scholarly journals Expression of Fas and Fas ligand protein and mRNA in mouse oocytes and embryos

Reproduction ◽  
2003 ◽  
pp. 791-799 ◽  
Author(s):  
RL Kelkar ◽  
SJ Dharma ◽  
TD Nandedkar
Keyword(s):  
Fas Ligand ◽  
Death Receptor ◽  
Preimplantation Embryos ◽  
Nuclear Antigen ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Proliferation And Apoptosis ◽  
Mrna And Protein Expression ◽  
Fas L ◽  
Apoptotic Pathways

During mammalian embryonic development, abnormal eggs and embryos are eliminated by apoptosis; however, the precise apoptotic pathways remain as yet unidentified. In the present study, expression of Fas and Fas ligand - the proximal members of the death receptor pathway, was evaluated in mouse preimplantation embryos by immunofluorescence and in situ hybridization techniques. Ovulated oocytes were collected from oviducts of cyclic mice on the day of oestrus (day 0), and one-cell, two-cell embryos and eight-cell morulae were collected from oviducts of mated animals on days 1, 2 and 3 of pregnancy, respectively. Blastocysts were flushed from the uterine horns on day 4. Expression of Fas and Fas L mRNAs and proteins was absent from embryos at days 0, 1 and 2. A marked increase in Fas and Fas L mRNA and protein expression was detected in all morphologically normal embryos on day 3 and day 4. In addition, embryos on days 3 and 4 were positive for terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) staining; however, absence of caspase 8 and 3 and intense localization of proliferating cell nuclear antigen confirmed the proliferative status of these embryos. Furthermore, TUNEL staining was absent in postimplantation embryonic sections obtained on day 6. The results of the present studies thus indicate an equilibrium between proliferation and apoptosis in the preimplantation embryo.

scholarly journals Fas and Fas ligand protein and mRNA in normal and atretic mouse ovarian follicles

Reproduction ◽  
2003 ◽  
pp. 783-789 ◽  
Author(s):  
SJ Dharma ◽  
RL Kelkar ◽  
TD Nandedkar
Keyword(s):  
In Situ Hybridization ◽  
Fas Ligand ◽  
Death Receptor ◽  
Underlying Mechanism ◽  
Ovarian Follicles ◽  
Chorionic Gonadotrophin ◽  
Follicular Atresia ◽  
Immature Female ◽  
Apoptotic Pathways

Apoptosis is the underlying mechanism of follicular atresia in the mammalian ovary. However, the apoptotic pathways governing this ovarian process are not completely elucidated. In the present study, expression of Fas and Fas ligand, the proximal members of the death receptor pathway, was evaluated in mouse ovarian follicles using immunofluorescence and in situ hybridization. Normal or atretic follicles were obtained from immature female Swiss mice after administration of 10 iu equine chorionic gonadotrophin for 48 or 72 h, respectively. Expression of both Fas and Fas ligand mRNA and protein was observed in granulosa cells of normal and atretic follicles. Although the oocytes of normal follicles failed to show any staining, those of atretic follicles stained intensely for Fas, indicating that the presence of Fas in the oocyte determines the fate of the follicle.

Modulation of apoptotic pathways in intestinal mucosa during hibernation

2005 ◽  
Vol 289 (2) ◽  
pp. R586-R595 ◽  
Author(s):  
Courtney C. Fleck ◽  
Hannah V. Carey
Keyword(s):  
Intestinal Mucosa ◽  
Specific Activity ◽  
P53 Protein ◽  
Chromatin Condensation ◽  
Fold Increase ◽  
Ground Squirrels ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Apoptotic Pathways ◽  
Nuclear Levels

Mammalian hibernation is associated with several events that can affect programmed cell death (apoptosis) in nonhibernators, including marked changes in blood flow, extended fasting, and oxidative stress. However, the effect of hibernation on apoptosis is poorly understood. Here, we investigated apoptosis and expression of proteins involved in apoptotic pathways in intestinal mucosa of summer and hibernating ground squirrels. We used terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) to identify possible apoptotic enterocytes in small intestine of summer squirrels and hibernating squirrels throughout the winter. Nuclear TUNEL staining increased as hibernation progressed, but the staining pattern was diffuse and not accompanied by chromatin condensation or apoptotic bodies. Electrophoresis of mucosal DNA revealed no ladders typical of apoptosis. Nuclear levels of proapoptotic p53 protein were fourfold less in hibernators compared with summer squirrels. A 12-fold increase in anti-apoptotic Bcl-xL compared with a 2-fold increase in proapoptotic Bax suggested a balance in favor of antiapoptotic signaling in hibernators. There was no change in Bcl-2 protein expression but phospho-Bcl-2 increased in mucosa of hibernators. Hibernation had minimal effects on expression of active caspase-8 or -9, whereas caspase-3-specific activity was lower in hibernators during an interbout arousal compared with summer squirrels. Expression of the prosurvival protein Akt increased 20-fold during hibernation, but phospho-Akt was not altered. These data provide evidence for enhanced expression of antiapoptotic proteins during hibernation that may promote enterocyte survival in a pro-oxidative, proapoptotic environment.

