335. X-LINKED INHIBITOR OF APOPTOSIS PROTEIN (XIAP) EXPRESSION PATTERNS IN THE SHEEP OVARY

2010 ◽  
Vol 22 (9) ◽  
pp. 135 ◽  
Author(s):  
H. R. Douglas ◽  
I. C. Kokay ◽  
D. R. Grattan ◽  
P. R. Hurst
Keyword(s):  
Inverse Relationship ◽  
Caspase 3 ◽  
Expression Patterns ◽  
Cysteine Proteases ◽  
Follicular Atresia ◽  
Study Objective ◽  
Antral Follicles ◽  
Xiap Expression ◽  
Apoptosis Protein ◽  
Active Caspase

At reproductive age, the ovary undergoes continual cyclicity of follicles due to multiple positive and negative signals that promote follicle growth and development, selection for ovulation, or atresia. The majority of follicles undergo atresia, a degenerative process involving granulosa cell apoptosis. This process is executed by caspases, which are cysteine proteases. Caspases are potently inhibited by XIAP providing a potential mechanism to control follicular atresia. The study objective was to test the hypothesis that XIAP will show elevated expression in healthy antral follicles compared to atretic antral follicles and will show an inverse relationship with active caspase-3 immunoreactivity in the same antral follicle during the sheep estrous cycle. Reproductively mature Romney ewes (n = 9) had estrous cycles synchronized with a prostaglandin F2α analogue, then tissue was collected on days 14, 15 and 16 of the subsequent natural cycle. Analysis involved determining the presence and localization of XIAP using in situ hybridization histochemistry and immunohistochemistry then comparing XIAP expression patterns with distribution of active caspase-3 protein. XIAP mRNA was not detected in primordial, primary and secondary follicles. In contrast, XIAP protein was present from the primary stage onwards. Antral follicles showed positive XIAP mRNA and protein expression in both granulosa and thecal cell layers and antral follicles on the same tissue section showed variable expression. All day 14 antral follicles were positive for XIAP mRNA expression irrespective of the level of active caspase-3 immunoreactivity, whereas an inverse relationship between active caspase-3 and XIAP was apparent in the majority of day 15 and 16 antral follicles. XIAP protein was widely expressed in active caspase-3 negative antral follicles and indicated a negative correlation with the onset of active caspase-3 expression in the majority of follicles. These results indicate that XIAP may regulate follicular atresia and act as an indicator of follicular health in the sheep ovary.

scholarly journals X-linked inhibitor of apoptosis protein and active caspase-3 expression patterns in antral follicles in the sheep ovary

Reproduction ◽  
2011 ◽  
Vol 142 (6) ◽  
pp. 855-867 ◽  
Author(s):  
Hollian R Phillipps ◽  
Ilona C Kokay ◽  
David R Grattan ◽  
Peter R Hurst
Keyword(s):  
Mrna Expression ◽  
Caspase 3 ◽  
Expression Patterns ◽  
Inhibitor Of Apoptosis ◽  
Mrna Levels ◽  
Follicular Atresia ◽  
Inhibitor Of Apoptosis Protein ◽  
Antral Follicles ◽  
Apoptosis Protein ◽  
Active Caspase

X-linked inhibitor of apoptosis protein (XIAP) interacts with caspases to inhibit their activity, thereby providing a potential mechanism for regulation of granulosa cell apoptosis occurring during follicular atresia. The aim of this study was to determine the presence and localization of XIAP mRNA and protein content in the sheep ovary and compare these expression patterns with active caspase-3 protein in the same antral follicles. Romney ewe estrous cycles (n=25) were synchronized with 2–3 Estrumate injections and ovarian tissue collected during the luteal and follicular phases of the cycle. The presence ofXIAPmRNA was confirmed by RT-PCR using laser capture microdissected ovarian cell samples.XIAPmRNA was subsequently localized byin situhybridization histochemistry and XIAP and active caspase-3 protein visualized by immunohistochemistry. In antral follicles extensive XIAP localization was evident in both granulosa and thecal cells. In contrast, mRNA expression was widespread in granulosa cells and only detected in thecal tissue from a small proportion of antral follicles. Active caspase-3 and XIAP comparative expression analysis showed positiveXIAPmRNA expression in all late luteal phase (day 14) follicles, despite varying levels of active caspase-3 protein. A proportion of follicular phase (days 15 and 16) follicles, however, showed an inverse expression relationship at the protein and mRNA levels in both granulosa and thecal tissue, as did XIAP protein in day 14 follicles. These results suggest high XIAP may prevent activation of caspase-3, thereby regulating follicular atresia in antral follicles and could potentially be utilized as a marker of follicular health.

