized; in 93% of the cultures examined, the angular relationship between these groups was 180-270 degrees. After 5 or more days in vitro, LHRH cells were found in directional tracks similar to those observed in vivo forming a continuum from the olfactory pit out onto the substratum, where many bipolar LHRH cells were discretely located. The maximum distance away from the olfactory pit that LHRH cells were detected was 0.9 mm, which is compatible with the distance traversed by LHRH cells through the nasal region in vivo. The reproducible spatiotemporal appearance of LHRH cells was unrelated to the concentric spread of the fibronectin and laminin producing cells of the explant. Taken together, these data indicate that LHRH cells migrated directionally in these explants and that the molecular cues governing the initial migration of these neurons remained intact in this system in the absence of brain tissue. Double-label immunocytochemistry indicated that at least three populations of neuronal fibers existed in the explants: N-CAM-positive, peripherin-positive, and N-CAM/peripherin-positive. Although all three fibers groups showed highly organized spatiotemporal distribution patterns, only peripherin-positive fibers correlated with the location of LHRH cells. LHRH cells were always preceded by, and in close association with, peripherin-positive fibers. We hypothesize that signals arising from the peripherin-positive axons provide the appropriate guidance cues to LHRH cells as they emigrate from the olfactory pit. (C) 1994 Academic Press, Inc.