Measurement of epididymal sperm motility as a test variable in the rat

1986 ◽  
Vol 36 (1) ◽  
pp. 317-324 ◽  
Author(s):  
Ralph E. Linder ◽  
Lillian F. Strader ◽  
W. Keith McElroy
2017 ◽  
Vol 4 (2) ◽  
pp. 16
Author(s):  
Angelica C. Bumanlag ◽  
Hannah Lei M. Harada ◽  
Cynthia C. Divina ◽  
Marlon B. Ocampo ◽  
Lerma C. Ocampo

Appropriate holding conditions for post mortem testicles of goat to yield quality epididymal sperm (ES) as a source of genetic material for cryobanking and fertilization studies are lacking. In this study, the effect of storage time on the motility of ES from post mortem testicles maintained at 5°C was evaluated. In the laboratory, the cauda epididymides were excised from the testicles after 4 hr (G-1) and 24 hr (G-2) of holding time before collecting the sperm in a Tris-citrate buffered solution and evaluated using a CASA. Sperm motility profiling revealed a subpopulation of static, slow, motile and progressive ES. The proportion of static sperm in the control (16.97±6.21) and G-1 (21.53±5.60) were lower significantly than G-2 (36.13±5.05). The proportion of slow moving sperm was lower significantly than G-1 (23.31±3.57) and G-2 (25.45±3.32). The proportion of motile and progressive motile sperm decreases significantly (P<0.05) as the holding time increases at 78.46±4.64% (G-1) to 63.85±4.06 (G-2) and 45.53±8.89 (G-1) to 25.46±8.42 (G-2), respectively. The results showed that prolonged storage of post mortem testicles at 5°C could result to a reduced percentage of motile and progressively motile ES. Nevertheless, this considerable proportion of ES remained useful both for cryobanking and fertilization studies.


1996 ◽  
Vol 15 (9) ◽  
pp. 736-738 ◽  
Author(s):  
N. Pant ◽  
R. Shankar ◽  
SP Srivastava

The spermatotoxic effect of carbaryl in adult and young male rats has been examined. Carbaryl 50 and 100 mg/kg b.wt. Male fed 5 d/week for 60 days, caused dose and age- dependent decline in epididymal sperm count and sperm motility, an increase in sperm with abnormal morphology. The dose of 25 mg/kg/d was a 'No observed effect level' for the indices studied. Young animals in comparison to adults exhibited pronounced spermatotoxic effects.


2009 ◽  
Vol 21 (1) ◽  
pp. 102
Author(s):  
J. D. Hourcade ◽  
M. Perez-Crespo ◽  
B. Pintado ◽  
A. Gutiérrez-Adán

