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

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).

scholarly journals 92THE ADDITION OF SEMINAL PLASMA FROM INDIVIDUAL BOARS TO FREEZING EXTENDER CAN IMPROVE THE POST-THAW SPERM SURVIVAL

2004 ◽  
Vol 16 (2) ◽  
pp. 167 ◽  
Author(s):  
T. Cremades ◽  
G. Carvajal ◽  
M. Hernandez ◽  
J.M. Vazquez ◽  
E.A. Martinez ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Potential Effect ◽  
Conclusive Evidence ◽  
Sperm Cryopreservation ◽  
Low Concentrations ◽  
Sperm Survival ◽  
And Control ◽  
Boar Spermatozoa

Contradictory results have been reported about the effect of seminal plasma (SP) on the freezability of mammalian spermatozoa. In pigs, current methods for sperm cryopreservation involve removing seminal plasma. Therefore, no conclusive evidence of the potential effect of SP on the freezability of boar spermatozoa has been reported. In this study, we evaluate the effect of the addition of low concentrations of SP from individual boars to the freezing extender on post-thaw sperm survival. Sperm cryopreservation procedure included: dilution of sperm-rich fraction in Beltsville Thaw Solution extender (BTS), cooling to 17°C for 16h, centrifugation at 2400g for 3min, dilution in lactose/egg-yolk/glycerol/Equex Stem (freezing extender) to a final concentration of 1×109 spermmL−1, dispensing into 0.5-mL straws, and freezing in a programmable cell freezer at 20°Cmin−1. Thawing was carried out in a waterbath at 37°C for 20s. Post-thaw sperm survival was assessed by progressive sperm motility (PSM) using a CASA system (SCA); plasma membrane integrity (PMI) and acrosome membrane integrity (AMI) were assessed by flow cytometric procedures (SYBR-14/PI and FITC-PNA/PI, respectively) at 30 and 150min post-thawing in BTS-diluted thaw spermatozoa held in a waterbath at 37°C. Four individual seminal plasma donors (SP1 to SP4) were selected in a preliminary study in which 48 ejaculates from 12 boars (4 ejaculates/boar) were cryopreserved. Then the boars were classified into 3 groups (good, moderate and bad freezers) based on their post-thaw sperm survival. SP1 and SP2 were good freezers (&gt;60% PSM and PMI), SP3 was a moderate freezer (40–60% PSM and PMI) and SP4 was a bad freezer (&lt;40% PSM and PMI). In the main experiment, pooled sperm-rich fractions collected from 9 mature hybrid boars were divided into five aliquots and each was diluted with freezing extender supplemented with 0% (control) or 10% of SP (1–4). Data from eight replicates were analyzed as a split plot design using a PROMIXED model. The addition of SP to freezing extender had a significant effect (P&lt;0.05) on post-thaw sperm survival compared to control. Moreover, there were significant differences (P&lt;0.05) between SP donors. PSM, PMI and AMI were significantly (P&lt;0.05) higher in SP1 (56.71±4.30; 57.16±4.01 and 57.22±4.01, respectively) and SP2 (59.48±4.30; 60.17±4.01 and 60.05±4.01, respectively) compared to control (50.39±4.30; 49.98±4.01 and 49.54±4.01, respectively). There were no differences (P&gt;0.05) between SP3, SP4 and control. These results indicate that the addition of SP from particular boars (good freezers) to freezing extender may improve post-thaw sperm survival. Individual differences in the SP composition should explain the above results. Supported by INIA (RZ01-019) and MCYT (AGL2001-0471).

scholarly journals The Re-Addition of Seminal Plasma after Thawing Does Not Improve Buck Sperm Quality Parameters

Animals ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3452
Author(s):  
Uchechi Linda Ohaneje ◽  
Uchebuchi Ike Osuagwuh ◽  
Manuel Alvarez-Rodríguez ◽  
Iván Yánez-Ortiz ◽  
Abigail Tabarez ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Quality Parameters ◽  
Mitochondrial Activity ◽  
Dna Integrity ◽  
Vitro Fertilization

