scholarly journals Acrosome integrity and mitochondrial membrane potential of frozen thawed buffalo semen treated with heparin binding protein

2020 ◽  
Vol 8 (3) ◽  
pp. 2438-2443
Author(s):  
Yanben M Kikon ◽  
K Loganathasamy ◽  
VS Gomathy ◽  
K Vijayarani ◽  
M Vengatachalam
2021 ◽  
Vol 69 (3) ◽  
pp. 291-297
Author(s):  
Muhammed Enes İnanç ◽  
Şükrü Güngör ◽  
Emir Gül ◽  
Barış Atalay Uslu ◽  
Ayhan Ata

Abstract The aim of this study was to determine the effects of gallic acid (GA) on frozen-thawed goat spermatozoa. Four Honamli goat bucks were used at their breeding season, and ejaculates were collected by an electroejaculator. Mixed semen was divided into the following four groups: control (0 mM), low (L; 1 mM), medium (M; 2 mM), and high (H; 4 mM) concentration of GA. All the groups were frozen and thawed in a water bath for spermatological evaluation. The lowest motility was observed in the control group (47.60 ± 5.70%) (P < 0.05), while the highest viability (62.45 ± 1.68%), plasma membrane and acrosome integrity (44.81 ± 4.57%), and high mitochondrial membrane potential (35.96 ± 2.50%) were observed in the low GA group (P < 0.05). Also, the lowest hypo-osmotic swelling test (HOS +) value was found in the high GA group (47.60 ± 4.82%) (P < 0.05). In conclusion, supplementing a low concentration (1 mM) of GA to the Tris-based semen extender had a positive effect on spermatological parameters after freeze-thawing of Honamli goat semen. Further studies should be continued in other species with different doses and combinations using commercial and/or homemade semen extenders.


Author(s):  
Amit Kumar ◽  
Sujata Pandita ◽  
N. Anand Laxmi ◽  
Mukesh Bhakat ◽  
T. K. Mohanty

Prostasomes are extracellular vesicles that fuse with sperms thereby improving its functional parameters. Present study aimed to isolate and characterise prostasomes from semen of KF bulls, and to investigate prostasomes effects on functional parameters of KF bull spermatozoa. Isolated prostasomes were characterized with respect to the binding of FITC-conjugated CD 26 antibodies, as well as protein, cholesterol and phospholipids content. Subsequently, effects of prostasomes supplementation (1mg/ml) were investigated on ROS production, Ca2+ signalling, mitochondrial membrane potential and acrosome integrity of fresh and cryopreserved-thawed spermatozoa. Isolated prostasomes were immunostained positively. Prostasomes showed higher proportion of both protein and cholesterol as compared to phospholipids. When sperm samples were supplemented with prostasomes, ROS production decreased, while all other functional parameters improved.


Reproduction ◽  
2020 ◽  
Vol 160 (5) ◽  
pp. 783-792
Author(s):  
Maria S Ferrer ◽  
Roberto Palomares ◽  
David Hurley ◽  
Anna-Claire Bullington ◽  
Alejandro Hoyos-Jaramillo ◽  
...  

Bovine antisperm antibodies (ASAs) have been associated with teratospermia and asthenospermia. It was hypothesized here that scrotal insulation induces the formation of ASAs and deterioration of sperm function. Scrotal insulation bags were placed in 10 bulls for 8 days. Semen was collected on days −29, −22 and −2, twice weekly from days 5 to 54, and thereafter weekly until day 96 (day 0 = first day of scrotal insulation). On each collection day, scrotal circumference, sperm motility, morphology, membrane integrity, acrosome integrity, apoptosis, lipid peroxidation, mitochondrial membrane potential, ASA binding and DNA integrity were evaluated. The percentage of IgG- and IgA-bound sperm increased between days 12 and 96 (P < 0.0001), in association with poor motility (days 19–30, P < 0.005) and morphology (days 8–40, P < 0.0001). Mean scrotal circumference decreased between days 15 and 75 (P < 0.0001). There was also a deterioration in sperm membrane integrity (days 19–40, P < 0.0001), acrosome integrity (days 26–89, P < 0.0001), lipid peroxidation (days 5–12, P < 0.0001), and mitochondrial membrane potential (days 12–96, P = 0.001). In contrast, a decrease in apoptotic cells (days 37–83, P = 0.0002) and lipid peroxidation (days 19–96, P < 0.0001) was noticed. Most bulls recovered normospermia by day 96. However, the persistence of ASAs, acrosomal damage and dysfunctional mitochondria suggest a long term effect of scrotal insulation on sperm function and the homeostasis of the reproductive immune system.


