Effects of Seminal Plasma Components on Motility and Acrosomal Integrity of Meishan Boar Spermatozoa after Cooling Treatments

Hyperactivity, a form of sperm motility characterized by vigorous flagellar movements, has been proposed as essential for fertilization in mammals. The objective of the present study was to establish a method for inducing hyperactivityin vitroin boar spermatozoa and to define threshold values to differentiate between hyperactive and non-hyperactive spermatozoa by computer-assisted sperm analysis (CASA) as a prerequisite for analyzing the energy metabolism during hyperactivity. In TALP-HEPES medium, non-frozen boar spermatozoa were stimulated to hyperactivity by 50 μmol l−1Ca2+within 15 min at 37 °C if 5 μmol l−1of the Ca2+ionophore A23187 was present. If 25% seminal plasma was present, boar spermatozoa required higher Ca2+concentrations (about 700 μmol l−1) for hyperactivity. Under both conditions, immobilization and head-to-head agglutination were low so that hyperactive spermatozoa could be analyzed for at least 40 min. The transition from normal to hyperactive movement was characterized by an increase in flagellar beat angle from 49° ± 12° to 200° ± 36° (n= 32) and a decrease in flagellar curvature ratio from 0.89 ± 0.04 to 0.47 ± 0.11 (n= 32). For quantification of hyperactive boar sperm, kinematic parameters of hyperactive and non-hyperactive spermatozoa were measured by CASA and statistically evaluated (receiver operating characteristic (ROC) curve analysis). The threshold values of the following four parameters were well suited for differentiating between hyperactive and non-hyperactive boar spermatozoa (ROC curve analysis: >50% specificity at 100% sensitivity). Hyperactive boar spermatozoa showed mean lateral head displacement >3.5 μm, curvilinear velocity >97 μm s−1, linearity <32% and wobble <71%. According to this multiparametric definition, induction of hyperactivity increased significantly (P< 0.0001) the fraction of hyperactive spermatozoa in semen samples from 5.1 ± 4.3% (n= 13) to 48.3 ± 6.6% (n= 7) in the absence and to 44.2 ± 7.6% (n= 10) in the presence of 25% seminal plasma, while the overall percentage of motile spermatozoa did not change significantly.

Download Full-text

Effects of post-thaw incubation on motility, acrosomal integrity and in vitro fertilizing capacity of boar spermatozoa cryopreserved in 0.5 and 5ml straws

Reproductive Biology ◽

10.1016/j.repbio.2013.03.005 ◽

2013 ◽

Vol 13 (2) ◽

pp. 166-168

Author(s):

Alejandro Córdova ◽

Rocío Hernández-Gil ◽

Cristian Alejandro Córdova-Jiménez ◽

Diogo José Cardilli ◽

Kellen de Sousa Oliveira ◽

...

Keyword(s):

Boar Spermatozoa ◽

Acrosomal Integrity

Download Full-text

Effect of a transcervical infusion of seminal plasma prior to insemination on the fertilising competence of low numbers of boar spermatozoa at controlled AI-ovulation intervals

Animal Reproduction Science ◽

10.1016/0378-4320(96)01553-9 ◽

1996 ◽

Vol 44 (3) ◽

pp. 165-173 ◽

Cited By ~ 6

Author(s):

D. Waberski ◽

J.A.G. Soares ◽

E. Bandeira^de Arruda ◽

K.F. Weitze

Keyword(s):

Seminal Plasma ◽

Boar Spermatozoa

Download Full-text

Adsorption of seminal plasma proteins by boar spermatozoa

Theriogenology ◽

10.1016/0093-691x(90)90024-n ◽

1990 ◽

Vol 34 (4) ◽

pp. 691-700 ◽

Cited By ~ 26

Author(s):

K.W. Metz ◽

Trish Berger ◽

E.D. Clegg

Keyword(s):

