scholarly journals Is there a role for endocannabinoids in sperm–oviduct interaction?

Reproduction ◽  
2010 ◽  
Vol 140 (2) ◽  
pp. 247-257 ◽  
Author(s):  
R Talevi ◽  
V Barbato ◽  
S De Iorio ◽  
V Mollo ◽  
T Capriglione ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Phospholipase D ◽  
Zona Pellucida ◽  
Endocannabinoid System ◽  
Sperm Storage ◽  
Sperm Viability ◽  
Progressive Motility ◽  
Bovine Spermatozoa ◽  
Sperm Reservoir

The endocannabinoid system (ECS) has been found in reproductive cells and tissues in several mammals. Spermatozoa are able to respond to anandamide, and the oviduct is able to synthesize and modulate the concentration of this endocannabinoid along the isthmic and ampullary regions. The main aim of this study was to understand whether the ECS has a role during sperm storage and release within the oviduct in cattle. Data showed that 1) the endocannabinoid receptors 1 and 2 (CB1 and CB2) are present in bovine spermatozoa both in the initial ejaculate and in spermatozoa bound to the oviduct in vitro; 2) CB1 receptor is still detectable in spermatozoa released from the oviduct through penicillamine but not in those released through heparin; 3) arachidonylethanolamide (AEA) does not affect sperm viability, whereas it depresses sperm progressive motility and kinetic values; 4) sperm–oviduct binding and release in vitro are not influenced by AEA; 5) AEA depresses sperm–zona pellucida (ZP) binding; 6) binding of heparin-capacitated spermatozoa to the ZP is not affected by AEA; 7) N-acylphosphatidylethanolamine-selective phospholipase D, the main enzyme involved in anandamide synthesis, is expressed in oviductal epithelial cells. In conclusion, secretion of AEA from epithelial cells might contribute to the oviduct sperm-reservoir function, prolonging the sperm fertile life through the depression of motility and capacitation. Capacitation signals, such as heparin, that promote sperm release, might remodel the sperm surface and cause a loss of the sperm sensitivity to AEA.

scholarly journals Effect of fruit-juice on spermatozoa viability and lipid peroxidation of Red Sokoto bucks during liquid storage

2021 ◽  
Vol 41 (1) ◽  
pp. 16-27
Author(s):  
J. O. Daramola ◽  
T. A. Sorongbe ◽  
O. M. Onagbesan ◽  
A. V. Jegede ◽  
A. O. Ladokun ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Cell Damage ◽  
Egg Yolk ◽  
Protective Effects ◽  
Sperm Viability ◽  
Progressive Motility ◽  
Liquid Storage ◽  
Watermelon Juice ◽  
And Control

Antioxidants are linked with sperm viability because of their protective effects against cell damage during preservation. In order to enhance the life span of refrigerated buck semen, this study was carried out to determine the effect of fruit-rich antioxidants on spermatozoa viability and lipid peroxidation (LPO) of buck semen during liquid storage. Pooled semen from five Red Sokoto bucks was diluted with Tris-egg yolk based extender and supplemented each with juices from pawpaw tomato and watermelon at 0, 2.5, 5, 7.5 and 10/ 100 ml respectively. Following dilution, the semen samples were assessed subjectively after in vitro storage at 5°C for 24, 48, 72 and 96 hours as regards sperm motility, abnormalities, and acrosome status using a phase-contrast microscope. The concentration of malondialdehyde (MDA) as indices of lipid peroxidation (LPO) in the stored semen was measured in thiobarbituric acid reactive substances (TBARS) at 24, 48, 72 and 96 hours. The results showed highest progressive motility in watermelon juice at 2.5% (P<0.05) during the first 24 hours of storage while the lowest progressive motility was recorded at various levels of pawpaw juice (P<0.05). After 48 hours of storage, extender supplemented with watermelon and tomato juices had better progressive motility compared to control except 7.5% and 10%% of tomato juice (P<0.05). Irrespective of level of juice in the extender, the percentage of intact acrosome was similar among the various juices and control. The results showed that spermatozoa extended with watermelon juice had the lowest (P<0.05) percentage abnormality compared to other extenders at 24, 48, 72 and 96 hours of storage. Higher (P<0.05) percent spermatozoa abnormality compared to other fruit juices and control was observed at 72 and 96 hours of storage in spermatozoa extended with pawpaw juice. Significant reductions of MDA concentrations were achieved by addition of fruit-rich antioxidants to Tris-egg yolk based extender during the first 72 hours and the reduction was much pronounced in extender supplemented with pawpaw juice compared to control (P<0.05). The findings reveal that fruit-rich antioxidants from watermelon and tomato have protective ability to maintain sperm viability and to reduce concentration MDA of buck semen during liquid storage.

