Comparison of four fluorochromes for the detection of the inner mitochondrial membrane potential in human spermatozoa and their correlation with sperm motility.
BACKGROUND
Sperm motility evaluation is associated with fertility in IVF programmes. The visual estimation of sperm motility is extremely subjective. Hence, alternative methods are required. Among them, determination of mitochondrial membrane potential (Deltapsi(m)) changes of spermatozoa using potentiometric dyes may be a reliable test to determine sperm quality. However, the use of the potentiometric dyes in sperm samples has not been compared.


METHODS
We have studied sperm samples from 28 infertile patients enrolled in an IVF programme in flow cytometry after staining of spermatozoa with four commonly used potentiometric dyes. Sperm motility was evaluated visually.


RESULTS
As expected, JC-1 seems to detect specifically Deltapsi(m) changes, CMX-Ros, DiOC(6)(3) and TMRE fluorescence is easily analysed and the latter three fluorochromes are particularly suitable for multiparametric staining. Irrespective of the Deltapsi(m)-dependent fluorochromes used to stain spermatozoa, a positive correlation was found between the percentage of Deltapsi(m)(high) cells and forward motility and also with high fertilization rates after IVF.


CONCLUSIONS
The four fluorochromes may be useful for evaluation of sperm samples from infertile patients. The choice of the potentiometric dyes will depend on their fluorescence characteristics in order to use them in combination with other fluorescent markers.
Marchetti
Carole
C
INSERM U459, Laboratoire d'Histologie, Faculté de Médecine 1, Place Verdun, 59045 Lille Cedex, France.
Jouy
Nathalie
N
Leroy-Martin
Brigitte
B
Defossez
André
A
Formstecher
Pierre
P
Marchetti
Philippe
P
D000328:Adult; D001562:Benzimidazoles; D002232:Carbocyanines; D002470:Cell Survival; D005306:Fertilization; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D007248:Infertility, Male / Q000175:diagnosis* / Q000503:physiopathology*; D008297:Male; D008564:Membrane Potentials; D008928:Mitochondria; D009930:Organic Chemicals; D009942:Organometallic Compounds; D012662:Semen Preservation; D012680:Sensitivity and Specificity; D013081:Sperm Motility; D013094:Spermatozoa; D013194:Staining and Labeling
D001562:Benzimidazoles; D002232:Carbocyanines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D009942:Organometallic Compounds; C107472:red dye CMXRos; C110932:tetramethyl rhodamine ethyl ester; C068624:5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine; C007392:3,3'-dihexyl-2,2'-oxacarbocyanine