Seminal Plasma ◽

Plasma Proteins ◽

Seminal Plasma Proteins ◽

Boar Spermatozoa

Download Full-text

BIOCHEMICAL STATUS OF BOAR SPERMATOZOA and SEMINAL PLASMA BEFORE and AFTER A 10-DAY DEPLETION TEST (DT)

Reproduction in Domestic Animals ◽

10.1111/j.1439-0531.1995.tb00035.x ◽

1995 ◽

Vol 31 (1) ◽

pp. 245-246 ◽

Cited By ~ 1

Author(s):

J. Strzezek ◽

W. Demianowicz ◽

W. Kordan ◽

J. Torska ◽

P. Wysocki ◽

...

Keyword(s):

Seminal Plasma ◽

Biochemical Status ◽

Before And After ◽

Boar Spermatozoa

Download Full-text

Slow cooling prevents cold-induced damage to sperm motility and acrosomal integrity in the black-footed ferret (Mustela nigripes)

Reproduction Fertility and Development ◽

10.1071/rd06096 ◽

2007 ◽

Vol 19 (5) ◽

pp. 652 ◽

Cited By ~ 18

Author(s):

R. M. Santymire ◽

P. E. Marinari ◽

J. S. Kreeger ◽

D. E. Wildt ◽

J. G. Howard

Keyword(s):

Sperm Motility ◽

Seminal Plasma ◽

Artificial Insemination ◽

Sperm Cryopreservation ◽

Slow Cooling ◽

Rapid Cooling ◽

Mustela Nigripes ◽

Factors Influencing ◽

Rate Of Cooling ◽

Acrosomal Integrity

The endangered black-footed ferret (Mustela nigripes) has benefited from artificial insemination; however, improved sperm cryopreservation protocols are still needed. The present study focused on identifying factors influencing gamete survival during processing before cryopreservation, including: (1) the presence or absence of seminal plasma; (2) temperature (25°C v. 37°C); (3) type of medium (Ham’s F10 medium v. TEST yolk buffer [TYB]); (4) cooling rate (slow, rapid and ultra-rapid); and (5) the presence or absence of glycerol. Seminal plasma did not compromise (P > 0.05) sperm motility or acrosomal integrity. Sperm motility traits were maintained longer (P < 0.05) at 25°C than at 37°C in Ham’s or TYB, but temperature did not affect (P > 0.05) acrosomal integrity. Overall, TYB maintained optimal (P < 0.05) sperm motility compared with Ham’s medium, but Ham’s medium maintained more (P < 0.05) intact acrosomes than TYB. Slow cooling (0.2°C min–1) was optimal (P < 0.05) compared to rapid cooling (1°C min–1), and ultra-rapid cooling (9°C min–1) was found to be highly detrimental (P < 0.05). Results obtained in TYB with 0% or 4% glycerol were comparable (P > 0.05), indicating that 4% glycerol was non-toxic to ferret sperm; however, glycerol failed to ameliorate the detrimental effects of either rapid or ultra-rapid cooling. The results of the present study demonstrate that the damage observed to black-footed ferret spermatozoa is derived largely from the rate of cooling.

Download Full-text

Effect of Two Cryopreservation Methods on Seminal Quality and Pregnancy Rate in Alpacas Induced to Ovulation with Seminal Plasma

SPERMOVA ◽

10.18548/aspe/0008.11 ◽

2020 ◽

Vol 10 (2) ◽

pp. 74-80

Author(s):

Wilber Garcia ◽

◽

Edwar Maxi ◽

Veronica Macedo ◽

Elizabeth Mendoza ◽

...

