Influence of IGF-I on buffalo (Bubalus bubalis) spermatozoa motility, membrane integrity, lipid peroxidation and fructose uptake in vitro

2009 ◽  
Vol 113 (1-4) ◽  
pp. 60-70 ◽  
Author(s):  
S. Selvaraju ◽  
I.J. Reddy ◽  
S. Nandi ◽  
S.B.N. Rao ◽  
J.P. Ravindra
Keyword(s):  
Lipid Peroxidation ◽  
Membrane Integrity ◽  
Bubalus Bubalis ◽  
Spermatozoa Motility ◽  
Igf I

scholarly journals Cooling of equine semen at 16°C for 36 hours with addition of different glutathione concentrations

2015 ◽  
Vol 36 (6) ◽  
pp. 3699
Author(s):  
Rodrigo Arruda de Oliveira ◽  
Marco Antônio De Oliveira Viu ◽  
Maria Lúcia Gambarini
Keyword(s):  
Lipid Peroxidation ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Membrane Lipid ◽  
Sperm Cells ◽  
Sperm Viability ◽  
Membrane Lipid Peroxidation ◽  
Acrosomal Membrane ◽  
In Vitro Effects

Handling equine semen during the refrigeration process reduces sperm viability, and consequently causes membrane lipid peroxidation, among other challenges. The present study aimed to evaluate the in vitro effects of glutathione (control, 1. 0, 1. 5, and 2. 5 mM) on equine semen in a refrigeration protocol of 16ºC for 36 hours. The following variables were evaluated after 0, 12, 24, and 36 hours refrigeration: total sperm motility, vigor, viability, and plasma and acrosomal membrane integrity. Motility was higher with 2. 5mM of glutathione (57. 8 ± 7. 3) after 12 hours of refrigeration compared to the control (53. 2 ± 8. 3) (P < 0. 05). After 36 hours of refrigeration, motility was higher with 1. 5 mM (43. 4 ± 12. 7) and 2. 5mM glutathione (45. 5 ± 6. 2), than it was with 1mM glutathione (38. 2 ± 9) and the control (35. 5 ± 18. 4) (P < 0. 05), respectively. Vigor was highest with 1. 5mM glutathione (3. 7 ± 0. 3) after 36 hours compared to the control (3. 2 ± 1. 1), (P < 0. 05). Viability differed between control and 1mM treatments (79. 5 ± 1. 8) only after 24 hours (75. 5 ± 9. 7) (P < 0. 05). Throughout the investigation, no significant differences were noted in plasma and acrosomal membrane integrity (P > 0. 05). The 1. 5 and 2. 5mM glutathione levels were more efficient in protecting sperm cells and yielded higher total motility values after 36 hours of refrigeration.

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.

109 EFFECTS OF VITRIFICATION PROCEDURES ON SUBSEQUENT DEVELOPMENT AND ULTRASTRUCTURE OF IN VITRO-MATURED SWAMP BUFFALO (BUBALUS BUBALIS) OOCYTES

2007 ◽  
Vol 19 (1) ◽  
pp. 172
Author(s):  
D. Boonkusol ◽  
T. Faisaikarm ◽  
A. Dinnyes ◽  
Y. Kitiyanant
Keyword(s):  
Membrane Integrity ◽  
Subsequent Development ◽  
Bubalus Bubalis ◽  
Ultrastructural Changes ◽  
Cortical Granules ◽  
Chi Square ◽  
Developmental Capacity ◽  
Swamp Buffalo ◽  
Chi Square Analysis

The purpose of this study was to investigate the effects of 2 vitrification procedures on the developmental capacity and ultrastructural changes of matured swamp buffalo (Bubalus bubalis) oocytes. In vitro-matured (IVM) oocytes were vitrified by using 35% and 40% ethylene glycol (EG) as vitrification solution (VS) for solid surface vitrification (SSV) and in-straw vitrification (ISV), respectively. Survival rate of vitrified–warmed oocytes was evaluated on the basis of homogeneous cytoplasm, membrane integrity, and complete zona pellucida. All developmental data were analyzed using chi-square analysis. P &lt; 0.05 was considered significant. The blastocyst rates of parthenogenetic vitrified–warmed oocytes were significantly higher with SSV (89.3% and 13.6%, respectively) than with ISV (81.8% and 5.5%, respectively). However, they were still significantly lower than those of control (100% and 34.2%, respectively). For examining the ultrastructural changes, fresh VS-exposed (ISV and SSV), and vitrified–warmed oocytes were processed for transmission electron microscopy. In VS-exposed oocytes, reduction of microvilli abundance and damage of mitochondrial membrane were found only in the ISV group. In vitrified–warmed oocytes, however, it was clear that both methods of vitrification induced profound ultrastructural modifications to microvilli, mitochondria, oolemma, and cortical granules as well as to the size and position of vesicles. Damaged mitochondria were, however, more abundant in ISV vitrified oocytes than in SSV vitrified oocytes, which correlated with the developmental data, showing the superiority of the SSV method. This study demonstrated for the first time the feasibility of vitrification of IVM swamp buffalo oocytes.

