这项工作是由美国国立卫生研究院授予（HL 098256），由国家和指导研究科学发展奖（K01 AR052840）和独立科学家奖（K02 HL105799）由美国国立卫生研究院授予太平绅士Konhilas和跨学科培训格兰特在心血管科学（HL007249支持）。支持从萨弗心脏中心在亚利桑那大学，距离史蒂芬M. Gootter基金会收到。

所有的实验中，住房和爱护动物中使用该坚持的准则和批准的机构动物护理和使用委员会在亚利桑那大学，并为护理实验动物和使用2011年美国国立卫生研究院准则的协议执行。 1。治疗与4 - 乙烯基环己烯二环氧化物<ol><li> VCD给药方案:在通风橱中，添加0.587毫升VCD到一个干净的瓶中，使终体积10毫升香油，再经反相轻轻地，盖组合和用封口膜密封。将该溶液转移到一个剂量小瓶中，用Parafilm密封，并在4℃下储存长达7天。确保所有试剂均为无菌。 <ol><li>用麻油作为车辆控制，VCD溶于香油。存储VCD（MW =140.2克/摩尔，密度= 0.00克/毫升）在-20℃下</li><li> VCD注射方案:在第2个月的年龄，体重和管理室脾气每日腹腔注射ATURE VCD在160毫克/公斤，连续20天的剂量。在与车辆的溶液，香油类似的方式注射的对照小鼠，在2.5毫升/千克的量。 注:在注射期间，尽量交替注射的部位，以减少局部感染的机会。使用下面的公式来计算所需要的注射量:体重（克）×公斤/ 1,000 GX2.5毫升/公斤=体积（ml）溶液，注入导致大约50毫升/注射用27号针头。由于针的尺寸并不需要局部麻醉剂。 </li><li>发情周期监测:注射周期结束后，通过获得阴道涂片日常监测动情周期。使用背颈背方法抑制小鼠。插入吸管用0.2ml无菌生理盐水到阴道口，以0.5厘米的深度。磨碎盐水（3次），然后将1滴在空白幻灯片。立即评估显微镜使用标准光源microsc样本氧化聚乙烯在100X幅度。 注:阴道脱落细胞学检查是决心确保自行车的损失，因此验证卵巢早衰图2A说明在整个发情周期的不同阶段，典型的阴道细胞学检查的小鼠，即发情前期，发情期，发情后期，间情期17-19。在动情前期，细胞包括有核上皮细胞（ 图2A中，箭头表示的有核细胞）;发情期期间，将细胞主要角化鳞状上皮细胞;动情后期的特征是细胞类型的混合，与白细胞占优势;动情间期是由白细胞为主。焦平面的调整可能需要在整个样本可视化细胞和细胞核。 如图2B所示 ，一个典型的鼠发情周期为4-5天。小鼠经过连续15天的持续性动情考虑非循环。连续地遍及每对研究期间监测动情周期IOD确认缺乏VCD治疗小鼠动情周期。 </li></ol></li></ol> 2。自愿笼轮练习 近交系C57BL / 6和B6C3F1雌性小鼠，年龄7个月，则暴露在自愿笼轮运行。房子个别动物在笼中（47厘米×26厘米×14.5厘米）与不受阻碍的访问到笼子轮28天。 <ol><li>设备设置:笼轮装置包括与5.0厘米宽的行驶面配备了数字磁计数器，该计数器是由轮子转动激活11.5厘米直径的车轮。各车轮行驶试验之前，根据制造商的指导校准笼轮。测量跑轮的内径，计算出圆周，并输入该数值到数字计数器的轮毂尺寸。 <ol><li>附加的数字磁记录计数器的计算机部向轮支架利用胶带。 </li><li>将计算机连接端口离子数字磁计数器车轮支架使用磁带。 </li><li>附加的数字磁记录计数器的自由磁体在相同侧上的中心杆的磁铁通过电缆连接到计算机上，再绑扎充分保护磁铁从咀嚼损坏。磁体的位置应使车轮每旋转记录由计算机没有任何干扰或阻力来完成车轮的转动。 注意:胶带已被成功地用于数字磁计数器的部件附加到传动轮。绑扎应足以完全覆盖磁铁，并防止损坏从由动物咀嚼。磁铁的位置应尽可能接近到车轮而不会妨碍其运动。然而，该设置也应该每天咀嚼伤害监测和修复是必要的。 </li><li>通过滑动轮站通过动物笼子的顶部布线安装车轮的笼子。固定轮架到t用长尾夹他笼接线。 注意:该夹应该是在布线的顶部，远离动物。通过保持器的布线喂计算机部和任何额外的电缆。将电脑和多余电缆动物接触不到的地方。这可通过将一个中等大小的塑料称量在轿厢的布线的顶盘以保持计算机和多余的电缆来实现。 </li></ol></li><li>练习:对于给定的垃圾，随机将小鼠无论是久坐控制或运动养生。给所有的动物和水标准硬盘啮齿动物饲料自由采食。 <ol><li>手动的距离跑（公里/天）和用于运行总时间（小时/天），由磁计数器给定的记录每天的值。记录距离和时间，每天在整个演习期间28天时间准确和一致的价值观相同的1个小时的时间框架内的值。清除（零）磁计数器后，每一天的记录，以确保准确的读数后续ING的日子。 </li><li>计算距离和速度的每个组的平均值。计算速度每天根据距离和时间的给定的值对每个小鼠。数据可以为每个测量参数作为每天或每周的平均值显示。 </li></ol></li><li>组织收获:在具体行使期结束时，安乐死内下吸入麻醉去除鸡笼颈椎脱位的30分钟行使和久坐对照组动物。称量体重，并迅速消费心，骨骼肌和卵巢。 <ol><li>在组织收获的时候，装载和弄脏收集卵巢与苏木精和伊红（H &amp;E; 图2C）。在放大镜下，计数和每个VCD治疗和控制动物20归类为毛囊原始，初级，中级，或胃窦部。甲原始卵泡包含一个小的初级卵母细胞，扁平或鳞状颗粒细胞密切相对于卵母细胞的单个层，以及基底椎板。一个主要的卵泡是由一个或多个立方形颗粒细胞的存在下被布置在围绕卵母细胞的单层限定。次级卵泡包含周围形成多个卵母细胞层和卵泡膜颗粒细胞。一个窦状卵泡的特征在于，含有流体的空腔或"腔"。 </li><li>切除的心脏，使内侧切口的腹侧和打开隔膜后可视化的心脏。在该基地并将其放置在冷磷酸盐缓冲液充分洗出残留心室的血液迅速切除心脏。修剪多余的组织的心脏，权衡整个心脏和捕捉冻结在液氮中。 </li><li>用改性冰冷的磷酸盐缓冲盐水溶液（PBS）洗涤所有其他切除的组织，和扣冻结在异戊烷在液氮中冷却。 </li></ol></li></ol> 3。强迫跑步机练习<ol><li>当采用强迫运动范式吨Ø测量运动能力，使用一台跑步机，并设置如下: <ol><li>将电击电网在跑步机上斜坡的底部。产生这种冲击作为一个刺激的运动。的电刺激的强度和频率是手动的使用者控制。网格可以启用或禁用单独为每个车道。所有数据收集是手动完成的。 </li><li>周期性地，根据制造商的指导进行校准的跑步机。 <ol><li>确保在跑步机控制器前面板切换设置为"STOP"。 </li><li>上跑步机带泳道1地点转速传感器和安全地与螺母拧紧在如下图所示的位置。 </li><li>使用拨号的跑步机控制面板上，调整速度887米/分钟之后，跑步机控制器面板上设置的电刺激频率为2 Hz的频率。 </li><li>按"里程表复位"按钮来初始化校准程序。 <strong&gt;的注意:久坐或nonexercise对照组必须包含在实验设计。久坐组为年龄匹配的运动组。之前我们已经表明，个别房屋的小鼠久坐在同样大小的笼子到行使小鼠相比，安置与兄弟姐妹久坐小鼠没有影响所需的参数。因此，久坐老鼠被安放在同一个房间内行使小鼠的兄弟姐妹。 </li></ol></li></ol></li><li>驯化:之前每天的实验3-7天驯化老鼠的跑步机。要做到这一点，地方的小鼠与带不动的震撼和电网断开，但对（噪声），带电机的跑步机。离开静置至少15分钟的小鼠。 </li><li>热身:热身如本协议中使用如下的步骤: <ol><li>将每只小鼠在跑步机上。禁用电网的冲击，提高带速4米/分钟持续15分钟。 </li><li>使电网的冲击，保持皮带速度4米/分钟的皮带速度为10分钟。 </li></ol></li><li>运动训练方案<ol><li>以下热身，不断冲击电网和增加皮带速度5米/分钟持续15分钟。 </li><li>提高带速为15米/分钟额外15分钟。 </li><li>下面的训练，转移小鼠回到自己的笼子。 注:在本研究中，运动训练1周每日执行。 </li><li>急性运动疗法:急性运动养生的目的是确定极端运动负荷的短期和长期影响。 <ol><li>以下预热，增加皮带速度为5米/分钟到10，15，20，25，和30米/分钟在10分钟的间隔。 注:终止运动，如果老鼠是无法继续疲惫和痛苦的运行或迹象。在我们的研究中，大多数动物都停在25米/分钟，无动物通过了30米/分的阶段。 </li><li>测试每个鼠标三次，让每个测试之间的2-3天。所有埃克塞计算平均值精确的会话每只小鼠。 </li></ol></li><li>运动养生耐力:耐力运动协议旨在培养老鼠的长期暴露在跑步机上运动的刺激。这种类型的协议测试容忍适度的运动应力的长期影响的能力。 <ol><li>首先在热身期间，4米/分钟概述的皮带速度。 </li><li>逐步提高皮带速度1m/min卷材，直到20米的最终速度/分钟为止。一个典型的研究方案的总时间为50分钟。 </li><li>运动养生后，小鼠转回到自己的笼子。 注意:运行小鼠消耗体力是可以避免的。如果在实验过程中的任何一点鼠标显示疲惫的迹象，停用电网的冲击，让鼠标休息和恢复。 </li></ol></li></ol></li></ol> 4。数据和统计分析<ol><li>本研究结果为平均值±标准差，并使用双因子ANOVA（治疗组和exercise为主要因素），其次是学生，纽曼Keuls检验事后检验或学生的t检验来比较平均值之间的差异。 </li><li>由每个行使动物的质量进行比较，以久坐组的平均心脏质量测定心脏重量与运动变化百分比。心脏质量差被表示为久坐不动的动物的百分比变化为每个相应的动物。的&lt;0.05的p值被认为是统计学显著。 </li></ol>