scholarly journals The effect of resorcinol on bovine spermatozoa parameters in vitro

2020 ◽  
pp. 675-686
Author(s):  
M Massányi ◽  
M Halo ◽  
L Strapáková ◽  
T Slanina ◽  
P Ivanič ◽  
...  
Keyword(s):  
Cultivation Temperature ◽  
Negative Effects ◽  
Mtt Test ◽  
Progressive Motility ◽  
Time Period ◽  
Entire Duration ◽  
Bovine Spermatozoa ◽  
Time Periods ◽  
Motility Parameters

The goal of this study was to observe the effect of resorcinol on motility, viability and morphology of bovine spermatozoa. The semen was used from six randomly chosen breeding bulls. Ejaculate was diluted by different solutions of resorcinol in 1:40 ratio. Samples were divided into 7 groups with different concentrations of resorcinol (Control, RES1 – 4 mg/ml, RES2 – 2 mg/ml, RES3 – 1 mg/ml, RES4 – 0.5 mg/ml, RES5 – 0.25 mg/ml and RES6 – 0.125 mg/ml). Motility of spermatozoa was detected using CASA method at temperature of 37 °C in time periods 0, 1, 2, 3, 4 hours from the start of the experiment. Significant motility differences between all groups except control and RES6 with difference of 5.58 %, as well as between RES1 and RES2 groups with difference of 2.17 % were found. Progressive motility had the same significant differences. Spermatozoa viability (MTT test) decreased compared to control in all experimental groups during the entire duration of experiment. Observing morphologically changed spermatozoa, no significant changes were observed and a higher percentage of spermatozoa with separated flagellum in all experimental resorcinol groups compared to control were detected. Also, increased number of spermatozoa with broken flagellum, acrosomal changes and other morphological forms in the group with the highest concentration of resorcinol (RES1) were found. Results of our study clearly show negative effects on motility parameters of spermatozoa which depend on concentration, cultivation temperature and time period.

scholarly journals Differential influence of ampullary and isthmic derived epithelial cells on zona pellucida hardening and in vitro fertilization in ovine

2016 ◽  
Vol 16 (1) ◽  
pp. 61-69 ◽  
Author(s):  
Navid Dadashpour Davachi ◽  
Ahmad Zare Shahneh ◽  
Hamid Kohram ◽  
Mahdi Zhandi ◽  
Helia Shamsi ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Epithelial Cells ◽  
Zona Pellucida ◽  
Vitro Fertilization

scholarly journals Bovine sperm-oviduct interactions are characterized by specific sperm behaviour, ultrastructure and tubal reactions which are impacted by sex sorting

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Miguel Camara Pirez ◽  
Heather Steele ◽  
Sven Reese ◽  
Sabine Kölle
Keyword(s):  
Beat Frequency ◽  
Ciliary Beat Frequency ◽  
Sperm Binding ◽  
Tail Movement ◽  
Bovine Spermatozoa ◽  
Sperm Reservoir ◽  
And Function ◽  
The Impact