scholarly journals Immunohistochemical localization of active caspase-3 in the mouse ovary: growth and atresia of small follicles

Reproduction ◽  
2002 ◽  
pp. 659-665 ◽  
Author(s):  
MA Fenwick ◽  
PR Hurst
Keyword(s):  
Granulosa Cells ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Active Form ◽  
Cysteine Proteases ◽  
Immunohistochemical Localization ◽  
Mouse Ovary ◽  
Physiological Factors ◽  
Cell Suicide ◽  
Active Caspase

Caspase-3 belongs to a family of highly conserved cysteine proteases that mediate the course of apoptotic cell suicide. It is recognized that ovarian follicular atresia is associated with apoptosis, a process that has been characterized mainly in larger antral follicles. The aims of this study were to investigate the expression of caspase-3 in the mouse ovary, and determine whether active caspase-3 is present within smaller follicles, which may constitute the resting pool. The inactive enzyme was expressed as a 32 kDa band on a western blot of tissue extracts, whereas the active form was localized immunohistochemically. Bromodeoxyuridine (BrdU) was administered to mice (n = 7) during a 12 h period and subsequently localized to identify potentially quiescent follicles. Measurements of BrdU-positive cells in the mouse ovary were extrapolated with data obtained by morphometric analyses of small follicles using the nucleator technique. BrdU was incorporated into the granulosa cells of follicles regardless of size and the number of cells they contained, but was absent in a large proportion (89%) of small, single layered follicles. Active caspase-3 was localized to both the oocyte and granulosa cells of follicles that were considered to be undergoing atresia, but was not localized to the granulosa cells of any small, single layered follicles. The results of this study indicate that, in small follicles, granulosa cell proliferation occurs independently of the size of follicles and the number of constituent cells, and that follicles of this type may be inherently less susceptible to the normal physiological factors that induce atresia.

Prenatally administered dexamethasone impairs folliculogenesis in spiny mouse offspring

2016 ◽  
Vol 28 (7) ◽  
pp. 1038 ◽  
Author(s):  
Monika Hułas-Stasiak ◽  
Piotr Dobrowolski ◽  
Ewa Tomaszewska
Keyword(s):  
Caspase 3 ◽  
Follicular Development ◽  
Treated Group ◽  
Spiny Mouse ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Follicular Atresia ◽  
Dexamethasone Treatment ◽  
Primordial Follicles ◽  
Acomys Cahirinus ◽  
Active Caspase

This study was designed to determine whether prenatal dexamethasone treatment has an effect on follicular development and atresia in the ovary of spiny mouse (Acomys cahirinus) offspring. Dexamethasone (125 µg kg–1 bodyweight per day) was administered to pregnant spiny mice from Day 20 of gestation to parturition. The processes of follicle loss were analysed using classical markers of apoptosis (terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling reaction, active caspase-3) and autophagy (Lamp1). The present study indicated that dexamethasone reduced the pool of healthy primordial follicles. Moreover, the oocytes from these follicles showed intensive caspase-3 and Lamp1 staining. Surprisingly, dexamethasone caused an increase in the number of secondary follicles; however, most of these follicles were characterised by extensive degeneration of the oocyte and caspase-3 and Lamp1 labelling. Western-blot analysis indicated that the glucocorticoid receptor as well as apoptosis and autophagy markers were more strongly expressed in the DEX-treated group than in the control. On the basis of these findings, we have concluded that dexamethasone impairs spiny mouse folliculogenesis and enhances follicular atresia through induction of autophagy or combined autophagy and apoptosis.

scholarly journals Short photoperiod-induced ovarian regression is mediated by apoptosis in Siberian hamsters (Phodopus sungorus)

Reproduction ◽  
2006 ◽  
Vol 131 (4) ◽  
pp. 771-782 ◽  
Author(s):  
C S Moffatt-Blue ◽  
J J Sury ◽  
Kelly A Young
Keyword(s):  
Caspase 3 ◽  
Ovarian Function ◽  
Corpora Lutea ◽  
Control Data ◽  
Antral Follicles ◽  
Siberian Hamsters ◽  
Estradiol Secretion ◽  
Induced Changes ◽  
Active Caspase