<ol>
	<li>Lindheim, S. 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=Comparison+of+estimates+of+insulin+sensitivity+in+pre-+and+postmenopausal+women+using+the+insulin+tolerance+test+and+the+frequently+sampled+intravenous+glucose+tolerance+test.">Comparison of estimates of insulin sensitivity in pre- and postmenopausal women using the insulin tolerance test and the frequently sampled intravenous glucose tolerance test.</a> J. Soc. Gynecol. Invest. 1, 150-154 (1994).</li><li>Dubnov-Raz, G., Pines, A., Berry, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diet+and+lifestyle+in+managing+postmenopausal+obesity.">Diet and lifestyle in managing postmenopausal obesity.</a> Climacteric. 10 Suppl 2, 38-41 (2007).</li><li>Grundy, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolic+syndrome:+connecting+and+reconciling+cardiovascular+and+diabetes+worlds.">Metabolic syndrome: connecting and reconciling cardiovascular and diabetes worlds.</a> J. Am. Coll. Cardiol. 47, 1093-1100 (2006).</li><li>Sharma, S., Firoozi, S., McKenna, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Value+of+exercise+testing+in+assessing+clinical+state+and+prognosis+in+hypertrophic+cardiomyopathy.">Value of exercise testing in assessing clinical state and prognosis in hypertrophic cardiomyopathy.</a> Cardiol. Rev. 9, 70-76 (2001).</li><li>Guyatt, G. H., Devereaux, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+heart+failure+treatment.">A review of heart failure treatment.</a> Mt. Sinai. J. Med. 71, 47-54 (2004).</li><li>Mayer, L. P., 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=Long-term+effects+of+ovarian+follicular+depletion+in+rats+by+4-vinylcyclohexene+diepoxide.">Long-term effects of ovarian follicular depletion in rats by 4-vinylcyclohexene diepoxide.</a> Reprod. Toxicol. 16, 775-781 (2002).</li><li>Mayer, L. P., Devine, P. J., Dyer, C. A., Hoyer, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+follicle-deplete+mouse+ovary+produces+androgen.">The follicle-deplete mouse ovary produces androgen.</a> Biol. Reprod. 71, 130-138 (2004).</li><li>Lohff, J. C., Christian, P. J., Marion, S. L., Hoyer, P. B. <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+duration+of+dosing+on+onset+of+ovarian+failure+in+a+chemical-induced+mouse+model+of+perimenopause.">Effect of duration of dosing on onset of ovarian failure in a chemical-induced mouse model of perimenopause.</a> Menopause. 13, 482-488 (2006).</li><li>Wright, L. 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=Comparison+of+skeletal+effects+of+ovariectomy+versus+chemically+induced+ovarian+failure+in+mice.">Comparison of skeletal effects of ovariectomy versus chemically induced ovarian failure in mice.</a> J. Bone Miner Res. 23, 1296-1303 (2008).</li><li>Ogawa, S., Chan, J., Gustafsson, J. A., Korach, K. S., Pfaff, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estrogen+increases+locomotor+activity+in+mice+through+estrogen+receptor+alpha:+specificity+for+the+type+of+activity.">Estrogen increases locomotor activity in mice through estrogen receptor alpha: specificity for the type of activity.</a> Endocrinology. 144, 230-239 (2003).</li><li>Roy, E. J., Wade, G. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+estrogens+in+androgen-induced+spontaneous+activity+in+male+rats.">Role of estrogens in androgen-induced spontaneous activity in male rats.</a> J. Comp. Physiol. Psychol. 89, 573-579 (1975).</li><li>Fahrbach, S. E., Meisel, R. L., Pfaff, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preoptic+implants+of+estradiol+increase+wheel+running+but+not+the+open+field+activity+of+female+rats.">Preoptic implants of estradiol increase wheel running but not the open field activity of female rats.</a> Physiol. Behav. 35, 985-992 (1985).</li><li>Morgan, M. A., Pfaff, D. W. <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+estrogen+on+activity+and+fear-related+behaviors+in+mice.">Effects of estrogen on activity and fear-related behaviors in mice.</a> Horm. Behav. 40, 472-482 (2001).</li><li>Gorzek, J. F., 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=Estradiol+and+tamoxifen+reverse+ovariectomy-induced+physical+inactivity+in+mice.">Estradiol and tamoxifen reverse ovariectomy-induced physical inactivity in mice.</a> Med. Sci. Sports Exerc. 39, 248-256 (2007).</li><li>Kadi, F., 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=The+effects+of+physical+activity+and+estrogen+treatment+on+rat+fast+and+slow+skeletal+muscles+following+ovariectomy.">The effects of physical activity and estrogen treatment on rat fast and slow skeletal muscles following ovariectomy.</a> J. Muscle Res. Cell Motil. 23, 335-339 (2002).</li><li>Konhilas, J. P., 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=Sex+modifies+exercise+and+cardiac+adaptation+in+mice.">Sex modifies exercise and cardiac adaptation in mice.</a> Am. J. Physiol. Heart Circ. Physiol. 287, 2768-2776 (2004).</li><li>Caligioni, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+reproductive+status/stages+in+mice.">Assessing reproductive status/stages in mice.</a> Curr. Protoc. Neurosci. 4, (2009).</li><li>Goldman, J. M., Murr, A. S., Cooper, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+rodent+estrous+cycle:+characterization+of+vaginal+cytology+and+its+utility+in+toxicological+studies.">The rodent estrous cycle: characterization of vaginal cytology and its utility in toxicological studies.</a> Birth Defects Res. B. Reprod. Toxicol. 80, 84-97 (2007).</li><li>McLean, A. C., Valenzuela, N., Fai, S., Bennett, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performing+vaginal+lavage,+crystal+violet+staining,+and+vaginal+cytological+evaluation+for+mouse+estrous+cycle+staging+identification.">Performing vaginal lavage, crystal violet staining, and vaginal cytological evaluation for mouse estrous cycle staging identification.</a> J. Vis. Exp. , (2012).</li><li>McKee, L. A., 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=Sexually+dimorphic+myofilament+function+and+cardiac+troponin+I+phosphospecies+distribution+in+hypertrophic+cardiomyopathy+mice.">Sexually dimorphic myofilament function and cardiac troponin I phosphospecies distribution in hypertrophic cardiomyopathy mice.</a> Arch. Biochem. Biophys. 535, 39-48 (2013).</li><li>Konhilas, J. P., 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=Exercise+can+prevent+and+reverse+the+severity+of+hypertrophic+cardiomyopathy.">Exercise can prevent and reverse the severity of hypertrophic cardiomyopathy.</a> Circ. Res. 98, 540-548 (2006).</li><li>Perez, N. J., Chen, H., Regan, J. A., Emert, A., Constantopoulos, E., Lynn, M., Konhilas, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+chemically-induced+ovarian+failure+on+voluntary+cage+wheel+exercise+and+cardiac+adaptation+in+mice.">The impact of chemically-induced ovarian failure on voluntary cage wheel exercise and cardiac adaptation in mice.</a> Compar. Med. In press, (2013).</li></ol>