Renal cellular response to ureteral obstruction: role of maturation and angiotensin II

1999 ◽  
Vol 277 (1) ◽  
pp. F41-F47 ◽  
Author(s):  
Robert L. Chevalier ◽  
Barbara A. Thornhill ◽  
Jennifer T. Wolstenholme
Keyword(s):  
Angiotensin Ii ◽  
Ureteral Obstruction ◽  
Cellular Response ◽  
Cellular Proliferation ◽  
Urinary Tract Obstruction ◽  
Renin Angiotensin System ◽  
Nuclear Antigen ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Ang Ii ◽  
Proliferation And Apoptosis

Renal angiotensin II (ANG II) is increased as a result of unilateral ureteral obstruction (UUO), and angiotensin AT2 receptors predominate over AT1 receptors in the early postnatal period. To examine the renal cellular response to 3-day UUO in the neonatal and adult rat, AT1and AT2 receptors were inhibited by losartan and PD-123319, respectively. Additional rats received exogenous ANG II, 0.5 mg ⋅ kg−1 ⋅ day−1. Renal cellular proliferation and apoptosis were quantitated by proliferating cell nuclear antigen and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling technique, respectively. In the neonate, UUO reduced proliferation and increased tubular apoptosis. Losartan had no detectable cellular effect, whereas PD-123319 increased cellular proliferation and suppressed apoptosis, and exogenous ANG II stimulated apoptosis. In the adult, UUO increased cellular proliferation as well as apoptosis, whereas losartan, PD-123319, and exogenous ANG II did not alter the cellular response. In conclusion, UUO impairs renal growth in the neonate by reducing proliferation and stimulating apoptosis, at least in part through angiotensin AT2 receptors. UUO stimulates both renal cellular proliferation and apoptosis in the adult, but these effects are independent of ANG II. We speculate that the unique early responses of the developing kidney to urinary tract obstruction are mediated by a highly activated renin-angiotensin system and preponderance of AT2 receptors.

Role of Fas (CD95) in tubulointerstitial disease induced by unilateral ureteric obstruction

1999 ◽  
Vol 277 (1) ◽  
pp. F26-F32 ◽  
Author(s):  
Jeremy Hughes ◽  
Richard J. Johnson
Keyword(s):  
Cell Fate ◽  
Cell Apoptosis ◽  
Fas Ligand ◽  
Interstitial Cell ◽  
Death Receptor ◽  
Tubular Cell ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tubulointerstitial Injury ◽  
Double Labeling

Murine renal tubular epithelial cells and interstitial fibroblasts may express both Fas (CD95) death receptor and Fas ligand and are vulnerable to Fas-mediated death in vitro. We therefore hypothesized that an absence of renal Fas may protect resident cells from undergoing apoptosis. We performed unilateral ureteric ligation [producing unilateral ureteral obstruction (UUO)] in 6-wk-old normal control mice and C57Bl6/ lpr mice, which express a nonfunctional Fas receptor. Obstructed kidneys were removed at days 3, 7, and 14 ( n= 6 per group). Tubular cell apoptosis at day 7 was significantly reduced in lpr mice [21.8 ± 5.8 vs. 45.7 ± 7.6 cells/10 high-power fields (hpf), P < 0.02]. Importantly, there was no difference in tubular cell proliferation between normal and lpr mice at any time point studied. Interestingly, double labeling with terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) and the proximal tubule-specific antibody Fx1A indicated that the absence of Fas reduced distal but not proximal tubular death at day 7. In addition, there was no difference in interstitial cell apoptosis or proliferation, suggesting that Fas does not play a significant role in interstitial cell death. Importantly, inflammatory macrophage infiltration and ultimate collagen I deposition was unchanged in lprmice. In conclusion, the absence of functional cell surface Fas in UUO provides distal tubular cells with partial protection from apoptosis but does not affect interstitial cell fate in this model of tubulointerstitial injury.

scholarly journals Apoptotic cell death in rat lung following mustard gas inhalation

