Yixun Liu

Chinese Academy of Sciences, Peping, Beijing, China

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Publications (24)46.97 Total impact

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    ABSTRACT: The acrosome is a specialized organelle that covers the anterior part of the sperm nucleus and plays an essential role in mammalian fertilization. However, the regulatory mechanisms controlling acrosome biogenesis and acrosome exocytosis during fertilization are largely unknown. Equatorin (Eqtn) is a membrane protein that is specifically localized to the acrosomal membrane. In the present study, the physiological functions of Eqtn were investigated using a gene knockout mouse model. We found that Eqtn(-/-) males were subfertile. Only approximately 50% of plugged females were pregnant after mating with Eqtn(-/-) males, whereas more than 90% of plugged females were pregnant after mating with control males. Sperm and acrosomes from Eqtn(-/-) mice presented normal motility and morphology. However, the fertilization and induced acrosome exocytosis rates of Eqtn-deficient sperm were dramatically reduced. Further studies revealed that the Eqtn protein might interact with Syntaxin1a and SNAP25, but loss of Eqtn did not affect the protein levels of these genes. Therefore, our study demonstrates that Eqtn is not essential for acrosome biogenesis but is required for the acrosome reaction. Eqtn is involved in the fusion of the outer acrosomal membrane and the sperm plasma membrane during the acrosome reaction, most likely via an interaction with the SNARE complex.
    Biochemical and Biophysical Research Communications 01/2014; · 2.28 Impact Factor
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    ABSTRACT: The Wt1 gene encodes a nuclear transcription factor that is specifically expressed in ovarian granulosa cells. However, the physiological significance of Wt1 in ovarian follicle development remains elusive. In this study, we found that Wt1(+/R394 W) mice were grossly normal, however, the females displayed severe reproductive defects. Only approximately 15% of the Wt1(+/R394 W) females became pregnant after mating with wild type males, compared with 88.2% of control females. Further study revealed that the subfertility of Wt1(+/R394 W) females was caused by aberrant ovarian follicle development. Compared with control females, the ovary size and the number of developing follicles was significantly decreased in Wt1 mutant ovaries which was very similar to Premature Ovarian Failure (POF) in human patients. The results of in vitro studies demonstrated that the expression of FSHR, 3β-HSD, and Aromatase was inhibited by Wt1 in granulosa cells, and mutation of Wt1 resulted in the upregulation of these genes and in the premature differentiation of granulosa cells. We also found that Wt1 was likely involved in granulosa cell development via the regulation of E-cadherin and Par6b expression. Mutation in Wt1 caused defects in polarity establishment in granulosa cells, which also likely contributed to the observed aberrant follicle development. The results of this study provide new mechanisms for understanding the regulation of ovarian follicle development and potential pathological cause of POF in human patients.
    Human Molecular Genetics 09/2013; · 6.68 Impact Factor
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    ABSTRACT: The steroidogenic acute regulatory protein (StAR) is the key regulatory protein of steroidogenesis.De novo synthesis of StAR protein is required for intramitochondrial translocation of cholesterol to the cytochrome P450 side chain cleavage enzyme which is located on the matrix side of the inner mitochondrial membrane. This is the rate-limiting step of steroid biosynthesis. Usingin situ hybridization and immunohistochemistry we studied StAR expression in various stages of the corpora luteal and its regulation by interferon-gamma (IFNγ) in the adult pseudopregnant rat. The results indicated that expression of StAR in the corpora luteal was correlated with progesteron production and IFNγ was capable of inhibiting its expression.
    Chinese Science Bulletin 04/2012; 45(23):2152-2157. · 1.37 Impact Factor
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    ABSTRACT: The distribution of mRNAs of tissue type (t) and urokinase type (u) plasminogen activator (PA) plus their corresponding inhibitors, type-1 (PAI-1) and type-2 (PAI-2) have been studied in the tissues of human first and second trimester placentae byin situ hybridization. The results show that: (i) All the molecules, tPA, uPA, PAI-1 and PAI-2, were identified in the blood vessels, the majority of extravillous trophoblastic cells of the decidual layer between Rohr’s and Nitabuch’s stria and in the trophoblast cells lining the chorionic plate, basal plate, intercotyledonary septae and cytotrophoblast cells of the chorionic villous tree. (ii) No expression of such probes was observed in the basal and chorionic plate, glandular cells of the decidua, the septal tissues or the villous core mesenchyme. The co-distribution of the molecules observed suggests that the co-ordinated expression of the activators and inhibitors in various cells of the placental tissue may play a role in angiogenesis related to conversion of spiral arteries into utero-placental arteries and establishment of a chorio-decidual blood flow during early stages of placentation. Keywordsplasminogen activator-plasminogen activator inhibitor-placentation-angiogenesis- in situ hybridization
    Chinese Science Bulletin 04/2012; 45(22):2056-2062. · 1.37 Impact Factor
  • Haizhen Liu, Yixun Liu
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    ABSTRACT: Byin situ hybridization, the localization of orphan receptor TR3 mRNA has been observed in early developmental follicles. TR3 mRNA is first expressed in the ovarian interstitial cells on day 2 after birth, and then in granulosa cells (GC) in primary follicles on day 4. The expression level of TR3 mRNA in GC increases following the follicular development. Its higher expression can be observed in the outer layer of GC and inner layer of theca cells (TC) on day 6, where the cells present active proliferation and differentiation. The expression of TR3 is in an increasing manner until the large antral follicles on day 30. The mRNA is only expressed in the healthy, but not atretic follicles in adult rat ovaries. Injection of epidermal growth factor (EGF) has dramatically enhanced its expression in the early stage of developmental follicles. It is therefore suggested that TR3 may play a role in regulating growth and differentiation of ovarian somatic cells in the early stage, and its expression is regulated by EGF.