scholarly journals Aquaporin-3 regulates endosome-to-cytosol transfer via lipid peroxidation for cross presentation

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0238484
Author(s):  
Sam C. Nalle ◽  
Rosa Barreira da Silva ◽  
Hua Zhang ◽  
Markus Decker ◽  
Cecile Chalouni ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Lipid Peroxidation ◽  
Mhc Class I ◽  
Cd8 T Cells ◽  
Membrane Integrity ◽  
Protein Antigens ◽  
Aquaporin 3 ◽  
Viral Response ◽  
Cross Presentation

Antigen cross presentation, whereby exogenous antigens are presented by MHC class I molecules to CD8+ T cells, is essential for generating adaptive immunity to pathogens and tumor cells. Following endocytosis, it is widely understood that protein antigens must be transferred from endosomes to the cytosol where they are subject to ubiquitination and proteasome degradation prior to being translocated into the endoplasmic reticulum (ER), or possibly endosomes, via the TAP1/TAP2 complex. Revealing how antigens egress from endocytic organelles (endosome-to-cytosol transfer, ECT), however, has proved vexing. Here, we used two independent screens to identify the hydrogen peroxide-transporting channel aquaporin-3 (AQP3) as a regulator of ECT. AQP3 overexpression increased ECT, whereas AQP3 knockout or knockdown decreased ECT. Mechanistically, AQP3 appears to be important for hydrogen peroxide entry into the endosomal lumen where it affects lipid peroxidation and subsequent antigen release. AQP3-mediated regulation of ECT was functionally significant, as AQP3 modulation had a direct impact on the efficiency of antigen cross presentation in vitro. Finally, AQP3-/- mice exhibited a reduced ability to mount an anti-viral response and cross present exogenous extended peptide. Together, these results indicate that the AQP3-mediated transport of hydrogen peroxide can regulate endosomal lipid peroxidation and suggest that compromised membrane integrity and coordinated release of endosomal cargo is a likely mechanism for ECT.

IGF-I Prevents Fas Ligand-Induced Apoptosis in Granulosa Cells of Buffalo (Bubalus bubalis) Ovarian Follicles In Vitro.

2011 ◽  
Vol 85 (Suppl_1) ◽  
pp. 657-657
Author(s):  
Ravindra P. Janivara ◽  
Sunil K. Patil ◽  
Jamuna V. Kolatalu ◽  
Dhanya Joseph ◽  
Raghavendra Subbarao ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Fas Ligand ◽  
Bubalus Bubalis ◽  
Ovarian Follicles ◽  
Igf I ◽  
Induced Apoptosis

136 EFFECTS OF INSULIN-LIKE GROWTH FACTOR I (IGF-1) ON THE DEVELOPMENT AND APOPTOSIS OF PREIMPLANTATION BUFFALO (BUBALUS BUBALIS) EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 215
Author(s):  
F. Lu ◽  
T. Luo ◽  
H. Sun ◽  
N. Li ◽  
X. Liu ◽  
...  
Keyword(s):  
Growth Factor ◽  
Mrna Level ◽  
Bubalus Bubalis ◽  
Apoptotic Index ◽  
Mrna Expression Level ◽  
Blastocyst Development ◽  
Factor I ◽  
Igf I ◽  
Bax Gene

