scholarly journals Mouse sperm delivery via cauda epididymis without transport medium

2016 ◽  
Author(s):  
Man-Xi Jiang ◽  
Mei-Shan Wang ◽  
Xiang-Hong Ou ◽  
Xue-Jin Chen ◽  
Yan Zhu
Keyword(s):  
Embryo Development ◽  
Sperm Motility ◽  
Room Temperature ◽  
Sperm Quality ◽  
Mouse Sperm ◽  
Spermatozoa Motility ◽  
Transport Medium ◽  
Fertilization Rates ◽  
Progressive Motility

This study was aimed to investigate the effects of room temperature (RT, 20-25 oC) and absence of medium during cauda epididymis transport on spermatozoa quality, fertility and embryo development. In the first experiment, fresh sperm from one side of cauda epididymis was used for in vitro fertilization, and another side was delivered at RT or 4-8 oC either with or without M2. In the second experiment, each side of cauda epididymis obtained from the same mouse was individually delivered at RT or 4 oC with or without M2. Finally, sperm motility, progressive motility scores and fertility of fresh spermatozoa or those from transported cauda epididymis, and IVF embryo development were evaluated. Progressive motility scores and fertilization rates were higher in fresh spermatozoa than transported sperm; sperm motility of transported cauda epididymis at 4-8 oC was comparable to fresh spermatozoa, but spermatozoa motility of transported cauda epididymis at RT was inferior to fresh spermatozoa. Spermatozoa motillty of transported cauda epididymis at 4-8 oC with transport medium was much higher than that without transport medium; absence of transport medium did not affect sperm motility of transported cauda epididymis at 4-8 oC but affected sperm motility of transported cauda epididymis at RT. Sperm quality from transported cauda epididymis can be efficiently kept at 4-8 oC, and cauda epididymis transport at 4-8 oC without M2 is more beneficial on keeping their fertility. Moreover cauda epididymis transport at RT without medium could sufficiently produce embryos for obtainning live offsprings.

scholarly journals Mouse sperm delivery via cauda epididymis without transport medium

2016 ◽  
Author(s):  
Man-Xi Jiang ◽  
Mei-Shan Wang ◽  
Xiang-Hong Ou ◽  
Xue-Jin Chen ◽  
Yan Zhu
Keyword(s):  
Embryo Development ◽  
Sperm Motility ◽  
Room Temperature ◽  
Sperm Quality ◽  
Mouse Sperm ◽  
Spermatozoa Motility ◽  
Transport Medium ◽  
Fertilization Rates ◽  
Progressive Motility

This study was aimed to investigate the effects of room temperature (RT, 20-25 oC) and absence of medium during cauda epididymis transport on spermatozoa quality, fertility and embryo development. In the first experiment, fresh sperm from one side of cauda epididymis was used for in vitro fertilization, and another side was delivered at RT or 4-8 oC either with or without M2. In the second experiment, each side of cauda epididymis obtained from the same mouse was individually delivered at RT or 4 oC with or without M2. Finally, sperm motility, progressive motility scores and fertility of fresh spermatozoa or those from transported cauda epididymis, and IVF embryo development were evaluated. Progressive motility scores and fertilization rates were higher in fresh spermatozoa than transported sperm; sperm motility of transported cauda epididymis at 4-8 oC was comparable to fresh spermatozoa, but spermatozoa motility of transported cauda epididymis at RT was inferior to fresh spermatozoa. Spermatozoa motillty of transported cauda epididymis at 4-8 oC with transport medium was much higher than that without transport medium; absence of transport medium did not affect sperm motility of transported cauda epididymis at 4-8 oC but affected sperm motility of transported cauda epididymis at RT. Sperm quality from transported cauda epididymis can be efficiently kept at 4-8 oC, and cauda epididymis transport at 4-8 oC without M2 is more beneficial on keeping their fertility. Moreover cauda epididymis transport at RT without medium could sufficiently produce embryos for obtainning live offsprings.

scholarly journals Effects of cryopreservation on sperm quality, nuclear DNA integrity, in vitro fertilization, and in vitro embryo development in the mouse

Reproduction ◽  
2007 ◽  
Vol 133 (3) ◽  
pp. 585-595 ◽  
Author(s):  
Cengiz Yildiz ◽  
Palma Ottaviani ◽  
Napoleon Law ◽  
Renise Ayearst ◽  
Ling Liu ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Embryo Development ◽  
Membrane Integrity ◽  
Fertilization Rate ◽  
Development Rate ◽  
Dna Integrity ◽  
Plasma Membrane Integrity ◽  
Mouse Sperm ◽  
Progressive Motility

