Manee-In, S.
(Department of Clinical Sciences and Public Health Medicine, Faculty of Veterinary Science, Mahidol University)
,
Parmornsupornvichit, S.
(Faculty of Veterinary Science, Mahidol University)
,
Kraiprayoon, S.
(Faculty of Veterinary Science, Mahidol University)
,
Tharasanit, T.
(Faculty of Veterinary Science, Chulalongkorn University)
,
Chanapiwat, P.
(Department of Clinical Sciences and Public Health Medicine, Faculty of Veterinary Science, Mahidol University)
,
Kaeoket, K.
(Department of Clinical Sciences and Public Health Medicine, Faculty of Veterinary Science, Mahidol University)
Cryopreservation of epididymal sperm is an effective technique to preserve genetic materials of domestic cats and wild felids when they unexpectedly die. However, this technique inevitably causes detrimental changes of cryopreserved-thawed spermatozoa, for example, by physical damage and excessive o...
Cryopreservation of epididymal sperm is an effective technique to preserve genetic materials of domestic cats and wild felids when they unexpectedly die. However, this technique inevitably causes detrimental changes of cryopreserved-thawed spermatozoa, for example, by physical damage and excessive oxidative stress. L-carnitine is an antioxidant that has been used to improve sperm motility in humans and domestic animals. This study aimed to investigate the effects of L-carnitine on cat epididymal sperm quality following cryopreservation and thawing. After routine castration, cauda epididymides were collected from 60 cat testes. The epididymal spermatozoa from 3 cauda epididymides were pooled as 1 replicate. Spermatozoa samples (16 replicates) were examined for spermatozoa quality and then randomly divided into 4 groups: 0 mM L-carnitine (control), 12.5 mM, 25 mM and 50 mM L-carnitine. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, plasma membrane integrity, DNA integrity and acrosome integrity were evaluated. The 25 mM L-carnitine significantly improved sperm motility compared with a control group (p<0.05), although this was not significantly different among other concentrations. In conclusion, supplementation of 25 mM L-carnitine in freezing extender improves cauda epididymal spermatozoa motility. The effects of L-carnitine on the levels of oxidative stress during freezing and thawing remains to be examined.
Cryopreservation of epididymal sperm is an effective technique to preserve genetic materials of domestic cats and wild felids when they unexpectedly die. However, this technique inevitably causes detrimental changes of cryopreserved-thawed spermatozoa, for example, by physical damage and excessive oxidative stress. L-carnitine is an antioxidant that has been used to improve sperm motility in humans and domestic animals. This study aimed to investigate the effects of L-carnitine on cat epididymal sperm quality following cryopreservation and thawing. After routine castration, cauda epididymides were collected from 60 cat testes. The epididymal spermatozoa from 3 cauda epididymides were pooled as 1 replicate. Spermatozoa samples (16 replicates) were examined for spermatozoa quality and then randomly divided into 4 groups: 0 mM L-carnitine (control), 12.5 mM, 25 mM and 50 mM L-carnitine. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, plasma membrane integrity, DNA integrity and acrosome integrity were evaluated. The 25 mM L-carnitine significantly improved sperm motility compared with a control group (p<0.05), although this was not significantly different among other concentrations. In conclusion, supplementation of 25 mM L-carnitine in freezing extender improves cauda epididymal spermatozoa motility. The effects of L-carnitine on the levels of oxidative stress during freezing and thawing remains to be examined.
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문제 정의
Until the present, the cryoprotective effect of L-carnitine during freezing and thawing remains unclear. Therefore, this study was performed to investigate the effects of different L-carnitine concentrations in the freezing extender on post-thaw quality of feline epididymal spermatozoa.
We therefore hypothesized that L-carnitine supplementation in a freezing extender would improve cat epididymal spermatozoa quality during cryopreservation. This is the first study to investigate the effect of L-carnitine supplementation in the freezing extender for cat epididymal spermatozoa.
가설 설정
, 2003). We therefore hypothesized that L-carnitine supplementation in a freezing extender would improve cat epididymal spermatozoa quality during cryopreservation. This is the first study to investigate the effect of L-carnitine supplementation in the freezing extender for cat epididymal spermatozoa.
제안 방법
Concentrations of L-carnitine, times and interaction between concentrations of L-carnitine and times were regarded as a fixed effect. Least-squares means were obtained from each class of the factors; and were compared by using least significant different test with Tukey-Kramer adjustment for multiple comparisons. All data was expressed as mean±SD.
The spermatozoa were then conventionally cryopreserved. Spermatozoa motility, plasma membrane integrity, acrosome integrity and DNA integrity were evaluated at 0, 2 and 4 hours after thawing.
대상 데이터
The epididymal spermatozoa from 3 cauda epididymides were randomly pooled as 1 replicate in order to increase the sperm numbers and also to reduce individual variations. A total of 16 replicates were performed in this study. To obtain the spermatozoa, cauda epididymides were cut into few pieces and then placed in thawing medium for 10 min at 37°C (Tittarelli et al.
Testes were collected from 24 healthy cats of various breeds, aged between 1-7 years old by routine closed-technique castration. They were immediately kept in 0.
이론/모형
Normal distribution of residuals from the statistical models was tested using UNIVARIATE procedure option NORMAL PLOT. Two-way analysis of variance was performed using PROC GLM procedure to compare the difference among mean numbers of percentage of spermatozoa motility, intact plasma membrane spermatozoa, intact acrosome spermatozoa and normal DNA integrity spermatozoa. Concentrations of L-carnitine, times and interaction between concentrations of L-carnitine and times were regarded as a fixed effect.
성능/효과
In conclusion, the 25 mM L-carnitine supplementation in cryopreserved sperm extender improves motility of cat epididymal spermatozoa during freezing thawing. The interactions between ROS and antioxidants supplemented into freezing extender in terms of subtle changes of plasma membrane and fertility should be further investigated.
Our results demonstrated that cryopreservation markedly reduced the spermatozoa quality, although the post-thawed spermatozoa quality was in an acceptable range (Tables 2 to 4). This result was also similar to previous reports (Axner et al.
The percentages of motility, plasma membrane integrity, DNA integrity and acrosome integrity fresh spermatozoa were 80.75±4.87, 78.40±5.24, 94.98±2.81, and 73.57±5.22, respectively.
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