The Effect of Genistein on the Plasma Membrane Integrity of Frozen Ongole Grade Bull Semen Based on Skim Milk – Soy Lecithin Extender

This study was designed to compare the kinetics of sperm survival in different types of bull semen. Fresh ejaculates from four bulls were pooled, diluted in Tris-citric acid-egg yolk-glycerol extender, cooled to 4°C, frozen in LN2 and thawed at 37°C. Fresh, diluted, cooled and frozen–thawed semen were incubated at 37°C, and evaluated at 0, 2, 4, 6, 12 and 24 h after the beginning of incubation. In Experiment 1, progressive sperm motility, normal acrosomes and plasma membrane integrity and asymmetry were determined. In Experiment 2, generation of superoxide anion (O2•) along with plasma membrane permeability and generation of hydrogen peroxide (H2O2) along with plasma membrane integrity were assessed. In Experiment 1, frozen–thawed semen had shorter survival times for progressive sperm motility, and spermatozoa with intact plasma membranes and acrosomes (IPM-IACR) as compared with other types of semen (P < 0.05). Fresh spermatozoa underwent a necrotic pathway, diluted and cooled spermatozoa underwent an apoptosis-like pathway and frozen–thawed spermatozoa underwent both necrotic and apoptosis-like pathways. In Experiment 2, spermatozoa in all four types of semen exhibited O2•– generation and increased plasma membrane permeability, and became necrotic without H2O2 generation during incubation (P < 0.05). In conclusion, frozen–thawed semen had shorter sperm longevity, which has important implications relating to the timing of artificial insemination. Different types of semen followed different death pathways. During incubation, spermatozoa in all types of semen generated O2•–, which increased the permeability and compromised the integrity of the plasma membrane.

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Effects of the platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen (AI) with reduced sperm motility

Polish Journal of Veterinary Sciences ◽

10.1515/pjvs-2016-0019 ◽

2016 ◽

Vol 19 (1) ◽

pp. 147-158 ◽

Cited By ~ 2

Author(s):

M. Lecewicz ◽

W. Kordan ◽

A. Majewska ◽

S. Kamiński ◽

A. Dziekońska ◽

...

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Platelet Activating Factor ◽

Quality Parameters ◽

Sperm Viability ◽

Plasma Membrane Integrity ◽

Positive Effects ◽

Bull Semen

Abstract The aim of the study was to determine the effects of platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen with reduced sperm motility used for artificial insemination. The aim of experiment 1 was to identify the optimal concentration of the phospholipid able to preserve sperm viability. Cryopreserved semen was treated with different PAF concentrations: 1×10-5M, 1×10-6M, 1×10-7M, 1×10-8M and 1×10-9M. The experiment demonstrated that PAF at concentration 1×10-9M increased most the sperm viability parameters (motility parameters, plasma membrane integrity and mitochondrial function) after 120 min of incubation of thawed semen at 37°C. Cryopreserved bull semen with reduced sperm motility (below 70%) was supplemented with PAF in a concentration of 1×10-9M. A statistically significant increase in sperm motility, percentage of linear motile spermatozoa and VSL value was observed after 120 min incubation of sperm with 1×10-9M PAF. Sperm supplementation with PAF also had positive effects on plasma membrane integrity and percentage of spermatozoa with preserved mitochondrial transmembrane potential, but the differences were not statistically significant. The results indicated positive effects of PAF supplementation at a concentration of 1×10-9M on the selected sperm quality parameters in cryopreserved bull semen with reduced motility.

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Polymyxin B changes the plasma membrane integrity of cryopreserved bull semen

Asian Pacific Journal of Reproduction ◽

10.1016/j.apjr.2016.07.008 ◽

2016 ◽

Vol 5 (5) ◽

pp. 411-415 ◽

Cited By ~ 1

Author(s):

Mojtaba Rashedi ◽

Mohammad Hashem Fazeli ◽

Mohammad Bahreini

Keyword(s):

Plasma Membrane ◽

Membrane Integrity ◽

Polymyxin B ◽

Plasma Membrane Integrity ◽

Bull Semen

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Effect of α-tocopherol supplementation in diluents on the motility, viability and plasma membrane integrity of Simmental bull spermatozoa after cooling

Ovozoa Journal of Animal Reproduction ◽

10.20473/ovz.v9i1.2020.1-6 ◽

2020 ◽

Vol 9 (1) ◽

pp. 1

Author(s):

Sarah Azura ◽

Hermin Ratnani ◽

Suherni Susilowati ◽

Mas'ud Hariadi ◽

Abdul Samik ◽

...