The aim of this study was to explore the effects of insulin-like growth factor-I (IGF-1) on the development and apoptosis of preimplantation buffalo (Bubalus bubalis) embryos derived from IVF or somatic cell nuclear transfer (SCNT) in order to improve the quality of in vitro embryo culture (IVC). Buffalo oocytes collected from ovaries at slaughter were cultured in the maturation medium (TCM-199 + 26.2 mmol L–1 NaHCO3 + 5 mmol L–1 HEPES + 5% FBS) for 22–24 h, and fertilized in vitro, or enucleated and reconstructed for SCNT. Embryos were then cultured in the culture medium (CM: TCM-199 + 3% FBS) supplemented with different concentrations of IGF-1. Blastocyst development was evaluated after 7 days of culture. A total of 1566 oocytes were used in this study. The experimental data were analyzed using two-way ANOVA, P < 0.05 was considered to be statistically significant. The results showed that the cleavage rates of IVF or SCNT embryos cultured with 0, 10, 50, or 100 ng mL–1 IGF-I, were not significantly different (P > 0.05). However, the blastocyst rate of IVF embryos cultured with 50 ng mL–1 IGF-1 was significantly increased compared to the 0 ng mL–1 group (35.1 v. 23.0%; P < 0.05), but not significantly different among the 0, 10, and 100 ng mL–1 groups (23.0 v. 28.2 and 26.5%; P > 0.05). In the same line, more SCNT embryos could develop to the blastocyst stage when cultured in the CM supplemented with 50 ng mL–1 IGF-I by comparison with the 0 ng mL–1 group (32.3 v. 20.2%; P < 0.05), but the blastocyst development decreased with 100 ng mL–1 (32.3 v. 21.4%; P < 0.05). Apoptosis and total cell number (TCN) of IVF/SCNT blastocysts were respectively detected by TUNEL or Hoechst 33342 staining. By comparison with the 0 ng mL–1 group, the TCN of IVF/SCNT blastocysts was significantly increased (IVF: 91.7 ± 6.9 v. 108.7 ± 3.9, SCNT: 76.3 ± 5.6 v. 92.8 ± 3.9; P < 0.05) and the apoptotic index was obviously decreased (IVF: 3.9 ± 0.7 v. 2.5 ± 0.7; 7.2 ± 0.5 v. 2.9 ± 0.5; P < 0.05) when the embryos were cultured in the CM with 50 ng mL–1 IGF-I. The result of RT-qPCR analysis showed that the mRNA expression level of the anti-apoptotic bcl-2 gene was distinctly enhanced, while the mRNA expression level of the pro-apoptotic bax gene was remarkably reduced in IVF/SCNT embryos cultured with 50 ng mL–1 IGF-I by comparison with the 0 ng mL–1 group (P < 0.05). These results demonstrated that supplementing CM with 50 ng mL–1 IGF-1 could improve the developmental competence of buffalo embryos, increase the TCN of blastocysts and decrease their apoptotic index, probably by down-regulating the mRNA level of pro-apoptotic bax gene and up-regulating the mRNA level of anti-apoptotic bcl-2 gene. This work was funded by the China High Technology Development Program (2011AA100607), China Natural Science Foundation (31072033), and Guangxi Science Foundation (2012GXNSFFA060004).

Lipid peroxidation in bull semen influences sperm traits and oxidative potential of Percoll®-selected sperm

Zygote ◽  
2021 ◽  
pp. 1-8
Author(s):  
Vivian Cardoso Castiglioni ◽  
Adriano Felipe Perez Siqueira ◽  
Luana de Cássia Bicudo ◽  
Tamie Guibu de Almeida ◽  
Thais Rose dos Santos Hamilton ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Lipid Peroxidation ◽  
Membrane Integrity ◽  
Oxidative Status ◽  
Oxidative Potential ◽  
Mitochondrial Potential ◽  
Bull Semen ◽  
Peroxidation Index ◽  
Sperm Traits

Summary Although bovine embryo in vitro production (IVP) is a common assisted reproductive technology, critical points warrant further study, including sperm traits and oxidative status of sperm for in vitro fertilization (IVF). Our aim was to evaluate whether the lipid peroxidation index of commercial bull semen is influenced by sperm traits and oxidative status of sperm populations selected using Percoll® gradient. Semen straws from 48 batches from 14 Nelore bulls were thawed individually, analyzed for motility and subjected to Percoll selection. After Percoll, the lipid peroxidation index of the extender was evaluated, whereas selected sperm were analyzed for motility, acrosome and membrane integrity, mitochondrial membrane potential, chromatin resistance and oxidative potential under IVF conditions. Batches were divided retrospectively in four groups according to lipid peroxidation index. Sperm from Group 4 with the lowest index of lipid peroxidation had, after Percoll selection, greater plasma membrane integrity (81.3%; P = 0.004), higher mitochondrial potential (81.1%; P = 0.009) and lower oxidative potential (135.3 ng thiobarbituric acid reactive substances (TBARS)/ml; P = 0.026) compared with Group 1 with highest lipid peroxidation index (74.3%, 73% and 213.1 ng TBARS/ml, respectively). Furthermore, we observed negative correlations for the lipid peroxidation index with motility, membrane integrity and mitochondrial potential, and positive correlations with oxidative potential. In conclusion, oxidative stress in semen straws, as determined using lipid peroxidation in the extender, is associated with sperm traits and their oxidative potential under IVF conditions. These results provided further insights regarding the importance of preventing oxidative stress during semen handling and cryopreservation, as this could affect sperm selected for IVF. Finally, Percoll selection did not completely remove sperm with oxidative markers.

