作者要感谢来自洛桑大学医院 (CHUV) 机械车间的达尼洛·古比安和斯蒂芬·阿勒图斯帮助创建了低氧装置。作者还要感谢黛安·马卡布里和梅勒妮·西皮翁在训练动物方面的帮助。

当地国家动物护理委员会 (瑞士洛桑省 VÉTÉRINAIRES 协会 (SCAV) 批准了所有实验 (授权 VD3224; 01.06.2017), 所有实验都是根据相关的准则和法规。
1. 动物住房和准备
<ol><li>在实验开始前, 6 至8周龄的 C57BL/6J 雄性老鼠在动物设施中至少有1周的时间, 以便这些老鼠适应新的住房条件。出于实际原因, 同一实验组的小鼠通常被安置在一起。</li>
	<li>将小鼠保持在温控的房间 (22±1°c), 有12小时的光/暗周期, 并能获得食物和水。</li>
</ol>2. 跑步机增量试验确定最大速度和性能改进的标准评定
请注意:以下步骤对于完成培训协议至关重要。
<ol><li>为老鼠使用机动跑步机, 老鼠可以在多个车道上并排, 有0°的倾斜度, 并安装在车道后面设置为 0.2 mA 的电网, 以鼓励老鼠奔跑。</li>
	<li>
在第一次测试之前, 按照以下程序将小鼠提交到4天的适应跑步机。
	<ol>
<li>在第一天, 让老鼠以 4.8/分钟的速度跑10分钟。</li>
		<li>第2天, 让老鼠在 6点/分钟跑10分钟。</li>
		<li>第3天, 让老鼠在 7.2 m/min 处跑10分钟。</li>
		<li>第4天, 让老鼠在 8.4 m/min 处跑10分钟。</li>
	</ol>
</li>
	<li>
第5天, 根据下面的协议, 将老鼠提交到一个增量测试中, 以耗尽。
	<ol>
<li>让小鼠在 4.8 m/min (在0°倾角下) 加热5分钟。</li>
		<li>每3分钟将速度提高 1.2 m/min (例如, 4.8 m/min 5分钟, 6 m/min 3分钟, 7.2 m/min 3分钟, 8.5分钟, 8.4/min 等 3分钟), 直到耗尽, 当鼠标在电网上连续花费3秒或受到100次冲击 (由仪器显示)。</li>
		<li>记下实现的速度 (被视为 V最大值)、持续时间、距离、冲击次数和在网格上花费的总时间。 请注意:通常情况下, V最大值为 28.8±3.7 m/2 分钟。</li>
		<li>在训练中期, 将小鼠重新提交到此测试, 以便重新调整训练速度, 使其达到更新后的最大 V值(例如, 如果训练协议持续 8周, 然后在4周内进行训练中期增量测试。在这种情况下, 用测试取代其中一项预定的培训), 并在研究结束时再次这样做, 以评估业绩的改善情况。</li>
		<li>在此测试前后实施48小时的休息时间。 请注意:所有的增量测试都是在上午进行的。</li>
	</ol>
</li>
</ol>3. 缺氧环境
<ol><li>对于缺氧的训练课程, 将跑步机放在连接到气体混合器的缺氧箱 (图 1) 中。使用校准的血氧仪定期控制盒子中氧气的环境分数 (fio 2 [即缺氧水平])。</li>
	<li>将气体混合器设置在100% 的氮气 (n2) 上, 并使用血氧仪验证缺氧水平。一旦 Fio2 = 0.13, 将气体混合器的参数从 100% n2 改为13% o 2.</li>
	<li>为了避免长时间被动暴露在缺氧环境中, 将小鼠放在一个临时较小的笼子里, 有垃圾和浓缩, 一旦达到 fio2 = 0.13, 就迅速将其放在盒子里。在将保持架放入时, 验证环境仍处于 13% o2;如果没有, 请重新调整它。</li>
	<li>在训练过程中定期验证 o2 的水平, 以确保它保持在 Fi i2= 0.13±0.002. </li>
</ol>4. 规范环境
<ol><li>对于诺莫夏的训练课程, 请将跑步机放在低氧箱中, 但请卸下手套, 以便有环境空气 (Fio2 = 0.21)。其目的是在缺氧的情况下重建与老鼠相同的训练环境。</li>
</ol>5. 自杀强度训练
<ol><li>
将小鼠放置在跑步机中的单个车道上 (在0°倾角下), 并将其提交给以下协议。
	<ol>
<li>让小鼠在 4.8 m/min 热身 5分钟, 然后在 9 m/min 加热5分钟。</li>
		<li>将冲刺速度设置为先前确定的 V最大值的150%。 请注意:通常情况下, 冲刺速度为 42.1±5.5 m/min。</li>
		<li>训练老鼠进行四套 5x10 s 冲刺, 每次冲刺之间休息20秒。联组休息时间为 5分钟 (图 2)。 请注意:如果培训课程的总工作量需要与其他培训组的工作量相匹配, 请添加冷却时间。</li>
	</ol>
</li>
	<li>每周进行3次培训, 最好在两次培训之间进行48小时。</li>
	<li>用棉签作为电击的补充方法, 以鼓励老鼠奔跑。将棉签放在车道顶部的缝隙中, 在鼠标和电网之间, 当鼠标到达跑步机的背面时, 轻轻地推动它。这将避免电击的传递, 并刺激老鼠以更柔软的方式运行。</li>
</ol>6. 低强度训练
<ol><li>
将小鼠放置在跑步机中的单个车道上 (在0°倾角下), 并将其提交给以下协议。
	<ol>
<li>让小鼠在 4.8 m/min 热身 5分钟, 然后在 7.2 m/min 加热5分钟。</li>
		<li>将连续运行会话的速度设置为先前确定的 V最大值的40%。 请注意:通常情况下, 连续运行速度为 9.9 m/min。</li>
		<li>训练老鼠40分钟。</li>
		<li>每周进行3次培训, 最好在两次培训之间进行48小时。</li>
		<li>用棉签作为电击的补充方法, 以鼓励老鼠奔跑。</li>
	</ol>
</li>
</ol>7. 小鼠安乐死和器官摘除
<ol><li>在训练协议结束时和最后一次增量试验后至少 24小时, 使用异氟醚 (在 O2中为 4%-5% 诱导麻醉, 在 100% o2 中引入 1%-2%, 以维持麻醉), 将小鼠麻醉至诱导室.使用爪缩回反射确认适当的麻醉 (牢固地捏动物的爪子; 当动物对刺激没有反应时, 麻醉被认为是适当的)。</li>
	<li>使用 25 G 针, 进行经皮心脏穿刺, 以收集最大血量, 如前面所述19。</li>
	<li>通过用圆尖剪刀切开腹部第一层皮肤, 并拉扯切口两侧 (朝向头部和尾部), 进行颈椎脱位并去除小鼠的皮肤。</li>
