INTRACELLULAR CA2+ OSCILLATIONS IN LHRH NEURONS FROM EMBRYONIC OLFACTORY PLACODE

胚胎嗅觉板 LHRH 神经元的细胞内 CA2 振荡

• 批准号: 6277677
• 负责人: Ei Terasawa-Grilley
• 金额: $ 6.06万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至 1999-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6277677
• 关键词: Primates; animal tissue; clinical research; endocrine gland /system; growth /development; microscopy; mother /infant health care; nervous system; women's health

OBJECTIVE: To determine the oscillatory pattern of LHRH neurons in culture. RESULTS Previously, we have shown that 1) cultured LHRH neurons derived from the olfactory placode of monkey embryos release LHRH in a pulsatile manner at approximately 50-min intervals, and 2) individual LHRH cells exhibit [Ca2+]i oscillations. In order to understand the mechanism of LHRH-pulse generation, we further characterized the pattern of [Ca2+]i oscillations in LHRH neurons. The olfactory placode and the migratory pathway of LHRH neurons from monkey embryos at E35-37 were dissected out and cultured on glass coverslips in a growth medium. Two to five weeks later cultured cells were labeled with the calcium dye, fura-2, and examined for [Ca2+]i oscillations. [Ca2+]i changes were recorded every 10 seconds for 30 to 152 min using a video imaging system. After recording, cells were fixed and immunostained for LHRH. In 16 cultures, 359 LHRH cells (4-51 cells/culture) exhibiting [Ca2+]i oscillations were recorded. In 13 of 16 cultures, [Ca2+]i oscillations occurred synchronously in 50-100% of the individual cells in a culture, whereas [Ca2+]i oscillations in cells in the remaining 3 cultures did not synchronize. Strikingly, in 8 of 13 cultures the synchronization of [Ca2+]i oscillations repeatedly occurred in complete unison at 47.3q3.4 min, which is similar to the period observed for LHRH release, whereas in 5 of 13 cultures the less tight synchronization of [Ca2+]i oscillations repeatedly occurred at 23.4q4.2 min. Interpulse intervals (IPI, 6.9q0.8 min, n=134) of [Ca2+]i oscillations in cells with tight synchronization and IPI (9.2q1.0 min, n=130) in cells with less tight synchronization did not differ significantly from IPI (8.5q0.4 min, n=95) in cells without synchronization. Neither age of cultures, nor type of chambers (with or without media perifusion) was related to synchronization of oscillations. Results from this study indicate that LHRH neurons derived from monkey olfactory placode possess an endogenous mechanism for synchronization of [Ca2+]i oscillations, which may be related to the periodicity of LHRH release. FUTURE DIRECTIONS We will examine the mechanism of the synchronization. KEY WORDS LHRH neurons, Ca2+ oscillation, LHRH-pulse generation, olfactory placode

目的：确定LHRH神经元的振荡模式 在文化中。 结果：1）培养的LHRH 猴胚胎嗅板神经元释放 以约50分钟的间隔以脉动方式给予LHRH，以及2） 单个LHRH细胞显示[Ca 2 +]i振荡。 为了 了解LHRH脉冲产生的机制，我们进一步 特征的[Ca ~（2+）]i振荡的模式在LHRH神经元。 嗅板和LHRH神经元的迁移通路 将E35-37的猴胚胎解剖出来并在玻璃上培养 盖玻片在生长培养基中。 两到五周后培养细胞 用钙染料fura-2标记，并检测[Ca 2 +]i 振荡 [Ca2+]i变化每10秒记录一次，持续30 至152分钟，使用视频成像系统。 记录后， 固定并免疫染色LHRH。 在16个培养物中，359个LHRH细胞 (4-51细胞/培养物）显示[Ca 2 +]i振荡。 在16个培养物中，有13个培养物的[Ca ~（2+）]i振荡同步发生， 50-100%的单个细胞在培养物中，而[Ca 2 +]i 其余3种培养物中细胞的振荡不同步。 引人注目的是，在13个培养物中的8个中，[Ca 2 +]i 振荡在47.3 ± 3.4分钟完全一致地重复发生， 这与观察到的LHRH释放期相似，而在5 13个培养物的[Ca ~（2+）]i振荡同步性较差 重复发生在23.4 q4.2 min。脉冲间期（IPI， 6.9q0.8min，n=134）的[Ca 2 +]i振荡在细胞内的紧密 同步化和IPI（9.2q1.0 min，n=130）在细胞中， 同步与IPI没有显著差异（8.5q0.4分钟， n=95）。 无论是文化时代，还是 腔室类型（有或无介质灌注）与 振荡的同步 本研究结果表明 猴嗅板LHRH神经元具有一种特异性， [Ca 2 +]i振荡同步化的内源性机制， 这可能与LHRH释放的周期性有关。 未来 方向我们将研究同步的机制。 关键 LHRH神经元，Ca 2+振荡，LHRH脉冲产生，嗅觉 placode