scholarly journals Time and Dose-Dependent Effects of Viscum Album Quercus on Rabbit Spermatozoa Motility and Viability in Vitro

2019 ◽  
pp. 955-972
Author(s):  
M. Halo ◽  
P. Massanyi ◽  
A. Gren ◽  
A. Lasak ◽  
T. Slanina ◽  
...  
Keyword(s):  
Membrane Integrity ◽  
Viscum Album ◽  
Significant Deterioration ◽  
Spermatozoa Motility ◽  
Time Dependent Effect ◽  
Dose Dependent Decrease ◽  
Casa System ◽  
Dose Dependent ◽  
Rabbit Spermatozoa

The target of this study was to evaluate the effect of extract of the European mistletoe – Viscum album quercus L. on spermatozoa motility and viability in vitro. The CASA system was used to determine the spermatozoa motility parameters at different time intervals (0, 1, 2 and 3 h) and spermatozoa viability was determined in five different doses of Viscum album quercus L [10 (QA), 6.6 (QB), 3.3 (QC), 2.5 (QD) and 2 (QE) mg/ml]. Results in experimental groups detected a significant deterioration on rabbit spermatozoa after 1, 2 and 3 hours, compared to the control. The initial total spermatozoa motility showed increased value for all doses of Viscum album quercus in comparison to control. After in vitro culture a dose–dependent decrease (QA: reduction of 69.7 %, QB: reduction of 40.9 %) was found. For the progressive spermatozoa most significant decrease (86.8 % for QA vs. 48.5 % for QB) was detected compared to the control after 3 hours of culture. Spermatozoa viability (MTT test) was decreased in all experiment groups at the end of experiment, but the differences were not significant. Significant alterations of membrane integrity were found in groups with the highest Viscum album quercus concentration (QA, QB), but acrosome integrity showed no significant changes. Results suggest negative dose– and time–dependent effect of Viscum album quercus at higher doses on spermatozoa motility and viability parameters in vitro.

scholarly journals Aquaporin-3 regulates endosome-to-cytosol transfer via lipid peroxidation for cross presentation

2020 ◽  
Author(s):  
Sam C. Nalle ◽  
Rosa Barreira da Silva ◽  
Hua Zhang ◽  
Markus Decker ◽  
Cecile Chalouni ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Lipid Peroxidation ◽  
Mhc Class I ◽  
Cd8 T Cells ◽  
Membrane Integrity ◽  
Protein Antigens ◽  
Aquaporin 3 ◽  
Viral Response ◽  
Cross Presentation

ABSTRACTAntigen cross presentation, whereby exogenous antigens are presented by MHC class I molecules to CD8+ T cells, is essential for generating adaptive immunity to pathogens and tumor cells (1). Following endocytosis, it is widely understood that protein antigens must be transferred from endosomes to the cytosol where they are subject to ubiquitination and proteasome degradation prior to being translocated into the endoplasmic reticulum (ER), or possibly endosomes, via the TAP1/TAP2 complex (2, 3). Revealing how antigens egress from endocytic organelles (endosome-to-cytosol transfer, ECT), however, has proved vexing. Here, we used two independent screens to identify the hydrogen peroxide-transporting channel aquaporin-3 (AQP3) as a regulator of ECT. AQP3 overexpression increased ECT, whereas AQP3 knockout or knockdown decreased ECT. Mechanistically, AQP3 appears to be important for hydrogen peroxide entry into the endosomal lumen where it affects lipid peroxidation and subsequent antigen release. AQP3-mediated regulation of ECT was functionally significant, as AQP3 modulation had a direct impact on the efficiency of antigen cross presentation in vitro. Finally, AQP3-/- mice exhibited a reduced ability to mount an anti-viral response and cross present exogenous extended peptide. Together, these results indicate that the AQP3-mediated transport of hydrogen peroxide can regulate endosomal lipid peroxidation and suggest that compromised membrane integrity and coordinated release of endosomal cargo is a likely mechanism for ECT.

Sign in / Sign up

Export Citation Format

Share Document