Identification of the neuropeptide content of individual rat neurohypophysial terminals
Identification of the neuropeptide content of individual rat neurohypophysial terminals
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DOI：
10.1016/j.jneumeth.2007.03.006
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发表时间：
2007-07-30
影响因子：
3
通讯作者：
Lemos, Jose R.
中科院分区：
文献类型：
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作者：
Custer, Edward E.;Ortiz-Miranda, Sonia;Lemos, Jose R.
本研究的目的是建立一种方法，可以可靠地确定精氨酸加压素（AVP）和/或催产素（OT）含量的个别大鼠神经垂体终末（NHT）直径≥ 5 μ m，用于电生理记录的大小。我们使用了一种市售的，高灵敏度的酶联免疫测定（ELISA）试剂盒的灵敏度为0.25 pg的AVP和1.0 pg的OT。AVP的NHT含量（2.21 +/- 0.10 pg）大于OT（1.77 +/- 0.08 pg），并随终末大小而增加。AVP阳性终末（10.2 +/- 0.21 μ m）直径大于OT阳性终末（9.1 +/- 0.24 μ m）。免疫细胞化学技术表明，一个较高的百分比（58%）的较小的终端包含OT，和一个较高的百分比（42%）的较大的NHTs被collabeled。当NHTs单独检测AVP时，免疫细胞化学（73%）和ELISA（72%）方法之间获得的AVP阳性末端百分比相似，但使用免疫细胞化学（43%）时OT末端的百分比高于ELISA（26%）。当NHT同时检测AVP和OT时，AVP阳性终末（60%）和OT阳性终末（18%）的百分比下降。因此，可靠鉴定AVP阳性NHT的最佳方法是仅测定AVP，因为这允许得出AVP阴性末端仅含有OT的结论。(c)2007 Elsevier B. V.保留所有权利。
The objective of this study was to develop a method that could reliably determine the arginine vasopressin (AVP) and/or oxytocin (OT) content of individual rat neurohypophysial terminals (NHT) >= 5 mu m in diameter, the size used for electrophysiological recordings. We used a commercially available, highly sensitive enzyme-linked immunoassay (ELISA) kit with a sensitivity of 0.25 pg to AVP and of 1.0 pg to OT. The NHT content of AVP (2.21 +/- 0.10 pg) was greater than OT (1.77 +/- 0.08 pg) and increased with terminal size. AVP-positive terminals (10.2 +/- 0.21 mu m) were larger in diameter than OT-positive terminals (9.1 +/- 0.24 mu m). Immunocytochemical techniques indicated that a higher percentage (58%) of smaller terminals contained OT, and that a higher percentage (42%) of larger NHTs were colabeled. Similar percentages of AVP-positive terminals were obtained between immunocytochemical (73%) and ELISA (72%) methods when NHTs were assayed for AVP alone, but there was a higher percentage of OT terminals when using immunocytochemistry (43%) compared to ELISA (26%). The percent of AVP-positive (60%) and OT-positive (18%) terminals decreased when NHT were assayed for both AVP and OT. Therefore, the best method to reliably identify AVP-positive NHTs is to assay only for AVP, since this allows the conclusion that AVP-negative terminals contain only OT. (c) 2007 Elsevier B.V. All rights reserved.