scholarly journals Enhancing liquid-chilled storage and cryopreservation capacities of ram spermatozoa by supplementing the diluent with different additives

2020 ◽  
Vol 33 (7) ◽  
pp. 1068-1076 ◽  
Author(s):  
Sherif A. Rateb ◽  
Marwa A. Khalifa ◽  
Ibrahim S. Abd El-Hamid ◽  
Hesham A. Shedeed
Keyword(s):  
Previous Treatment ◽  
Computer Assisted ◽  
Omega 3 Fatty Acids ◽  
Omega 3 ◽  
Vitro Fertilization ◽  
Sperm Analysis ◽  
Treatment Groups ◽  
Chilled Storage ◽  
Computer Assisted Sperm Analysis

Objective: In the present study, we determined efficiency of incorporating caffeine, melatonin or omega-3 polyunsaturated fatty acid in the diluent on mitigating consequences of (a) liquid chilled- and (b) cryo-storage of ram spermatozoa.Methods: In the first experiment, ejaculates (n = 30) were collected from 5 adult rams and were pooled, diluted (1:10) with Tris-citric acid (base diluent) and were split into 4 aliquots assigned for: control (untreated), caffeine (0.1 mM), melatonin (0.3 mM) or omega-3 fatty acids (0.3 mM) (T0). The diluted specimens were stored at 4°C for 48 h, during which sperm physical and cytological properties were evaluated along with oxidative stress indices (T24, T48). In the second experiment, 15 ejaculates (3 per male) were pooled, diluted with glycerolized base diluent (4% glycerol, v/v) and were split corresponding to the same previous treatment groups before being processed for cryopreservation. Post-thaw physical and kinematic sperm properties were assessed by a computer-assisted sperm analysis system.Results: The results clarified superiority of both melatonin and omega-3 supplementation on maintaining (p<0.05) sperm properties, while reducing (p<0.05) lipid peroxidase reaction and enzymatic activities of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase in preservation medium, compared to caffeine either during liquid-chilled storage or cryopreservation of spermatozoa.Conclusion: Melatonin and omega-3 are regarded efficient alternatives to caffeine when processing ram spermatozoa for application of artificial insemination or in vitro fertilization.

scholarly journals Effects of chilled storage and pH of activating solution on different motility parameters in burbot (Lota lota) sperm

2018 ◽  
Vol 63 (No. 11) ◽  
pp. 429-434
Author(s):  
Zoltán Bokor ◽  
Balázs Csorbai ◽  
Levente Várkonyi ◽  
Zsolt Szári ◽  
Ferenc Fodor ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Chilled Storage ◽  
Straight Line ◽  
H 26 ◽  
Computer Assisted Sperm Analysis ◽  
Motility Parameters ◽  
Activating Solution

The effects of a simple saline solution prepared using two different pH (4.4 and 8.5) on sperm motility in burbot were investigated. Results were recorded during a 96-hour chilled storage (4°C) in 24-hour intervals. Measurements were focused on the detailed characteristics of motility using 12 parameters obtained from the Computer-assisted Sperm Analysis (CASA). Significantly higher progressive motility (pMOT), distance average path (DAP), distance curved line, distance straight line (DSL), average path velocity (VAP), curvilinear velocity, straight line velocity, and beat cross frequency (BCF) were observed with the activating solution buffered at pH 8.5 in comparison with pH 4.4. Already after 24 h a significant reduction was measured in pMOT (0 h: 49 ± 24%, 24 h: 12 ± 7%). Similar decreasing tendency was recorded only after 72 h in DAP (0 h: 26 ± 4 µm/s, 72 h: 19 ± 9 µm/s), DSL (0 h: 21 ± 5 µm/s, 72 h: 17 ± 8 µm/s), VAP (0 h: 59 ± 9 µm/s, 72 h: 43 ± 21 µm/s), and BCF (0 h: 28 ± 2 Hz, 72 h: 18 ± 10 Hz). The response of different investigated CASA parameters to different treatments varied in our experiments. According to our studies, numerous burbot sperm motility parameters are sensitive to chilled storage and to low pH of the activating solution. Our results could support the effective sperm quality assessment and successful artificial propagation process in burbot.

scholarly journals Exosomes derived from HEK293T cells interact in an efficient and noninvasive manner with mammalian sperm in vitro

