Laboratorio de Ecofisiología de Organismos Acuáticos, Departamento de Biotecnología Marina, Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE), Ensenada, Baja California, México.
Departamento de Fisiopatología Cardio-Renal, Instituto Nacional de Cardiología "Ignacio Chávez", Mexico City, Mexico.
PLoS One. 2022 Aug 29;17(8):e0273554. doi: 10.1371/journal.pone.0273554. eCollection 2022.
Mitochondrial respirometry is key to understand how environmental factors model energetic cellular process. In the case of ectotherms, thermal tolerance has been hypothesized to be intimately linked with mitochondria capability to produce enough adenosine triphosphate (ATP) to respond to the energetic demands of animals in high temperatures. In a recent study made in Octopus maya was proposed the hypothesis postulating that high temperatures could restrain female reproduction due to the limited capacity of the animals' heart to sustain oxygen flow to the body, affecting in this manner energy production in the rest of the organs, including the ovarium Meza-Buendia AK et al. (2021). Unfortunately, until now, no reports have shown temperature effects and other environmental variables on cephalopod mitochondria activity because of the lack of a method to evaluate mitochondrial respiratory parameters in those species' groups. In this sense and for the first time, this study developed a method to obtain mitochondrial respirometry data of adult Octopus maya's heart. This protocol illustrates a step-by-step procedure to get high yield and functional mitochondria of cephalopod heart and procedure for determining the corresponding respiratory parameters. The procedure described in this paper takes approximately 3 to 4 hours from isolation of intact mitochondria to measurement of mitochondrial oxygen consumption.
线粒体呼吸测量是理解环境因素如何模拟能量细胞过程的关键。对于变温动物，热耐受性被假设与线粒体产生足够三磷酸腺苷（ATP）的能力密切相关，以满足动物在高温下的能量需求。在最近对章鱼的一项研究中，提出了一个假设，即高温可能会由于动物心脏维持氧气流向身体的能力有限而限制雌性繁殖，从而影响包括卵巢在内的其他器官的能量产生。Meza-Buendia AK 等人。（2021）。不幸的是，到目前为止，由于缺乏评估这些物种组中线粒体呼吸参数的方法，还没有报告显示温度影响和其他环境变量对头足类动物线粒体活性的影响。从这个意义上说，这是首次开发了一种方法来获得成年章鱼心脏的线粒体呼吸测量数据。该方案说明了获得头足类动物心脏高产和功能线粒体的分步程序，以及确定相应呼吸参数的程序。本文所述的程序从完整线粒体的分离到线粒体耗氧量的测量大约需要 3 到 4 个小时。
