Evaluation of Human Spermatozoa Mitochondrial Membrane Potential Using the JC‐1 Dye

膜电位 线粒体 线粒体内膜 电容 顶体反应 线粒体凋亡诱导通道 运动性 生物物理学 细胞生物学 生物 线粒体载体 化学 生物化学 细菌外膜 体外 基因 大肠杆菌
作者
David F. Carrageta,Laís Freire‐Brito,Pedro F. Oliveira,Marco G. Alves
出处
期刊:Current protocols [Wiley]
卷期号:2 (9) 被引量:21
标识
DOI:10.1002/cpz1.531
摘要

Mitochondria are fundamental for human spermatozoa motility and fertilizing ability. Mitochondria participate not only in ATP production, but also in reactive oxygen species production, redox equilibrium, and calcium regulation, all of which are central for human spermatozoa motility, capacitation, acrosome reaction, and ultimately, oocyte fertilization. Mitochondrial membrane potential is a key indicator of mitochondrial health and activity. Most commonly used methods for the study of mitochondrial membrane potential, however, cannot be applied to human spermatozoa due to their unique characteristics, including high motility and time-dependent decay of quality, limiting the study of this important parameter in these cells. Here, we describe an easy, fast, and cheap protocol for the quantitative evaluation of human spermatozoa mitochondrial membrane potential, using the fluorescent cationic dye 5,5,6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimi-dazoylcarbocyanine iodide (JC-1). JC-1 is a sensitive marker for mitochondrial membrane potential, exhibiting a potential-dependent accumulation in the mitochondria. At high mitochondrial membrane potential, JC-1 forms J-aggregates, which emit red fluorescence, whereas at low mitochondrial membrane potential, JC-1 remains at its monomer state, which emits green fluorescence. We first describe how to evaluate human spermatozoa mitochondrial membrane potential using JC-1 and a fluorescence plate reader, for high-throughput studies. The calculation of the JC-1 ratio (indicative of the J-aggregates/monomers ratio) is then used to quantitatively evaluate mitochondrial health and activity. In addition, we describe an imaging protocol for the qualitative evaluation of human spermatozoa mitochondrial membrane potential using a fluorescence microscope. This allows for a visual analysis of the results that can complement the quantitative data. These protocols can be used to study the effects of spermatozoa exposure to compounds of interest, and alterations due to diseases or different conditions. While these protocols are illustrated with human spermatozoa, they can be adapted and used on spermatozoa of different species. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Quantitative evaluation of human spermatozoa mitochondrial membrane potential using the JC-1 dye and a fluorescence plate reader Basic Protocol 2: Qualitative evaluation of human spermatozoa mitochondrial membrane potential using the JC-1 dye and fluorescence microscopy Support Protocol: Preparation of the JC-1 working solution.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
刚刚
2秒前
小马甲应助Lam采纳,获得10
2秒前
JJXIONG完成签到,获得积分10
3秒前
河马发布了新的文献求助10
4秒前
4秒前
可爱的函函应助肖肖采纳,获得10
5秒前
CodeCraft应助平常的紫蓝采纳,获得10
5秒前
好宝宝完成签到,获得积分10
5秒前
3237924531发布了新的文献求助10
6秒前
hdy331完成签到,获得积分10
6秒前
完美世界应助123采纳,获得10
7秒前
7秒前
怕孤独的忆南完成签到,获得积分10
9秒前
追风完成签到 ,获得积分10
10秒前
10秒前
yao完成签到,获得积分10
10秒前
常芹发布了新的文献求助10
11秒前
ED应助科研路漫漫采纳,获得10
11秒前
12秒前
布鲁克完成签到,获得积分10
13秒前
13秒前
量子星尘发布了新的文献求助10
14秒前
今后应助朴实的绣连采纳,获得30
14秒前
<小天才>发布了新的文献求助10
16秒前
16秒前
17秒前
Smy完成签到 ,获得积分10
17秒前
在水一方应助梁晓雯采纳,获得10
18秒前
Yy123发布了新的文献求助10
18秒前
tao发布了新的文献求助10
18秒前
3237924531完成签到,获得积分10
18秒前
健忘小霜完成签到,获得积分10
19秒前
20秒前
scholar完成签到,获得积分10
21秒前
wei发布了新的文献求助10
21秒前
鳗鱼灵阳完成签到,获得积分20
22秒前
22秒前
23秒前
无情的聋五完成签到 ,获得积分10
23秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
‘Unruly’ Children: Historical Fieldnotes and Learning Morality in a Taiwan Village (New Departures in Anthropology) 400
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
基于可调谐半导体激光吸收光谱技术泄漏气体检测系统的研究 350
Robot-supported joining of reinforcement textiles with one-sided sewing heads 320
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3988838
求助须知:如何正确求助?哪些是违规求助? 3531250
关于积分的说明 11252914
捐赠科研通 3269838
什么是DOI,文献DOI怎么找? 1804820
邀请新用户注册赠送积分活动 881943
科研通“疑难数据库(出版商)”最低求助积分说明 809028