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Mol. Hum. Reprod. Advance Access originally published online on February 11, 2005
Molecular Human Reproduction 2005 11(3):161-166; doi:10.1093/molehr/gah146
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Molecular Human Reproduction Vol. 11 No. 3 © European Society of Human Reproduction and Embryology 2005; all rights reserved

HCG up-regulates survivin mRNA in human granulosa cells

Yukiyo Kumazawa1,3, Kazuhiro Kawamura1, Toshiharu Sato1, Naoki Sato1, Yoshitomo Konishi1, Yosushi Shimizu1, Jun Fukuda1, Hideya Kodoma2 and Toshinobu Tanaka1

1Division of Obstetrics and Gynecology, Department of Reproductive and Developmental Medicine, Akita University School
of Medicine, 1-1-1 Hondo, Akita, 010-8543, and 2Faculty of Health and Science, Department of Reproductive and Developmental Medicine, Akita University School of Medicine, 1-1-1 Hondo, Akita, 010-8543, Japan

3 To whom correspondence should be addressed at: Division of Obstetrics and Gynecology, Department of Reproductive and Developmental Medicine, Akita University School of Medicine, 1-1-1 Hondo, Akita, 010-8543, Japan. Email: kakki{at}bk9.so-net.ne.jp


   Abstract
Top
 Abstract
Introduction
 Materials and methods
 Results
 Discussion
 References
 
The apoptosis of granulosa cells is involved in follicular atresia and degeneration of the corpus luteum. The mechanisms that regulate follicular atresia and luteal degeneration remain obscure. Survivin is a member of the family of inhibitors of apoptosis protein that is expressed during fetal development and in cancer tissues. The present study investigates the expression of survivin, as well as its regulation and function in granulosa cells. We identified survivin at the protein level in granulosa cells and detected not only survivin but also splice-variant transcripts in human and mouse granulosa-luteal cells. One-step real-time PCR analysis revealed that HCG increases the amount of survivin mRNA expressed in cultured human granulosa cells. These results suggest that survivin is involved in supporting luteal function, and that HCG contributes to this role.

Key words: gonadotrophin/granulosa cells/inhibitors of apoptosis protein/ovary/survivin


   Introduction
Top
 Abstract
 Introduction
Materials and methods
 Results
 Discussion
 References
 
Apoptosis is a physiological process that is essential to the normal embryonic development and adult tissue homeostasis. Survivin is an inhibitor of apoptosis protein (Ambrosini et al., 1997Go) that directly interacts with several caspases to inhibit apoptosis (Shin et al., 2001Go) and also functions as a chromosomal passenger protein during chromosomal segregation (Adams et al., 2001Go). The expression of human survivin is regulated in a strict cell cycle-dependent manner with levels reaching a maximum during the G2/M phase and its anti-apoptotic function is mediated both by its BIR domain and by the interaction of its C-terminal coiled-coil domain with microtubules of the mitotic spindle (Adida et al., 1998aGo,bGo; Ambrosini et al., 1998Go; Li et al., 1998aGo,bGo). Survivin probably plays a prominent role in regulating apoptosis during normal development because several human fetal tissues express this protein (Adida et al., 1998aGo,bGo).

The mouse survivin gene is transcribed into three alternatively spliced variants, named survivin 40, survivin 121, and survivin 140 (Conway et al., 2000Go). Survivin 140 consists of all four exons. Survivin 40 lacks sequences derived from exon 2, leading to an in-frame stop codon, and survivin 121 is derived from exons 1 to 3 and retains part of intron 3. Each of these splice-variants has different antiapoptotic properties against apoptotic stimuli and is differentially expressed during fetal development and in adult tissues (Conway et al., 2000Go). The four splice-variants in humans are survivin, survivin-2B, survivin-{Delta}Ex3 and survivin-3B (Mahotka et al., 1999Go; Badran et al., 2004Go). Survivin consists of all four exons, and survivin-2B has an additional exon (exon 2B) inserted between exons 2 and 3. Survivin-{Delta}Ex3 lacks exon 3 as well as a frame shift with an extension of the reading frame into the open reading frame of the 3' untranslated region (Mahotka et al., 1999Go). Survivin-3B consists of all four exons plus the novel exon 3B that is flanked by exons 3 and 4 (Badran et al., 2004Go). These splice-variants have different functions and properties in the regulation of apoptosis.