2017 ◽  
Vol 312 (6) ◽  
pp. L959-L968 ◽  
Author(s):  
Devon K. Andres ◽  
Brian M. Keyser ◽  
Ashley A. Melber ◽  
Betty J. Benton ◽  
Tracey A. Hamilton ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Fas Ligand ◽  
Control Level ◽  
Inhalation Injury ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Caspase 8 ◽  
Rat Lung ◽  
Caspase 9 ◽  
Unexposed Control

To investigate apoptosis as a mechanism of sulfur mustard (SM) inhalation injury in animals, we studied different caspases (caspase-8, -9, -3, and -6) in the lungs from a ventilated rat SM aerosol inhalation model. SM activated all four caspases in cells obtained from bronchoalveolar lavage fluid (BALF) as early as 6 h after exposure. Caspase-8, which is known to initiate the extrinsic Fas-mediated pathway of apoptosis, was increased fivefold between 6 and 24 h, decreasing to the unexposed-control level at 48 h. The initiator, caspase-9, in the intrinsic mitochondrial pathway of apoptosis as well as the executioner caspases, caspase-3 and -6, all peaked ( P < 0.01) at 24 h; caspase-3 and -6 remained elevated, but caspase-9 decreased to unexposed-control level at 48 h. To study further the Fas pathway, we examined soluble as well as membrane-bound Fas ligand (sFas-L and mFas-L, respectively) and Fas receptor (Fas-R) in both BALF cells and BALF. At 24 h after SM exposure, sFas-L increased significantly in both BALF cells ( P < 0.01) and BALF ( P < 0.05). However, mFas-L increased only in BALF cells between 24 and 48 h ( P < 0.1 and P < 0.001, respectively). Fas-R increased only in BALF cells by 6 h ( P < 0.01) after SM exposure. Apoptosis in SM-inhaled rat lung specimens was also confirmed by both immunohistochemical staining using cleaved caspase-3 and -9 antibodies and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining as early as 6 h in the proximal trachea and bronchi, but not before 48 h in distal airways. These findings suggest pathogenic mechanisms at the cellular and molecular levels and logical therapeutic target(s) for SM inhalation injury in animals.

scholarly journals Expression of Fas ligand by human gastric adenocarcinomas: a potential mechanism of immune escape in stomach cancer

Gut ◽  
1999 ◽  
Vol 44 (2) ◽  
pp. 156-162 ◽  
Author(s):  
M W Bennett ◽  
J O’Connell ◽  
G C O’Sullivan ◽  
D Roche ◽  
C Brady ◽  
...  
Keyword(s):  
Cell Death ◽  
Stomach Cancer ◽  
Fas Ligand ◽  
Immune Escape ◽  
Immune Privilege ◽  
Lymphoid Cells ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Gastric Adenocarcinomas

BackgroundDespite being immunogenic, gastric cancers overcome antitumour immune responses by mechanisms that have yet to be fully elucidated. Fas ligand (FasL) is a molecule that induces Fas receptor mediated apoptosis of activated immunocytes, thereby mediating normal immune downregulatory roles including immune response termination, tolerance acquisition, and immune privilege. Colon cancer cell lines have previously been shown to express FasL and kill lymphoid cells by Fas mediated apoptosis in vitro. Many diverse tumours have since been found to express FasL suggesting that a “Fas counterattack” against antitumour immune effector cells may contribute to tumour immune escape.AimTo ascertain if human gastric tumours express FasL in vivo, as a potential mediator of immune escape in stomach cancer.SpecimensThirty paraffin wax embedded human gastric adenocarcinomas.MethodsFasL protein was detected in gastric tumours using immunohistochemistry; FasL mRNA was detected in the tumours using in situ hybridisation. Cell death was detected in situ in tumour infiltrating lymphocytes using terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL).ResultsPrevalent expression of FasL was detected in all 30 resected gastric adenocarcinomas examined. In the tumours, FasL protein and mRNA were co-localised to neoplastic gastric epithelial cells, confirming expression by the tumour cells. FasL expression was independent of tumour stage, suggesting that it may be expressed throughout gastric cancer progression. TUNEL staining disclosed a high level of cell death among lymphocytes infiltrating FasL positive areas of tumour.ConclusionsHuman gastric adenocarcinomas express the immune downregulatory molecule, FasL. The results suggest that FasL is a prevalent mediator of immune privilege in stomach cancer.