Physiological bases of the sperm selection processes within the female reproductive tract before they meet and fertilize the oocyte are unknown. The aim of this work was to determine if one of the keys of spermatozoa selection could be DNA integrity. It has been reported that sperm DNA damage does not impair in vitro fertilization (IVF). However, it has been suggested that the zona pelucida (ZP) is able to select spermatozoa with unfragmented DNA (Liu and Baker 2007 Hum. Reprod. 22, 1597–1602). In this work, DNA damage of spermatozoa was artificially induced by scrotal heat treatment (HT) (42°C, 30 min). Twenty-one days after the HT, spermatozoa were recovered from the epididymis caudae of CD1 mice and from the uterine horns near the cervix (Uc), from the uterine horns near the oviducts (Uo), and from the oviducts (Ov) of CD1 females 1–2 h after mating with HT and control males. In each region we determined numbers of spermatozoa, individual motility and sperm DNA integrity by COMET assay (% DNA in tail, tail length, and COMET moment was calculated). Also, females naturally mated either with HT or control males were killed at Day 14 of pregnancy, and number of foetuses and resorptions was recorded. Additionally, IVF was performed with epididymal sperm from HT or control males, Two hours after IVF attached and un-attached spermatozoa to the ZP were recovered and samples were evaluated for sperm motility (CASA), sperm zona-binding, and sperm DNA fragmentation (COMET). Also cleavage rate of fertilized oocytes with sperm from HT or control males was analyzed. One-way ANOVA was used to compare the results form each group. Epididymal sperm count (12*106 and 4.4*106 for control and HT respectively), sperm motility (75 and 21% respectively) and testis weight (133.90 and 68.76 mg, respectively) were significantly reduced after heat treatment (P < 0.001). For the heat treatment, COMET values decreased significantly during the transit from Uc to Uo and from Uo to Ov (Tail DNA: 25.7, 23.5, and 14.4% respectively, P < 0.01; Tail length: 38.4, 29.4, and 11.2 pixels, P < 0.001; COMET Moment: 12.5, 8.5, and 2 respectively, P < 0.001). Heat treatment reduced numbers of foetuses (7 ± 0.5 v. 5 ± 0.49, control and HT group, respectively), but number of resorptions was not altered. Spermatozoa bound per ZP in IVF experiments (55 ± 7 and 13 ± 6, control and HT, respectively) and cleavage rate (61 ± 1 v. 15 ± 6, control and HT, respectively) were significantly reduced in the HT group. Two hours after IVF, spermatozoa attached to the ZP in HT group showed a significant decrease in COMET parameters as in tail length (59.46 ± 2.895 v. 34.66 ± 3.531), and in tail moment compared with unattached spermatozoa. Our results indicate that DNA integrity sperm selection mechanisms are present in both the female tract and the ZP. We suggest that genital tract and sperm-ZP binding process plays an important role in selection of sperm with normal chromatin DNA.


2000 ◽  
Vol 12 (6) ◽  
pp. 337 ◽  
Author(s):  
H. Lambrechts ◽  
F. E. van Niekerk ◽  
S. W. P. Cloete ◽  
W. A. Coetzer ◽  
G. van der Horst

Microscopically evaluated sperm parameters, as well as computer-aided sperm motility analysis (CASMA), were used to assess sperm quality and the effect of cryopreservation on ram semen obtained from two genetically diverse Merino lines. These lines were divergently selected on maternal ranking values for multiple rearing ability from the same base population since 1986. Replacements in the high (+) line were preferentially the progeny of ewes rearing >1 lamb per joining. Progeny of ewes rearing <1 lamb per joining was preferred as replacements in the low (–) line. Sperm quality, as assessed by percentages of live, abnormal and acrosome-intact spermatozoa as well as by motility, was independent (P≤0.20) of line, time of sampling and their interaction in ejaculated samples obtained from the eight rams used as sires in 1995. Sperm quality of frozen–thawed samples was adversely affected (P≤0.01) by cryopreservation and thawing at 35˚C for 30 s relative to fresh ejaculated samples. No consistent differences between lines were found in epididymal sperm samples obtained from 12 slaughtered rams (6 from each line). The adverse effect (P≤0.05) of cryopreservation and thawing at 35˚C for 30 s on sperm viability and motility was also demonstrated for these samples.


2014 ◽  
Vol 27 (6) ◽  
pp. 791-796 ◽  
Author(s):  
S. Manee-in ◽  
S. Parmornsupornvichit ◽  
S. Kraiprayoon ◽  
T. Tharasanit ◽  
P. Chanapiwat ◽  
...  

1998 ◽  
Vol 41 (1) ◽  
pp. 23-26 ◽  
Author(s):  
A. G. Del Rio ◽  
A. M. Blanco ◽  
H. Niepomniscze ◽  
C. Carizza ◽  
F. Parera

1990 ◽  
Vol 68 (2) ◽  
pp. 528-535 ◽  
Author(s):  
Nongnuj Tanphaichitr ◽  
Jacqueline Smith ◽  
Morris Kates