	<li>用薄点剪刀穿过老鼠左侧胸腔下的腹膜, 到达脾脏, 并在需要时提取。 请注意:必要时解剖出肌肉。</li>
	<li>
解剖出肺动脉。
	<ol>
<li>使用小剪刀和钳子, 取出胸腔笼, 清除心肺区域。</li>
		<li>用 "自闭" 推子, 尽可能靠近先端, 轻轻拉, 拉长主动脉弓和肺动脉的底部。</li>
		<li>用右手, 在肺动脉和主动脉下插入弯曲的推子, 然后将推子向后移动一点, 只容纳肺动脉 (图 3)。</li>
		<li>用左手插入另一对子, 用右手代替拿着的子。</li>
		<li>在右手用锋利的直剪刀, 一边尽可能靠近心脏, 一边尽可能远的地方解剖肺动脉。 请注意:哪只手拿着哪只乐器并不重要, 尽管我们发现用右手切割比用左手切割更容易。</li>
		<li>将其放入一个2毫升的管与冷磷酸盐缓冲盐 (PBS) 缓冲和保持在冰上。</li>
	</ol>
</li>
	<li>
进行全身灌注。
	<ol>
<li>在小鼠右下肢的顶部, 使用推子清除内侧右胫骨动脉下到右股动脉 (腹股沟韧带下)。使用锋利的直剪刀, 使股动脉充分切割。</li>
		<li>在心脏左心室插入装有冷 PBS 的5毫升 25 g 注射器, 然后轻轻注射冷 PBS, 从血管中取出剩余的血液。 请注意:由于肺动脉的提取, 有可能是 PBS 不循环的所有方式到切口。</li>
	</ol>
</li>
	<li>使用推拿器, 尽可能彻底地将主动脉周围的软组织从左腹股沟和右腹股沟韧带取出到心脏。 请注意:如有必要, 可以提取心脏进行进一步分析。</li>
	<li>使用推拿子和微剪刀, 解剖心脏到臀部外动脉的最低点 (左和右两腿), 并将完全解剖的部分在一个10厘米直径的盘子与冷 PBS。</li>
	<li>使用推子和/或微剪刀, 通过轻轻地将主动脉和动脉周围的脂肪从血管中轻轻拉或割开, 完成对其的清洁。</li>
	<li>用微剪刀, 在左、右的胫骨动脉分叉处割断左胫骨动脉, 并将其存放作进一步分析。</li>
	<li>使用微剪刀, 切断左肾动脉下的腹主动脉, 并将提取的血管放置在冷 PBS 缓冲液在冰上 (图 4)。</li>
	<li>将剩余的清洁血管, 从主动脉弓到左肾动脉右上方, 存放, 以便进一步分析。</li>
</ol><img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/58708/58708fig4.jpg"> 图 4: 解剖船只的图片.从腹主动脉顶部 (左肾动脉下方) 提取血管至右胫骨动脉末端, 准备放置在冰上的冷 PBS 缓冲液中。(1) 腹主动脉。(2) 右髋关节动脉。(3) 外侧胫骨动脉。(4) 胫骨内动脉。(5) 股动脉。<a href="https://www.jove.com/files/ftp_upload/58708/58708fig4large.jpg" target="_blank">请点击这里查看此图的较大版本.</a>
8. 体内血管功能评估
请注意:洗涤对应于空腔和加满房间与 Krebs。
<ol><li>根据前面描述的协议20, 将孤立的肺动脉、腹主动脉和右胫骨动脉段切割成 1.0-2.0 mm 长的血管环, 并将每个环安装在两个直径 0.1 mm 的马刺上, 通过管腔。</li>
	<li>将血管环悬浮在垂直器官腔中, 其中装有10毫升改性的克瑞格-林格碳酸氢盐溶液 (1118.3 mM Ncl, 4.7 mM KCl, 2.5 mM Ccl2, 1.2 mm MgSO4, 1.2 mm kh h 2 po 4,25.0mm nahco3和 11.1 mm葡萄糖) 保持在 37°c, 并加气与 95% o 2-2% 二氧化碳 (ph 7.4 ).一个马刺锚定在器官室的底部, 另一个被连接到应变片, 以克为指标测量等距力。</li>
	<li>使血管达到最佳的静息张力: 肺动脉的环拉伸至 0.5 g, 腹腔动脉1.5 克, 腹主动脉2克, 20分钟后进行平衡清洗。重复拉伸等隔水洗步骤1x。</li>
	<li>为了测试船只的可行性, 将环与235μl 的 KCl (10-1 m ) 收缩 10分钟, 再清洗 10分钟, 并再次与235Μl 的 kcl (10-1 m) 收缩约 20分钟,直到到达高原。</li>
	<li>再次清洗血管 10分钟, 并添加588.4μl 的消炎痛 (10-5 m) (环氧合酶活性抑制剂) 至少 20分钟, 以避免可能干扰内源性前列腺素。</li>
	<li>添加 10-9 (10Μl) 至10-4 m (或10-9 至 10-5 m 的总剂量----苯乙胺 (phe) ; 10-9 m 以上所有浓度为 9μl ) 以收缩血管。</li>
	<li>最后剂量后, 等待约 1小时, 直到血管达到相对稳定的收缩状态 (高原)。</li>
	<li>添加与内皮依赖性的血管扩张剂乙酰胆碱 (ACh) 的累积剂量, 从10-9到 10-4 m (10-9 m 为 588.4条. l, 所有浓度超过 10-9 m 的累积剂量为12.6μl 和 40μl), 以诱导出一氧化氮氧化物 (NO) 介导的放松。</li>
	<li>在松弛曲线的末端, 清洗容器 10分钟, 并添加 588.4条 Xl 的消炎痛 (10-5 m), 以及 184μl 的 Ng-nit-l-精氨酸 (nla, 10-4 m),这是 nos 的抑制剂, 至少20分钟。</li>
	<li>将血管与独特剂量为10μl 的 Phe (肺动脉为 10-5和10-4 m, 腹主动脉和腹腔动脉为 10-4 m) 再收缩 1小时, 以诱导相对稳定的收缩.</li>
	<li>加入独特剂量为40μl 的 ach (10-4 m), 直到到达高原。</li>
	<li>再次清洗血管 10分钟, 然后加入588.4μl 的消炎痛 (10-5 m) 和184ΜL 的 nla (10-4 m) 20分钟。</li>
	<li>用10μl 的 Phe (10-5和10-4 m) 将船舶承包1小时。</li>
	<li>添加 no 供体二乙胺 (dea)/no的累积剂量 (10-9 [584条-l] 至 10-4 m [所有浓度超过 10-9M]), 以评估与内皮独立的 no诱导松弛. </li>
	<li>在实验结束时, 将容器储存在液氮中, 以便将来根据需要进行分析。</li>
</ol>