Magnolol Reduces Renal Ischemia and Reperfusion Injury via Inhibition of Apoptosis

2017 ◽  
Vol 45 (07) ◽  
pp. 1421-1439 ◽  
Author(s):  
Chia-Yu Tang ◽  
Chang-Chi Lai ◽  
Po-Hsun Huang ◽  
An-Han Yang ◽  
Shu-Chiung Chiang ◽  
...  
Keyword(s):  
Serum Levels ◽  
Interleukin 10 ◽  
Renal Ischemia ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Ischemia And Reperfusion ◽  
Ischemia And Reperfusion Injury ◽  
Histological Damage ◽  
Underlying Mechanisms ◽  
Apoptotic Pathways

Magnolol, a constituent of the bark of Magnolia officinalis, has been reported to decrease myocardial stunning and infarct size. In this study, we investigated whether magnolol can reduce renal ischemia and reperfusion (I/R) injury. Renal I/R, induced by a 60-min occlusion of bilateral renal arteries and a 24-h reperfusion, significantly increased blood urea nitrogen (BUN) and creatinine levels, and caused histological damage to the kidneys of rats. Apoptosis, as evaluated by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining and caspase-3 activation, was significantly increased in the kidneys. Furthermore, serum levels of tumor necrosis factor-[Formula: see text] (TNF-[Formula: see text]), interleukin-1β (IL-1β), and interleukin-6 (IL-6) were significantly elevated, while the interleukin-10 (IL-10) level was suppressed. However, intravenous pretreatment with magnolol at doses of 0.003[Formula: see text]mg/kg and 0.006[Formula: see text]mg/kg 10[Formula: see text]min before renal I/R significantly limited the increases of BUN, creatinine, the histological damage, and apoptosis in the kidneys. The increases in TNF-[Formula: see text], IL-1β, and IL-6, and the decrease in IL-10 were also significantly inhibited. Additionally, magnolol increased Bcl-2 and decreased Bax in the kidneys. Phosphorylation of the prosurvival kinases, including Akt and extracellular signal-regulated kinases 1 and 2 (ERK1/2), was elevated, while phosphorylation of the pro-apoptotic mitogen-activated protein kinases, including p38 and c-Jun N-terminal kinase (JNK), was suppressed. In conclusion, magnolol reduces renal I/R injury. The underlying mechanisms for this effect might be related to the prevention of apoptosis, possibly via the inhibition of both extrinsic and intrinsic apoptotic pathways, including the reduction of TNF-[Formula: see text] production and the modulation of pro- and anti-apoptotic signaling elements.

scholarly journals TRAIL Mediates Neuronal Death in AUD: A Link between Neuroinflammation and Neurodegeneration

2021 ◽  
Vol 22 (5) ◽  
pp. 2547
Author(s):  
Liya Qin ◽  
Jian Zou ◽  
Alexandra Barnett ◽  
Ryan Vetreno ◽  
Fulton Crews ◽  
...  
Keyword(s):  
Neuronal Death ◽  
Neuronal Apoptosis ◽  
Neuronal Cell Death ◽  
Death Receptor ◽  
Neuronal Cell ◽  
Fold Increase ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Tunel Staining ◽  
Slice Culture ◽  
Cortical Regions

Although the cause of progressive neurodegeneration is often unclear, neuronal death can occur through several mechanisms. In conditions such as Alzheimer’s or alcohol use disorder (AUD), Toll-like receptor (TLR) induction is observed with neurodegeneration. However, links between TLR activation and neurodegeneration are lacking. We report a role of apoptotic neuronal death in AUD through TLR7-mediated induction of death receptor signaling. In postmortem human cortex, a two-fold increase in apoptotic terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining in neurons was found in AUD versus controls. This occurred with the increased expression of TLR7 and tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) death receptors. Binge ethanol treatment in C57BL/6 mice increased TLR7 and induced neuronal apoptosis in cortical regions that was blocked by TLR7 antagonism. Mechanistic studies in primary organotypic brain slice culture (OBSC) found that the inhibition of TLR7 and its endogenous ligand let-7b blocked ethanol-induced neuronal cell death. Both IMQ and ethanol induced the expression of TRAIL and its death receptor. In addition, TRAIL-neutralizing monoclonal antibodies blocked both imiquimod (IMQ) and ethanol induced neuronal death. These findings implicate TRAIL as a mediator of neuronal apoptosis downstream of TLR7 activation. TLR7 and neuronal apoptosis are implicated in other neurodegenerative diseases, including Alzheimer’s disease. Therefore, TRAIL may represent a therapeutic target to slow neurodegeneration in multiple diseases.