Abstract To date sperm-oviduct interactions have largely been investigated under in vitro conditions. Therefore we set out to characterize the behaviour of bovine spermatozoa within the sperm reservoir under near in vivo conditions and in real-time using a novel live cell imaging technology and a newly established fluorescent sperm binding assay. Sperm structure and tubal reactions after sperm binding were analysed using scanning and transmission electron microscopy and histochemistry. As a model to specify the impact of stress on sperm-oviduct interactions, frozen-thawed conventional and sex-sorted spermatozoa from the same bulls (n = 7) were co-incubated with oviducts obtained from cows immediately after slaughter. Our studies revealed that within the oviductal sperm reservoir agile (bound at a tangential angle of about 30°, actively beating undulating tail), lagging (bound at a lower angle, reduced tail movement), immotile (absence of tail movement) and hyperactivated (whip-like movement of tail) spermatozoa occur, the prevalence of which changes in a time-dependent pattern. After formation of the sperm reservoir, tubal ciliary beat frequency is significantly increased (p = 0.022) and the epithelial cells show increased activity of endoplasmic reticula. After sex sorting, spermatozoa occasionally display abnormal movement patterns characterized by a 360° rotating head and tail. Sperm binding in the oviduct is significantly reduced (p = 0.008) following sexing. Sex-sorted spermatozoa reveal deformations in the head, sharp bends in the tail and a significantly increased prevalence of damaged mitochondria (p < 0.001). Our results imply that the oviductal cells specifically react to the binding of spermatozoa, maintaining sperm survival within the tubal reservoir. The sex-sorting process, which is associated with mechanical, chemical and time stress, impacts sperm binding to the oviduct and mitochondrial integrity affecting sperm motility and function.

Characterisation of an in vitro system to study maternal communication with spermatozoa

2012 ◽  
Vol 24 (7) ◽  
pp. 988 ◽  
Author(s):  
Ahmed Aldarmahi ◽  
Sarah Elliott ◽  
Jean Russell ◽  
Thomas Klonisch ◽  
Sabine Hombach-Klonisch ◽  
...  
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
In Vitro Model ◽  
Epithelial Cell Line ◽  
Prostaglandin E ◽  
Sperm Viability ◽  
Cell Passage ◽  
In Vitro System ◽  
Vitro Model

In vivo, gamete maturation, fertilisation and early embryonic development take place inside the oviduct. Several studies have indicated that local responses towards gametes and embryos are generated by the maternal reproductive tract. However, no defined in vitro model currently exists to allow detailed and systematic investigation of maternal communications with gametes and embryos. Therefore, we characterised an in vitro model based on the interaction of boar spermatozoa with an immortalised porcine oviduct epithelial cell line to evaluate different factors that may affect this model. The factors tested were sperm viability, source of spermatozoa, cell passage effect and the effect of reproductive and non-reproductive epithelial cells in the interaction with spermatozoa. After 24 h of co-incubation, RNA was extracted and used to synthesise cDNA for quantitative real-time PCR. Alteration in the expression of genes such as adrenomedullin, heat-shock 70-kDa protein 8 and prostaglandin E synthase was considered as the end point of this assay. The results showed that sperm viability and cell passage number had an effect on oviductal gene expression in response to spermatozoa. Oviductal cells showed significant alterations in gene expression when compared with non-reproductive epithelial cells. The simple in vitro system described here has potential application for further studies in our understanding of mechanisms involved in maternal interactions with spermatozoa.

ID: 112: ROLE OF PHOSPHOLIPASE D IN IDIOPATHIC PULMONARY FIBROSIS

2016 ◽  
Vol 64 (4) ◽  
pp. 964.1-964
Author(s):  
V Suryadevara ◽  
T Royston ◽  
E Berdyshev ◽  
L Huang ◽  
V Natarajan ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Idiopathic Pulmonary Fibrosis ◽  
Pulmonary Fibrosis ◽  
Phospholipase D ◽  
Human Lung ◽  
Lung Fibroblasts ◽  
Human Lung Fibroblasts