In order to achieve a higher post-thaw buck sperm quality, an approach in the thawing protocol of cryopreserved sperm doses under in vitro capacitation conditions mimicking the in vivo female environment was studied. Therefore, functional and kinetic characteristics of buck thawed sperm from males of different ages, the season of collection, and melatonin implanted males in the non-breeding season were assessed after 3 h of incubation in an in vitro fertilization (IVF) media with 20% of buck seminal plasma (SP). Previously, fresh ejaculates were collected via artificial vagina from eight males of the Cabra Blanca de Rasquera breed during two consecutive years in breeding and non-breeding periods. Prior to semen collection in non-breeding seasons, males were split into two groups: one group was implanted with melatonin, while the other was not. In each group, semen samples were pooled, centrifuged, and diluted in an extender containing 15% powdered egg yolk and 5% glycerol before freezing. After thawing, sperm were washed and incubated in three different media: (a) control media (modified phosphate-buffered saline (PBS), (b) IVF commercial media, and (c) IVF media + 20% SP. Sperm motility was evaluated by CASA, while plasma and acrosome membrane integrity, mitochondria activity, and DNA fragmentation were analysed by flow cytometer at 0 h and after 3 h incubation. A significant reduction in motility, mitochondrial activity, plasma, and acrosome membrane integrity were observed after incubation in the presence of SP, although similar to that observed in IVF media alone. DNA integrity was not affected under in vitro capacitation conditions, regardless of SP addition. In conclusion, the addition of SP failed to improve post-thaw buck sperm quality under in vitro conditions irrespective of male age, the season of collection, and melatonin implant.

scholarly journals Improvement of the in vitro fertilization and embryo development using frozen–thawed spermatozoa of microminipigs

2021 ◽  
Vol 64 (1) ◽  
pp. 265-271
Author(s):  
Zhao Namula ◽  
Yasuhiro Isumi ◽  
Yoko Sato ◽  
Quynh Anh Le ◽  
Qingyi Lin ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Sperm Quality ◽  
Blastocyst Formation ◽  
Large White ◽  
Vitro Fertilization ◽  
Beneficial Effects ◽  
Penetration Ability ◽  
Development In Vitro ◽  
Boar Spermatozoa

Abstract. This study aimed to compare the quality and the penetration ability of frozen–thawed spermatozoa from three microminipigs and Large White boars and to evaluate the effects of caffeine and heparin as well as the sperm–oocyte co-incubation length on the fertilization and embryonic development in vitro. Results showed that the fertilization rates of spermatozoa from three microminipig boars were significantly lower than those of a Large White boar. In the post-thaw spermatozoa from one of three microminipig boars, the sperm quality, penetration ability, and the oocyte development after in vitro fertilization were significantly lower than those of the spermatozoa from other boars. The caffeine supplementation in the fertilization media increased the rates of fertilization and blastocyst formation for the microminipig spermatozoa with low sperm quality. In addition to caffeine supplementation, the rates of fertilization and blastocyst formation after using microminipig spermatozoa were significantly higher with a 10 h sperm–oocyte co-incubation than with 3 h of co-incubation length. Our results indicate that the differences between the males and the breed influence the quality and fertility of frozen–thawed boar spermatozoa. In conclusion, the presence of caffeine in the in vitro fertilization (IVF) medium and adequate length of sperm–oocyte co-incubation may have beneficial effects for improving IVF results when using microminipig spermatozoa with low quality.

The relationship between membrane status and fertility of boar spermatozoa after flow cytometric sorting in the presence or absence of seminal plasma

1998 ◽  
Vol 10 (5) ◽  
pp. 433 ◽  
Author(s):  
W. M. C. Maxwell ◽  
C. R. Long ◽  
L. A. Johnson ◽  
J. R. Dobrinsky ◽  
G. R. Welch
Keyword(s):  
Seminal Plasma ◽  
Hoechst 33342 ◽  
Cell Sorter ◽  
Vitro Fertilization ◽  
Flow Cytometric ◽  
Collection Medium ◽  
Flow Cytometric Sorting ◽  
Boar Spermatozoa ◽  
The Relationship

The motility, viability (percent live), capacitation status and in vitro fertility of boar spermatozoa were examined, after staining with Hoechst 33342 and flow cytometric sorting in the absence or presence of seminal plasma. Viability was higher in unstained controls and when seminal plasma was present in the medium used to collect spermatozoa from the cell sorter than when seminal plasma was absent or in the staining extender only, but motility was highest when seminal plasma was included in the extender only, compared with the controls and other treatments. The proportions of capacitated spermatozoa were increased by sorting, but were lower when seminal plasma was present, rather than absent, from the staining extender and the collection medium. Compared with unstained controls, extension and staining without sorting only increased the proportion of capacitated spermatozoa after washing in preparation for in vitro fertilization. The percentages of polyspermic, penetrated and cleaved oocytes were lower when inseminated with unsorted (stained) than control (unstained) spermatozoa, regardless of the presence or absence of seminal plasma. These parameters were higher for sorted than for control spermatozoa in the absence of seminal plasma, but in its presence penetration and cleavage were substantially lower. The proportions of capacitated spermatozoa were lower when seminal plasma was present in the collection medium only than in the staining extender or when it was absent altogether, but the former treatment substantially reduced the proportions of polyspermic, penetrated and cleaved oocytes, and the proportion of blastocysts. These findings indicate that sperm capacitation associated with flow cytometric sorting can be reduced by the inclusion of seminal plasma in the collection medium, but this treatment reduces the ability of spermatozoa to fertilize oocytes in vitro under these conditions.

scholarly journals Seminal Plasma: Relevant for Fertility?

