A transgenic mouse line for assaying tissue-specific changes in endoplasmic reticulum proteostasis.
A transgenic mouse line for assaying tissue-specific changes in endoplasmic reticulum proteostasis.
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DOI：
10.1007/s11248-023-00349-7
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发表时间：
2023-06
影响因子：
3
通讯作者：
Harvey, Brandon K.
中科院分区：
文献类型：
--
作者：
Svarcbahs, Reinis;Blossom, Sarah M.;Baffoe-Bonnie, Helena S.;Trychta, Kathleen A.;Greer, Lacey K.;Pickel, James;Henderson, Mark J.;Harvey, Brandon K.
钙稳态的维持对内质网的正常功能至关重要。当细胞应激条件耗尽内质网中的高浓度钙时，内质网驻留蛋白就会分泌到细胞外空间，这一过程被称为外源性。监测外源性疾病提供了内质网稳态和蛋白质稳态变化的见解，这些变化是由内质网钙失调相关的细胞应激引起的。为了监测完整动物的细胞类型特异性外溢，我们创建了一个转基因小鼠系，该系具有基于Gaussia荧光素酶（GLuc）的分泌内质网钙调节蛋白SERCaMP，其前面是LoxP-STOP-LoxP （LSL）序列。将cre依赖性LSL-SERCaMP小鼠与白蛋白(Alb)-Cre和多巴胺转运体(DAT)-Cre小鼠杂交。葡萄糖- sercamp在小鼠器官和细胞外液中的表达被表征，葡萄糖- sercamp的分泌在内质网钙药物耗竭后被监测。在LSL-SERCaMP × Alb-Cre小鼠中，仅在肝脏和血液中观察到GLuc活性，而在LSL-SERCaMP × DAT-Cre小鼠中，在中脑多巴胺能神经元和多巴胺能投射神经支配的组织样本中观察到GLuc活性。在钙缺失后，我们发现Alb-Cre和DAT-Cre杂交收集的血浆和脑脊液中GLuc信号分别升高。该小鼠模型可用于研究疾病发病过程中特定细胞和组织类型er驻留蛋白的分泌，并可能有助于确定疾病的治疗方法和生物标志物。在线版本包含补充材料，下载地址：10.1007/s11248-023-00349-7。
Maintenance of calcium homeostasis is important for proper endoplasmic reticulum (ER) function. When cellular stress conditions deplete the high concentration of calcium in the ER, ER-resident proteins are secreted into the extracellular space in a process called exodosis. Monitoring exodosis provides insight into changes in ER homeostasis and proteostasis resulting from cellular stress associated with ER calcium dysregulation. To monitor cell-type specific exodosis in the intact animal, we created a transgenic mouse line with a Gaussia luciferase (GLuc)—based, secreted ER calcium-modulated protein, SERCaMP, preceded by a LoxP-STOP-LoxP (LSL) sequence. The Cre-dependent LSL-SERCaMP mice were crossed with albumin (Alb)-Cre and dopamine transporter (DAT)-Cre mouse lines. GLuc-SERCaMP expression was characterized in mouse organs and extracellular fluids, and the secretion of GLuc-SERCaMP in response to cellular stress was monitored following pharmacological depletion of ER calcium. In LSL-SERCaMP × Alb-Cre mice, robust GLuc activity was observed only in the liver and blood, whereas in LSL-SERCaMP × DAT-Cre mice, GLuc activity was seen in midbrain dopaminergic neurons and tissue samples innervated by dopaminergic projections. After calcium depletion, we saw increased GLuc signal in the plasma and cerebrospinal fluid collected from the Alb-Cre and DAT-Cre crosses, respectively. This mouse model can be used to investigate the secretion of ER-resident proteins from specific cell and tissue types during disease pathogenesis and may aid in the identification of therapeutics and biomarkers of disease. The online version contains supplementary material available at 10.1007/s11248-023-00349-7.
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DOI：
10.1016/j.jphotobiol.2018.04.050
发表时间：
2018-06-01
影响因子：
5.4
作者：
Larionova, Marina D.;Markova, Svetlana V.;Vysotski, Eugene S.
通讯作者：
Vysotski, Eugene S.
影响因子：
64.8
作者：
BURNS, K;DUGGAN, B;MICHALAK, M
通讯作者：
MICHALAK, M
DOI：
10.1126/science.1257521
发表时间：
2014-11-07
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Jan CH;Williams CC;Weissman JS
通讯作者：
Weissman JS
影响因子：
16.2
作者：
Agarwal A;Wu PH;Hughes EG;Fukaya M;Tischfield MA;Langseth AJ;Wirtz D;Bergles DE
通讯作者：
Bergles DE
影响因子：
8
作者：
Dijkema, Fenne Marjolein;Nordentoft, Matilde Knapkoien;Winther, Jakob R.
通讯作者：
Winther, Jakob R.