作者什么都没有透露。

两个笼子轮运动和跑步机上跑步运动疗法已被用于在我们以前的研究16,21,22。所有的动物都保存在专门的运动室，以尽量减少对环境或人类的干扰。动物的任何干扰"生物节律可以显著影响动物的运动性能。因此，整个动物保健，性能监控和记录，以及运动疗法应在每天相同的时间框架内完成。调查人员应尽量避免额外的压力对动物通过展示的预防措施，如限制应激暴露的持续时间和覆盖的笼子在适当的时候。 凯奇转轮运动 当准备笼子轮装置，车轮应该是方便的鼠标，安装并退出。应在笼的底部最小的床上用品，以防止老鼠从嵌套围绕轮盘和阻碍频率Ë轮运动。磁传感器被放置在轮的固定部分和固定到所述自由轮的磁体，以便计数轮每转一圈的鼠标行使。附着在磁传感器记录装置应放置在上述保持器的电线盖，及所有裸露的电线必须要么胶带或保持够不着从小鼠中，以便提供对咀嚼损伤的屏障。磁体和磁传感器也被贴在适当的结构以防止二者损坏和不希望的运动。在日常的数据收集，所有的录音线必须密切检查，以确保电线保持完好，并应retaped是必要的。因为它可能需要数天的小鼠，以适应新的环境，并开始定期锻炼，数据记录应针对每个个体动物。虽然动物往往要花费许多人力资源上的车轮，大部分的运行发生在夜间。因此，一些动物可能揭示HIGH性能的活动，即使他们出现在白天蛰伏。此外，它的情况并不少见，有些动物会表现出有限的笼子轮运行。通常情况下，笼轮的活动，相当于平均1公里/天以下是随机的笼子里活动的结果，并没有与任何显著运行的刺激有关。因此，动物的运行参数为1千米/天或1小时/天或更少应该被排除在最终的数据分析nonrunners。 取决于个体的研究，涉及小鼠的菌株的具体目的，运动的长度可以改变。我们以前的研究表明，对于涉及两个小鼠品系，平均轮运行速度和运动上笼轮的持续时间逐渐增加随着时间的推移和4周趋于​​稳定。 凯奇轮运转是在动物模型中一种自愿性的锻炼，这是一般取为压力较小的情况下被执行。 UNL其中某些参数可预置IKE跑步机上锻炼，在测试笼子里所有正在运行的是自愿的，容易受环境或行为线索。因此，通常可以避免不同的实验设置之间的比较，除非使用大型实验组号码。其他潜在的研究领域，可以从这种类型的活动范式的受益是把行为反应与生理后果，如认知神​​经科学的研究。对于利用自愿笼轮子范式的研究中，最关键的步骤是笼设置和监测工作的活动。 跑步机上锻炼 驯化步骤是必须的准备工作动物在跑步机上锻炼的协议。取决于个体研究的具体目的，驯化期间的持续时间可以变化。在这项研究中，为期一周的被选择让动物acclim吃了跑步机设备和电机的噪声在笼子里，热身期间是运动训练方案的一个重要步骤。类似人类同行，老鼠必须进入任何剧烈运动疗法，以避免受伤和潜在的数据工件前热身。在这项研究中，我们开始带以尽可能低的速度（4米/分钟）与电网的冲击关闭15分钟。所涉及的所有小鼠开始运行和关闭的皮带。震荡网格然后接通和皮带速度​​保持在4米/分钟。为了避免过度强调小鼠，我们设置了电击在温和的水平（1赫兹）。在几天内，所有小鼠适应了跑步机皮带。 行驶的持续时间以及加速度，也可以针对每个实验设计。推荐的启动加速度为1米/分钟。小鼠呈现枯竭的迹象的跑步机通道，必须立即关闭。疲惫的迹象通常由英里展出行政长官在跑步机上载列如下: <ol><li>连续暴露在冲击电网&gt; 5-10秒， </li><li>运行能力造成冲击电网曝光&gt; 5倍单运动养生期间和损失</li><li>运行能力造成冲击电网暴露的损失单一的运动处方中&gt; 2秒的至少3倍。 </li></ol>上面列出的值将被用于主要是作为一个特定的实验指导方针。这些参数需要为每一个练习的协议确定，因为它从不同小鼠品系小鼠品系，实验进行实验，并调查调查。 不像笼轮运动，跑步机的研究必须重复几次，以消除对环境和生物因素上运行的性能的影响。这是并不少见的，即使是同一种动物的测试运行之间的性能波动。由于每个运动养生自曝运行小鼠的STREβ，测试之间的适当的休息间隔是必要的。根据我们的经验，2-3天每次测试之间的休息时间为足以让老鼠恢复。 如同人类的跑步机上锻炼时，速度耐力运动的心脏率或血氧饱和度，一个技术上的困难的壮举与小鼠的​​研究是最好的决定。跑步研究中最关键的步骤是判定用尽，这通常单个小鼠之间变化的，并且被设置为80％的最大速度。在我们自己的经验结果的记录可能个别研究人员之间的显著差异，因为标准的疲惫大多是主观的。因此，相同的标准在记录过程中一致的应用是至关重要的。

自然发病更年期的特点是卵巢原始卵泡通过闭锁的枯竭，最终导致卵巢衰老。在美国，一个妇女一生中超过30％将用于绝经后。为应对卵巢功能下降，一些生理和心理后果可能发生，包括增加肥胖，血管不稳定，情绪或睡眠障碍。因为肥胖/肥胖的增加而绝经前对口1,2相比，绝经后的妇女特别易受影响代谢综合征和相关的并发症，包括心血管疾病（CVD）3。此外，它正变得越来越明显的是，成功的心脏康复患者的CVD包括规律的有氧运动，而且运动可以降低心血管疾病发病率和死亡率在这些科目4,5。然而，如何运动能力茶在售前到售后，绝经过渡期民营企业还没有得到很好的研究。更重要的是，运动作为一种预防性措施，在绝经后妇女心血管疾病风险的作用仍然相当充分研究。 用于妇女绝经模仿最常用的诱导模型是卵巢切除（OVX），啮齿目动物。近日，职业性化学性4 -乙烯基环己烯二环氧化物（VCD）已被证明通过加快闭锁的自然过程，最终导致与在其他组织中6未发现明显毒性卵巢功能衰竭专门针对小规模小学和原始卵泡。 VCD治疗的小鼠的动情周期，这是类似于一个人的月经周期，完成VCD注射停止后2-3个月内，这反过来又导致雌激素的逐渐撤离，模仿围与绝经后过渡。因此，卵泡消耗臭氧层，卵巢完好的动物十分接近人类自然通过发展预以近绝经后过渡7-9。此外，它提供了一种非手术的替代OVX模型减少手术的并发症，如感染。在这项研究中，我们采用VCD诱发更年期小鼠来研究运动对更年期小鼠心脏适应的影响。 有迹象表明，建议在男女双方的鼠类10-13雌激素和保持架转轮运动之间的关联几个实例。雌激素耗竭手术去势降低小鼠和大鼠14,15自愿演习活动。练习用两种方法在这项研究中，测试VCD诱发更年期小鼠的运动能力。凯奇轮运转被普遍认为是在动物模型中一种自愿性的锻炼和表现，想必，压力较小的情况下。然而，笼子轮运行并不表示动物的相对运动能力，这就要求对动物的健身活动SE在更高的层次。强迫跑步锻炼是用来确定运动能力，衡量一个宽容运动强度和耐力。此外，我们先前已经表明，笼转轮运动提供了一个刺激心肌肥厚和增生性这是成长性依赖16。所以，我们还测量了心脏适应于VCD诱发更年期笼的小鼠转轮运动。