Siberian hamster reproduction is mediated by photoperiod-induced changes in gonadal activity. However, little is known about how photoperiod induces cellular changes in ovarian function. We hypothesized that exposing female hamsters to short (inhibitory) as opposed to long (control) photoperiods would induce an apoptosis-mediated disruption of ovarian function. Ovaries and plasma from hamsters exposed to either long (LD, 16 h light:8 h darkness) or short (SD, 8 h light:16 h darkness) days were collected during diestrus II after 3, 6, 9 and 12 weeks and processed for histology or RIA respectively. Apoptosis was assessed byin situTUNEL and active caspase-3 protein immunolabeling. No significant differences were observed among LD hamsters for any parameter; therefore, these control data were pooled. SD exposure induced a decline in preantral follicles (P< 0.05), early antral/antral follicles (P< 0.01) and corpora lutea (P< 0.01) by week 12 as compared with LD. Terminal atretic follicles appeared by SD week 9; by week 12, these had become the predominant ovarian structures. Estradiol concentrations decreased by weeks 9 and 12 SD when compared with both LD and week-3 SD hamsters (P< 0.05); however, no changes were observed for progesterone. TUNEL-positive follicles in SD ovaries increased at week 3 and subsequently declined by week 12 as compared with LD ovaries (P< 0.01). Active capsase-3 protein immunostaining peaked at SD week 3 as compared with all other groups (P< 0.01). TUNEL and capsase-3 immunolabeling were localized to granulosa cells of late-preantral and early-antral/antral follicles. These data indicate that SD exposure rapidly induces follicular apoptosis in Siberian hamsters, which ultimately disrupts both estradiol secretion and folliculogenesis, resulting in the seasonal loss of ovarian function.

scholarly journals Nrdp1 increases neuron apoptosis via downregulation of Bruce following intracerebral haemorrhage

2019 ◽  
Vol 16 (1) ◽  
Author(s):  
Changlong Zhou ◽  
Qingjun Liu ◽  
Wang Zhao ◽  
Ling Yang ◽  
Zhongyan Huang ◽  
...  
Keyword(s):  
Brain Edema ◽  
Cortical Neurons ◽  
Neuronal Apoptosis ◽  
Caspase 3 ◽  
Neurological Injury ◽  
Apoptosis Protein ◽  
And Oxidative Stress ◽  
Active Caspase ◽  
Pro Inflammatory Cytokines

Abstract Background Neuregulin receptor degradation protein-1 (Nrdp1) is an E3 ubiquitin ligase that plays an important role in regulating cell growth, apoptosis and oxidative stress. However, the data regarding its expression and exact mechanism in neuronal injury following ICH has not been well identified. Methods In this study, primary cortical neurons from C57BL/6 mice were subjected to erythrocyte lysates. Nrdp1 expression, cell apoptosis, caspase-3 and BRUCE levels were detected. In addition, inflammatory response, brain edema, and neurological injury in ICH mice were also assessed. Results We found that the expression of Nrdp1 was significantly increased in neuron cells accompanied by up-regulation of active caspase-3 and decreased expression of BRUCE (an inhibitor of apoptosis protein). However, inhibiting Nrdp1 levels of neurons reduced caspase-3 activity but induced up-regulation of BRUCE. In vivo, inhibiting Nrdp1 levels increased pro-inflammatory cytokines, brain edema, and neurological injury following ICH. Conclusions Taken together, the data suggested that Nrdp1 might play a crucial role in neuronal apoptosis via inhibiting BRUCE following ICH.

Survivin Inhibition of p34Cdc2 Via Protection of Wee1 Correlates with Survival.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3352-3352
Author(s):  
Javier Rivera Guzman ◽  
Seiji Fukuda ◽  
Louis Pelus
Keyword(s):  
Cell Cycle ◽  
Caspase 3 ◽  
Survivin Expression ◽  
Dominant Negative ◽  
Dependent Manner ◽  
Hematopoietic Stem ◽  
Protein Levels ◽  
Apoptosis Protein ◽  
Apoptotic Activity ◽  
Active Caspase