Keyword(s):

Seminal Plasma ◽

Egg Yolk ◽

Final Concentration ◽

Quality Analysis ◽

Freezing Process ◽

Vaginal Fluid ◽

Gnrh Analog ◽

Freezing Method ◽

Semen Collection ◽

Acrosomal Integrity

The objective was to evaluate the effect of two methods of freezing on seminal quality and effect of seminal plasma as an inducer of ovulation in pregnancy percentages in inseminated alpacas. The semen collection was made post copula. After the collection of the ejaculates, motility and volume were assessed. The ejaculates with volume (≥1 ml) and motility (≥ 60%) were mixed (pool), 80 % was used (3 samples/pool) and 20 % (2 samples/pool) during the experiment. Then and diluted in Tris-base with 20% egg yolk. The samples were cooled 1.5 hours at 5 °C. At that temperature it was combined with the basic dilutor plus glycerol, obtaining a final concentration of 5% glycerol, then they were packed in 0.5 mL (13 x 106 spem/straw) straws to be frozen by horizontal and vertical methods. The seminal (semen + vaginal fluid) quality analysis was performed fresh and after cooling and thawing. Insemination was performed in two groups with thawed semen from two straws of the vertical method, the first group 20 females induced to ovulate with GnRH analog, the second group 20 females induced to ovulate with seminal plasma. When comparing, the results obtained of motility, viability, HOST and acrosomal integrity of fresh sperm, after the freezing process, decreased (p<0.05) compared to fresh and refrigerated samples. On the other hand, when comparing the freezing methods, the sperm values frozen by the vertical method were higher than those obtained by the horizontal method, with (p<0.05) in motility and HOST without (p>0.05) in acrosomal integrity and viability. The vertical semen freezing method can replace the horizontal method to obtain pregnancy.

Download Full-text

269IN VITRO FERTILITY OF BOAR SPERMATOZOA PRESERVED AT 10^°C FOR 22 DAYS

Reproduction Fertility and Development ◽

10.1071/rdv16n1ab269 ◽

2004 ◽

Vol 16 (2) ◽

pp. 255

Author(s):

H. Funahashi

Keyword(s):

Oxidative Stress ◽

Functional Status ◽

Pregnancy Rate ◽

Seminal Plasma ◽

Artificial Insemination ◽

Fluorescence Assay ◽

Live Cells ◽

Sperm Viability ◽

Boar Spermatozoa

Fertility of boar spermatozoa as determined following artificial insemination seems to be maintained during liquid preservation at 10–15°C for several days, although prolonged liquid preservations reduce the pregnancy rate rapidly. However, it is not clear if spermatozoa can penetrate into oocytes in an IVF system even after a prolonged liquid preservation. Oxidative stress could also be one of the possible detrimental factors in liquid preservation of spermatozoa. In the present study, fertility of liquid-preserved spermatozoa was examined using an IVM-IVF system. Whether cysteine can improve the fertility was also determined. Spermatozoa (from four Berkshires) was resuspended at 1×108 cells mL−1 in Modena solution containing 15% (v/v) boar seminal plasma and 0 or 5mM cysteine after washing 3 times. Sperm suspensions (1mL) were then preserved at 10°C for 22 days following a program for cooling down (to 15°C for 4h, keeping at 15°C for 12h and then to 10°C for 6h). At Days 1, 8, 15 and 22 after the start of preservation, spermatozoa (5×105 cells mL−1) were co-cultured with IVM oocytes in an IVM/IVF system (Funahashi et al., 1997 Biol Reprod 57, 49–53). Viability and functional status of spermatozoa were also examined at Days 8 and 15 of preservation by using LIVE/DEAD sperm viability kit and CTC fluorescence assay. Data (mean±SEM) from 4–6 replicates were analyzed by ANOVA and Fisher’s protected LSD test. When spermatozoa that had been preserved without cysteine (Cys−) were used, penetration rates were not different (P>0.05) from those with cysteine (Cys+) at Day 8 of preservation (91.4±3.4% in Cys− and 99.3±0.7% in Cys+), but lower (P<0.02) at Days 15 and 22 (72.6±13.6% and 33.8±8.4% in Cys−; 94.8±2.1% and 71.1±10.8% in Cys+, respectively). Both viability and proportion of uncapacitated live cells were higher (P<0.05) in Cys+ than Cys− at Days 8 and 15. These results demonstrate that boar spermatozoa can penetrate into oocytes in vitro even after a liquid preservation at 10°C for 22 days and that cysteine can improve the viability and penetrability in vitro of spermatozoa during liquid preservation. Supported by the Ito Foundation.