<table><tbody><tr><td>4-Vinylcyclohexene diepoxide (VCD)</td><td>Sigma</td><td>V-3630</td><td></td></tr><tr><td>11.5 cm Diameter wheel with a 5.0 cm wide running surface</td><td>Petsmart</td><td>model 6208</td><td></td></tr><tr><td>Digital magnetic counter</td><td>Sigma Sport</td><td>model BC 600</td><td></td></tr><tr><td>Treadmill exercise</td><td>Columbus Instruments</td><td>model 1055SDRM</td><td></td></tr></tbody></table>

a method to study the impact of chemically-induced ovarian failure on exercise capacity and cardiac adaptation in mice

心血管疾病（CVD）的风险增加了绝经后的妇女，但是，锻炼的作用，作为一种预防性措施，在绝经后妇女心血管疾病的风险并没有得到充分研究。因此，我们研究了自愿笼轮运动和心脏适应更年期小鼠强迫跑步机上锻炼的影响。用于妇女绝经模仿最常用的诱导模型是卵巢切除（OVX），啮齿目动物。然而，去势模型从更年期在人类几相异性。在这项研究中，我们给予4 - 乙烯基环己烯二环氧化物（VCD）到雌性小鼠，从而加速卵巢功能衰竭的替代更年期模型来研究在绝经期小鼠运动的影响。 VCD选择性地加速了初级和原始卵泡导致的内分泌状态密切从模仿自然进程的损失预以近至绝经后的人类。要确定运动对运动的影响能力和心脏适应VCD治疗的雌性小鼠，两种方法被使用。首先，我们曝光了一组视像处理和未经处理的小鼠对自愿笼轮。第二，我们被迫使用跑步机运动，以确定测量公差运动强度和耐力独立成团VCD处理和未经处理的小鼠运动能力。

