[Evaluation of sperm mitochondrial membrane potential by JC-1 fluorescent staining and flow cytometry].
OBJECTIVE
To investigate the feasibility and clinical significance of detecting sperm mitochondrial membrane potential (MMP) by JC-1 fluorescent staining and flow cytometry, and to explore the relationship between the results of JC-1 staining and seminal parameters.


METHODS
Sixty-three semen samples were divided into a fertile (n = 31) and an infertile group (n = 32) and underwent computer-assisted semen analysis (CASA). All the samples were washed, followed by JC-1 staining and evaluation of sperm MMP by flow cytometry. The percentage of normal sperm MMP was indicated as the percentage of sperm emitting orange-red fluorescence (JC-1 + %).


RESULTS
The JC-1 + % was significantly higher in the fertile group than in the infertile one ([75.89 +/- 15.69]% vs [54.04 +/- 22.21] %, P = 0.000), correlated positively with sperm motility (r = 0.610, P = 0.000) and the percentage of grade a + b sperm (r = 0.614, P = 0.000) and negatively with grade d sperm (r = -0.504, P = 0.000). There was a significant positive correlation between the results of JC-1 staining (JC-1 + %) and that of Rh123 /PI dual fluorescent staining (Rh123 + / PI (-)%) (r = 0.938, P = 0.000).


CONCLUSIONS
JC-1 staining and flow cytometry could readily and quickly detect sperm MMP and the sperm JC-1 + % could be an auxiliary marker for the diagnosis of male infertility.
Xia
Xin-Yi
XY
Dept. of Reproduction & Genetics, PLA Research Institute of Clinical Laboratory Medicine, School of Clinical Medicine, Nanjing University Medical College/Nanjing General Hospital of Nanjing Military Region, PLA, Jiangsu 210002, China. xiaxy@androl.cn
Wu
Yong-Ming
YM
Hou
Bao-Shan
BS
Yang
Bin
B
Pan
Lian-Jun
LJ
Shi
Yi-Chao
YC
Jin
Bao-Fang
BF
Shao
Yong
Y
Cui
Ying-Xi
YX
Huang
Yu-Feng
YF
D000328:Adult; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000502:physiology*; D008875:Middle Aged; D015203:Reproducibility of Results; D013081:Sperm Motility / Q000502:physiology; D013094:Spermatozoa / Q000502:physiology*; D013194:Staining and Labeling / Q000379:methods
D005456:Fluorescent Dyes