Bioluminescence imaging of β cells and intrahepatic insulin gene activity under normal and pathological conditions.
Bioluminescence imaging of β cells and intrahepatic insulin gene activity under normal and pathological conditions.
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DOI：
10.1371/journal.pone.0060411
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Takahashi S
中科院分区：
文献类型：
--
作者：
Katsumata T;Oishi H;Sekiguchi Y;Nagasaki H;Daassi D;Tai PH;Ema M;Kudo T;Takahashi S
在糖尿病研究中，生物发光成像（BLI）已应用于β细胞损伤、发育和胰岛移植的研究。为了开发能够对β细胞进行非侵入性成像的小鼠模型，我们产生了细菌人工染色体（BAC）转基因小鼠，其中包含胰岛素I基因的小鼠200-kbp基因组片段驱动荧光素酶表达（Ins 1-luc BAC转基因小鼠）。腹腔注射后，使用IVIS Spectrum系统进行小鼠的BLI，并分析来自胰腺区域的生物发光信号。当与在9.2-kbp小鼠胰岛素I启动子（MIP）控制下表达荧光素酶的MIP-Luc-VU小鼠[FVB/N-Tg（Ins 1-luc）VUPwrs/J]相比时，Ins 1-luc BAC转基因小鼠的生物发光发射增强约4倍。链脲佐菌素处理的Ins 1-luc BAC转基因小鼠发生了严重的糖尿病，伴随着胰腺中BLI信号强度的急剧下降。相反，喂食高脂肪饮食8周的小鼠显示信号增加，反映了β细胞质量的减少或增加。尽管胰岛的生物发光强度与体外分离的胰岛数量相关性良好，但从活体小鼠体内获得的强度不一定反映病理条件下β细胞质量的绝对定量。另一方面，在Ins 1-luc BAC转基因小鼠中，腺病毒介导的β细胞相关转录因子的基因转导产生了来自肝脏区域的发光超过1周。这些结果表明，Ins 1-luc BAC转基因小鼠中的BLI提供了在正常和病理条件下对肝脏中的胰岛β细胞和胰岛素基因的胰腺外活性进行成像的非侵入性方法。
In diabetes research, bioluminescence imaging (BLI) has been applied in studies of β-cell impairment, development, and islet transplantation. To develop a mouse model that enables noninvasive imaging of β cells, we generated a bacterial artificial chromosome (BAC) transgenic mouse in which a mouse 200-kbp genomic fragment comprising the insulin I gene drives luciferase expression (Ins1-luc BAC transgenic mouse). BLI of mice was performed using the IVIS Spectrum system after intraperitoneal injection of luciferin, and the bioluminescence signal from the pancreatic region analyzed. When compared with MIP-Luc-VU mice [FVB/N-Tg(Ins1-luc)VUPwrs/J] expressing luciferase under the control of the 9.2-kbp mouse insulin I promoter (MIP), the bioluminescence emission from Ins1-luc BAC transgenic mice was enhanced approximately 4-fold. Streptozotocin-treated Ins1-luc BAC transgenic mice developed severe diabetes concomitant with a sharp decline in the BLI signal intensity in the pancreas. Conversely, mice fed a high-fat diet for 8 weeks showed an increase in the signal, reflecting a decrease or increase in the β-cell mass. Although the bioluminescence intensity of the islets correlated well with the number of isolated islets in vitro, the intensity obtained from a living mouse in vivo did not necessarily reflect an absolute quantification of the β-cell mass under pathological conditions. On the other hand, adenovirus-mediated gene transduction of β-cell-related transcription factors in Ins1-luc BAC transgenic mice generated luminescence from the hepatic region for more than 1 week. These results demonstrate that BLI in Ins1-luc BAC transgenic mice provides a noninvasive method of imaging islet β cells and extrapancreatic activity of the insulin gene in the liver under normal and pathological conditions.
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影响因子：
7.7
作者：
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通讯作者：
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作者：
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DOI：
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发表时间：
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影响因子：
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DOI：
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发表时间：
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期刊：
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影响因子：
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通讯作者：
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影响因子：
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