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E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+damage,+metabolism,+and+oxidative+stress+in+mdx+mice:+Impact+of+aerobic+running.">Muscle damage, metabolism, and oxidative stress in mdx mice: Impact of aerobic running.</a> Muscle &amp; Nerve. 54 (1), 110-117 (2016).</li><li>Cho, J., Kim, S., Lee, S., Kang, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+Training+Intensity+on+Nonalcoholic+Fatty+Liver+Disease.">Effect of Training Intensity on Nonalcoholic Fatty Liver Disease.</a> Medicine &amp; Science in Sports &amp; Exercise. 47 (8), 1624-1634 (2015).</li><li>Sabatier, M. J., Redmon, N., Schwartz, G., English, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treadmill+training+promotes+axon+regeneration+in+injured+peripheral+nerves.">Treadmill training promotes axon regeneration in injured peripheral nerves.</a> Experimental Neurology. 211 (2), 489-493 (2008).</li><li>Rolim, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aerobic+interval+training+reduces+inducible+ventricular+arrhythmias+in+diabetic+mice+after+myocardial+infarction.">Aerobic interval training reduces inducible ventricular arrhythmias in diabetic mice after myocardial infarction.</a> Basic Research in Cardiology. 110 (4), 44 (2015).</li><li>Lab Animal Research. Blood Withdrawal I. JoVE Science Education Database Available from: <a target="_blank" href="https://www.jove.com/science-education/10246/blood-withdrawal-i">https://www.jove.com/science-education/10246/blood-withdrawal-i</a> (2018)</li><li>Peyter, A. -. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscarinic+receptor+M1+and+phosphodiesterase+1+are+key+determinants+in+pulmonary+vascular+dysfunction+following+perinatal+hypoxia+in+mice.">Muscarinic receptor M1 and phosphodiesterase 1 are key determinants in pulmonary vascular dysfunction following perinatal hypoxia in mice.</a> American Journal of Physiology-Lung Cellular and Molecular Physiology. 295 (1), 201-213 (2008).</li><li>Faiss, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Significant+Molecular+and+Systemic+Adaptations+after+Repeated+Sprint+Training+in+Hypoxia.">Significant Molecular and Systemic Adaptations after Repeated Sprint Training in Hypoxia.</a> PLOS ONE. 8 (2), (2013).</li><li>Faiss, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repeated+Double-Poling+Sprint+Training+in+Hypoxia+by+Competitive+Cross-country+Skiers.">Repeated Double-Poling Sprint Training in Hypoxia by Competitive Cross-country Skiers.</a> Medicine &amp; Science in Sports &amp; Exercise. 47 (4), 809-817 (2015).</li><li>Billat, V. L., Mouisel, E., Roblot, N., Melki, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inter-+and+intrastrain+variation+in+mouse+critical+running+speed.">Inter- and intrastrain variation in mouse critical running speed.</a> Journal of Applied Physiology. 98 (4), 1258-1263 (2005).</li><li>Ferguson, S. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+living+at+moderate+altitude+on+pulmonary+vascular+function+and+exercise+capacity+in+mice+with+sickle+cell+anemia.">Effects of living at moderate altitude on pulmonary vascular function and exercise capacity in mice with sickle cell anemia.</a> The Journal of Physiology. , (2018).</li><li>Lightfoot, J. T., Turner, M. J., Debate, K. A., Kleeberger, S. 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作者没有什么可透露的。