一个典型的实验方案，如在本研究中使用的，示于图1。继VCD治疗连续20天，阴道细胞学检查是用来确定在VCD注入小鼠体内是否存在周期性的;媒介物注射的小鼠也进行监测。在发情周期的阶段是由上皮细胞，角化上皮细胞，白细胞和在阴道涂片（ 图2A）的比例确定。仅角质化上皮细胞的存在所指示的鼠标是在发情期，因此仍然骑自行车。动情周期小鼠一般持续4天，因此VCD注射的动物被认为是后无环执着动情，15天中，阴道细胞学检查发现白细胞的大部分（ 图2A）。 VCD治疗的小鼠的动情周期变得不规则，再VCD注射开始（ 图2B）后2-3个月内停止</STRONG&gt;。 VCD卵巢组织呈显著萎缩，与对照组相比。在高倍镜下，我们看到小学和原始卵泡（ 图2C）的完全耗尽。 为了测试VCD诱发卵巢早衰的运动表现的影响，我们随机7个月旧VCD和媒介物处理的小鼠（双株，C57BL / 6和B6C3F1）要么久坐或笼轮运动组4-6周后停止循环（ 图1）。整个4周的运动期间，记录每天的锻炼价值的时间和距离对于每个行使动物。日常运行中值的距离，时间和速度的每周的平均值示于图3为与车辆或VCD处理的C57BL / 6小鼠。我们没有看到由平均每天的时间，距离和速度上在任C57BL / 6或B6C3F1鼠标VCD和车辆处理组之间的笼测量车轮运动表现显著差异菌株（ 图4A）。计算出的车轮运行的速度超过先前16所示的4个星期的运行周期逐渐增多，但并没有实验组之间的显著差异。这些研究表明VCD引起的卵巢功能衰竭自愿笼转轮运动的影响超越了至少2个小鼠品系。 继笼自愿转轮运动协议的期限，身体形态测量，记录和心脏被迅速切除，称重。之前我们已经表明，自愿笼转轮运动诱发增加心脏重量16。在这里，我们也显示心肌肥厚响应笼转轮运动是绝对的心脏重量（HW）测量和硬件标准化，以胫骨长度（TL）（ 表1）。相比同行久坐的绝对心脏质量和HW / TL比值均显著更大的车辆和VCD治疗的小鼠。然而，没有measurable差异控制和VCD引起的卵巢功能衰竭的小鼠下笼自愿转轮运动之间的心肌肥厚。 接下来，我们使用非自愿（强制）跑步机上运行，​​以确定运动能力。我们首先进行小鼠对高强度的协议，增加平均速度逐步每10分钟直到筋疲力尽。平均来说，载体处理的小鼠的最大速度为26.7±0.95米/分钟和VCD处理的小鼠的最大速度为用尽前28±1.4米/分钟。我们在使用运行20米/分钟（约80％最大），以测定对续航能力较低强度。 如图4B所示 ，在高或低强度的运动能力无显著差异车辆和视像处理组之间被发现，表明VCD诱导的卵巢功能衰竭，对小鼠的运动能力没有影响。 总之，更年期引起的VCD疗法对无论是自愿或被迫R无影响<html> unning容量，以及在心脏的适应性反应来锻炼。 <img alt="图1" fo:content-width="5in" fo:src="/files/ftp_upload/51083/51083fig1highres.jpg" src="/files/ftp_upload/51083/51083fig1.jpg" /> 图1:VCD-管理和实验方案。小鼠施用VCD（160毫克/千克）或香油（车辆控制）开始于2月龄，连续20天。以下VCD或车辆注射，动情周期监测和旋回分别如在方法部分中描述的确定。卵巢功能丧失发生在60-90天，是由阴道细胞学检查确诊。 7个月的年龄，老鼠被暴露在自愿笼转轮运动4周或遭受跑步机运动方案。 oad/51083/51083fig2highres.jpg"SRC ="/ files/ftp_upload/51083/51083fig2.jpg"/&gt; 。小鼠图2 VCD诱导卵巢功能衰竭答:从车辆注入小鼠表现出发情周期的四个阶段未染色阴道涂片细胞学:1）发情前期，主要包括有核上皮细胞2）发情期，其特点是未-成核的角化细胞; 3）发情后期，包括有核上皮细胞（箭头），未核角化上皮细胞（箭头）和白细胞; 4）动情，白细胞主要包括;和B:车辆和VCD注射组的发情周期平均长度（天）。一个周期是从在发情期的第一天测到在发情期下一期的第一天。阴道细胞学开始大约6周后注射的发作。 VCD-注射的动物的动情周期变得不规律，比注射赋形剂的动物s，而12后的发情周期，所有的VCD细胞学注射的动物不再显示循环C:H＆E在8月龄卵巢组织染色。横截面为5μm的厚度做了车辆处理（左图）和VCD处理（右图）小鼠20​​X（上图）和200X（下图）的放大倍率。在放大200倍的卵巢组织显示了一些在车辆相比，VCD治疗的小鼠卵泡。 <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/51083/51083fig2highres.jpg" target="_blank">请点击这里查看这个数字的放大版本。</a> <img alt="图3" fo:content-width="5in" fo:src="/files/ftp_upload/51083/51083fig3highres.jpg" src="/files/ftp_upload/51083/51083fig3.jpg" /> 图3:在车辆或VCD的处理C57BL/6J雌性小鼠自愿笼轮的表现。左图:平均跑动距离（公里/天），每1周的时间超过4周的研究期间中间面板:平均运行时间（小时/天），每1周的时间超过4周的研究期间右面板:平均运行速度（公里/小时），每1周时间内超过4周的研究期间， <a href="https://jove.unicartagenaproxy.elogim.com/files/ftp_upload/51083/51083fig3highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" fo:content-width="5in" fo:src="/files/ftp_upload/51083/51083fig4highres.jpg" src="/files/ftp_upload/51083/51083fig4.jpg" /> 图4。自愿笼车轮和车辆与VCD治疗的小鼠跑步机的性能。答:（上图）的C57BL / 6平均行驶距离（公里/日）和时间，每24小时超过4周的研究期间花在轮（小时/天）。 （ 下图）＆＃160; B6C3F1平均行驶距离（公里/日）和时间，每24小时超过4周的研究期间 B花在轮（小时/天）:C57BL / 6跑步机运行参数。最大使用时间的方法部分的详细方案（ 上图）和最大速度（Botoom面板 ）。

Watch this Scientific Journal Video about A Method to Study the Impact of Chemically-induced Ovarian Failure on Exercise Capacity and Cardiac Adaptation in Mice at JoVE.com

A Method to Study the Impact of Chemically-induced Ovarian Failure on Exercise Capacity and Cardiac Adaptation in Mice

两个运动范式是一个新开发的化学诱导更年期小鼠模型测试，以检查更年期对运动能力和心脏的适应锻炼的影响。

一个方法研究的化学诱导卵巢早衰的运动能力和适应心脏的影响在小鼠

The risk of cardiovascular disease (CVD) increases in post-menopausal women, yet, the role of exercise, as a preventative measure for CVD risk in ...

a-method-to-study-impact-chemically-induced-ovarian-failure-on

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

15.3K 次观看.  University of Arizona. 两个运动范式是一个新开发的化学诱导更年期小鼠模型测试，以检查更年期对运动能力和心脏的适应锻炼的影响。 

视频: A Method to Study the Impact of Chemically-induced Ovarian Failure on Exercise Capacity and Cardiac Adaptation in Mice

Culture and Co-Culture of Mouse Ovaries and Ovarian Follicles

The mammalian ovary is composed of ovarian follicles, each follicle consisting of a single oocyte surrounded by somatic granulosa cells, enclosed together within a basement membrane. A finite pool of follicles is laid down during embryonic development, when oocytes in meiotic arrest form a close association with flattened granulosa cells, forming primordial follicles. By or shortly after birth, mammalian ovaries contain their lifetime&#8217;s supply of primordial follicles, from which point onwards there is a steady release of follicles into the growing follicular pool.
The ovary is particularly amenable to development in vitro, with follicles growing in a highly physiological manner in culture. This work describes the culture of whole neonatal ovaries containing primordial follicles, and the culture of individual ovarian follicles, a method which can support the development of follicles from an immature through to the preovulatory stage, after which their oocytes are able to undergo fertilization in vitro. The work outlined here uses culture systems to determine how the ovary is affected by exposure to external compounds. We also describe a co-culture system, which allows investigation of the interactions that occur between growing follicles and the non-growing pool of primordial follicles.

This protocol describes the primary culture/co-culture of mouse ovarian tissue, using ovaries from neonatal mice and individual ovarian follicles from prepubertal mice. The culture techniques support development in a highly physiological manner, allowing investigation of the effect of extrinsic agents on the ovary, and of interactions between ovarian follicles.

文化和共同培养小鼠的卵巢和卵巢卵泡

The mammalian ovary is composed of ovarian follicles, each follicle consisting of a single oocyte surrounded by somatic granulosa cells, enclosed together ...

科研

发育生物学

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 ...