We previously showed that survivin mRNA and protein are expressed in mouse pre-implantation embryos and that it could protect the embryos from apoptosis by inhibiting a caspase-mediated apoptotic pathway (Kawamura et al., 2003Go). A recent study has demonstrated that survivin mRNA and protein are expressed in hen granulosa and theca tissue, and that the levels of survivin mRNA during follicular development are highest in granulosa cells from the prehierarchal follicles (Johnson et al., 2002Go).

Most ovarian follicles undergo atresia, a hormonally controlled apoptotic process (Kaipia and Hsueh, 1997Go). Follicles become atretic, the corpus luteum degenerates via apoptosis (Tilly et al., 1992Go), Ca2+/Mg2+-dependent endonuclease activity is activated and apoptotic DNA undergoes fragmentation in ovarian granulosa and luteal cells (Zeleznik et al., 1989Go; Dharmarajan et al., 1994Go).

The present study establishes the presence of survivin in human and mouse granulosa cells and determines whether gonadotrophins stimulate survivin levels in these cells.


   Materials and methods
Top
 Abstract
 Introduction
 Materials and methods
Results
 Discussion
 References
 
Animals and sample collection
Immature 4-week-old female IVCS mice (Institute for Animal Reproduction, Ibaragi, Japan) were injected with 10 IU of pregnant mare serum gonadotrophin (Sigma, St Louis, MO) followed by 10 IU of HCG (Sigma) 48 h later. Twenty-four hours after the HCG injection, the ovaries were removed and the granulosa-luteal cells were excised under a microscope using a 26-gauge needle as described (Thordarson et al., 1997Go; Corambula et al., 2002Go). All procedures involving the care and use of animals were approved by the animal research committee at Akita University School of Medicine.

Culture of human granulosa-luteal cells
Women who underwent follicle aspiration for IVF provided written informed consent to participate in all aspects of the study according to the guidelines of our Institutional Review Board. Follicular growth was stimulated with 150~300 IU of HMG (HMG-Nikken; Nikken, Japan) daily, which was administered from the second day of menstruation together with 900 µg of a GnRH analogue (GnRHa; Buserelin: Sprecur, Mochida, Japan) administered daily from the mid-luteal phase of the previous cycle as described (Tasdemir et al., 1993Go). An i.m. injection of 10 000 IU of HCG was administered 35 h before follicular puncture. Pooled follicular fluid obtained from several IVF patients was separated by centrifugation at 3000 g for 20 min, and then pellets containing granulosa-luteal cells were suspended in phosphate-buffered saline (PBS; Sigma) containing 0.1% hyaluronidase. After incubation at 37 °C for 20 min, the cells were placed on a 50% (v/v) Percoll cushion (Pharmacia LKB Biotechnology, Piscataway, NJ) and sedimented by centrifugation at 3000 g for 30 min. The cell pellets were washed twice with PBS and suspended in serum-free GIT medium (Wako Chemicals, Osaka, Japan) containing 50 IU/ml Penicillin G and 50 µg/ml streptomycin. Approximately, 4 x 104 cells/well in 200 µl of serum-free GIT medium were placed in collagen-coated 48-well plates (CelltightC1 Plate 48F; Sumitomo Bakelite Co, Tokyo, Japan) and incubated at 37 °C under 5% CO2 in air for 48 h. Adherent cells were washed with serum free GIT medium and HCG (0, 0.01, 0.1, 1, 10 or 100 IU/ml) was added to the wells.