Keyword(s):

Plasma Membrane ◽

Membrane Integrity ◽

Skim Milk ◽

Egg Yolk ◽

Oxygen Species ◽

Cold Temperatures ◽

Plasma Membrane Integrity ◽

Semen Storage ◽

Simmental Cattle ◽

Antioxidant Supplement

Semen storage in cold temperatures might cause an increase in reactive oxygen species (ROS) production. This condition resulted in spermatozoa damage and quality decrease. This study was conducted to investigate the effects of α-tocopherol supplementation in diluents on the motility, viability, and plasma membrane integrity of Simmental bull spermatozoa after cooling. Semen samples were diluted in skim milk egg yolk supplemented with 0, 0.5, 1.0, and 1.5 mM α-tocopherol respectively for control, Tl, T2, and T3. Spermatozoa were evaluated for their motility, viability, and membrane integrity in cooling temperature (5°C). The daily evaluation showed that 1.5 mM α-tocopherol was the best in maintaining motility, viability, and plasma membrane integrity, while 1.0 mM α-tocopherol was only good for maintaining viability. Therefore, it can be concluded that α-tocopherol at the concentration of 1.5 mM was an efficient antioxidant supplement for Simmental cattle semen in skim milk egg yolk diluent.

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Cryopreservation of bull semen shipped overnight and its effect on post-thaw sperm motility, plasma membrane integrity, mitochondrial membrane potential and normal acrosomes

Animal Reproduction Science ◽

10.1016/j.anireprosci.2011.04.018 ◽

2011 ◽

Vol 126 (1-2) ◽

pp. 23-31 ◽

Cited By ~ 25

Author(s):

M. Anzar ◽

T. Kroetsch ◽

L. Boswall

Keyword(s):

Plasma Membrane ◽

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Sperm Motility ◽

Mitochondrial Membrane ◽

Membrane Integrity ◽

Plasma Membrane Integrity ◽

Bull Semen

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12 FERTILITY OF FROZEN EQUINE SPERM IN SYSTEMS FOR CRYOPRESERVATION

Reproduction Fertility and Development ◽

10.1071/rdv24n1ab12 ◽

2012 ◽

Vol 24 (1) ◽

pp. 117

Author(s):

R. R. D. Maziero ◽

P. N. Guasti ◽

I. D. P. Blanco ◽

I. Martin ◽

G. A. Monteiro ◽

...

Keyword(s):

Plasma Membrane ◽

Liquid Nitrogen ◽

Membrane Integrity ◽

Skim Milk ◽

Egg Yolk ◽

Automated System ◽

Epifluorescence Microscopy ◽

Computer Assisted ◽

Plasma Membrane Integrity ◽

Sperm Analysis

Optimizing cryopreservation of equine sperm will facilitate genetic banking and propagation of important horse strains through assisted reproduction. This study aimed to evaluate the motility pattern using computer-assisted sperm analysis (CASA) and plasma membrane integrity by epifluorescence microscopy of equine semen frozen in 0.5 mL straws at different freezing rates; also, a fertility trial was performed according to the freezing protocol. Three ejaculates from four stallions of various breeds (Mangalarga Marchador, Westfallen, Hanovarian and Arabian) and ages (5 to 20 years) were collected and processed for cryopreservation. The stallions were housed at the CERBEQ, Reproduction Centre of the Department of Animal Reproduction and Veterinary Radiology, UNESP. The ejaculates were filtered and submitted to analysis by CASA (HTM IVOS 12, Hamilton Thorne Research, USA). In addition, the plasma membrane integrity was determined by fluorescent probes. After evaluation, the ejaculates were diluted at 1:1 (extender:semen) with skim milk extender Botu-Semen™ and centrifuged at 600 × g for 10 min. The supernatant was removed and the pellet resuspended to a final concentration of 100 × 106 sperm mL–1 with milk-egg yolk freezing extender (Botu-Crio™). Semen was packaged in 0.5-mL straws (IMV, LAigle, France) and was placed in nitrogen for 20 min and then from room temperature to 5°C and then frozen in two different cooling systems: an isothermic box (42 cm × 28 cm × 12.5 cm) was placed upon racks suspended 6 cm above liquid nitrogen or other 20 min then immersed into nitrogen and automated system Mini Digitcool™ (IMV Technologies, France), cooling at a –40°C min–1 rate. All straws were stored in liquid nitrogen until thawing and analysis. The straws were thawed in a water bath at 46°C for 20 s and the samples were evaluated for progressive motility, angular progressive velocity, progressive velocity, track speed, percentage of rapid sperm and percentage of sperm with plasma membrane integrity. For the fertility trial, 65 clinically healthy mares had their oestrous cycle monitored by ultrasound and inseminated postovulation with sperm into the uterus. Ovulation was induced with 1 mL of deslorelin acetate (GnRH) injected IM when a 35-mm follicle was detected. Thirty-six hours later, mares were monitored every 6 h until ovulation was detected. When it was detected, mares were inseminated with 800 × 106 total sperm. Pregnancy was confirmed via ultrasound examination 15 days after ovulation. Pregnancy rate was 52.2% using the isothermic box and 60% using the automated machine. Statistical analysis from the frozen–thawed semen evaluated parameters was performed using the statistics software Proc. MIXED of SAS 9.1 and for the fertility trial, logistic regression using the Proc GENMOD from SAS 9.1. The conventional method using the isothermic box was similar to the automated machine with a fast freezing rate. Additionally, AI with 800 × 106 sperm frozen in the isothermic box or automated system resulted in similarly acceptable conception rates.