这项研究的第一个目的是评估小鼠低氧高强度训练的可行性, 并确定该协议的适当特征, 这些特征对小鼠的耐受性很高。有目的, 由于没有数据使用超大鼠 (即超过 V 最大值) 的小鼠强度训练, 我们不得不根据以前与运动员开发的协议进行试验, 其中包括四到五套五套全能冲刺 (约为V最大值), 穿插了20秒的活动恢复, 交错活动恢复为 5分21秒,22...

在缺氧时, 氧 (o2) 的启发分数减少会导致低氧血症 (缺氧时动脉压力降低) 和 o2 输运能力改变1。急性缺氧诱导针对骨骼肌2的交感神经血管收缩活性增加, 并导致相反的 "补偿性" 血管舒张。
在缺氧的亚最大强度下, 相对于正常条件下相同水平的运动, 这种 "补偿性" 血管舒张是公认的3。这种血管扩张是必不可少的, 以确保增加血液流动和维持 (或限制改变) 氧气输送到活跃的肌肉。腺苷被证明在这一反应中没有独立的作用, 而一氧化氮 (NO) 似乎是主要的内皮源, 因为在缺氧期间, 一氧化氮合酶 (NOS) 抑制的增强血管舒张反应明显减弱练习4。在低氧运动中, 其他几种血管活性物质可能在补偿性血管舒张中起到一定作用。
这种增强的低氧运动充血与低氧引起的动脉 o2 含量下降成正比,随着运动强度的增加而增大, 例如在缺氧的剧烈增量运动中。
补偿性血管舒张的 no 介导的成分通过不同的途径调节, 运动强度增加 3: 如果β-肾上腺素能受体-刺激 no 成分在低强度低氧运动中显得最重要, 随着运动强度的增加, NO 的来源对β肾上腺素能机制的依赖程度似乎较低。在高强度低氧运动中, 还有其他刺激 no 释放的候选药物, 如从红细胞和内皮衍生前列腺素释放的 ATP。
缺氧中的上脑组织运动 (在运动生理学文献中称为缺氧重复短跑训练) 是最近的训练方法5 , 为团队或机架运动运动员提供了性能提升。这种方法不同于在最大速度6 (v 最大) 或接近最大速度 6 (v 最大) 时进行的低氧间歇训练, 因为 rsh 在最大强度下进行的训练会导致更大的肌肉灌注和氧合7以及特定的肌肉转录反应 8。有几种机制可以解释 RSH 的有效性: 在缺氧的冲刺过程中, 补偿性血管扩张和相关的较高血流比慢抽搐纤维更有利于快速抽搐纤维。因此, RSH 效率很可能是纤维类型的选择性和强度的依赖。我们推测, 血管系统反应能力的提高在 RSH 中是最重要的。
运动训练已在小鼠中进行了广泛的研究, 包括健康个体和病理小鼠模型9,10.训练老鼠最常见的方法是使用啮齿类动物跑步机, 传统上使用的方案是低强度训练, 最大 v 的 40%-60% (使用增量跑步机测试11确定), 为 30-60分钟 12,13 ,14,15。在16,17小鼠中, 最大强度间歇训练及其对病理的影响得到了广泛的研究;因此, 为小鼠开发了间歇训练运行协议。这些协议通常包括大约10回合运行在80%–100% 的 v 最大在啮齿类动物电动跑步机上, 1-4分钟, 穿插与主动或被动休息16,18。
缺氧中小鼠对在超甲强度 (即在 V最大值以上) 运动的兴趣来自于先前的研究结果, 即微血管血管舒张补偿和间歇性运动性能在以下情况下都有更大的提高。超大比在最大或中等强度。然而, 据我们所知, 无论是在诺莫夏还是缺氧, 都没有关于小鼠超脑组织训练方案的报告。
本研究的第一个目的是测试小鼠超乙酰强度训练的可行性, 以及确定一个可容忍和适当的协议 (强度、冲刺持续时间、恢复等)。第二个目的是评估诺莫夏和缺氧不同训练方案对血管功能的影响。因此, 我们测试的假设是: (1) 小鼠在缺氧时耐受良好的超脑运动, (2) 该协议在诺莫夏的血管功能得到了比运动更大的改善, 但也比低强度的低氧运动更大。