Nanomedicine ◽  
2020 ◽  
Vol 15 (20) ◽  
pp. 1965-1980
Author(s):  
Teresa Vilanova-Perez ◽  
Celine Jones ◽  
Stefan Balint ◽  
Rebecca Dragovic ◽  
Michael L Dustin ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Mitochondrial Membrane ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Sperm Function ◽  
Sperm Analysis ◽  
Mammalian Sperm ◽  
Boar Sperm ◽  
Computer Assisted Sperm Analysis

Aim: To investigate exosomes as a noninvasive delivery tool for mammalian sperm. Materials & Methods: Exosomes were isolated from HEK293T cells and co-incubated with boar sperm in vitro. Results: Internalized exosomes were detected within 10 min of co-incubation. Computer-assisted sperm analysis and flow cytometry demonstrated that even after 5-h of exposure to exosomes, there were no significant deleterious effects with regard to sperm motility, viability, membrane integrity and mitochondrial membrane potential (p > 0.05), thus indicating that exosomes did not interfere with basic sperm function. Conclusion: HEK293T-derived exosomes interacted with boar sperm without affecting sperm function. Exosomes represent a versatile and promising research tool for studying sperm biology and provide new options for the diagnosis and treatment of male infertility.

329 COMPARING CHANGES IN MOTION PARAMETERS IN EPIDIDYMAL AND EJACULATED BOAR SPERMATOZOA UNDER THREE DIFFERENT TREATMENTS

2007 ◽  
Vol 19 (1) ◽  
pp. 280
Author(s):  
M. Sansegundo ◽  
J. C. Gardon ◽  
F. Garcia-Vazquez ◽  
J. Gadea ◽  
C. Matas
Keyword(s):  
Seminal Plasma ◽  
Objective Assessment ◽  
Mammalian Species ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Motion Parameters ◽  
Computer Assisted Sperm Analysis ◽  
Boar Spermatozoa ◽  
Curvilinear Trajectory

The motion ability of mammalian spermatozoa is acquired during their epididymal transit but observed only upon dilution with seminal plasma (SP) at the time of ejaculation (Yanahimachi 1994 in The Physiology of Reproduction, New York: Raven Press). The bicarbonate present in seminal plasma activates multiple sperm functions, some of which are essential for the initiation of motility. Sperm hyperactivity has been observed in vitro in various mammalian species, especially if capacitation of spermatozoa was induced with Ca2+ and bicarbonate media, such as TALP (Harrison et al. 1996 Mol. Reprod. Dev. 45, 378–391). Computer-assisted sperm analysis (CASA) is a tool for the objective assessment of sperm motility. The aim of this study was to determine if there are differences in motility parameters of ejaculated (EJ) and epididymal (EP) boar spermatozoa under different treatments. Ejaculated and epididymal sperm cells from 10 different boars in each group were used. The sperm treatments were: washed in Dulbecco&apos;s PBS supplemented with 0.1&percnt; BSA (PBS-BSA), washed on a Percoll gradient (PG), and unwashed (UW: Control); the sperm samples were incubated in TALP medium at 38.5&deg;C and 5&percnt; CO2 during the analysis. Motion parameters were determined using a computer-assisted sperm analysis (CASA) system. A 7-&micro;L drop of the sample was placed on a warmed (37&deg;C) slide. At least 4 fields per sample were evaluated, with a minimum of 100 spermatozoa counted per sub-sample. The CASA-derived motility characteristics studied were motility (MOT, &percnt;), progressive motility (PM, &percnt;), curvilinear velocity (VCL, &micro;m s&minus;1), straight-line velocity (VSL, &micro;m s&minus;1), average path velocity (VAP, &micro;m s&minus;1), linearity of the curvilinear trajectory (LIN, ratio of VSL/VCL, &percnt;), straightness (STR, ratio of VSL/VAP, &percnt;), amplitude of lateral head displacement (ALH, &micro;m), wobble of the curvilinear trajectory (WOB, ratio of VAP/VCL, &percnt;), and beat cross-frequency (BCF, Hz). Data were analyzed by ANOVA. If we evaluated all of the data together (EJ vs. EP), EP sperm after treatment showed a higher motility (PM: 38.20&percnt;; MOT: 74.23&percnt;) than EJ sperm (PM: 29.27&percnt;; MOT: 63.24&percnt;), and all of the motion parameters related to velocities and ALH were higher in EP (VCL: 86.02; VSL: 41; VAP: 57.94; ALH: 3.21) than in EJ (VCL: 69.70; VSL: 34.67; VAP: 48.16; ALH: 2.54). No differences were found for LIN, STR, WOB, and BCF. The treatments significantly affected the VCL and ALH, with lower values for the PG treatment. When VCL was lower and the VSL and VAP were similar, consequently the LIN and WOB were significantly higher for the PG group. STR also was higher for the PG group. In conclusion, when both groups of sperm were incubated in TALP medium, the EJ sperm showed a decrease in the majority of motion parameters when compared with EP sperm. This work was supported by MEC (AGL2006-03495/GAN) and Fundaci&oacute;n S&eacute;neca (03018/PI/05).