    Chinese Science Bulletin 04/2012; 45(12):1122-1127. · 1.37 Impact Factor
  • YiXun Liu, XiXia Li
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    ABSTRACT: Artificial cryptorchidism or local testicular heat treatment can induce reversible oligospermia or azoospermia in monkeys and rats via germ cell apoptosis. Local warming of monkey testes in water at 43°C for 2 consecutive days (30 min per day) decreased the number of sperm in the semen by up to 80% on d 28, and the effect was completely reversed on d 144. Germ cells rely heavily on Sertoli cells for structural and nutritional support. Specialized junctions that play a pivotal role in spermatogenesis occur at sites of Sertoli-Sertoli and Sertoli-germ cell contact in the seminiferous epithelium. We demonstrated that expression of tight junction (TJ)-associated molecules, such as occludin and zonula occludens-1 (ZO-1), were greatly reduced 24-48 h after heat treatment, while the permeability of the blood-testis barrier (BTB) was simultaneously increased, but recovered 10 d later. These results indicate a reversible disruption of the BTB associated with transient inductions of transforming growth factor (TGF) β2 and β3 expression, p38 mitogen-activated protein kinase and extracellular signal-regulated kinase activation, and concomitant loss of occludin and ZO-1. This suggests that expression of TJ-associated molecules and the BTB was reversibly perturbed by mild testicular hyperthermia, and that the heat-induced induction of TGF-β might be involved in downregulating TJ-associated proteins, leading to cell junction reduction. This review discusses the changes in total gene expression patterns after experimental cryptorchidism in adult mouse testes, and the cloning of several novel, physiologically significant spermatogenesis-specific genes.
    Science China. Life sciences 11/2010; 53(11):1274-83. · 1.51 Impact Factor
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    ABSTRACT: Inhibin has long been considered as a suppresser of follicle-stimulating hormone (FSH) secretion from anterior pituitary through pituitary-gonad negative feedback to regulate follicle development. We demonstrated that addition of inhibin A could significantly suppress FSH-induced FSHR mRNA level in cultured rat granulosa cells (GCs) measured by real-time PCR. The inhibin A exerted its action mainly by inhibiting FSHR promoter activity. Furthermore, exogenous inhibin A could dramatically decrease FSH-induced P450arom and P450scc level and suppress progesterone and estradiol production in the cultured GCs, but it did not decrease forskolin-induced steroidogenesis, indicating that the inhibitory effect of inhibin A on FSH action may be upstream of cAMP signaling. Inhibin A was also capable of suppressing FSH-induced expression of steroidogenic factor 1 (SF-1) and androgen receptor, but stimulating DAX-1 expression in the culture. Our study has provided new evidence to show that inhibin A is capable of feedback antagonizing FSH action on GCs by reducing FSHR expression at ovarian level via a short feedback loop. Transcriptional factor receptors, such as SF-1, AR and DAX-1 were involved in this regulation.
    Molecular and Cellular Endocrinology 11/2008; 298(1-2):48-56. · 4.24 Impact Factor
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    ABSTRACT: Unexplained pregnancy loss and recurrent miscarriage seriously impair human fecundity. However, the underlying molecular mechanisms remain elusive. Recent studies suggest that the adhesion molecule CD146 may be involved in unexplained recurrent miscarriage. Here, we investigate the effect of CD146 on early pregnancy. Using in situ hybridization and immunohistochemistry, we found that CD146 was specifically expressed in the receptive maternal uteri and invasive embryonic trophoblasts during the early stages of pregnancy, but it was completely absent in the non-pregnant uterus. Our in vitro studies demonstrated that blocking CD146 with a function-perturbation antibody AA98 significantly inhibited the attachment of blastocysts onto the receptive uterine luminal epithelial monolayer, the trophoblastic outgrowth of blastocysts and ectoplacental cones, and the secretion of matrix metalloproteinases. Animal experiments showed that applying this antibody before embryo implantation caused pregnancy failure in mice. Our data present direct evidence for the role of CD146 in mediating embryonic attachment and trophoblastic invasion, and provide new insight into the molecular mechanism underlying unexplained pregnancy loss and recurrent miscarriage.
