Appearance of cholinergic myenteric neurons during enteric nervous system development: comparison of different ChAT fluorescent mouse reporter lines.
Appearance of cholinergic myenteric neurons during enteric nervous system development: comparison of different ChAT fluorescent mouse reporter lines.
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DOI：
10.1111/nmo.12343
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发表时间：
2014-06
影响因子：
3.5
通讯作者：
Gosain A
中科院分区：
文献类型：
--
作者：
Erickson CS;Lee SJ;Barlow-Anacker AJ;Druckenbrod NR;Epstein ML;Gosain A
胆碱能神经元已被乙酰胆碱合成酶胆碱乙酰转移酶（ChAT）识别。然而，ChAT 很难定位于新分化的周围神经元，这使得胆碱能神经元发育的研究存在问题。因此，研究人员使用小鼠报告系来表明 ChAT 的存在。我们的目标是确定哪个 ChAT 报告系是 ChAT 表达最敏感的指标。我们利用两种不同的荧光 ChAT 报告系（ChAT-GFP 和 ChAT-Cre;R26R:floxSTOP:tdTomato）以及 ChAT 蛋白的免疫定位 (ChAT-IR) 来表征整个 ENS 发育过程中肌间神经元 ChAT 的空间和时间表达。 ChAT-IR 细胞首次在 E10.5 时在肠道中发现，甚至在神经前体的迁移波前内。远端小肠 (dSI) 和近端结肠内的肌间神经元首先用 ChAT-IR 标记，然后用 ChAT-GFP 标记，最后用 ChAT-Cre tdTomato 标记。 ChAT-IR 神经元的百分比相当于 E13.5 中 dSI 中的成人水平以及 P0 中近端结肠中的成人水平。在这些阶段之后，尽管神经元密度发生了巨大变化，但在整个发育过程中，百分比仍保持相对恒定。这些观察结果表明神经递质表达发生得很早，并且神经发生和神经递质表达之间仅存在短暂的间隙。我们发现，ChAT 肌间神经元的比例在胚胎发育过程中达到了成年水平，这表明胆碱能神经元的命运受到严格调控，并且它们的分化可能会影响神经元的进一步发育。 ChAT-GFP 是比 ChAT-Cre;R26R:floxSTOP:tdTomato 报告小鼠更准确的早期 ENS 胆碱能神经元分化指标。
Cholinergic neurons have been identified with the acetylcholine synthetic enzyme choline acetyltransferase (ChAT). However, ChAT is difficult to localize in newly differentiated peripheral neurons making the study of cholinergic neuronal development problematic. Consequently researchers have used mouse reporter lines to indicate the presence of ChAT. Our objective was to determine which ChAT reporter line was the most sensitive indicator of ChAT expression. We utilized two different fluorescent ChAT reporter lines (ChAT-GFP and ChAT-Cre;R26R:floxSTOP:tdTomato) together with immunolocalization of ChAT protein (ChAT-IR) to characterize the spatial and temporal expression of ChAT in myenteric neurons throughout ENS development. ChAT-IR cells were first seen in the intestine at E10.5, even within the migration wavefront of neural precursors. Myenteric neurons within the distal small intestine (dSI) and proximal colon were first labeled by ChAT-IR, then ChAT-GFP, and finally ChAT-Cre tdTomato. The percentage of ChAT-IR neurons is equivalent to adult levels in the dSI by E13.5 and proximal colon by P0. After these stages, the percentages remained relatively constant throughout development despite dramatic changes in neuronal density. These observations indicate that neurotransmitter expression occurs early and there is only a brief gap between neurogenesis and neurotransmitter expression. Our finding that the proportion of ChAT myenteric neurons reached adult levels during embryonic development suggests that the fate of cholinergic neurons is tightly regulated and that their differentiation might influence further neuronal development. ChAT-GFP is a more accurate indicator of early ENS cholinergic neuronal differentiation than the ChAT-Cre;R26R:floxSTOP:tdTomato reporter mouse.
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DOI：
10.1523/jneurosci.6684-10.2011
发表时间：
2011-06-15
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Li Z;Chalazonitis A;Huang YY;Mann JJ;Margolis KG;Yang QM;Kim DO;Côté F;Mallet J;Gershon MD
通讯作者：
Gershon MD
影响因子：
2.5
作者：
PHAM, TD;GERSHON, MD;ROTHMAN, TP
通讯作者：
ROTHMAN, TP
影响因子：
5.3
作者：
Hao MM;Young HM
通讯作者：
Young HM
影响因子：
2.5
作者：
Bergner, Annette J.;Stamp, Lincon A.;Gonsalvez, David G.;Allison, Margaret B.;Olson, David P.;Myers, Martin G., Jr.;Anderson, Colin R.;Young, Heather M.
通讯作者：
Young, Heather M.
影响因子：
4.6
作者：
Lei, Jun;Howard, Marthe J.
通讯作者：
Howard, Marthe J.