A vascular smooth muscle-specific integrin-α8 Cre mouse for lymphatic contraction studies that allows male-female comparisons and avoids visceral myopathy.
A vascular smooth muscle-specific integrin-α8 Cre mouse for lymphatic contraction studies that allows male-female comparisons and avoids visceral myopathy.
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DOI：
10.3389/fphys.2022.1060146
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发表时间：
2022
影响因子：
4
通讯作者：
Zawieja, Scott D.
中科院分区：
文献类型：
--
作者：
Davis, Michael J.;Kim, Hae Jin;Li, Min;Zawieja, Scott D.
关键词：
前言：广泛使用的他莫昔芬诱导的平滑肌特异性CRE，MYH11-CreERT2有两个缺点：1)它携带在Y染色体上，因此只对雄性小鼠的基因缺失有效；2)它在血管和非血管SM中重组，可能导致不需要的或混淆的胃肠道表型。在这里，我们测试了一种新的基于整合素α8启动子(ITGA8-CRERT2)的SM特异性Cre的有效性，它是最近开发和表征的，以评估Cav1.2缺失对小鼠淋巴SM功能的影响。方法：CAV1.2(L型电压门控钙通道)是淋巴起搏和收缩所必需的，MYH11-CreERT2或ItGa8-CreERT2的缺失可阻断自发性淋巴管收缩。用两种体外方法测定小鼠淋巴管收缩功能。结果：MYH11-CreERT2；Cav1.2f/f小鼠在第一次注射他莫昔芬后20天内死于胃肠梗阻，随后几天健康状况逐渐变差，症状包括体重减轻、打扮不佳、驼背姿势和整体活动减少。相比之下，Itga8-CreERT2；Cav1.2f/f小鼠在诱导后存活80天，并处于正常健康状态，直到为实验研究牺牲时为止。Cav1.2缺失在雄性和雌性小鼠中同样有效。讨论：我们的结果表明，Itga8-Creer T2可以有效地删除淋巴平滑肌中的基因，同时避免潜在的致命内脏肌病，并允许对雄性和雌性小鼠的淋巴收缩功能进行比较研究。
Introduction: The widely-used, tamoxifen-inducible, smooth muscle (SM)-specific Cre, Myh11-CreERT2 , suffers from two disadvantages: 1) it is carried on the Y-chromosome and thus only effective for gene deletion in male mice, and 2) it recombines in both vascular and non-vascular SM, potentially leading to unwanted or confounding gastrointestinal phenotypes. Here, we tested the effectiveness of a new, SM-specific Cre, based on the integrin α8 promoter (Itga8-CreERT2 ), that has been recently developed and characterized, to assess the effects of Cav1.2 deletion on mouse lymphatic SM function. Methods: Cav1.2 (the L-type voltage-gated calcium channel) is essential for lymphatic pacemaking and contraction and its deletion using either Myh11-CreERT2 or Itga8-CreERT2 abolished spontaneous lymphatic contractions. Mouse lymphatic contractile function was assessed using two ex vivo methods. Results: Myh11-CreERT2 ; Cav1.2 f/f mice died of gastrointestinal obstruction within 20 days of the first tamoxifen injection, preceded by several days of progressively poor health, with symptoms including weight loss, poor grooming, hunched posture, and reduced overall activity. In contrast, Itga8-CreERT2 ; Cav1.2 f/f mice survived for >80 days after induction and were in normal health until the time of sacrifice for experimental studies. Cav1.2 deletion was equally effective in male and female mice. Discussion: Our results demonstrate that Itga8-CreER T2 can be used to effectively delete genes in lymphatic smooth muscle while avoiding potentially lethal visceral myopathy and allowing comparative studies of lymphatic contractile function in both male and female mice.
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影响因子：
4.9
作者：
Cromer W;Wang W;Zawieja SD;von der Weid PY;Newell-Rogers MK;Zawieja DC
通讯作者：
Zawieja DC
影响因子：
5.2
作者：
Grcevic, Danka;Pejda, Slavica;Matthews, Brya G.;Repic, Dario;Wang, Liping;Li, Haitao;Kronenberg, Mark S.;Jiang, Xi;Maye, Peter;Adams, Douglas J.;Rowe, David W.;Aguila, Hector L.;Kalajzic, Ivo
通讯作者：
Kalajzic, Ivo
影响因子：
3.4
作者：
Imtiaz, Mohammad S.;Zhao, Jun;van Helden, Dirk F.
通讯作者：
van Helden, Dirk F.
影响因子：
4.6
作者：
Jamalian, Samira;Jafarnejad, Mohammad;Moore, James E., Jr.
通讯作者：
Moore, James E., Jr.
影响因子：
1.7
作者：
Mellor, Russell H.;Tate, Naomi;Mortimer, Peter S.
通讯作者：
Mortimer, Peter S.