Abstract The inhibitor of apoptosis protein Survivin and p34Cdc2 (the cyclin-dependent kinase, Cdk1) are involved in cell cycle progression and apoptosis. Cdc2 and Survivin are tightly regulated in normal cells but deregulated in cancer. We have previously shown that Survivin regulates entry of hematopoietic stem cells into cell cycle. It has been established that Cdc2 phosphorylates Survivin at threonine 34 (T34), which is believed to be a stabilizing event necessary for normal Survivin function and anti-apoptotic activity. Activation of Cdc2 is required for its apoptotic activity and inhibition of the proapoptotic activity of Cdc2 can be achieved by phosphorylation at tyrosine 15 (Tyr15). Stable overexpression of a wild-type (wt) mouse Survivin-IRES-GFP construct in IL-3-dependent mouse BaF3 cells resulted in increased Tyr15 phosphorylation of Cdc2 and enhanced cell proliferation, while cells transduced with a threonine 34 to alanine (T34A) dominant negative (DN) Survivin construct showed reduced Cdc2 Tyr15 phosphorylation and accelerated apoptosis. Furthermore, when apoptosis was induced in stably transduced BaF3 cells by IL-3 withdrawal, Survivin expression and phospho-Tyr15 levels correlated with enhanced survival and decreased survival and low phospho-Tyr15 levels correlated with Survivin disruption by T34A-DN-Survivin. Since Survivin does not contain intrinsic kinase activity we sought to determine what kinase(s) could be involved in stabilization or increase in phosphoTyr-15-Cdc2 by Survivin. Since Survivin inhibits caspase 3 activity and the Cdc2-targeting kinase Wee1 is a degradation target of active caspase 3, we performed immunoblotting assays on Survivin wt and T34A-transduced BaF3 cells and showed that cells over-expressing wt Survivin contain 2-fold higher levels of Wee1 protein as compared to those expressing vector or DN-Survivin. In addition, cells overexpressing wt Survivin contain >2-fold higher intact PARP protein levels which is another target of active caspase 3, as compared to cells transduced with MIEG vector. In order to test whether mimicking wt Survivin inhibition of caspase 3 could lead to increased Cdc2-Tyr15 phosphorylation, we evaluated the specific caspase 3 inhibitor Ac-DEVD-CHO. Inhibition of caspase 3 in BaF3 cells produced an increase in Cdc2Tyr15 phosphorylation in a dose-dependent manner, with 25 mM giving >4-fold higher intact PARP and >6-fold higher phospho-Tyr15 protein levels. Taken together, these results suggest that the antiapoptotic effect of Survivin is mediated through inactivation of Cdc2 by phosphorylation on Tyr-15 as a consequence of protection of the Wee1 kinase from degradation by caspase 3.

scholarly journals Chemerin Suppresses Ovarian Follicular Development and Its Potential Involvement in Follicular Arrest in Rats Treated Chronically With Dihydrotestosterone

Endocrinology ◽  
2013 ◽  
Vol 154 (8) ◽  
pp. 2912-2923 ◽  
Author(s):  
Ji Young Kim ◽  
Kai Xue ◽  
Mingju Cao ◽  
Qi Wang ◽  
Jia-yin Liu ◽  
...  
Keyword(s):  
Granulosa Cell ◽  
Rat Model ◽  
Caspase 3 ◽  
Growth Arrest ◽  
Cytoskeletal Proteins ◽  
Inhibitor Of Apoptosis ◽  
Follicular Growth ◽  
Inhibitor Of Apoptosis Protein ◽  
Antral Follicles ◽  
Apoptosis Protein

Abstract In the present study, we have investigated the cellular mechanisms of androgen-induced antral follicular growth arrest and the possible involvement of chemerin and its receptor chemokine-like receptor 1 (CMKLR1) in this process, using a chronically androgenized rat model. We hypothesize that hyperandrogenism induces antral follicle growth arrest via the action of chemerin and ovarian structural changes, resulting from granulosa cell and oocyte apoptosis and theca cell survival. Dihydrotestosterone (DHT) treatment resulted in increased expression of chemerin and CMKLR1 in antral follicles, absence of corpus luteum, and increased atypical follicles. Addition of chemerin to follicle cultures induced granulosa cell apoptosis and suppressed basal, FSH- and growth differentiation factor-9-stimulated follicular growth. DHT down-regulated aromatase expression and increased active caspase-3 content and DNA fragmentation in granulosa cells in vivo. These changes were accompanied by higher phosphatase and tensin homolog and lower phospho-Akt (Ser473) content in antral follicles and higher calpain expression and down-regulation of cytoskeletal proteins in atypical follicles, which were constituted predominantly of theca cells. DHT also activated granulosa cell caspase-3, decreased X-linked inhibitor of apoptosis protein, poly(ADP-ribose) polymerase, and phospho-Akt contents and induced apoptosis in vitro, responses readily attenuated by forced X-linked inhibitor of apoptosis protein expression. These findings are consistent with our hypothesis that antral follicular growth arrest in DHT-treated rats results from increased chemerin expression and action, as well as changes in follicular cell fate and structure, which are a consequence of dysregulated interactions of pro-survival and pro-apoptotic modulators in a cell-specific manner. Our observations suggest that this chronically androgenized rat model may be useful for studies on the long-term effects of androgens on folliculogenesis and may have implications for the female reproductive disorders associated with hyperandrogenism.