RT–PCR
Total RNA was extracted from mouse and human granulosa-luteal cells using the RNeasy Mini Kit (Qiagen, Valencia, CA) according to the manufacturer's instructions. The quality of RNA in the extracts was assayed and the concentration was determined using a SmartSpec spectrophotometer (Bio-Rad Laboratories, Hercules, CA). The RNA samples were reverse-transcribed into cDNA and PCR proceeded as described in the legends to Table I. The primers for mouse and human survivin were based on the published sequences shown in Table I. Simultaneously amplified mouse and human placenta cDNA served as the positive control. Water was substituted for mRNA in the negative control. The PCR products were separated by 2% agarose gel electrophoresis (Agarose-LE, Nacalai Tesque, Inc., Kyoto, Japan) in the presence of ethidium bromide (Sigma) and visualized using an ultra-violet transilluminator (Funakoshi, Tokyo, Japan). To confirm the identity of the products, DNA bands were eluted from the gels using the QIAquick Gel Extraction Kit (Qiagen KK, Tokyo, Japan), ligated into the pDrive Cloning vector (Qiagen KK), and cloned according to standard protocols. Plasmid DNA was recovered using a Quantum Prep Plasmid Miniprep Kit (Bio-Rad, Hercules, CA), cycle sequenced using the BigDye Terminator Cycle Sequencing Ready Reaction Kit (Applied Biosystems, Tokyo, Japan) and examined in an ABI PRISM 3100 Genetic Analyzer (Applied Biosystems) using T7 or SP6 site-specific primers.


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  		Table I. Primers used for RT–PCR, PCR cycles, and temperatures for amplification of the different cDNA

 
One-step real-time quantitative RT–PCR
Real-time quantitative RT–PCR proceeded in a LightCycler System (Roche Diagnostics, Tokyo, Japan) using the QuantiTect Probe RT-PCR Kit (Qiagen, Valencia, CA) and the one-step method based on TaqMan as described (Konishi et al., 2004Go). The reaction mixtures contained 10 µl of 2xQuantiTect Probe RT–PCR Master Mix, 0.2 µl Quantitect Probe RT Mix, 0.5 µl of 40 µM primer, 0.5 µl of 8 µM TaqMan probe and 2 µM template RNA adjusted to 20 µl with DNase/RNase-free water. The reaction mix was loaded into glass capillary tubes. Table II shows the primers and TaqMan probes for 18S ribosomal RNA and human survivin mRNA designed using Primer Express V1.0 software (Applied Biosystems, Foster City, CA). Conditions for the one-step real-time quantitative RT–PCR were 20 min at 50 °C (reverse transcription) and 15 min at 95 °C (RT inactivation and initial activation) followed by 35 cycles of amplification for 0 s at 95 °C (denaturation) and 1 min at 60 °C (annealing and extension). All heating and cooling steps proceeded at a slope of 20 °C/s. A single fluorescence reading at 530 nm was taken from each sample during extension.


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  		Table II. Sequences of primers and TaqMan probes used in quantitative RT–PCR for survivin and 18S

 
Western blotting to detect survivin protein
Protein extracts from human and mouse granulosa-luteal cells were subjected to western blot analysis essentially as described previously (Sato et al., 2000Go) and separated by SDS-PAGE using 16% polyacrylamide gels (Tefco, Tokyo, Japan). Resolved proteins were transferred to a polyvinylidene difluoride (PVDF) membrane using a Tefco electroblot apparatus and then incubated with anti-human survivin antibody overnight at 4 °C, or with horse–radish peroxidase-conjugated anti-rabbit immunoglobulin G secondary antibody for 1 h at room temperature. Immunoreactive bands were detected using Enhanced Chemiluminescence Plus Western blotting detection reagents (Amersham Biosciences, NJ).