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			<td height="21" style="height:21px;width:260px;">Cotton swab</td>
			<td style="width:191px;">Q-tip</td>
			<td style="width:119px;"></td>
			<td style="width:165px;"></td>
		</tr>
		<tr height="67">
			<td height="67" style="height:67px;width:260px;">Gas mixer Sonimix 7100</td>
			<td style="width:191px;">LSI Swissgas, Geneva, Switzerland</td>
			<td style="width:119px;"></td>
			<td style="width:165px;">Gas-flow: 10 L/min and 1 L/min for O2 and CO2, respectively</td>
		</tr>
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			<td height="21" style="height:21px;width:260px;">Hypoxic Box&#160;</td>
			<td style="width:191px;">Homemade</td>
			<td style="width:119px;"></td>
			<td>Made in Plexiglas</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Motorized rodents treadmill Panlab LE-8710</td>
			<td style="width:191px;">Bioseb, France</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:260px;">Oximeter Greisinger GOX 100</td>
			<td style="width:191px;">GREISINGER electronic Gmbh, Regenstauf, Germany</td>
			<td style="width:119px;"></td>
			<td></td>
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			<td height="21" style="height:21px;width:260px;">Sedacom software</td>
			<td style="width:191px;">Bioseb, France</td>
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			<td height="42" style="height:42px;width:260px;">Strain gauge</td>
			<td style="width:191px;">PowerLab/8SP; ADInstruments</td>
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			<td></td>
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supramaximal intensity hypoxic exercise and vascular function assessment in mice

运动训练是保持健康、预防多种慢性病的重要策略。这是国际指南为患有心血管疾病的患者推荐的一线治疗方法, 更具体地说, 是下肢动脉疾病患者的行走能力发生了很大的变化, 影响了患者的行走能力。生活质量。
传统上, 无论是低连续锻炼还是间歇训练都使用过。最近, 海马上训练也被证明可以通过血管适应等机制来提高运动员的表现。这种类型的训练与缺氧的结合可以带来额外的和/或协同作用, 这可能是对某些病理感兴趣的。在这里, 我们描述如何执行在缺氧中的健康小鼠以150% 的最大速度, 使用电动跑步机和低氧盒。我们还展示了如何解剖小鼠, 以检索感兴趣的器官, 特别是肺动脉, 腹主动脉, 和胫骨动脉。最后, 我们展示了如何对被回收的血管进行体外血管功能评估, 并利用等距张力研究。

据我们所知, 本研究首次描述了小鼠诺莫夏和缺氧的超脑度强度训练方案。在这个协议中, 老鼠在每次冲刺之间跑了四组 5 1 s 冲刺, 恢复了 2 0。这些套被穿插在5分钟的恢复期。目前尚不清楚这些老鼠是否有能力维持这样的协议并适当地完成它。然而, 根据图 5, 接受上血强度训练的小鼠体重增加与诺莫夏和缺氧小鼠的体重增加相似。
<p class="jove_conte...

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Supramaximal Intensity Hypoxic Exercise and Vascular Function Assessment in Mice

低氧高强度训练是一种方案, 已被证明能诱导一些患者可能有益的血管适应, 提高运动员的反复冲刺能力。在这里, 我们测试使用该方案训练小鼠的可行性, 并使用体外血管功能评估确定这些血管适应。

小鼠低血压运动的下位强度及血管功能评估

Exercise training is an important strategy for maintaining health and preventing many chronic diseases. It is the first line of treatment recommended by ...

我们的协议意义重大，因为它首次描述了如何在小鼠中进行上部毒性强度训练，特别是在缺氧，如果在诱导特定的血管内皮适应。在小鼠身上使用这种技术的主要优点是，它进一步研究了血管机理。运动训练是维持健康、预防许多慢性病的重要策略， 运动与缺氧相结合，可进行特定的细胞和分子适应，具有潜在的治疗应用。例如，在周围动脉疾病或高血压患者。证明这个程序将是杰西卡·拉维尔，一个博士生的心理学系实验室，斯蒂夫·门

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8.3K 次观看.  Lausanne University Hospital (CHUV). 我们的协议意义重大，因为它首次描述了如何在小鼠中进行上部毒性强度训练，特别是在缺氧，如果在诱导特定的血管内皮适应。在小鼠身上使用这种技术的主要优点是，它进一步研究了血管机理。运动训练是维持健康、预防许多慢性病的重要策略， 运动与缺氧相结合，可进行特定的细胞和分子适应，具有潜在的治疗应用。例如，在周围动脉疾病或高血压患者。证明这个程?...

视频: Supramaximal Intensity Hypoxic Exercise and Vascular Function Assessment in Mice

Assessment of Sensorimotor Function in Mouse Models of Parkinson's Disease

Sensitive and reliable behavioral outcome measures are essential to the evaluation of potential therapeutic treatments in preclinical trials for many neurodegenerative diseases. In Parkinson's disease, sensorimotor tests sensitive to varying degrees of nigrostriatal dysfunction are fundamental for testing the efficacy of potential therapeutics. Reliable and quite elegant sensorimotor measures exist for rats, however many of these tests measure sensorimotor asymmetry within the rat and are not entirely suitable for the newer genetic mouse models of PD. We have put together a battery of sensorimotor tests inspired by the sensitive tests in rats and adapted for mice. The test battery highlighted in this study is chosen for a) its sensitivity in a wide variety of mouse models of PD, b) its ease in implementing into a study, and c) its low expense. These tests have proven useful in characterizing novel genetic mouse models of PD as well as in testing potential disease-modifying therapies.