Efficient freezing, archiving, and thawing of sperm are essential techniques to support large scale research programs using mouse models of human disease. The purpose of this study was to investigate the effects of variable combinations and concentrations of cryoprotectants on sperm-assessment parameters of frozen–thawed mouse sperm in order to optimize cryopreservation protocols. Sperm was frozen using combinations of 3% skim milk + 0.2 or 0.3 M nonpermeating raffinose with either permeating glucose, fructose, propylene glycol, ethylene glycol, glycerol, or sodium pyruvate in CD-1, C3FeB6F1/J, B6129SF1, C57BL/6NCrIBR, 129S/SvPaslco, and DBA/2NCrIBR mice. Sperm-assessment parameters included progressive motility, plasma membrane integrity (SYBR-14 + PI),in vitrofertilization rate, andin vitroembryo development rate to blastocyst. DNA content analysis of sperm was measured by the sperm chromatin structure assay (SCSA). 0.3 M raffinose with 0.1 M fructose significantly improved post-thaw sperm-assessment parameters for CD-1, C3B6F1, B6129SF1 mice (P< 0.05–0.01), whereas 0.2 M raffinose with 0.1 M glycerol or 0.1 M fructose enhanced sperm assessment values for C57BL/6 and 129S mice (P< 0.01), compared to 0.3 M raffinose alone. DNA fragmentation during cryopreservation was significantly increased in all strains evaluated when compared with fresh control sperm in a strain-dependent manner (P< 0.01). Supplementation with permeating glycerol or fructose to the cryoprotectant (CPA) solution showed a significant protective effect to DNA integrity when cryopreserving sperm from C57BL/6 and 129S mice. Damage to sperm DNA significantly decreased the rate ofin vitroembryo development to blastocyst in C57BL/6 mice. The type of monosaccharide sugar or polyols, CPA molarity, and combination of permeating and nonpermeating cryoprotectant are significant factors for improving progressive motility, plasma membrane integrity, DNA integrity,in vitrofertilization rate, andin vitroembryo development rate to blastocyst in cryopreserved mouse sperm.

Embryo development capacity of oocytes fertilized by immature sperm and sperm treated with motility stimulants

1994 ◽  
Vol 6 (1) ◽  
pp. 113 ◽  
Author(s):  
O Lacham-Kaplan ◽  
AO Trounson
Keyword(s):  
Embryonic Development ◽  
Embryo Development ◽  
Sperm Motility ◽  
Cumulus Cells ◽  
Progressive Motility ◽  
Development Capacity ◽  
Fertilizing Ability ◽  
Embryonic Arrest ◽  
Human And Mouse

The fertilizing ability of mouse spermatozoa develops during maturation and coincides with the acquisition of motility. The lack of progressive motility of spermatozoa from the testis and precaudal segments of the epididymis interferes with their ability to fertilize oocytes after insemination in vitro. The removal of cumulus cells for insemination in vitro and the use of subzonal injection of a single spermatozoon resulted in a higher number of oocytes fertilized by immature caput and corpus spermatozoa. High rates of embryonic arrest and retarded development were observed in oocytes fertilized by caput and corpus spermatozoa when compared with oocytes fertilized by cauda spermatozoa. However, when the oocytes were enclosed in their cumulus cells or microinjected with a single spermatozoon, these effects were reduced. A block in embryonic development was also observed after human and mouse oocytes were exposed to the sperm motility stimulants pentoxifylline (PTF) and 2-deoxyadenosine (DOA). These observations suggest that exposure of oocytes to PTF and DOA should be avoided.

Supplementation of sperm media with zinc, D-aspartate and co-enzyme Q10 protects bull sperm against exogenous oxidative stress and improves their ability to support embryo development

Zygote ◽  
2017 ◽  
Vol 25 (2) ◽  
pp. 168-175 ◽  
Author(s):  
Vincenza Barbato ◽  
Riccardo Talevi ◽  
Sabrina Braun ◽  
Anna Merolla ◽  
Sam Sudhakaran ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Dna Fragmentation ◽  
Embryo Development ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Reproductive Technologies ◽  
Developmental Competence ◽  
Sperm Dna Fragmentation ◽  
Bull Sperm