    Journal of Cellular Physiology 07/2008; 215(3):621-6. · 3.87 Impact Factor
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    ABSTRACT: RING finger proteins play important roles in spermatogenesis. Here, we report that a novel RING finger protein RNF151, with a C3HC4-type RING finger domain, a putative nuclear localization signal (NLS), and a TRAF-type zinc finger domain, was exclusively expressed in the mouse testis and developmentally regulated during spermatogenesis. While RNF151 mRNA was present in round spermatids, its protein was expressed in elongating spermatids of the stage VIII-IX seminiferous tubules. The NLS together with the RING domain were necessary and sufficient for the nuclear localization of RNF151-EGFP in transfected cells. Yeast two-hybrid screening identified the physical interaction of mouse RNF151 and dysbindin, which was confirmed by the co-immunoprecipitation of the proteins and by their co-localization in intact cells. As dysbindin has lately been shown to be involved in membrane biogenesis and fusion, a key process for acrosome formation, we propose that RNF151 may play a role in acrosome formation.
    Archives of Biochemistry and Biophysics 10/2007; 465(1):157-63. · 3.04 Impact Factor
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    ABSTRACT: The mRNA of the mitochondrial uncoupling protein 2 (UCP2) was up-regulated by cryptorchidism, a testicular hyperthermic condition under which germ cells undergo severe apoptosis. We investigated whether UCP2 was able to protect germ cells from hyperthermia-induced apoptosis. UCP2 was predominantly present in elongate spermatids under normal conditions, and was detected in all germ cells with its level significantly increased if the testes were exposed to 43 degrees C for 5 min. Such a short heat exposure was non-lethal and enabled the preconditioned cells to be resistant to apoptosis induced by a longer hyperthermic treatment (15 min). While hyperthermia resulted in oxidative stress in mouse testes, it did not change the total anti-oxidative capacity. Indeed, overexpression of UCP2 in the GC-2 germ cell line protected the cells from radical oxygen species (ROS)-induced apoptosis. Taken together, we propose that UCP2 may represent an effective weaponry used by germ cells to combat ROS-induced apoptosis.
    Biochemical and Biophysical Research Communications 09/2007; 360(2):327-32. · 2.28 Impact Factor
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    ABSTRACT: The corpus luteum (CL) is a transient endocrine organ that secretes progesterone to support early pregnancy. If implantation is unsuccessful, luteolysis is initiated. Extensive tissue remodeling occurs during CL formation and luteolysis. In this study, we have studied the possible involvement of MMP-2, -9, -14, and their inhibitors, TIMP-1, -2, -3 in the CL of cycling rhesus monkey at various stages by in situ hybridization, immunohistochemistry and microscopic assessment. The results showed that the MMP-2 mRNA and protein were mainly expressed in the endothelial cells at the early and middle stages of the CL development, while their expressions were observed in the luteal cells at the late stage during luteal regression. MMP-9 protein was detected in the CL at the early and middle stages, and obviously increased at the late stage. The expressions of MMP-14 and TIMP-1 mRNA were high at the early and late stages, and low at the middle stage. TIMP-2 mRNA was high throughout all the stages, the highest level could be observed at the late stage. The TIMP-3 production was detected throughout all the stages, but obviously declined during CL regression. MMP-9, -14 and TIMP-1, -2, -3 were mainly localized in the cytoplasm of the steroidogenic cells. The results suggest that the MMP/TIMP system is involved in regulation of CL development in the primate, and the coordinated expression of MMP-2, -14 and TIMP-1, -3 may have a potential role in the CL formation and the functional maintaining, while the interaction of MMP-2, -9, -14 and TIMP-1, -2, -3 might also play a role in CL regression at the late stage of CL development in the primate.