2021 ◽  
Vol 22 (9) ◽  
pp. 4368
Author(s):  
Heriberto Rodriguez-Martinez ◽  
Emilio A. Martinez ◽  
Juan J. Calvete ◽  
Fernando J. Peña Vega ◽  
Jordi Roca
Keyword(s):  
Seminal Plasma ◽  
Current Knowledge ◽  
Inorganic Ions ◽  
Vitro Fertilization ◽  
Dna And Rna ◽  
Model Animal ◽  
Sexual Glands ◽  
Species Specific

Seminal plasma (SP), the non-cellular component of semen, is a heterogeneous composite fluid built by secretions of the testis, the epididymis and the accessory sexual glands. Its composition, despite species-specific anatomical peculiarities, consistently contains inorganic ions, specific hormones, proteins and peptides, including cytokines and enzymes, cholesterol, DNA and RNA—the latter often protected within epididymis- or prostate-derived extracellular vesicles. It is beyond question that the SP participates in diverse aspects of sperm function pre-fertilization events. The SP also interacts with the various compartments of the tubular genital tract, triggering changes in gene function that prepares for an eventual successful pregnancy; thus, it ultimately modulates fertility. Despite these concepts, it is imperative to remember that SP-free spermatozoa (epididymal or washed ejaculated) are still fertile, so this review shall focus on the differences between the in vivo roles of the SP following semen deposition in the female and those regarding additions of SP on spermatozoa handled for artificial reproduction, including cryopreservation, from artificial insemination to in vitro fertilization. This review attempts, including our own results on model animal species, to critically summarize the current knowledge of the reproductive roles played by SP components, particularly in our own species, which is increasingly affected by infertility. The ultimate goal is to reconcile the delicate balance between the SP molecular concentration and their concerted effects after temporal exposure in vivo. We aim to appraise the functions of the SP components, their relevance as diagnostic biomarkers and their value as eventual additives to refine reproductive strategies, including biotechnologies, in livestock models and humans.

scholarly journals Association between HOX Transcript Antisense RNA Single-Nucleotide Variants and Recurrent Implantation Failure

2021 ◽  
Vol 22 (6) ◽  
pp. 3021
Author(s):  
Jeong Yong Lee ◽  
Eun Hee Ahn ◽  
Hyeon Woo Park ◽  
Ji Hyang Kim ◽  
Young Ran Kim ◽  
...  
Keyword(s):  
Antisense Rna ◽  
Coagulation Factors ◽  
Healthy Controls ◽  
Implantation Failure ◽  
Single Nucleotide Variants ◽  
Recurrent Implantation Failure ◽  
Single Nucleotide ◽  
Vitro Fertilization ◽  
And Control

Recurrent implantation failure (RIF) refers to the occurrence of more than two failed in vitro fertilization–embryo transfers (IVF-ETs) in the same individual. RIF can occur for many reasons, including embryo characteristics, immunological factors, and coagulation factors. Genetics can also contribute to RIF, with some single-nucleotide variants (SNVs) reported to be associated with RIF occurrence. We examined SNVs in a long non-coding RNA, homeobox (HOX) transcript antisense RNA (HOTAIR), which is known to affect cancer development. HOTAIR regulates epigenetic outcomes through histone modifications and chromatin remodeling. We recruited 155 female RIF patients and 330 healthy controls, and genotyped HOTAIR SNVs, including rs4759314, rs920778, rs7958904, and rs1899663, in all participants. Differences in these SNVs were compared between the patient and control groups. We identified significant differences in the occurrence of heterozygous genotypes and the dominant expression model for the rs1899663 and rs7958904 SNVs between RIF patients and control subjects. These HOTAIR variants were associated with serum hemoglobin (Hgb), luteinizing hormone (LH), total cholesterol (T. chol), and blood urea nitrogen (BUN) levels, as assessed by analysis of variance (ANOVA). We analyzed the four HOTAIR SNVs and found significant differences in haplotype patterns between RIF patients and healthy controls. The results of this study showed that HOTAIR is not only associated with the development of cancer but also with pregnancy-associated diseases. This study represents the first report showing that HOTAIR is correlated with RIF.

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