Anti-apoptotic and pro-survival effects of exercise training on hypertensive hearts

2012 ◽  
Vol 112 (5) ◽  
pp. 883-891 ◽  
Author(s):  
Chih-Yang Huang ◽  
Ai-Lun Yang ◽  
Yueh-Min Lin ◽  
Fan-Ni Wu ◽  
James A. Lin ◽  
...  
Keyword(s):  
Exercise Training ◽  
Caspase 3 ◽  
Fas Ligand ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Therapeutic Effects ◽  
Death Domain ◽  
Protein Levels ◽  
Survival Pathway ◽  
Cardiac Apoptosis ◽  
Apoptotic Pathways

Background: activated cardiac apoptosis was found in hearts from hypertensive animals, but little information regarding the effects of exercise training on cardiac apoptosis in hypertension is available. The purpose of this study was to evaluate the anti-apoptotic and pro-survival effects of exercise training on hypertensive hearts. Methods: 28 spontaneously hypertensive rats were divided into sedentary group (SHR) or underwent running exercise on treadmill for 1 h/day, 5 sessions/wk, for 12 wk (SHR-EX). Fourteen age-matched Wistar Kyoto rats served as a sedentary normotensive group (WKY). After exercise training or sedentary status, the excised hearts were measured by hemotoxylin and eosin staining, terminal deoxynucleotidyl transferase dUTP-mediated nick-end labeling (TUNEL) assay, and Western blotting. Results: fewer TUNEL-positive apoptotic cells were in SHR-EX groups than those in SHR. Protein levels of Fas ligand, Fas death receptor, tumor necrosis factor (TNF)-α, TNF receptor 1, Fas-associated death domain (FADD), activated caspase-8, and activated caspase-3 (Fas-dependent apoptotic pathways), as well as Bid, t-Bid, Bad, p-Bad, Bak, cytochrome c, activated caspase 9, and activated caspase-3 (mitochondria-dependent apoptotic pathways) were decreased in the SHR-EX group compared with the SHR group. Protein levels of IGF-1, IGF-1R, p-PI3K, p-Akt, p-Bad, and Bcl2 (cardiac pro-survival pathway) become more activated in SHR-EX groups than SHR and WKY. Conclusions: exercise training prevented hypertension-enhanced cardiac Fas-dependent and mitochondria-dependent apoptotic pathways and enhanced cardiac pro-survival pathway in rat models. Our findings demonstrate new therapeutic effects of exercise training on hypertensive hearts for preventing apoptosis and enhancing survival.

Apoptotic pathway in the rat small intestinal mucosa is different between fasting and ischemia-reperfusion

2006 ◽  
Vol 291 (1) ◽  
pp. G110-G116 ◽  
Author(s):  
Takehiro Fujise ◽  
Ryuichi Iwakiri ◽  
Bin Wu ◽  
Sadahiro Amemori ◽  
Takashi Kakimoto ◽  
...  
Keyword(s):  
Intestinal Mucosa ◽  
Fas Ligand ◽  
Ischemia Reperfusion ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Type I ◽  
Small Intestinal Mucosa ◽  
Apoptotic Pathway ◽  
Small Intestinal ◽  
Apoptotic Pathways ◽  
Induced Apoptosis

We have previously demonstrated that fasting and ischemia-reperfusion (I/R) induced apoptosis in rat intestinal mucosa. It is widely accepted that apoptosis is induced through two main pathways. This study aimed to compare apoptotic pathways following fasting and I/R. Rats were divided into two groups: the I/R group involved occlusion of the superior mesenteric artery for 60 min, followed by 60-min reperfusion, whereas the fasting group involved fasting for 24 or 48 h. Intestinal apoptosis was assessed as percentage of fragmented DNA, by electrophoresis and by a terminal deoxynucleotidyl transferase mediated dUDP-biotin nick- end labeling (TUNEL) assay. Apoptotic proteins including death ligands/receptors and caspases were evaluated by Western blot analysis. Small intestinal mucosal height and mitochondrial dehydrogenase function were assessed. Fasting and I/R significantly induced intestinal apoptosis. Mucosal height was significantly decreased in fasting rats, and mitochondrial dysfunction was induced only by I/R. Expressions of Fas, Fas ligand, and TNF-α type 1 receptor were enhanced in fasting and I/R rats. After I/R, expressions of cytochrome c and cleaved caspase-9 were significantly increased. In contrast, expressions of cleaved caspase-8 and cleaved caspase-3 increased in fasting rats. Fasting promoted mucosal apoptosis via a receptor-mediated type I apoptotic pathway in the rat small intestine, and I/R induced apoptosis via a mitochondria-mediated type II pathway.

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