Download Full-text

309 SEX SORTED BOAR SPERMATOZOA: TIME COURSE AND PROFILE OF THEIR IN VITRO PENETRATION ABILITY AFTER STORAGE IN THE PRESENCE OF HOMOLOGOUS SEMINAL PLASMA

Reproduction Fertility and Development ◽

10.1071/rdv17n2ab309 ◽

2005 ◽

Vol 17 (2) ◽

pp. 305

Author(s):

I. Parrilla ◽

J.M. Vazquez ◽

M.A. Gil ◽

I. Caballero ◽

C. Almiñana ◽

...

Keyword(s):

Seminal Plasma ◽

Time Course ◽

Egg Yolk ◽

Vitro Fertilization ◽

Penetration Ability ◽

And Control ◽

Pig Oocyte ◽

Boar Spermatozoa ◽

Penetration Time

Addition of seminal plasma (SP) to the collection medium has been shown to be beneficial for motility and viability of sex-sorted and stored spermatozoa. However, SP could not only delay but also decrease the in vitro fertilization rates of IVM pig oocytes. In the present study, the time-course of IVM pig oocyte penetration of sex sorted boar spermatozoa stored in the presence or absence of SP was evaluated. Spermatozoa were sex-sorted following the Beltsville sperm sexing technology (Johnson and Welch 1999 Theriogenology 52, 1323–1342) and collected in TEST-egg yolk buffer (2%) with (10%) or without (control) SP. Sex-sorted spermatozoa were stored at 20°C during 0, 2, 5, and 10 h after sorting. Oocytes were matured in vitro in NCSU23 (Peters and Wells 1663 J. Reprod. Fertil. 48, 61–73) for 44 h in 5% CO2 in air at 39°C. The in vitro penetration time-course was determined by co-incubating the sex-sorted and stored spermatozoa with IVM oocytes during 3, 6, and 18 h in modified TRIS-buffered medium (mTBM) (Abeydeera and Day 1997 Theriogenology 48, 537–544) at 39°C in an atmosphere of 5% CO2 in air. Penetration rates and number of spermatozoa per oocyte were assessed after fixation and staining of the oocytes. Statistical analyses were conducted by ANOVA. Presence of SP did not delay the onset of the oocyte penetration. Moreover, at 3 h of co-incubation, SP increased (P < 0.05) both penetration rates and mean number of spermatozoa per oocyte in sorted and stored boar spermatozoa when compared with control (45 vs. 20, 50 vs. 32, 38 vs. 23, 15 vs. 8, at 0, 2, 5, and 10 h of storage with SP and control, respectively). High penetration rates were reached after 6 h of co-incubation (82 vs. 51, 96 vs. 76, 83 vs. 48, 31 vs. 24, at 0, 2, 5, and 10 h of storage with SP and control, respectively) in sorted and stored samples, with no further increase at 18 h (70 vs. 63, 92 vs. 79, 87 vs. 53, 55 vs. 40, at 0, 2, 5, and 10 h of storage with SP and control, respectively). Spermatozoa stored 2 h in the presence of SP showed the best penetration rate and highest mean number of spermatozoa per oocyte. The mean number of spermatozoa per oocyte increased as the co-incubation time increased (ranging from 2.1 to 5.8 for sorted spermatozoa stored 2 h in the presence of SP at 3 h and 18 h of co-incubation, respectively). In conclusion, the presence of SP during the storage of sex-sorted spermatozoa improves their in vitro fertilizing ability without affecting the onset of the oocyte penetration time. This work was supported by DGICYT (AGL 2001-0471), Fundación Seneca (PB174/FS/02) and CTIC (2103SIU0040).

Download Full-text