Immunohistochemistry
Mouse ovaries were fixed in paraformaldehyde in PBS and embedded in paraffin blocks. Serial 3 µm sections were cut, and the histopathological diagnosis was confirmed in sections stained with haematoxylin and eosin. The sections were deparaffinized in xylene, rehydrated in a graded ethanol series, quenched in 3% hydrogen peroxidase, blocked and incubated with affinity-purified anti-human survivin antibody (20 µg/ml) overnight at 4 °C. After several PBS washes, the sections were incubated with peroxidase-conjugated goat anti-rabbit immunoglobulin G (DAKO, Carpinteria, CA) for 1 h at room temperature followed by 3',3'-diaminobenzidine and then counterstained with haematoxylin. The primary antibody in the negative control was incubated with human survivin control peptide for 1 h at 37 °C before further processing as described above.

Statistical analysis
The results are presented as means ± SEM. All data were analysed using the Mann–Whitney U test. P<0.001 was considered statistically significant.


   Results
Top
 Abstract
 Introduction
 Materials and methods
 Results
Discussion
 References
 
Expression of survivin mRNA and protein
We detected mRNA for survivin in mouse and human granulosa-luteal cells using RT–PCR (Figure 1) and two survivin bands in mouse granulosa-luteal cells. Sequencing the DNA of these bands revealed that the predicted survivin amplification product of 348 bp (survivin 140) was the larger, whereas the smaller was a 238 bp survivin fragment (survivin 40) lacking 110 bp of exon 2 between positions 3267 and 3377 as reported (Kawamura et al., 2003Go). We detected three survivin bands in human granulosa-luteal cells, the largest being identical to the survivin splice-variant, survivin-2B, with the additional exon 2B. The middle band was the predicted survivin amplification product of 428 bp. The smallest band was consistent with the survivin splice-variant, survivin-{Delta}Ex3 that lacks exon 3.



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  		Figure 1. Survivin detected in human and mouse granulosa cells by RT–PCR. Marker, {varphi}{chi} 174-HaeIII digest; posi, positive control survivin human and mouse placenta cDNA; neg, negative control without template cDNA; GC, granulosa cells; bp, base pair.

 
We identified a survivin protein of approximately 16.5 kDa in the total cellular proteins collected from human and mouse granulosa-luteal cells (Figure 2). Our RT–PCR revealed splice variants of survivin mRNA; however, by western blotting, we could not detect other bands that correspond to those variants. One of the reasons might be due to the small molecular weight of survivin protein and the extremely low levels of survivin proteins in granulosa cells (Adida et al., 1998aGo,bGo; Krieg et al., 2002Go).



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  		Figure 2. Western blots of survivin protein in extracts from human and mouse granulosa cells. Comparison with co-migrating size markers shows specific bands of about 16.5 kDa.

 
Survivin expression in mouse ovaries during development
Ovarian sections from immature female mice were incubated with pregnant mare's serum gonadotrophin (PMSG) for 0, 24 and 48 h, or with PMSG/HCG for 24 h. Figure 3 shows that ovaries from immature mice contained many preantral, early antral and preovulatory follicles. Immunohistochemical staining detected survivin in mouse granulosa cells and oocytes at all stages of development, but more of it was expressed in oocytes than in granulosa cells from follicles.



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  		Figure 3. Immunolocalization of survivin in mouse ovarian follicles at various developmental stages. Fixed adjacent sections from immature mice before and after PMSG and HCG exposure. (A–F) sections probed with anti-survivin antibody (G–K) negative control sections; O, oocyte; GC, granulosa cells; CL, corpus luteum (24 h post-HCG); (A,G) magnification x40; (B–F, H–L) magnification x20.

 
Human survivin mRNA levels are up-regulated by HCG
Granulosa-luteal cells were stimulated with various concentrations of HCG for 24 h to determine whether HCG enhances survivin mRNA expression. Figure 4 shows that survivin mRNA expression assessed by one-step real-time quantitative RT–PCR was significantly increased by HCG at both 10 and 100 IU/ml (5.4-fold, P<0.001), whereas concentrations of HCG below 10 IU/ml had no effect. We analysed total RNA extracted from human granulosa-luteal cells at different times after exposure to 10 IU/ml of HCG by quantitative RT–PCR. Survivin mRNA levels peaked at 8.8-fold within both 6 and 12 h of HCG incubation (P<0.001; Figure 5).