Idiopathic pulmonary fibrosis (IPF) is a deadly interstitial disease that leads to scarring and fibrosis of the lung tissue. In pulmonary fibrosis, there is injury and denudation of the alveolar epithelium, which further leads to activation of fibroblasts which differentiate into myofibroblasts. This includes several mechanisms including epithelial to mesenchymal transition (EMT). In this study, we investigated the role of phospholipase D (PLD) in IPF and also its underlying mechanism like EMT and fibroblast proliferation and differentiation. An in vivo murine model of bleomycin-induced pulmonary fibrosis (PF) and in vitro models of murine alveolar type-II epithelial cells (MLE-12) and human lung fibroblasts were used. C57BL/6 and genetically engineered PLD2−/− mice were intratracheally challenged with bleomycin (1.5 U/kg animal) for 14 days and markers of inflammation, EMT and fibrosis were determined. MLE-12 cells were treated with specific PLD1 or PLD2 inhibitors prior to bleomycin (10 mU/ml) challenge, and the role of PLD in EMT and apoptosis of alveolar epithelial cells was studied. Human lung fibroblasts were serum-starved (3h), pretreated with PLD1 or PLD2 inhibitors, and the effect of TGF-β (5 ng/ml) on differentiation of lung fibroblast to myofibroblast was determined. Intra-tracheal instillation of bleomycin in the mice for 14 days leads to the progression of fibrosis in the lung. The lung tissues of the bleomycin treated mice were found to have increased PLD2 protein expression, myofibroblast markers like α-SMA, fibronectin, mesenchymal markers like vimentin, inflammatory cytokines and collagen. Genetic deletion of PLD2 in mice attenuated bleomycin-induced lung inflammation and pulmonary fibrosis. In vitro, MLE-12 cells pretreated with either PLD1 or PLD2 inhibitor did not show a profound reduction either in apoptosis or the expression of transcription factors such as SNAIL, and other markers of EMT. However, MLE-12 cells pretreated with both PLD1 (250 nM) and PLD2 (500 nM) inhibitors were resistant to bleomycin-induced apoptosis, and exhibited reduced expression of SNAIL and mesenchymal markers. On the contrary, human lung fibroblasts pretreated with PLD1 and PLD2 inhibitors showed increased fibroblast to myofibroblast differentiation mediated by TGF-β. The present study suggests a role for PLD2 in bleomycin-induced PF. In vitro, inhibition of both PLD1 and PLD2 was necessary to attenuate bleomycin-induced EMT in epithelial cells and TGF-β mediated differentiation of fibroblasts to myofibroblasts. The in vivo and in vitro results identify the mechanism by which PLD regualtes PF and suggest PLD as a potential therapeutic target in pulmonary fibrosis. This work was supported by National Institutes of Health grant P01 HL98050 to VN.

Hyperosmolality and sperm storage in hibernating bats: prolongation of sperm life by dehydration

1994 ◽  
Vol 267 (5) ◽  
pp. R1363-R1370 ◽  
Author(s):  
E. G. Crichton ◽  
B. T. Hinton ◽  
T. L. Pallone ◽  
R. H. Hammerstedt
Keyword(s):  
Metabolic Rate ◽  
Water Transport ◽  
Sperm Storage ◽  
Myotis Lucifugus ◽  
In Vitro Studies ◽  
Progressive Motility ◽  
Epididymal Spermatozoa ◽  
Cell Storage

Osmolalities of epididymal fluids obtained by micropuncture from hibernating species of bats (Myotis lucifugus) rise during sperm storage periods to as high as 1,523 mmol/kgH2O (approximately 5 times that of plasma). In vitro studies establish that hyperosmolality can preserve viability and prevent initiation of progressive motility in bat epididymal spermatozoa as well as induce their quiescence by reducing respiration. Reduction of osmolality (to 500-600 mmol/kgH2O) induces swelling of sperm and allows the initiation of motility and increased metabolic rate; further reduction of osmolality to < 300 mmol/kgH2O compromises permeability barriers and causes loss of motility. We hypothesize that seasonal establishment of hyperosmotic conditions driven by those cells that constitute the limits of the epididymal lumen dehydrates the compliant spermatozoa and thereby minimizes their metabolic needs. A novel form of cell storage dependent on unique adaptations of the epididymal epithelium for solute and water transport is implicated. To date, the operative osmolyte or osmolytes responsible for elevating osmolality in this system remain elusive.