Effects of permethrin, cypermethrin and 3-phenoxybenzoic acid on rat sperm motility in vitro evaluated with computer-assisted sperm analysis

2010 ◽  
Vol 24 (2) ◽  
pp. 382-386 ◽  
Author(s):  
Chen Yuan ◽  
Cheng Wang ◽  
Shu-Qin Gao ◽  
Tian-Tian Kong ◽  
Lei Chen ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis

12 COMPUTER-ASSISTED SPERM ANALYSIS OF FRESH EPIDIDYMAL CAT SPERMATOZOA AND THE IMPACT OF COOLED STORAGE (4°C) ON SPERM QUALITY

2008 ◽  
Vol 20 (1) ◽  
pp. 86
Author(s):  
M. Filliers ◽  
T. Rijsselaere ◽  
P. Bossaert ◽  
V. De Causmaecker ◽  
J. Dewulf ◽  
...  
Keyword(s):  
Reference Values ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Epididymal Sperm ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
The Impact ◽  
Motility Parameters

Feline epididymal sperm is commonly used for in vitro fertilization. It also yields the opportunity to conserve genetic material from valuable males that suddenly die. Epididymal sperm quality parameters vary considerably among laboratories, implicating the need for objective evaluation methods. The aim of the present study was to describe reference values of computer-assisted sperm analysis (CASA) parameters of fresh epididymal cat sperm and to assess the effect of prolonged cooled storage (4�C) on various sample characteristics. Epididymides obtained from tomcats after routine orchiectomy (2–4 pairs/replicate) were sliced to release spermatozoa. The sperm suspension was placed on a 2-layer gradient and, after centrifugation, the sperm pellet was recovered. In Experiment 1 (20 replicates), sperm motility parameters were assessed immediately after retrieval (T0) using the Hamilton Thorne analyzer Ceros 12.1 (HTR; Hamilton Thorne Biosciences, Beverly, MA, USA). In Experiment 2, fresh (T0) sperm samples (4 replicates) were evaluated for motility parameters (HTR), acrosomal status (FITC-Pisum sativum agglutinin staining), morphology (eosin/nigrosin (E/N) staining), and membrane integrity (E/N and SYBR�-14-propidium iodide staining; Molecular Probes, Inc., Eugene, OR, USA). After addition (1:2) of a Tris-glucose-citrate diluent containing 20% egg yolk, samples were cooled and reassessed on Days 1 (T1), 3 (T3), 5 (T5), 7 (T7), and 10 (T10). Results were analyzed in a mixed linear model, with replicate as random factor and time as fixed effect (S-PLUS 7.0; Insightful Corp., Seattle, WA, USA). Results of Experiment 1 were as follows (mean � SD): motility (MOT): 80.8% � 23.5; progressive motility (PMOT): 69.9% � 23.2; velocity average pathway (VAP): 98.7 µm s–1 � 24.2; velocity straight line (VSL): 89.3 µm s–1 � 25.4; velocity curved line (VCL): 134.8 µm s–1 � 31.9; amplitude lateral head (ALH): 4.3 µm � 2.0; beat cross frequency (BCF): 34.6 Hz � 7.0; and straightness (STR): 89.6% � 6.6. In Experiment 2, MOT, PMOT, VAP, VSL, VCL, BCF, and the percentage of normal spermatozoa showed a decrease over time (P < 0.05) compared to fresh samples, starting from T1, T3, T5, T7, T5, T3, and T1, respectively. In contrast, STR, ALH, membrane integrity, and the percentage of acrosome-intact spermatozoa were not affected (P > 0.05) by cooled storage. To summarize, we have presented a set of reference values for CASA-parameters of fresh, epididymal cat spermatozoa. Cooled storage impaired most motility parameters and lowered the percentage of normal spermatozoa, but did not influence membrane integrity or acrosomal status. The effect of cooled storage on DNA fragmentation of sperm and its subsequent influence on in vitro embryo development require further investigation.