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  		Figure 4. HCG induces survivin mRNA in cultured human granulosa-luteal cells. After initial culture, cells were incubated in serum-free GIT medium containing indicated concentrations of HCG for 24 h and then total RNA was isolated. Expression levels of survivin mRNA analysed by one-step real-time quantitative RT–PCR and normalized against 18S ribosomal RNA levels. These data are calculated as means of six experiments. Bars represent SEM. *P<0.001 versus control.

 


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  		Figure 5. Levels of survivin mRNA in cultured human granulosa-luteal cells before (0 h) and after adding HCG. Total RNA was isolated from cells initially cultured and then incubated for 0 (before), 1, 3, 6, 12 and 24 h in serum-free GIT medium containing 10 IU/ml of HCG. Expression levels of survivin mRNA in total RNA determined by one-step real-time quantitative RT–PCR were normalized with 18S ribosomal RNA levels. These data are calculated as means of four experiments. Bar represents SEM. *, P<0.001 versus control.

 

   Discussion
Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
References
 
The present study demonstrated that survivin mRNA and protein are expressed in human and mouse granulosa-luteal cells. Survivin protein was expressed throughout all the developmental stages in mouse granulosa cells and embryos following gonadotrophin stimulation. Moreover, HCG significantly induced the up-regulation of survivin mRNA levels in cultured human granulosa-luteal cells.

Progesterone production by cultured cells was simultaneously and time-dependently stimulated by all tested concentrations of HCG (data not shown), findings that are consistent with those of other reports (Matsubara et al., 2000Go; Abraham et al., 2003Go). The binding of HCG to granulosa cell membrane receptors increases cAMP levels, phosphorylates proteins and activates the protein kinase A pathway, as well as steroidogenesis (Sugino et al., 2000Go). In contrast to progesterone production, survivin mRNA shows an increase in a time-dependent manner in the present study. Whether the up-regulation of survivin expression by HCG is a direct or indirect action via the protein kinase A pathway remains unknown. The intracellular signalling pathway of HCG in the up-regulation of survivin mRNA also remains obscure.

Survivin was detected by immunohistochemical staining in mouse granulosa cells and oocytes at all stages of development and the oocytes were intensely stained. As indicated in the introduction, survivin protects the embryo from apoptosis by inhibiting an apoptotic pathway (Kawamura et al., 2003Go). Furthermore, survivin mRNA is present at the earliest stages of Xenopus oocytes and it accumulates during oogenesis. It is also present in early Xenopus embryos and after the onset of zygotic transcripts, the amount of survivin mRNA rapidly declines to undetectable levels (Murphy et al., 2002Go). Survivin-null mouse embryos show embryonic lethality (Uren et al., 2000Go). These studies indicate that survivin is necessary for the oocytes and high levels should be expressed.

Several factors might control the apoptosis of granulosa cells. Gonadotrophins, epidermal growth factor/transforming growth factor-{alpha}, basic fibroblast growth factor, insulin-like growth factor I, estrogens and Fas ligand are known survival factors (Tilly et al., 1992Go; Chun et al., 1995Go; Quirk et al., 1995Go; Kaipia et al., 1996Go). The inhibitors of apoptosis proteins, Xiap and Hiap-2 are expressed in rat granulosa cells of preovulatory follicles after HCG stimulation (Leung and Steele, 1992Go). Gonadotrophin increases the Xiap and Hiap-2 protein content and high levels of antiapoptotic gene expression confer protection against apoptosis. Bcl-2 is also expressed in human and rat luteal cells and HCG significantly increases Bcl-2 mRNA and protein levels (Tilly et al., 1995Go; Yuan wei and Giudice, 1997Go). We found here that survivin is expressed at basal levels in granulosa cells and that HCG up-regulates survivin mRNA levels. HCG prevents the spontaneous apoptotic DNA fragmentation of cultured rat preovulatory follicles or rabbit corpus luteum (Chun et al., 1994Go; Dharmarajan et al., 1994Go). These observations suggest that HCG functions as an antiapoptotic factor in the human corpus luteum and that the role of survivin is consistent with its potential function in luteal regression.

