Single cell ganglioside catabolism in primary cerebellar neurons and glia.
Single cell ganglioside catabolism in primary cerebellar neurons and glia.
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DOI：
10.1007/s11064-012-0733-1
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发表时间：
2012-06
影响因子：
4.4
通讯作者：
Dovichi, Norman J.
中科院分区：
文献类型：
--
作者：
Essaka, David C.;Prendergast, Jillian;Keithley, Richard B.;Hindsgaul, Ole;Palcic, Monica M.;Schnaar, Ronald L.;Dovichi, Norman J.
通过分析大鼠小脑单个原代神经元和神经胶质中荧光四甲基罗丹明标记的GM 1（TMR-GM 1）的分解，确定神经节苷脂catalysts的细胞间异质性。将从5-6日龄大鼠小脑分离的细胞培养7天，然后与TMR-GM 1孵育14小时。通过温和的蛋白水解从培养物中回收完整的细胞，多聚甲醛固定，并进行单细胞分析。将单个细胞捕获在毛细管中，裂解，并将释放的单细胞内容物进行毛细管电泳，并对分解代谢产物进行定量激光诱导荧光检测。非神经元细胞平均比神经元细胞吸收更多的外源性TMR-GM 1，并且更广泛地分解代谢它。孵育14小时后，非神经元细胞仅保留14%的TMR产物作为GM 1和GM 2，而神经元细胞的TMR产物>50%。平均而言，非神经元细胞含有74%的TMR标记产物作为TMR-神经酰胺，而神经元仅为42%。非神经元细胞保留的TMR-GM 3（7%）是神经元细胞（1%）的7倍。为了证实所观察到的单细胞代谢组学，我们裂解并比较了来自混合神经元/神经胶质细胞培养物和来自通过用抗有丝分裂剂阿糖胞苷处理而耗尽非神经元细胞的培养物的TMR-GM 1分解代谢谱。整个培养物的分解代谢曲线与单个神经元和胶质细胞的平均曲线一致。我们的结论是，超灵敏的分析方法准确地反映了单细胞神经节苷脂catalysts在不同的细胞群从大脑。
Cell-to-cell heterogeneity in ganglioside catabolism was determined by profiling fluorescent tetramethylrhodamine-labeled GM1 (TMR-GM1) breakdown in individual primary neurons and glia from the rat cerebellum. Cells isolated from 5–6 day old rat cerebella were cultured for 7 days, and then incubated for 14 h with TMR-GM1. Intact cells were recovered from cultures by mild proteolysis, paraformaldehyde fixed, and subjected to single cell analysis. Individual cells were captured in a capillary, lysed, and the released single-cell contents subjected to capillary electrophoresis with quantitative laser-induced fluorescent detection of the catabolic products. Non-neuronal cells on average took up much more exogenous TMR-GM1 than neuronal cells, and catabolized it more extensively. After 14 h of incubation, non-neuronal cells retained only 14% of the TMR products as GM1 and GM2, compared to >50% for neurons. On average, non-neuronal cells contained 74% of TMR-labeled product as TMR-ceramide, compared to only 42% for neurons. Non-neuronal cells retained seven times as much TMR-GM3 (7%) compared to neuronal cells (1%). To confirm the observed single cell metabolomics, we lysed and compared TMR-GM1 catabolic profiles from mixed neuron/glial cell cultures and from cultures depleted of non-neuronal cells by treatment with the antimitotic agent cytosine arabinoside. The whole culture catabolic profiles were consistent with the average profiles of single neurons and glia. We conclude that the ultrasensitive analytic methods described accurately reflect single cell ganglioside catabolism in different cell populations from the brain.
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DOI：
10.1083/jcb.100.2.384
发表时间：
1985-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Hatten ME
通讯作者：
Hatten ME
影响因子：
3.5
作者：
Sobhani, Kimia;Michels, David A.;Dovichi, Norman J.
通讯作者：
Dovichi, Norman J.
毛细血管电泳中的六个数量级动态范围，具有超敏激光诱导的荧光检测。
影响因子：
6.1
作者：
Whitmore, Colin D.;Essaka, David;Dovichi, Norman J.
通讯作者：
Dovichi, Norman J.
影响因子：
4.2
作者：
Kolter, T;Sandhoff, K
通讯作者：
Sandhoff, K
DOI：
10.1016/j.bbagen.2007.10.008
发表时间：
2008-03-01
影响因子：
3
作者：
Todeschini, Adriane Regina;Hakomori, Sen-Itiroh
通讯作者：
Hakomori, Sen-Itiroh