scholarly journals Beclin 1 Interacts With Active Caspase-3 and Bax in Oocytes From Atretic Follicles in the Rat Ovary

2019 ◽  
Vol 67 (12) ◽  
pp. 873-889 ◽  
Author(s):  
María L. Escobar ◽  
Olga M. Echeverria ◽  
Sebastián Palacios-Martínez ◽  
Silvia Juárez-Chavero ◽  
Luis Sánchez-Sánchez ◽  
...  
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Beclin 1 ◽  
Follicular Atresia ◽  
Different Types ◽  
Old Rats ◽  
Rat Ovary ◽  
Apoptotic Proteins ◽  
Autophagic Proteins ◽  
Active Caspase

Oocyte cell death is a normal process in the mammalian ovary during follicular growth. Recent reports have demonstrated the presence of pro-apoptotic and pro-autophagic proteins during oocyte elimination. The goal of this study was to identify the interactions between proteins involved in different types of programmed cell death in the same oocyte during follicular atresia. We evaluated the presence of Beclin 1 and its interaction with the pro-apoptotic proteins active caspase-3, Bax, and Bak by means of histochemical observations, ultrastructural immunodetection, and immunoprecipitation techniques in ovaries from prepubertal (28- and 33-day-old), juvenile (40-day-old), and young adult (90-day-old) rats. In this study, we identified that oocyte elimination occurred with a high quantity of pro-autophagic protein Beclin 1 and increased the presence of the pro-apoptotic proteins active caspase-3, Bax, and Bak. Conversely, the antiapoptotic protein Bcl-2 was reduced in oocytes from atretic follicles. In addition, Beclin 1 was shown to interact with active caspase-3 and Bax. Our results suggest that the increase in Beclin 1 is directly related to the rise of pro-apoptotic proteins, which could promote the apoptotic process during oocyte elimination.

scholarly journals Transcriptome Analysis Reveals HgCl2 Induces Apoptotic Cell Death in Human Lung Carcinoma H1299 Cells through Caspase-3-Independent Pathway

2021 ◽  
Vol 22 (4) ◽  
pp. 2006
Author(s):  
Mi Jin Kim ◽  
Jinhong Park ◽  
Jinho Kim ◽  
Ji-Young Kim ◽  
Mi-Jin An ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Transcriptome Analysis ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Expression Patterns ◽  
Nitrogen Compound ◽  
Apoptotic Cell Death ◽  
Mercury Chloride ◽  
Rna Seq

Mercury is one of the detrimental toxicants that can be found in the environment and exists naturally in different forms; inorganic and organic. Human exposure to inorganic mercury, such as mercury chloride, occurs through air pollution, absorption of food or water, and personal care products. This study aimed to investigate the effect of HgCl2 on cell viability, cell cycle, apoptotic pathway, and alters of the transcriptome profiles in human non-small cell lung cancer cells, H1299. Our data show that HgCl2 treatment causes inhibition of cell growth via cell cycle arrest at G0/G1- and S-phase. In addition, HgCl2 induces apoptotic cell death through the caspase-3-independent pathway. Comprehensive transcriptome analysis using RNA-seq indicated that cellular nitrogen compound metabolic process, cellular metabolism, and translation for biological processes-related gene sets were significantly up- and downregulated by HgCl2 treatment. Interestingly, comparative gene expression patterns by RNA-seq indicated that mitochondrial ribosomal proteins were markedly altered by low-dose of HgCl2 treatment. Altogether, these data show that HgCl2 induces apoptotic cell death through the dysfunction of mitochondria.

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