SummaryHigh levels of reactive oxygen species in the semen of infertile patients or spontaneously generated during in vitro sperm handling may impair sperm quality, fertilization and embryo developmental competence. We recently reported that zinc, d-aspartate and co-enzyme Q10, contained in the dietary supplement Genadis® (Merck Serono), have protective effects on human and bull sperm motility, lipid peroxidation and DNA fragmentation in vitro; furthermore, in bovine, treated spermatozoa had an improved ability to support embryo development. However, only a few studies have investigated the protective role of antioxidants during in vitro sperm handling in the presence of an exogenous oxidative stress. Herein, to simulate such conditions in an animal model, we induced exogenous oxidative stress on spermatozoa through the xanthine–xanthine oxidase system and investigated its effects on sperm function and subsequent embryo developmental competence in the presence of zinc, d-Asp and CoQ10 protection. The main results showed that exogenous oxidative stress decreased sperm motility, increased sperm DNA fragmentation, and reduced fertilization and blastocyst rates and quality. Pre-treatment with zinc, d-aspartate and co-enzyme Q10 before exogenous oxidative stress was able to prevent these effects. Supplementation of sperm culture media with zinc, d-aspartate and co-enzyme Q10 could protect sperm from oxidative stress damage during in vitro handling in assisted reproductive technologies.

Paternal hypoxia exposure impairs fertilization process and preimplantation embryo development

Zygote ◽  
2021 ◽  
pp. 1-9
Author(s):  
Hai-Ping Tao ◽  
Gong-Xue Jia ◽  
Xiao-Na Zhang ◽  
Yu-Jun Wang ◽  
Bin-Ye Li ◽  
...  
Keyword(s):  
Embryo Development ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Early Embryo ◽  
Hypoxic Exposure ◽  
Male Mice ◽  
Early Embryo Development ◽  
Hypoxia Exposure ◽  
Pronuclear Formation

Summary Environmental hypoxia exposure causes fertility problems in human and animals. Compelling evidence suggests that chronic hypoxia impairs spermatogenesis and reduces sperm motility. However, it is unclear whether paternal hypoxic exposure affects fertilization and early embryo development. In the present study, we exposed male mice to high altitude (3200 m above sea level) for 7 or 60 days to evaluate the effects of hypoxia on sperm quality, zygotic DNA methylation and blastocyst formation. Compared with age-matched controls, hypoxia-treated males exhibited reduced fertility after mating with normoxic females as a result of defects in sperm motility and function. Results of in vitro fertilization (IVF) experiments revealed that 60 days’ exposure significantly reduced cleavage and blastocyst rates by 30% and 70%, respectively. Immunohistochemical staining of pronuclear formation indicated that the pronuclear formation process was disturbed and expression of imprinted genes was reduced in early embryos after paternal hypoxia. Overall, the findings of this study suggested that exposing male mice to hypoxia impaired sperm function and affected key events during early embryo development in mammals.

PSV-5 Relationship between sire conception rate, sperm motility, sperm SERPINA5 relative concentration, and in vitro produced embryos in dairy bulls

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 310-310
Author(s):  
Saulo Menegatti Zoca ◽  
Julie Walker ◽  
Taylor Andrews ◽  
Adalaide C Kline ◽  
Jerica J Rich ◽  
...  
Keyword(s):  
Embryo Development ◽  
Sperm Motility ◽  
Relative Concentration ◽  
Early Embryo ◽  
Conception Rate ◽  
Cleavage Rate ◽  
Blastocyst Rate ◽  
Positive Correlations ◽  
Sire Conception Rate

Abstract Sire conception rate (SCR) is a field measure of fertility among bulls, but it can be influenced by several factors (Sperm transport, sperm-egg binding, early embryo development, etc). The objective of this study was to evaluate the relationship between SCR, sperm motility, SERPINA5 concentrations, and in vitro embryo development. Measurements were performed in 19 bulls with SCR values ranging from -7.7 to 4.45. For each bull, an aliquot of frozen-thawed semen was used for analyses of total (TMOT) and progressive (PROG) motility. Remaining semen was fixed with 2% formaldehyde, and concentration of SERPINA5 was determined by immunolocalization (antibody SERPINA5/Dylight405; PA5-79976-Invitrogen / ab201798-Abcam). Mean fluorescence intensity was determined in ~200 sperm heads/bull. Approximately 149 oocytes/bull were fertilized in vitro for embryo development analysis (cleavage and blastocyst rates). Statistical procedures were performed in SAS (9.4) using the procedures CORR for correlations (SCR, TMOT, PROG, SERPINA5, cleavage and blastocyst) and GLIMMIX for comparison of “field-fertility” (SCR divided in HIGH or LOW) and “field-embryo-fertility” (LOW-SCR sires were divided based on blastocyst rate (HIGH or LOW) resulting in two classifications; LOW-HIGH≥31% and LOW-LOW≤26%, respectively). There were positive correlations (P &lt; 0.05) between cleavage-blastocyst (r=0.50), SERPINA5-cleavage (r=0.48), and TMOT-PROG (r=0.76). Sire SCR was not associated with SERPINA5, TMOT, PROG, cleavage and blastocyst rate (P &gt; 0.52). Among LOW-SCR sires, LOW-LOW sires (-4.83±0.60) tended to have a better SCR score than LOW-HIGH (-6.18±0.42) sires (P = 0.08), but there were no differences (P &gt; 0.43) between LOW-HIGH, LOW-LOW, and HIGH sires for SERPINA5, TMOT, PROG, and cleavage. In conclusion, some LOW SCR sires have good embryo development indicating a different mechanism for their low SCR; however, these differences in SCR could not be explained by TMOT, PROG, SERPINA5, cleavage and blastocyst. There were, however, positive correlations between cleavage-blastocyst rate, and SERPINA5-cleavage rate.