The purpose of this study was to determine whether sulfogalactosylglycerolipid (SGG) was desulfated during mouse sperm capacitation. Levels of [35S]SGG were determined in freshly retrieved caudal epididymal sperm, motile capacitated sperm, and immotile sperm, after feeding mature male mice with [35S]sulfate-laced chow for 32 days. Caudal epididymal sperm and coisolated epididymal cells were separated into pellet and interphase fractions by centrifugation through a two-step Percoll gradient (45 and 90%). Upon resuspension in Krebs–Ringer bicarbonate medium supplemented with 0.4% bovine serum albumin, the Percoll-gradient pellet fraction consisted mainly of motile capacitated sperm, whereas the interphase fraction comprised largely immotile sperm and fragmented epididymal epithelial cells. The level of [35S]SGG in the Percoll-gradient-pelleted sperm appeared to be much higher than that in the Percoll-gradient interphase sperm. Percoll-gradient-pelleted sperm were further incubated in the culture medium for 2 h. The level of [35S]SGG showed little or no change after 1 h, but was reduced appreciably after 2 h. At this time point, sperm motility was also decreased. Reduction of sperm SGG is correlated with sperm immotility and (or) senescence and may have no direct relation to the capacitation process.Key words: sulfogalactosylglycerolipid, sperm motility, sperm capacitation.


2013 ◽  
Vol 25 (1) ◽  
pp. 271
Author(s):  
G. A. Monteiro ◽  
C. P. Freitas-DellAqua ◽  
P. N. Guasti ◽  
Y. F. R. Sancler-Silva ◽  
C. Ramires-Neto ◽  
...  

The development of a reliable technique for freezing epididymal semen would provide a unique opportunity to preserve valuable genetic material from unexpectedly lost stallions. The semen analysis method with the best ability to predict fertility is an examination of the sperm chromatin structure. This test evaluates the susceptibility of spermatozoa DNA to denaturation. The ability of spermatozoal DNA to maintain an intact double-stranded configuration is determined by exposure to an acid environment. The aim of this study was to compare the DNA fragmentation index of sperm obtained from ejaculate (G1) and sperm from the cauda epididymis (G2). For G1, two ejaculates from each of seven stallions were collected and then subjected to cryopreservation using BotuCrioTM extender. One week after the last semen collection, the stallions underwent bilateral orchiectomy. Sperm from the cauda epididymis was harvested immediately after castration (G2) by retrograde flushing of the caudal portion of the epididymis using a skim milk-based extender (BotuSemenTM). The recovered sperm was then cryopreserved using BotuCrioTM extender. The sperm motility parameters were analysed by computer-assisted sperm analysis (HTM IVOS 12, Hamilton Thorne Inc., Beverly, MA, USA), and the DNA fragmentation index was estimated using acridine orange test epifluorescence microscopy. The samples were evaluated immediately (0 h) and 8 h after thawing. The total motility, progressive motility, and percentage of rapid cells of the G1 v. G2 samples at 0 h were, respectively, 62.3 ± 12.9a v. 72.6 ± 8.4a, 31.6 ± 9.2a v. 35.3 ± 10.32a, and 49.3 ± 14.33a v. 59.7 ± 13.59a. At 8 h, the results were 26.0 ± 21.6b v. 54.7 ± 12.2a, 6.1 ± 6.4b v. 17.4 ± 8.54a, and 13.7 ± 14.85b v. 37.6 ± 14.15a. Evaluation of the DNA fragmentation in the G1 and G2 samples yielded 6.7 ± 1.41a v. 5.7 ± 1.60a at 0 h and 8.3 ± 1.78b v. 7.2 ± 1.19b at 8 h for percentage of DNA fragmentation after thawing. At 0 h, no differences in the sperm parameters were observed between groups, but statistical differences were observed in the sperm motility parameters between the treatment groups after 8 h. For the DNA fragmentation index, no difference was found at 0 and 8 h between the groups. However, after thawing, a higher percentage of DNA fragmentation was observed in the ejaculated sperm (8 h) as compared with the epididymal sperm (0 h). On the basis of these results, we can conclude that frozen–thawed cauda epididymal sperm had similar or higher motion parameters than ejaculated sperm after thawing. In addition, incubating the sperm at 20°C for 8 h after thawing resulted in higher motion parameters and less DNA fragmentation of the epididymal sperm. This finding suggests that epididymal sperm are more resistant to the cold shock caused by cryopreservation. FAPESP for financial support and Botupharma for donation of BotuSemenTM and BotuCrioTM extender.


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