Similar to the findings of a previous study that demonstrated the time-dependent onset of apoptosis of the rabbit corpus luteum in vitro, DNA started to disintegrate as early as 30 min after culture initiation and reached maximal levels within 8 h of incubation (Dharmarajan et al., 1994Go). Our data indicated that survivin mRNA reached a maximal level within 6 h of the HCG incubation and continued for a further 24 h. We speculate that a significant increase in the amount of survivin affects the maintenance of granulosa-luteal cell survival.

The levels of survivin mRNA expression are higher in granulosa cells from undifferentiated, prehierarchal hen follicles compared with differentiated, preovulatory follicles, and decrease during the early stages of atresia (Johnson et al., 2002Go). The present study did not address the developmental expression of survivin. However, HCG rescues antral and preovulatory follicles from atresia (Kaipia and Hsueh, 1997Go) and up-regulates survivin expression, indicating that survivin is a survival factor for granulosa cells.

In summary, this study demonstrated that the survivin mRNA and protein are expressed in mouse and human granulosa cells during follicular development and that HCG induces their up-regulation. Therefore, survivin might help support luteal function by suppressing apoptosis and HCG might contribute to survivin expression. Further studies should clarify the molecular mechanisms of survivin underlying the action of gonadotrophin as a survival factor in suppressing granulosa cell apoptosis.


   References
Top
 Abstract
 Introduction
 Materials and methods
 Results
 Discussion
 References
 
Abraham A, Iris KT, Dorit A, Dantes A, Abigail LB, Eli R, Ravid S and Liron H (2003) Steroidogenesis and apoptosis in the mammalian ovary. Steroids 68, 861–867.[CrossRef][Web of Science][Medline]

Adams R, Camena M and Earnshaw WC (2001) Chromosomal passengers and the (aurora) ABCs of mitosis. Trends Cell Biol 11, 49–54.[CrossRef][Web of Science][Medline]

Adida C, Berrebi D, Peuchmaur M, Reyes-Mugica M and Altieri DC (1998a) Anti-apoptosis gene, survivin and prognosis of neuroblastoma. Lancet 351, 882–883.[CrossRef][Web of Science][Medline]

Adida C, Crotty PL, McGrath J, Berrebi D, Diebold J and Altieri DC (1998b) Developmentally regulated expression of novel cancer anti-apoptosis gene survivin in human and mouse differentiation. Am J Pathol 152, 43–49.[Abstract]

Ambrosini G, Adida C and Altieri DC (1997) A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma. Nat Med 39, 917–921.

Ambrosini G, Adida C, Sirugo G and Altieri DC (1998) Induction of apoptosis and inhibition of cell proliferation by surviving gene targeting. J Bio Chem 273, 11177–11182.[Abstract/Free Full Text]

Badran A, Yoshida A, Ishikawa K, Goi T, Yamaguchi A, Ueda T and Inuzuka M (2004) Identification of a novel spliced variants of the human anti-apoptosis gene survivin. Biochem Biophys Res Commun 314, 902–907.[CrossRef][Web of Science][Medline]

Carambula SF, Matikainen T, Lynch MP, Flavell RA, Goncalves PB, Tilly JL and Rueda BR (2002) Caspase-3 is a pivotal mediator of apoptosis during regression of the ovarian corpus luteum. Endocrinology 143, 1495–1501.[Abstract/Free Full Text]

Chun S-Y, Billig H, Tilly JL, Tsafriri A and Hsueh AJW (1994) Gonadotropin suppression of apoptosis in cultured preovulatory follicles: mediatory role of endogenous insulin-like growth factor I. Endocrinology 135, 1845–1853.[Abstract]