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Donkey Epididymal Transport for Semen Cooling and Freezing

Animals ◽

10.3390/ani10122209 ◽

2020 ◽

Vol 10 (12) ◽

pp. 2209

Author(s):

Yamilka Lago-Alvarez ◽

Giorgia Podico ◽

Lorenzo G. Segabinazzi ◽

Lais L. Cunha ◽

Leonardo Barbosa ◽

...

Keyword(s):

Plasma Membrane ◽

Mixed Models ◽

Membrane Integrity ◽

Skim Milk ◽

Egg Yolk ◽

Semen Parameters ◽

Sperm Activation ◽

Plasma Membrane Integrity ◽

Semen Extender ◽

Motility Parameters

The objectives of this study were to assess the cooling and freezing of donkey epididymal semen harvested immediately after castration (Experiment 1, n = 4) or after the shipment (24 or 48 h) of epididymides attached to testicles (Experiment 2, n = 14) or dissected apart (Experiment 3, n = 36). In each experiment, semen was frozen immediately (Non-Centrif) in an egg yolk-based semen extender (EY) or after processing through cushion-centrifugation (Centrif) while extended in a skim milk-based extender (SC). In all three experiments, cooled, pre-freeze, and post-thaw epididymal semen was assessed for total motility (TM), progressive motility (PM), plasma membrane integrity (PMI), and high mitochondrial membrane potential (HMMP). Data were analyzed with R using mixed models and Tukey’s test as posthoc. Results showed that the cooling of epididymal semen up to 24 h after harvesting did not affect motility parameters or plasma membrane integrity; furthermore, in Experiment 3, the post-thaw evaluation of both Centrif and Non-Centrif achieved similar TM and PM. Collectively, the post-thaw results revealed low motility parameters across groups; while, the PMI and HMMP did not reflect this trend, and the values remained high, suggesting that there was a lack of epididymal sperm activation with either centrifugation or extenders. In summary, freshly harvested and cooled-shipped and cooled semen had satisfactory semen parameters. Future studies need to address donkey epididymal semen fertility in mares and jennies.

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Comparision of DNA fragmentantion and plasma membrane integrity between chilled and frozen semen of bulls

Reproduction Abstracts ◽

10.1530/repabs.1.p248 ◽

2014 ◽

Author(s):

Mello Papa Patricia de ◽

Carlos Ramires Neto ◽

Priscilla Nascimento Guasti ◽

Rosiara Rosaria Dias Maziero ◽

Yame F R Sancler-Silva ◽

...

Keyword(s):

Plasma Membrane ◽

Membrane Integrity ◽

Plasma Membrane Integrity ◽

Frozen Semen

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Plasma membrane integrity in health and disease: significance and therapeutic potential

Cell Discovery ◽

10.1038/s41421-020-00233-2 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Catarina Dias ◽

Jesper Nylandsted

Keyword(s):

Plasma Membrane ◽

Therapeutic Potential ◽

Calcium Influx ◽

Membrane Integrity ◽

Cell Types ◽

Physical Parameters ◽

Membrane Repair ◽

Plasma Membrane Integrity ◽

Repair Mechanisms ◽

And Function

AbstractMaintenance of plasma membrane integrity is essential for normal cell viability and function. Thus, robust membrane repair mechanisms have evolved to counteract the eminent threat of a torn plasma membrane. Different repair mechanisms and the bio-physical parameters required for efficient repair are now emerging from different research groups. However, less is known about when these mechanisms come into play. This review focuses on the existence of membrane disruptions and repair mechanisms in both physiological and pathological conditions, and across multiple cell types, albeit to different degrees. Fundamentally, irrespective of the source of membrane disruption, aberrant calcium influx is the common stimulus that activates the membrane repair response. Inadequate repair responses can tip the balance between physiology and pathology, highlighting the significance of plasma membrane integrity. For example, an over-activated repair response can promote cancer invasion, while the inability to efficiently repair membrane can drive neurodegeneration and muscular dystrophies. The interdisciplinary view explored here emphasises the widespread potential of targeting plasma membrane repair mechanisms for therapeutic purposes.

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