行为学

The Treadmill Fatigue Test: A Simple, High-throughput Assay of Fatigue-like Behavior for the Mouse

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of current interventions to adequately treat fatigue in all patients leaves a need for new treatment options. Despite the therapeutic need and importance of preclinical research in helping identify promising novel treatments, few preclinical assays of fatigue are available. Moreover, the most common preclinical assay used to assess fatigue-like behavior, voluntary wheel running, is not suitable for use with some strains of mice, may not be sensitive to drugs that reduce fatigue, and has relatively low throughput. The current protocol describes a novel, non-voluntary preclinical assay of fatigue-like behavior, the treadmill fatigue test, and provides evidence of its efficacy in detecting fatigue-like behavior in mice treated with a chemotherapy drug known to cause fatigue in humans and fatigue-like behavior in animals. This assay may be a beneficial alternative to wheel running, as fatigue-like behavior and potential interventions can be assessed in a greater number of mice over a shorter time frame, thus permitting faster discovery of new therapeutic options.

Fatigue is a common, undertreated and frequently poorly-understood symptom in many diseases and disorders. New preclinical assays of fatigue may help to improve current understanding and future treatment of fatigue. To that end, the current protocol provides a novel means of measuring fatigue-like behavior in the mouse.

跑步机疲劳试验:疲劳类似的行为，为鼠标操作简单，高通量检测

Fatigue is a prominent symptom in many diseases and disorders and reduces quality of life for many people. The lack of clear pathogenesis and failure of ...

An In Vivo Estrogen Deficiency Mouse Model for Screening Exogenous Estrogen Treatments of Cardiovascular Dysfunction After Menopause

Postmenopausal women are at greater risk of developing cardiovascular diseases than premenopausal women. Female mice ovariectomized (OVX) at weaning display increased atherosclerotic lesions in the aorta compared with female mice with intact ovarian function. However, laboratory models involving estrogen-deficient mice with atherosclerosis-prone status are lacking. This deficit is crucial because clinical estrogen deficiency in menopausal women may aggravate the incidence of pre-existing or ongoing lipid disruption and atherosclerosis. In this study, we establish an in vivo estrogen-deficient mouse model by bilateral ovariectomy via a double dorsal-lateral incision in apolipoprotein E (apoE)-/- mice. We then compare the effects of 17&#946;-estradiol and pseudoprotodioscin (PPD) (a phytoestrogen) perorally administered via hazelnut spread. We find that although PPD exerts some effect on reducing final body weight and plasma TG in OVX apoE-/- mice, it has anti-atherosclerotic and cardiac-protective capacities comparable with its 17&#946;-estradiol counterpart. PPD is a phytoestrogen that has been reported to exert anti-tumor properties. Thus, the proposed method is applicable for screening phytoestrogens via peroral administration to substitute for traditional hormone replacement therapy in postmenopausal women, which has been reported to have potentially deleterious tumorigenetic capacity. Peroral administration via hazelnut spread is noninvasive, rendering it widely applicable to many patients. This article contains step-by-step demonstrations of bilateral ovariectomy via the double dorsal-lateral incision in apoE-/- mice and peroral 17&#946;-estradiol or phytoestrogen hormone replacement via hazelnut spread. Plasma lipid and cardiovascular function analyses using echocardiography follow.

Clinically, estrogen deficiency in menopausal women may aggravate the incidence of lipid disruption and atherosclerosis. We established an in vivo estrogen deficiency model by bilateral ovariectomy via a double dorsal-lateral incision in apoE-/- mice. The mouse model is applicable for screening exogenous estrogen treatments of cardiovascular dysfunction after menopause.

用于筛查绝经后心血管功能障碍的外源雌激素治疗的活体雌激素缺乏小鼠模型

Postmenopausal women are at greater risk of developing cardiovascular diseases than premenopausal women. Female mice ovariectomized (OVX) at weaning ...

生物化学

Orthotopic Ovarian Transplantation Procedures to Investigate the Life- and Health-span Influence of Ovarian Senescence in Female Mice

Ovarian transplantation was first conducted at Utah State University in 1963. In more recent work, heterochronic transplantation of mammalian ovaries is being used to investigate the health-protective effects of young ovaries in young females. The current procedures employ an orthotopic transplantation method, where allogenic ovaries are transplanted back to their original position in the ovarian bursa. This is in contrast to the more commonly used heterotopic transplantation of ovaries/ovarian tissue subcutaneously or under the kidney capsule. All three locations provide efficient revascularization of the transplanted tissues. However, orthotopic transplantation provides the ovary with the most natural signaling environment and is the only procedure that provides the opportunity for the animal to reproduce naturally post-operatively. One must take care to remove all endogenous ovarian tissue during the ovariectomy procedure. If any endogenous tissue remains or if only one ovary is removed, the transplanted tissue will remain dormant until the existing tissue becomes senescent. While revascularization of the transplanted ovaries occurs very quickly, the transplant recipient can take a considerable amount of time to adapt to a new hypothalamic/pituitary/gonadal/adrenal (HPG/A) axis signaling regime associated with the transplanted tissue. This normally takes about 100 days in the mouse. Therefore, transplantation experiments should be designed to accommodate this adaptation period. Typical results with ovarian transplantation will include changes in the health of the recipient that reflect the age of the transplanted ovary, rather than the chronological age of the recipient.

Described here are orthotopic ovarian transplantation protocols for modifying ovarian influence on the physiology of the recipient mouse. This surgical procedure is used to expose female mice to various types of ovarian influence and may include heterochronic transplantations and transplantation of ovaries that have undergone various ex vivo treatments.

原位卵巢移植术对雌性小鼠卵巢衰老的影响及寿命及健康范围研究

Ovarian transplantation was first conducted at Utah State University in 1963. In more recent work, heterochronic transplantation of mammalian ovaries is ...