Omega-3 Fatty Acids (O3FA) and Embryo Quality: Do Erythrocyte Levels Affect In Vitro Fertilization (IVF) Outcomes?

2014 ◽  
Vol 101 (2) ◽  
pp. e22-e23
Author(s):  
K.P. Comerford ◽  
M.H. Papadakis ◽  
M.L. Matthews ◽  
P.B. Marshburn ◽  
R.S. Usadi ◽  
...  
Keyword(s):  
Fatty Acids ◽  
In Vitro Fertilization ◽  
Embryo Quality ◽  
Omega 3 Fatty Acids ◽  
Omega 3 ◽  
Vitro Fertilization ◽  
Ivf Outcomes

scholarly journals Effects of zearalenone, α-zearalenol, and genistein on boar sperm motility in vitro

2017 ◽  
Vol 62 (No. 10) ◽  
pp. 435-445 ◽  
Author(s):  
A. Krejcárková ◽  
P. Folková ◽  
O. Šimoník ◽  
M. Šašková ◽  
R. Krejčířová ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Toxicological Research ◽  
Computer Assisted Sperm Analysis ◽  
In Vitro Effects ◽  
Dose Dependent Increase ◽  
Dose Dependent

Genistein (GEN) and zearalenone (ZEA), environmental oestrogens commonly present in feedstuff for pigs, are known for their effects on reproductive functions. The aim was to verify the in vitro effects of 0.5–20 µM concentrations of GEN, ZEA and its metabolite α-zearalenol (α-ZOL) on pig sperm motility. A dose-dependent increase of the immotile sperm amount against fast and medium-fast sperm clusters was observed with all three oestrogens from the lowest concentrations tested. Individual CASA (computer-assisted sperm analysis) parameters of motile sperms seemed to be less sensitive indicators. This should be considered especially in toxicological research on a sperm model. Background of inconsistencies in to date-published papers is discussed. The results shift the effective concentrations of ZEA, α-ZOL, and GEN to values achievable in vivo and raises the questions of risk assessment of these compounds in pig reproduction.

scholarly journals V-Set and Immunoglobulin Domain-Containing 1 (VSIG1), Predominantly Expressed in Testicular Germ Cells, Is Dispensable for Spermatogenesis and Male Fertility in Mice

Animals ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 1037
Author(s):  
Yena Jung ◽  
Hyewon Bang ◽  
Young-Hyun Kim ◽  
Na-Eun Park ◽  
Young-Ho Park ◽  
...  
Keyword(s):  
Male Fertility ◽  
Computer Assisted ◽  
Developmental Defects ◽  
Vitro Fertilization ◽  
Sperm Analysis ◽  
Reverse Transcription Pcr ◽  
Short Palindromic Repeat ◽  
Immunoglobulin Domain

To elucidate the functional role of V-set and immunoglobulin domain-containing 1 (VSIG1) in spermatogenesis and fertilization, we knocked out (KO) VSIG1 in a mouse embryo using CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9) -mediated genome editing. Reverse transcription PCR was performed using cDNA synthesized from VSIG1 KO testis RNA. Although Western blot analysis using a specific antibody to VSIG1 confirmed VSIG1 protein defects in the KO mice, hematoxylin-eosin staining analysis was similar in the KO and wild-type mice. Additionally, computer-assisted sperm analysis and in vitro fertilization experiments were conducted to confirm the activity and fertilization ability of sperm derived from the KO mouse. Mice lacking VSIG1 were viable and had no serious developmental defects. As they got older, the KO mice showed slightly higher weight loss, male mice lacking VSIG1 had functional testes, including normal sperm number and motility, and both male and female mice lacking VSIG1 were fertile. Our results from VSIG1 KO mice suggest that VSIG1 may not play essential roles in spermatogenesis and normal testis development, function, and maintenance. VSIG1 in sperm is dispensable for spermatogenesis and male fertility in mice. As several genes are known to possess slightly different functions depending on the species, the importance and molecular mechanism of VSIG1 in tissues of other species needs further investigation.

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