Chun SY, Eisenhauer KM, Kubo M and Hsueh AJW (1995) Interleukin-1ß suppresses apoptosis in rat ovarian follicles by increase nitric oxide production. Endocrinology 136, 3120–3127.[Abstract]

Conway EM, Pollefeyt S, Cornelissen J, DeBaere I, Steiner-Mosonyi M, Ong K and Baenes M (2000) Three differentially expressed survivin cDNA variants encode proteins with distinct antiapoptotic functions. Blood 95, 1435–1442.[Abstract/Free Full Text]

Dharmarajan AM, Goodman SB, Tilly KI and Tilly JL (1994) Apoptosis during functional corpus luteum regression: evidence of a role for chorinic gonadotropin in promoting luteal cell survival. Endocr J 2, 295–303.

Johnson AL, Langer JS and Bridgham JT (2002) Survivin as a cell cycle-related and antiapoptotic protein in granulosa cells. Endocrinology 143, 3405–3413.[Abstract/Free Full Text]

Kaipia A and Hsueh AJW (1997) Regulation of ovarian follicle atresia. Annu Rev Physiol 59, 349–364.[CrossRef][Web of Science][Medline]

Kaipia A, Chun SY, Eisenhauer K and Hsueh AJW (1996) Tumor necrosis factor-{alpha} and its second messenger, ceramide, stimulate apoptosis in cultured ovarian follicles. Endocrinology 137, 4864–4870.[Abstract]

Kawamura K, Sato N, Fukuda J, Kodama H, Kumagai J, Tanikawa H, Shimizu Y and Tanaka T (2003) Survivin acts as an antiapoptotic factor during the development of mouse preimplantation embryos. Dev Biol 256, 331–341.[CrossRef][Web of Science][Medline]

Konishi Y, Sato H and Tanaka T (2004) Anisomycin superinduces annexin V mRNA expression through the ERK1/2 but not the p38 MAP kinase pathway. Biochem Biophys Res Commun 313, 974–980.

Krieg A, Mahotka C, Krieg T, Grabsch H, Muller W, Takano S, Suschek CV, Heydthausen M, Gabbert HE and Gerharz CD (2002) Expression of different survivin variants in gastric carcinoma: first clues to a role of surviving-2B in tumor progression. Br J Cancer 86, 737–743.[CrossRef][Web of Science][Medline]

Leung PCK and Steele GL (1992) Intracellular signaling in the gonads. Endocr Rev 13, 476–498.[Abstract/Free Full Text]

Li F, Ambrosini G, Chun EY, Plescia J, Tognin S, Marchisio PC and Altieri DC (1998a) Control of apoptosis and mitotic spindle checkpoint by survivin. Nature 396, 580–584.[CrossRef][Medline]

Li J, Kim JM, Liston P, Li M, Miyazaki T, Mackenzie AE, Korneluk RG and Tsang BK (1998b) Expression of inhibitor of apoptosis proteins (IAPs) in rat granulosa cells during ovarian follicular development and atresia. Endocrinology 139, 1321–1328.[Abstract/Free Full Text]

Mahotka C, Wenzel M, Springer E, Gabbert HE and Gerharz CD (1999) Survivin-{Delta}Ex3 and survivin-2B: two novel splice variants of the apoptosis inhibitor survivin with different antiapoptotic properties. Cancer Res 59, 6097–6102.[Abstract/Free Full Text]

Matsubara H, Ikuta K, Ozaki Y, Suzuki Y, Suzuki Y, Suzuki N, Sato T and Suzumori K (2000) Gonadotropins and cytokines affect luteal function through control of apoptosis in human luteinized granulosa cells. J Clin Endocrinol Metab 85, 1620–1626.[Abstract/Free Full Text]

Murphy CR, Sabel JL, Sandler AD and Dagle JM (2002) Survivin mRNA is down-regulated during early Xenopuse laevis embryogenesis. Dev Dyn 225, 597–601.[CrossRef][Web of Science][Medline]

Quirk SM, Cowan RG, Joshi SG and Henrikson KP (1995) Fas antigen-mediated apoptosis in human granulosa/luteal cells. Biol Reprod 52, 279–287.[Abstract]