Supramaximal Intensity Hypoxic Exercise and Vascular Function Assessment in Mice

Exercise training is an important strategy for maintaining health and preventing many chronic diseases. It is the first line of treatment recommended by international guidelines for patients suffering from cardiovascular diseases, more specifically, lower extremity artery diseases, where the patients' walking capacity is considerably altered, affecting their quality of life. 
Traditionally, both low continuous exercise and interval training have been used. Recently, supramaximal training has also been shown to improve athletes' performances via vascular adaptations, amongst other mechanisms. The combination of this type of training with hypoxia could bring an additional and/or synergic effect, which could be of interest for certain pathologies. Here, we describe how to perform supramaximal intensity training sessions in hypoxia on healthy mice at 150% of their maximal speed, using a motorized treadmill and a hypoxic box. We also show how to dissect the mouse in order to retrieve organs of interest, particularly the pulmonary artery, the abdominal aorta, and the iliac artery. Finally, we show how to perform ex vivo vascular function assessment on the retrieved vessels, using isometric tension studies.

High-intensity training in hypoxia is a protocol that has been proven to induce vascular adaptations potentially beneficial in some patients and to improve athletes&#39; repeated sprint ability. Here, we test the feasibility of training mice using that protocol and identify those vascular adaptations using ex vivo vascular function assessment.

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 ...

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, ...

Accurate Follicle Enumeration in Adult Mouse Ovaries

Sexually reproducing female mammals are born with their entire lifetime supply of oocytes. Immature, quiescent oocytes are found within primordial follicles, the storage unit of the female germline. They are non-renewable, thus their number at birth and subsequent rate of loss largely dictates the female fertile lifespan. Accurate quantification of primordial follicle numbers in women and animals is essential for determining the impact of medicines and toxicants on the ovarian reserve. It is also necessary for evaluating the need for, and success of, existing and emerging fertility preservation techniques. Currently, no methods exist to accurately measure the number of primordial follicles comprising the ovarian reserve in women. Furthermore, obtaining ovarian tissue from large animals or endangered species for experimentation is often not feasible. Thus, mice have become an essential model for such studies, and the ability to evaluate primordial follicle numbers in whole mouse ovaries is a critical tool. However, reports of absolute follicle numbers in mouse ovaries in the literature are highly variable, making it difficult to compare and/or replicate data. This is due to a number of factors including strain, age, treatment groups, as well as technical differences in the methods of counting employed. In this article, we provide a step-by-step instructional guide for preparing histological sections and counting primordial follicles in mouse ovaries using two different methods: [1] stereology, which relies on the fractionator/optical dissector technique; and [2] the direct count technique. Some of the key advantages and drawbacks of each method will be highlighted so that reproducibility can be improved in the field and to enable researchers to select the most appropriate method for their studies.

Here, we describe, compare, and contrast two different techniques for accurate follicle counting of fixed mouse ovarian tissues.

成人小鼠卵巢中的精确卵泡图解

Sexually reproducing female mammals are born with their entire lifetime supply of oocytes. Immature, quiescent oocytes are found within primordial ...

Methods for Studying Uterine Contributions to Pregnancy Establishment in an Ovariectomized Mouse Model

For pregnancy to be established, a viable blastocyst must successfully interact with a receptive uterine lining (endometrium) to facilitate implantation and placenta formation and enable ongoing pregnancy. The limitations to pregnancy success caused by embryonic defects are well known and have been largely overcome in recent decades with the rise of in vitro fertilization (IVF) and assisted reproductive technologies. As yet, however, the field has not overcome the limitations caused by an inadequately receptive endometrium, thus resulting in stagnating IVF success rates. Ovarian and endometrial functions are closely intertwined, as hormones produced by the ovary are responsible for the endometrium's menstrual cyclicity. As such, when using rodent models of pregnancy, it can be difficult to ascertain whether an observed result is due to an ovarian or uterine deficit. To overcome this, an ovariectomized mouse model was developed with embryo transfer or artificial decidualization to allow the study of uterine-specific contributions to pregnancy. This article will provide instructions on how to perform ovariectomy and offer insights into various techniques for supplying exogenous hormones to support successful artificial decidualization or pregnancy following embryo transfer from healthy donors. These techniques include subcutaneous injection, slow-release pellets, and osmotic mini pumps. The key advantages and disadvantages of each method will be discussed, enabling researchers to choose the best study design for their specific research question.

Pregnancy establishment is a dynamic process involving complex embryo and uterine crosstalk. The precise contributions of the maternal uterine environment to these processes remain an active area of investigation. Here, detailed protocols are provided to aid in designing in vivo animal models to address these research questions.

研究卵巢切除小鼠模型中子宫对妊娠建立的贡献的方法

For pregnancy to be established, a viable blastocyst must successfully interact with a receptive uterine lining (endometrium) to facilitate implantation ...

<meta charset="utf8"></head><body>通过控制来曲唑释放诱导小鼠多囊卵巢综合征

A Modified Technique for Inducing Polycystic Ovary Syndrome in Mice

Polycystic ovary syndrome (PCOS) is one of the leading causes of infertility in women. Animal models are widely used to study the etiologic mechanisms of PCOS and for related drug development. Letrozole-induced mouse models replicate the metabolic and reproductive phenotypes of patients with PCOS. The traditional method of letrozole treatment in PCOS mice requires daily dosing over a certain period, which can be labor-intensive and cause significant stress to the mice. This study describes a simple and effective method for inducing PCOS in mice by implanting a controlled letrozole-releasing mini-pump. A mini-pump capable of stable, continuous release of a quantitative amount of letrozole was fabricated and implanted subcutaneously in mice under anesthesia. This study demonstrated that the mouse model successfully mimicked PCOS features after letrozole mini-pump implantation. The materials and equipment used in this study are readily available to most laboratories, requiring no special customization. Collectively, this article provides a unique, easy-to-perform method for inducing PCOS in mice.

<meta charset="utf8"></head><body>作者聚焦：研究多囊卵巢综合征的机制和诱导模型

This protocol introduces a technique for inducing polycystic ovary syndrome (PCOS) models in mice through controlled letrozole release using mini-pumps. Under adequate anesthesia, the mini-pump was implanted subcutaneously, and PCOS was successfully induced in the mice after a certain period of the mini-pump release.

诱导小鼠多囊卵巢综合征的改良技术

Polycystic ovary syndrome (PCOS) is one of the leading causes of infertility in women. Animal models are widely used to study the etiologic mechanisms of ...