scholarly journals Immune activation decreases sperm viability in both sexes and influences female sperm storage

2012 ◽  
Vol 279 (1742) ◽  
pp. 3577-3583 ◽  
Author(s):  
Preethi Radhakrishnan ◽  
Kenneth M. Fedorka
Keyword(s):  
Immune System ◽  
Immune Activation ◽  
Acute Infection ◽  
Sperm Storage ◽  
Sperm Viability ◽  
Immune Effector ◽  
Sexually Transmitted ◽  
And Storage ◽  
Pathogenic Infection

All animals are under the constant threat of pathogenic infection. However, little is known regarding the influence of acute infection on sperm viability, particularly in female insects. This information is crucial for our understanding of mating and immune system coevolution, considering that females store sperm and serve as the site of sperm competition. Using the fruitfly, Drosophila melanogaster , we examined the influence of infection on sperm viability and storage. Twenty-four hours after haemocoel inoculation with a pathogen mimic (peptidoglycan, PGN) both sexes exhibited reduced sperm viability, indicating that systemic immune activation played a significant role in gamete survival. Surprisingly, sperm death did not appear to result from a reproductive-immune system trade-off, considering that sperm survived 24 h in vitro once removed from their somatic resources. Instead, our results are most consistent with death owing to immune effector collateral damage. We also examined the potential for sexually transmitted pathogens to influence sperm storage. Females mated with ‘infected’ males (created by dipping genitalia into a PGN solution) exhibited a higher proportion of empty sperm stores 48 h after mating compared to their controls. Remarkably, these data indicate that females may increase their fitness by removing ‘infected’ ejaculates from storage over time.

scholarly journals Expression of endocannabinoid system components in human airway epithelial cells: impact of sex and chronic respiratory disease status

2020 ◽  
Vol 6 (4) ◽  
pp. 00128-2020
Author(s):  
Matthew F. Fantauzzi ◽  
Jennifer A. Aguiar ◽  
Benjamin J.-M. Tremblay ◽  
Michael J. Mansfield ◽  
Toyoshi Yanagihara ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Physiological Responses ◽  
Endocannabinoid System ◽  
Disease Status ◽  
Airway Epithelial Cells ◽  
Airway Epithelial ◽  
Human Airway ◽  
Human Airway Epithelial Cells

Cannabis smoking is the dominant route of delivery, with the airway epithelium functioning as the site of first contact. The endocannabinoid system is responsible for mediating the physiological effects of inhaled phytocannabinoids. The expression of the endocannabinoid system in the airway epithelium and contribution to normal physiological responses remains to be defined.To begin to address this knowledge gap, a curated dataset of 1090 unique human bronchial brushing gene expression profiles was created. The dataset included 616 healthy subjects, 136 subjects with asthma, and 338 subjects with COPD. A 32-gene endocannabinoid signature was analysed across all samples with sex and disease-specific analyses performed. Immunohistochemistry and immunoblots were performed to probe in situ and in vitro protein expression.CB1, CB2, and TRPV1 protein signal is detectable in human airway epithelial cells in situ and in vitro, justifying examining the downstream endocannabinoid pathway. Sex status was associated with differential expression of 7 of 32 genes. In contrast, disease status was associated with differential expression of 21 of 32 genes in people with asthma and 26 of 32 genes in people with COPD. We confirm at the protein level that TRPV1, the most differentially expressed candidate in our analyses, was upregulated in airway epithelial cells from people with asthma relative to healthy subjects.Our data demonstrate that the endocannabinoid system is expressed in human airway epithelial cells with expression impacted by disease status and minimally by sex. The data suggest that cannabis consumers may have differential physiological responses in the respiratory mucosa.

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