Assessment of Sensorimotor Function in Mouse Models of Parkinson&#039;s Disease

In Parkinson's disease and movement disorders in general, sensitive and reliable behavioral assays are essential for testing novel potential therapeutics. Here, we describe a manageable battery of sensorimotor tests for mice that are sensitive to varying degrees of injury to the nigrostriatal system and useful for preclinical studies.

帕金森氏症小鼠模型的感觉功能评估

Sensitive and  reliable behavioral outcome measures are essential to the evaluation of  potential therapeutic treatments in preclinical trials for many  ...

科研

行为学

Getting to Compliance in Forced Exercise in Rodents: A Critical Standard to Evaluate Exercise Impact in Aging-related Disorders and Disease

There is a major increase in the awareness of the positive impact of exercise on improving several disease states with neurobiological basis; these include improving cognitive function and physical performance. As a result, there is an increase in the number of animal studies employing exercise. It is argued that one intrinsic value of forced exercise is that the investigator has control over the factors that can influence the impact of exercise on behavioral outcomes, notably exercise frequency, duration, and intensity of the exercise regimen. However, compliance in forced exercise regimens may be an issue, particularly if potential confounds of employing foot-shock are to be avoided. It is also important to consider that since most cognitive and locomotor impairments strike in the aged individual, determining impact of exercise on these impairments should consider using aged rodents with a highest possible level of compliance to ensure minimal need for test subjects. Here, the pertinent steps and considerations necessary to achieve nearly 100% compliance to treadmill exercise in an aged rodent model will be presented and discussed. Notwithstanding the particular exercise regimen being employed by the investigator, our protocol should be of use to investigators that are particularly interested in the potential impact of forced exercise on aging-related impairments, including aging-related Parkinsonism and Parkinson&#8217;s disease.

Exercise may alleviate behavioral impairments associated with neurodegenerative diseases. Studies of the mechanisms underlying exercise-mediated effects have increased in disease models, but the neurobiological background accompanying these diseases, specifically aging, is likely not considered. Here, we illustrate measures to increase the likelihood of a successful exercise paradigm in aged rats.

让合规的强制锻炼鼠害：一项关键的标准在衰老相关的疾病和疾病评估运动的影响

There is a major increase in the awareness of the positive impact of exercise on improving several disease states with neurobiological basis; these ...

Assessment of Murine Exercise Endurance Without the Use of a Shock Grid: An Alternative to Forced Exercise

Using laboratory mouse models, the molecular pathways responsible for the metabolic benefits of endurance exercise are beginning to be defined. The most common method for assessing exercise endurance in mice utilizes forced running on a motorized treadmill equipped with a shock grid. Animals who quit running are pushed by the moving treadmill belt onto a grid that delivers an electric foot shock; to escape the negative stimulus, the mice return to running on the belt. However, avoidance behavior and psychological stress due to use of a shock apparatus can interfere with quantitation of running endurance, as well as confound measurements of postexercise serum hormone and cytokine levels. Here, we demonstrate and validate a refined method to measure running endurance in na&#239;ve C57BL/6 laboratory mice on a motorized treadmill without utilizing a shock grid. When mice are preacclimated to the treadmill, they run voluntarily with gait speeds specific to each mouse. Use of the shock grid is replaced by gentle encouragement by a human operator using a tongue depressor, coupled with sensitivity to the voluntary willingness to run on the part of the mouse. Clear endpoints for quantifying running time-to-exhaustion for each mouse are defined and reflected in behavioral signs of exhaustion such as splayed posture and labored breathing. This method is a humane refinement which also decreases the confounding effects of stress on experimental parameters.

A method to assess exercise endurance in laboratory mice without the use of a shock grid is demonstrated. This method is a humane refinement that can decrease the confounding effects of stress on experimental parameters.

小鼠运动耐力的评估没有一个冲击电网的使用：另一种强迫运动

Using laboratory mouse models, the molecular pathways responsible for the metabolic benefits of endurance exercise are beginning to be defined. The most ...

Assessment of Cocaine-induced Behavioral Sensitization and Conditioned Place Preference in Mice

It is thought that rewarding experiences with drugs create strong contextual associations and encourage repeated intake. In turn, repeated exposures to drugs of abuse make lasting alterations in the brain function of vulnerable individuals, and these persistent alterations likely serve to maintain the maladaptive drug seeking and taking behaviors characteristic of addiction/dependence2. In rodents, reward experience and contextual associations are frequently measured using the conditioned place preference assay, or CPP, wherein preference for a previously drug-paired context is measured. Behavioral sensitization, on the other hand, is an increase in a drug-induced behavior that develops progressively over repeated exposures. Since sensitized behaviors can often be measured after several months of drug abstinence, depending on the dose and length of initial exposure, they are considered observable correlates of lasting drug-induced plasticity. Researchers have found these assays useful in determining the neurobiological substrates mediating aspects of addiction as well as assessing the potential of different interventions in disrupting these behaviors. This manuscript describes basic, effective protocols for mouse CPP and locomotor behavioral sensitization to cocaine.

