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Relationship between sperm mitochondrial membrane potential, sperm motility, and fertility potential Tsuyoshi Kasai, Keigo Ogawa, Kaoruko Mizuno, Seiichiro Nagai, Yuzo Uchida, Shouji Ohta, Michiko Fujie, Kohta Suzuki, Shuji Hirata, Kazuhiko Hoshi Department of Obstetrics and Gynecology, Yamanashi Medical University, Yamanashi 409-3898, Japan Asian J Androl 2002 Jun; 4: 97-103 Keywords:
|
|
Mitochondrial
membrane potential |
P
value |
||
High |
Moderate |
Low |
||
No.
of patients |
8 |
5 |
13 |
|
CON
(M/mL) |
57.7
33.6 |
44.3
14.5 |
60.4
41.2 |
|
%MOT |
91.1
8.5 |
94.0
5.5 |
63.0
3.7 |
a |
%PMOT |
50.4
19.9 |
60.0
25.2 |
43.5
29.4 |
|
VAP
(mm/s) |
73.0
14.2 |
68.1
15.5 |
52.1
12.5 |
b,c |
VSL
(mm/s) |
54.8
13.9 |
55.9
17.5 |
42.6
12.4 |
|
VCL
(mm/s) |
127.0
28.1 |
106.9
15.0 |
87.0
22.6 |
b |
ALH
(mm) |
4.5
1.3 |
4.4
0.6 |
3.6
1.3 |
|
BCF
(Hz) |
25.0
6.4 |
17.9
1.5 |
24.5
8.6 |
|
STR
(%) |
75.4
10.3 |
75.5
6.5 |
81.7
7.3 |
|
LIN
(%) |
45.7
8.3 |
44.5
4.2 |
52.3
10.7 |
|
%HA |
27.3
23.6 |
13.8
7.2 |
7.2
9.0 |
b |
The clinical characteristics of patients and the comparative outcomes of IVF-ET are shown in Table 2. There were no significant differences in the demographic characteristics among the three groups. The fertilization rates and the proportions of fertilized oocytes are significantly higher in the high potential compared with the low potential group. Total fertilization failure occurred only in the low potential group. However, pregnancy rates were the same in the three groups.
Table2. Demographic characteristics and outcomes of IVF-ET corresponding to three mitochondrial membrane potentials. a: High vs. Low, P<0.05, b: Moderate vs. Low, P <0.01.
|
Mitochondrial
membrane potential |
P
value |
||
High |
Moderate |
Low |
||
No.
of patients |
8 |
5 |
13 |
|
Female
age (years) |
35.6
4.4 |
34.0
2.6 |
36.8
3.7 |
|
Infertility
(%) |
||||
primary |
62.5 |
60.0 |
76.9 |
|
secondary |
37.5 |
40.0 |
23.1 |
|
Duration
of infertility (months) |
46.1
14.5 |
58.2
34.6 |
69.5
33.3 |
|
Fertilization
rate (%) |
80.5
10.2 |
78.9
10.4 |
50.4
31.4 |
a |
Fertilized
oocytes (%) |
87.5(49/56) |
80.0(36/45) |
59.0(69/117) |
a,b |
Total
fertilization failure (%) |
0.0(0/8) |
0.0(0/5) |
15.4(2/13) |
|
Pregnancy
rate (%) |
37.5(3/8) |
40.0(2/5) |
23.1(3/13) |
|
4 Discussion
The importance of sperm motility during the fertilization process has received considerable attention over the past decades. Several researchers have reported the relationship of fertility potential in vitro and sperm motility parameters measured with CASA [21]. Objective analysis of sperm motility parameters resulted in significant correlations between the value of ALH [22], VCL [23-25], VAP [26], LIN [24] and the in vitro fertilization rates. In addition to VCL and VAP, sperm hyperactivation has been shown to be an important marker of fertilizing ability in the in vitro situation [27-30].
Energy is stored in the mitochondria as a proton concentration gradient and an electric potential gradient across the membrane. These gradients are generated by electron transport maintained by the inner mitochondrial membrane and drive the synthesis of ATP. Membrane permeable lipophilic cations accumulate in the mitochondria and exhibit a negative interior membrane potential. Recently, the lipophilic cationic fluorescent carbocyanine dye, JC-1, has been used to differentially label mitochondria with high and low membrane potential. When JC-1 forms monomers in mitochondria with low potential, the JC-1 stain emits a green fluorescence (510-520 nm), while the JC-1 form multimers known as J-aggregates after accumulation in mitochondria with high membrane potential, the JC-1 stain emits a bright red-orange fluorescence at 590 nm. Any changes in mitochondrial membrane potential could be a good indicator of sperm motility. Mitochondria membrane potential of spermatozoa has been evaluated with JC-1 in a variety of species [31-33], including human [34,35]. However, evaluation of the relationship between the mitochondria membrane potential and the sperm fertility potential has not yet been reported for human spermatozoa.
In the present study, JC-1 was used with flow cytometry to assess the sperm mitochondrial functional status. The use of flow cytomery allows the analysis of large populations of spermatozoa from a given sample. Our results show that there are significant differences in %MOT (P<0.05), VAP (P<0.01) and VCL (P<0.01) between the high and low mitochondrial membrane potential group. These results suggest that there might be significant correlations between the value of %MOT, VAP and VCL reflecting the fertility potential in vitro, and the mitochondrial membrane potential. Our results also show that there are significant correlation between the percentage of %HA and the mitochondrial membrane potential.
In conclusion, the results indicate that as the sperm mitochondrial membrane potential increases, sperm motility parameters, such as %MOT, VAP, VCL and %HA, also increase together with their fertility potential. When the sperm mitochondrial membrane potential is low, application of ICSI to couples with unexplained infertility should be considered. It is suggested that the JC-1 staining method is useful to predict of sperm fertility potential.
Acknowledgements
We thank Ms. K.L.K. Tamashiro, Department of Psychiatry, University of Cincinnati, for her assistance in the preparation of the manuscript.
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Correspondence
to: Prof. Kazuhiko HOSHI,
Department of Obstetrics and Gynecology, Yamanashi Medical University,
Shimokato 1110, Tamaho, Nakakoma, Yamanashi 409-3898, Japan.
Tel: +81-55-273-1111 Ext. 2360, Fax: +81-55-273-3746
E-mail: kazuho@res.yamanashi-med.ac.jp
Received 2002-04-29
Accepted 2002-05-08