2007 ◽  
Vol 292 (1) ◽  
pp. L165-L177 ◽  
Author(s):  
Dean A. Wiseman ◽  
Sandra M. Wells ◽  
Maryann Hubbard ◽  
Jonathan E. Welker ◽  
Stephen M. Black

Oxidative stress has been associated with multiple pathologies and disease states, including those involving the cardiovascular system. Previously, we showed that pulmonary artery endothelial cells (PAECs) undergo apoptosis after acute exposure to H2O2. However, the underlying mechanisms regulating this process remain unclear. Because of the prevalence of H2O2 in normal physiological processes and apparent loss of regulation in disease states, the purpose of this study was to develop a more complete understanding of H2O2-mediated adverse effects on endothelial cell survival. Acute exposure of PAECs to H2O2 caused a dose-dependent increase in cellular release of lactate dehydrogenase and a significant increase in production of superoxide ions, which appear to be generated within the mitochondria, as well as a significant loss of mitochondrial membrane potential and activity. Subsequent to the loss of mitochondrial membrane potential, PAECs exhibited significant caspase activation and apoptotic nuclei. We also observed a significant increase in intracellular free Zn2+ after bolus exposure to H2O2. To determine whether this increase in Zn2+ was involved in the apoptotic pathway induced by acute H2O2 exposure, we developed an adenoviral construct for overexpression of the Zn2+-binding protein metallothionein-1. Our data indicate that chelating Zn2+, either pharmacologically with N,N,N′, N-tetrakis(2-pyridylmethyl)ethylene diamine or by overexpression of the Zn2+-binding protein metallothionein-1, in PAECs conferred significant protection from induction of apoptosis and cell death associated with the effects of acute H2O2 exposure. Our results show that the acute toxicity profile of H2O2 can be attributed, at least in part, to liberation of Zn2+ within PAECs. We speculate that regulation of Zn2+ levels may represent a potential therapeutic target for cardiovascular disease associated with acute oxidative stress.


Author(s):  
А.А. Соколовская ◽  
Э.Д. Вирюс ◽  
В.В. Александрин ◽  
А.С. Роткина ◽  
К.А. Никифорова ◽  
...  

Цель исследования. Ишемические повреждения головного мозга, являются одной из наиболее частой причин инвалидности и смертности во всем мире. Недавно была установлена роль апоптоза тромбоцитов в патофизиологии инсульта, однако его механизмы до сих пор остаются невыясненными. Несмотря на различные экспериментальные модели, направленные на мониторинг апоптоза тромбоцитов, результаты, относительно изучения и выявления апоптоза тромбоцитов при ишемии головного мозга у крыс, весьма немногочисленны. Цель исследования - анализ апоптоза тромбоцитов с помощью метода проточной цитофлуориметрии на модели глобальной ишемии мозга у крыс. Методика. В экспериментах использовано 6 крыс-самцов Вистар в возрасте от 5 до 6 мес., разделенных на 2 группы: интактный контроль (К) и глобальная ишемия головного мозга. Модель глобальной ишемии головного мозга у крыс воспроизводилась путём билатеральной окклюзии общих сонных артерий на фоне гипотензии. Уровень системного артериального давления снижали посредством кровопотери до 40-45 мм рт. ст. Суспензию тромбоцитов крыс получали методом гельфильтрации с использованием сефарозы 2B. Для анализа экстернализации фосфатидилсерина (ФС) тромбоциты крыс инкубировали с Аннексином V-PE в связывающем буфере. Для оценки митохондриального мембранного потенциала (ММП) тромбоциты инкубировали с катионным красителем JC-1. После инкубации образцы немедленно анализировали на проточном цитофлуориметре FACSCalibur (Becton Dickinson, США). Результаты. Согласно полученным данным, экстернализация ФС на тромбоцитах крыс, перенесших инсульт, была значительно выше (53,45 ± 4,21%), чем в контрольной группе крыс (5,27 ± 2,40%). Данный эффект подтверждается выраженной деполяризацией митохондриальных мембран (DYm). После экспериментальной ишемии мозга почти 40% тромбоцитов было деполяризовано. Заключение. Использованный в работе подбор методов и маркеров обеспечивает понимание механизмов апоптоза тромбоцитов как в экспериментальных, так и в клинических условиях. Полученные данные позволяют сделать заключение, что апоптоз тромбоцитов является одним из факторов развития глобальной ишемии головного мозга у крыс. Результаты могут быть использованы для понимания механизмов, участвующих в развитии ишемического повреждения, что, в свою очередь, может быть использовано при разработке новых терапевтических стратегий. Aim. Stroke is one of the most common causes of disability and mortality worldwide. Multiple experimental models of stroke have focused on monitoring of platelet apoptosis. However, studies on and detection of platelet apoptosis in rats with ischemic stroke are very scarce. We investigated platelet apoptosis in rats with global brain ischemia using flow cytometry. Methods. Experiments were carried out on healthy, adult Wistar male rats weighing 300-350 g. The rats were divided into the following 2 groups: intact rats and rats with global brain ischemia. Global brain ischemia was induced by two-vessel (2-VO) carotid occlusion in combination with hypotension. Systemic blood pressure was reduced by 40-45 mm Hg by inducing haemorrhage. Platelets were isolated by gel filtration on Sepharose 2B. For evaluation of phosphatidylserine (PS) externalization, platelets were incubated with Annexin V-PE and analyzed on FACSCalibur (BD Biosciences). Mitochondrial membrane potential (DY) was measured during platelets apoptosis using JC-1, a mitochondrial membrane potential indicator. Platelets were analyzed by flow cytometry immediately after the incubation. Results. PS externalization on platelets was significantly greater after global brain ischemia (53.45 ± 4.21%) than in the control group (5.27 ± 2.40%). Pronounced depolarization of mitochondrial membrane potential (DYm) confirmed this finding. In the rat group with experimental brain ischemia, almost 40% (35.24 ± 5.21%) of platelets were depolarized. Conclusion. Our results provide insight into mechanisms involved in platelet apoptosis during ischemic stroke and can be used in further development of new therapeutic strategies.


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