Seminal plasma proteins protect flow-sorted ram spermatozoa from freeze - thaw damage

2009 ◽  
Vol 21 (4) ◽  
pp. 571 ◽  
Author(s):  
T. Leahy ◽  
J. I. Marti ◽  
G. Evans ◽  
W. M. C. Maxwell
Keyword(s):  
Seminal Plasma ◽  
High Speed ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Weight Fraction ◽  
Protein Supplementation ◽  
Freezing Process ◽  
Spermatozoa Motility ◽  
Functional Integrity

Seminal plasma improves the functional integrity of compromised ram spermatozoa but has been reported to be toxic to sorted spermatozoa. The present study attempted to clarify this paradoxical effect and improve the functional integrity of spermatozoa following sorting and cryopreservation. The in vitro function of sorted spermatozoa (motility characteristics and membrane integrity) was examined after supplementation with differing concentrations and protein fractions of seminal plasma at various stages of the sorting and freezing process. For all experiments, spermatozoa (two males, n = four ejaculates per male) were processed through a high-speed flow cytometer before cryopreservation, thawing and incubation for 6 h (37°C). Supplementation of crude seminal plasma (CP), its low molecular weight fraction (LP; <10 kDa) or protein-rich fraction (SPP; >10 kDa), immediately before freezing improved the functional integrity of sorted spermatozoa compared with no supplementation (control), whereas supplementation after thawing had no effect for CP and LP. The protective effect of seminal plasma was not altered by increasing the amount of protein supplementation. No toxic effect of CP, SPP or LP was evident even when supplemented at high protein concentrations. It is concluded that seminal plasma protein, if added to ram spermatozoa after sorting and before freezing, can improve post-thaw sperm quality and consequently the efficiency of sorting. This effect is most likely related to protection of the spermatozoa during freeze–thawing.

scholarly journals Vitamin C ameliorates tetrahydrocannabinol-induced spermatotoxicity in-vitro

2020 ◽  
Author(s):  
Abdullateef Isiaka Alagbonsi ◽  
Luqman Aribidesi Olayaki
Keyword(s):  
Vitamin C ◽  
Cannabinoid Receptors ◽  
Group Iv ◽  
Group Vi ◽  
Spermatozoa Motility ◽  
Progressive Motility ◽  
Straight Line ◽  
Lateral Head

Abstract Background: We investigated the in-vitro effects of vitamin C on delta-9-tetrahydrocannabinol (THC) -induced reduction in spermatozoa motility and kinematics. Methods: Six rats were used for the study. Semen from each of the 6 rats was randomly divided into 6 groups such that each rat’s semen was in all of the groups. Groups I-III received placebo, THC (1 mM), and vitamin C (5 mM) respectively. Group IV was pretreated with cannabinoid receptors’ blockers (CBs-) 1 and 2, followed by THC. Groups V and VI received THC and vitamin C, but group VI was additionally pre-treated with CBs-. Results: The spermatozoa progressive motility, average path velocity (VAP), curvilinear velocity (VCL), straight-line velocity (VSL), amplitude of lateral head (ALH) and beat cross frequency (BCF) were reduced by THC (6.08±1.16%; 5.64±0.82 µm/s; 6.96±0.74 µm/s; 2.75±0.23 µm/s; 0.31±0.02 µm; and 0.78±0.08 Hz respectively) but increased by vitamin C (51.20±1.32 %; 17.90±0.21 µm/s; 25.11±0.96 µm/s; 8.80±0.27 µm/s; 0.75±0.01 µm; and 3.15±0.03 Hz respectively) when compared to control (39.72±0.38 %; 13.70±0.29 µm/s; 18.04±0.58 µm/s; 7.54±0.34 µm/s; 0.65±0.02 µm; and 2.79±0.01 Hz respectively). Vitamin C inhibited the THC-induced reduction in these parameters (37.36±0.73 %; 10.98±0.45 µm/s; 13.58±0.30 µm/s; 7.11±0.22 µm/s; 0.58±0.01 µm; and 2.60±0.01 Hz respectively) in the absence of CBs- 1 and 2, and even caused additional increases in progressive motility (49.54±1.01 %), VAP (15.70±0.38 µm/s) and VCL (22.53±0.29 µm/s) above the control levels with CBs-.Conclusion: Vitamin C ameliorates the THC-induced reduction in spermatozoa motility in-vitro by modulation of their kinematics.