This protocol is intended to enable researchers to conduct experiments designed to test these aspects of addiction using the conditioned place preference and locomotor behavioral sensitization assays.

可卡因诱导行为敏和小鼠位置偏爱评估

It is thought that rewarding experiences with drugs create strong contextual associations and encourage repeated intake. In turn, repeated exposures to ...

An Objective and Child-friendly Assessment of Arm Function by Using a 3-D Sensor

Progressive and irreversible muscle atrophy characterizes Spinal Muscular Atrophy (SMA) and other similar muscle disorder diseases. Objective assessment of muscle functions is an essential and important, although challenging, prerequisite for successful clinical trials. Current clinical rating scales restrain the movement abnormalities to certain predefined coarse-grained individual items. The Kinect 3-D sensor has emerged as a low-cost and portable motion sensing technology used to capture and track people&#39;s movement in many medical and research fields. A novel approach using this 3-D sensor was developed and a game-like test was designed to objectively measure the upper limb function of patients with SMA. The prototype test targeted joint movement capability. While sitting in a virtual scene, the patient was instructed to extend, flex, and lift the whole arm in order to reach and place some objects. Both kinematic and spatiotemporal characteristics of upper limb movement were extracted and analyzed, e.g., elbow extension and flexion angles, hand velocity, and acceleration. The first study included a small cohort of 18 ambulant SMA patients and 19 age- and gender-matched healthy controls. A comprehensive analysis of arm movement was achieved; however, no significant difference between the groups were found due to the mismatch of patient&#39;s capability and the test difficulty. Based on this experience, a second version of the test consisting of a modified version of the first game with increased difficulties and a second game targeting muscle endurance were designed and implemented. The new test has not been conducted in any patient groups yet. Our work has demonstrated the potential capability of the 3-D sensor in assessing such muscle function and suggested an objective approach to complement the clinical rating scales.

An objective measure of muscle functions is challenging especially in children. Based on a commercially available digital 3-D sensor, a child-friendly gaming test was developed to assess upper limb function for clinical trials.

基于3维传感器的 Arm 功能的客观和儿童友好评估

Progressive and irreversible muscle atrophy characterizes Spinal Muscular Atrophy (SMA) and other similar muscle disorder diseases. Objective assessment ...

Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia

In keeping with the 3R Principle (Replacement, Reduction, Refinement) developed by Russel and Burch, scientific research should use alternatives to animal experimentation whenever possible. When there is no alternative to animal experimentation, the total number of laboratory animals used should be the minimum needed to obtain valuable data. Moreover, appropriate refinement measures should be applied to minimize pain, suffering, and distress accompanying the experimental procedure. The categories used to classify the degree of pain, suffering, and distress are non-recovery, mild, moderate, or severe (EU Directive 2010/63). To determine which categories apply in individual cases, it is crucial to use scientifically sound tools.
The well-being-assessment protocol presented here is designed for procedures during which general anesthesia is used. The protocol focuses on home cage activity, the Mouse Grimace Scale, and luxury behaviors such as burrowing and nest building behavior as indicators of well-being. It also uses the free exploratory paradigm for trait anxiety-related behavior. Fecal corticosterone metabolites as indicators of acute stress are measured over the 24-h post-anesthetic period.
The protocol provides scientifically solid information on the well-being of mice following general anesthesia. Due to its simplicity, the protocol can easily be adapted and integrated in a planned study. Although it does not provide a scale to classify distress in categories according to the EU Directive 2010/63, it can help researchers estimate the degree of severity of a procedure using scientifically sound data. It provides a way to improve the assessment of well-being in a scientific, animal-centered manner.

We developed a protocol to assess well-being in mice during procedures using general anesthesia. A series of behavioral parameters indicating levels of well-being as well as glucocorticoid metabolites were analyzed. The protocol can serve as a general aid to estimate the degree of severity in a scientific, animal-centered manner.

全身麻醉下小鼠幸福度的系统评价

In keeping with the 3R Principle (Replacement, Reduction, Refinement) developed by Russel and Burch, scientific research should use alternatives to animal ...

Short Session High Intensity Interval Training and Treadmill Assessment in Aged Mice

High intensity interval training (HIIT) is emerging as a therapeutic approach to prevent, delay, or ameliorate frailty. In particular short session HIIT, with regimens less than or equal to 10 min is of particular interest as several human studies feature routines as short as a few minutes a couple times a week. However, there is a paucity of animal studies that model the impacts of short session HIIT. Here, we describe a methodology for an individually tailored and progressive short session HIIT regimen of 10 min given 3 days a week for aged mice using an inclined treadmill. Our methodology also includes protocols for treadmill assessment. Mice are initially acclimatized to the treadmill and then given baseline flat and uphill treadmill assessments. Exercise sessions begin with a 3 min warm-up, then three intervals of 1 min at a fast pace, followed by 1 min at an active recovery pace. Following these intervals, the mice are given a final segment that starts at the fast pace and accelerates for 1 min. The HIIT protocol is individually tailored as the speed and intensity for each mouse are determined based upon initial anaerobic assessment scores. Additionally, we detail the conditions for increasing or decreasing the intensity for individual mice depending on performance. Finally, intensity is increased for all mice every two weeks. We previously reported in this protocol enhanced physical performance in aged male mice and here show it also increases treadmill performance in aged female mice. Advantages of our protocol include low administration time (about 15 min per 6 mice, 3 days a week), strategy for individualizing for mice to better model prescribed exercise, and a modular design that allows for the addition or removal of the number and length of intervals to titrate exercise benefits.