Sato H, Ogata H and De Luca LM (2000) Annexin V inhibits the 12-O-tetradecanoylphorbol-13-acetate-induced activation of Ras/extracellular signal regulated kinase (ERK) signaling pathway upstream of Shc in MCF-7 cells. Oncogene 19, 2904–2912.[CrossRef][Web of Science][Medline]

Shin S, Sung BJ, Cho YS, Kim HJ, Ha NC, Hwang JI, Chung CW, Jung YK and Oh BH (2001) An anti-apoptotic protein human survivin is a direct inhibitor of caspase-3 and -7. Biochemistry 40, 1117–1123.[CrossRef][Medline]

Sugino N, Suzuki T, Kashida S, Karube A, Takiguchi S and Kato H (2000) Expression of Bcl-2 and Bax in the human corpus luteum during the menstrual cycle and in early pregnancy: regulation by human chorionic gonadotropin. J Clin Endocrinol Metab 85, 4379–4386.[Abstract/Free Full Text]

Tasdemir M, Tasdemir I, Kodama H and Tanaka T (1993) Pentoxifylline-enhanced acrosome reaction correlates with fertilization in vitro. Hum Reprod 8, 2102–2107.[Abstract/Free Full Text]

Thordarson G, Galosy S, Gudmundsson GO, Newcomer B, Sridaran R and Talamantes F (1997) Interaction of mouse placental lactogens and androgens in regulating progesterone release in cultured mouse luteal cells. Endocrinology 138, 3236–3241.[Abstract/Free Full Text]

Tilly JL, Kowalski KI, Johnson AL and Hsueh AJW (1992) Involvement of apoptosis in ovarian folicular atresia and postovulatory regression. Endocrinology 129, 2799–2801.[Web of Science]

Tilly JL, Billing H, Kowalski KI and Hsueh AJW (1992) Epidermal growth factor and basic fibroblast growth factor suppress the spontaneous onset of apoptosis in cultured rat ovarian granulosa cells and follicles by a tyrosine kinase-dependent mechanism. Mol Endocrinol 6, 1942–1950.[Abstract/Free Full Text]

Tilly JL, Tilly KI, Kenton ML and Johnson AL (1995) Expression of members of the Bcl-2 gene family in the immature rat ovary: equine chorionic gonadotropin-mediated inhibition of granulosa cell apoptosis is associated with decreased Bax and constitutive Bcl-2 and Bcl-xlong messenger ribonucleic acid levels. Endocrinology 136, 232–241.[Abstract]

Uren AG, Wong L, Pakusch M, Fowler KJ, Burrows FJ, Vaux DL and Choo KH (2000) Survivin and the inner centromere protein INCENP show similar cell-cycle localization and gene knockout phenotype. Curr Biol 10, 1319–1328.[CrossRef][Web of Science][Medline]

Yuan W and Giudice LC (1997) Programmed cell death in human ovary is a function of follicle and corpus luteum status. J Clin Endocrinol Metab 82, 3148–3155.[Abstract/Free Full Text]

Zeleznik AJ, Ihring LL and Bassett SG (1989) Developmental expression of Ca2+/Mg2+-dependent endonuclease activity in rat granulosa and luteal cells. Endocrinology 125, 2218–2220.[Abstract/Free Full Text]

Submitted on October 12, 2004; accepted on December 18, 2004.


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A. Jasinska, Z. Strakova, M. Szmidt, and A. T. Fazleabas
Human Chorionic Gonadotropin and Decidualization in Vitro Inhibits Cytochalasin-D-Induced Apoptosis in Cultured Endometrial Stromal Fibroblasts
Endocrinology, September 1, 2006; 147(9): 4112 - 4121.
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S. Fukuda and L. M. Pelus
Survivin, a cancer target with an emerging role in normal adult tissues
Mol. Cancer Ther., May 1, 2006; 5(5): 1087 - 1098.
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