scholarly journals Short-term storage of tiger salamander (Ambystoma tigrinum) spermatozoa: The effect of collection type, temperature and time

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0245047
Author(s):  
Amanda B. Gillis ◽  
Emmet L. Guy ◽  
Andrew J. Kouba ◽  
Peter J. Allen ◽  
Ruth M. Marcec-Greaves ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Hormone Treatment ◽  
Effect Of Temperature ◽  
Initial Assessment ◽  
Ambystoma Tigrinum ◽  
Tiger Salamander ◽  
Storage Duration ◽  
Spermatozoa Motility ◽  
Over Time

The aims of this project were to characterize tiger salamander (Ambystoma tigrinum) spermatozoa motility over time, when excreted as either milt or spermic urine prior to packaging into a spermatophore, and to determine the effect of temperature on sperm motility. A split-plot design was utilized to assess the motility of the two pre-spermatophore sample types at two temperatures, 0°C and 20°C (n = 10 for each treatment). Spermiation was induced through exogenous hormone treatment of luteinizing hormone releasing hormone analog in order to collect both milt and spermic urine, which were evaluated for motility, divided into two separate aliquots, and subsequently stored in either an ice-bath (0°C) or on the benchtop (20°C). The decay rate of sperm motility was assessed by reevaluating subsamples at 0.5, 1, 2, 3, 5, 7, and 24 hours following the initial assessment. Results showed that sperm stored at 0°C had significantly higher progressive, non-progressive, and total motility for both sperm collection types over time. An interaction was found between collection type and time, with milt exhibiting lower initial motility that was more sustainable over time, compared to spermic urine. For both milt and spermic urine, motility decreased rapidly with storage duration, indicating samples should be used as soon as possible to maximize motility for in-vitro fertilization and cryopreservation. This is the first study to describe the differences in sperm motility between milt and spermic urine from an internally fertilizing caudate and demonstrates the benefits of near freezing temperatures on sperm longevity.

scholarly journals Antioxidant Effect of a Polyphenol-Rich Murtilla (Ugni molinae Turcz.) Extract and Its Effect on the Regulation of Metabolism in Refrigerated Boar Sperm

2019 ◽  
Vol 2019 ◽  
pp. 1-15 ◽  
Author(s):  
Ignacio Jofré ◽  
Magdalena Cuevas ◽  
Leticia Signori de Castro ◽  
João Diego de Agostini Losano ◽  
Mariana Andrade Torres ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Damage ◽  
Antioxidant Effect ◽  
Boar Sperm ◽  
Fertilization Capacity ◽  
Ugni Molinae

The production of reactive oxygen species (ROS) in boar spermatozoa increases in refrigeration; this can have an impact on sperm quality and fertilization capacity. We evaluated the effect of polyphenol-rich aqueous extract of murtilla (Ugni molinae Turcz) on boar sperm stored at 17°C in order to reduce oxidative stress and improve sperm quality in the long term. Five experiments were performed: first, characterization of the polyphenol content from five genotypes of murtilla; second, determination of the genotype with the best antioxidant effect (MT-Ex); third, the antioxidant capacity on O2- and lipid peroxidation; fourth, the influence of MT-Ex on motility, calcium movement, cAMP, and metabolic parameters; and fifth, analysis of long-term refrigeration. The average phenolic content was 344 ppm; gallic acid, catechin, quercetin, myricetin, and kaempferol were detected. All extracts evaluated presented a concentration-dependent antioxidant effect. MT-Ex reduces intracellular O2-/peroxides but low lipid peroxidation. MT-Ex in nonstimulated ROS conditions reduces sperm motility, mitochondrial membrane potential, cAMP, and ATP, but the succinate dehydrogenase activity remained normal; also, we observed a reduction in calcium movement in in vitro sperm capacitation. The long-term analyses showed that MT-Ex improved sperm motility decay and reduced membrane damage and ROS at 168 h. Based on this study, we propose MT-Ex as a supplement in semen extenders.

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