Short session (&#8804;10 min) high intensity interval training (HIIT) is emerging as an alternative to longer exercise modalities, yet the shorter variants are rarely modeled in animal studies. Here, we describe a 10 min, 3 day a week, uphill treadmill HIIT protocol that enhances physical performance in male and female aged mice.

老年小鼠短期高强度间隔训练与跑步机评价

High intensity interval training (HIIT) is emerging as a therapeutic approach to prevent, delay, or ameliorate frailty. In particular short session HIIT, ...

Assessment of Sexual Behavior of Male Mice

Sexual behavior is highly species-specific. Although rodents have slightly different sexual behaviors, mice and rats have a similar sexual behavioral pattern. The purpose of this article is to describe the hormone-induced estrus ovariectomized female model and the experimental procedure for the assessment of sexual behavior of male mice. The most important sexual behavioral elements are demonstrated in the video and illustrations. The critical steps, advantages, and limitations of the sexual behavior test are explained as well. Finally, the behavior parameters are presented, and mounting, intromission, and ejaculation processes in mating are distinguished. Behavioral parameters are assessed in terms of the occurred duration and counts during the test period.

This article describes how to perform sexual behavior tests in male mice.

雄性小鼠性行为评估

Sexual behavior is highly species-specific. Although rodents have slightly different sexual behaviors, mice and rats have a similar sexual behavioral ...

Impact of High-intensity Interval Exercise and Moderate-Intensity Continuous Exercise on the Cardiac Troponin T Level at an Early Stage of Training

An elevation in cardiac troponin T (cTnT), as a highly specific biomarker of cardiomyocyte damage, after moderate-intensity continuous exercise (MCE) has been described. The exercise-induced cTnT response distorts the diagnostic role of the cTnT assay. Although high-intensity interval exercise (HIE) is growing in popularity and concerns remain about its safety, available data related to cTnT release after HIE is limited, which hampers the use of HIE as a health intervention. Here, we present three representative HIE protocols [traditional HIE (repeated 4 min cycling at 90% V&#775;O2max interspersed with 3 min rest, 200 kJ/session); sprint interval exercise (SIE, repeated 1 min cycling at 120% V&#775;O2max interspersed with 1.5 min rest, 200 kJ/session); and repeated sprint exercise (RSE, 40 x 6 s all-out sprints interspersed with 9 s rest)] and one representative MCE protocol (continuous cycling exercise at an intensity of 60% V&#775;O2max, 200 kJ/session). Forty-seven sedentary, overweight young women were randomly assigned to one of four groups (HIE, SIE, RSE, and MCE). Six bouts of respective exercise were performed by every single group, with each being 48 h apart. Meanwhile, for four groups, the duration of the entire testing period was identical, being 10 days. Before and after the first and final exercise bouts, an assessment was conducted of cTnT. The current study provides a frame of reference giving a clear picture of how a specific exercise session affects the circulating cTnT concentration at the early stage of training. The information may assist with clinical interpretations of post-exercise cTnT elevation and guide the prescription of exercise, especially for HIE.

Here, we present protocols of high-intensity interval and moderate-intensity continuous exercise to observe the response of circulating cardiac troponin T (cTnT) concentration to acute exercise over 10 days. The information may assist with clinical interpretations of post-exercise cTnT elevation and guide the prescription of exercise.

高强度间歇锻炼和中强度连续运动对心脏肌钙蛋白T级在训练早期阶段的影响

An elevation in cardiac troponin T (cTnT), as a highly specific biomarker of cardiomyocyte damage, after moderate-intensity continuous exercise (MCE) has ...

Assessment of Memory Function in Pilocarpine-induced Epileptic Mice

Cognitive impairment is one of the most common comorbidities in temporal lobe epilepsy. To recapitulate epilepsy-associated cognitive decline in an animal model of epilepsy, we generated pilocarpine-treated chronic epileptic mice. We present a protocol for three different behavioral tests using these epileptic mice: novel object location (NL), novel object recognition (NO), and pattern separation (PS) tests to evaluate learning and memory for places, objects, and contexts, respectively. We explain how to set the behavioral apparatus and provide experimental procedures for the NL, NO, and PS tests following an open field test that measures the animals&#8217; basal locomotor activities. We also describe the technical advantages of the NL, NO, and PS tests with respect to other behavioral tests for assessing memory function in epileptic mice. Finally, we discuss possible causes and solutions for epileptic mice failing to make 30 s of good contact with the objects during the familiarization sessions, which is a critical step for successful memory tests. Thus, this protocol provides detailed information about how to assess epilepsy-associated memory impairments using mice. The NL, NO, and PS tests are simple, efficient assays that are appropriate for the evaluation of different kinds of memory in epileptic mice.

This article presents experimental procedures for assessing memory impairments in pilocarpine-induced epileptic mice. This protocol can be used to study the pathophysiologic mechanisms of epilepsy-associated cognitive decline, which is one of the most common comorbidities in epilepsy.

皮洛卡平诱导癫痫小鼠记忆功能评估

Cognitive impairment is one of the most common comorbidities in temporal lobe epilepsy. To recapitulate epilepsy-associated cognitive decline in an animal ...