    Science in China Series C Life Sciences 03/2006; 49(1):37-45. · 1.61 Impact Factor
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    ABSTRACT: In vitro cultured embryonic stem (ES) cells are derived from the inner cell mass (ICM) of pre-implantation embryos, and are capable of giving rise to all cell and tissue types of the three germ layers upon being injected back into blastocysts. These cells are therefore said to possess pluripotency that can be maintained infinitely in culture under optimal conditions. Such pluripotency maintenance is believed to be due to the symmetrical cleavage of the cells in an undifferentiated state. The pluripotency of ES cells is the basis for their various practical and potential applications. ES cells can be used as donor cells to generate knockout or transgenic animals, asin vitro models of mammalian development, and as cell resources for cell therapy in regenerative medicine. The further success in these applications, particularly in the last two, is dependent on the establishment of a culture system with components in the medium clearly defined and the subsequent procedures for controlled differentiation of the cells into specific lineages. In turn, elucidating the molecular mechanism for pluripotency maintenance of ES cells is the prerequisite. This paper summarizes the recent progresses in this area, focusing mainly on the LIF/STAT3, BMPs/Smads, canonical Wnt, TGFβ/activin/nodal, PI3K and FGF signaling pathways and the genes such asoct4, nanog that are crucial in ES cell pluripotency maintenance. The regulatory systems of pluripotency maintenance in both mouse and human ES cells are also discussed. We believe that the cross-talkings between these signaling pathways, as well as the regulatory system underlying pluripotency maintenance will be the main focus in the area of ES cell researches in the future. KeywordsES cells-pluripotency-signaling pathway-pluripotency maintenance associated genes
    Chinese Science Bulletin 10/2005; 50(19):2121-2131. · 1.37 Impact Factor
  • Yixun Liu
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    ABSTRACT: Embryo in uterine implantation is a complex and multifactor-related process and is a downstream and ideal point for woman fertility control. Understanding the cellular and molecular mechanism of implantation is a prerequisite for development of anti-implantation contraceptives. In spite of considerable accumulation of information from the laboratory animals that has been achieved, it is difficult to generate such information in human due to ethical restriction and experimental limitation, and the present knowledge for understanding the definitive mechanisms which control these events remains elusive. Embryo implantation can also occur outside uterus. Some women with abdominal pregnancies could successfully complete the processes of gestation and bear normal babies, implying that implantation itself may be not an endometrium-specific process. Reproductive biologists should cooperate with gynecologists to further comparatively study the molecular and cellular mechanisms of implantation normally occurring in endometrium and abnormally appearing outside uterine cavity. Such collaborative studies may generate new important information for developing anti-implantation contraceptive and for techniques of accurate diagnosis of ectopic pregnancy. A specially designed GnRH-2 analog and a combination use of low dose RU486 and gossypol as anti-implantation contraceptives have been suggested.
    Science in China Series C Life Sciences 09/2004; 47(4):293-302. · 1.61 Impact Factor
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    ABSTRACT: Using immunohistochemistry, in situ hybridization, Western blot and TUNEL methods, we have studied the expression of Fas/FasL, Bcl-2/Bax and caspase-3 in the corpora lutea (CL) at various stages of pseudopregnant rat induced by injection of PMSF/hCG. The results showed that no apoptotic signal could be observed until Day 14 after hCG injection. Fas weakly expressed in the CL at all the stages increased when luteolysis took place. FasL signal increased dramatically on Day 14 and reached the maximum level on Day 21. The expression of Bcl-2 and Bax was detected in a time-dependent manner. At the early stage of CL development, Bcl-2 expression was stronger, while Bax was low. The expression of Bcl-2 and Bax in the CL was completely reversed. Caspase-3 antigen could be detected throughout all the phases of CL development in a time-dependent fashion, low on Day 2 and reaching the maximum on Day 21. These results suggest that luteal regression at the late phases may be related to cell apoptosis.
    Science in China Series C Life Sciences 07/2003; 46(3):273-85. · 1.61 Impact Factor
  • Xuan Jin, Yixun Liu
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    ABSTRACT: Only limited numbers of primordial follicles in mammalian ovary grow and differentiate to reach the stage of dominate follicles and ovulate. 99% of the follicles in the ovary undergo atresia at various stages of development. Regulation of follicular growth, development and atresia is a complex process and involves interactions between endocrine factors and intraovarian regulators. This review summarized: i) FSH may not be a survival factor in regulating slow-growing preantral follicles. Some locally produced growth factors, activin and orphan receptors might play a more important role at this stage. ii) Estrogen, activin/ inhibin and follistatin coordinate with FSH to regulate and control follicle differentiation. iii) There are two types of follicular atresia induced by apoptosis which originates from GC or oocyte, respectively. Early translation of tPA mRNA into tPA protein in oocyte may be associated with oocyte apoptosis. Keywordsprimordial follicle-oocyte-granulose cell-differentiation-atresia
    Chinese Science Bulletin 01/2003; 48(17):1786-1790. · 1.37 Impact Factor
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    ABSTRACT: In order to investigate the relationship between the endometrial receptivity and matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-1,-3 (TIMP-1,-3) in the endometrium, we used early pregnant mice as the animal model and studied the expression of MMP-2, TIMP-1,-3 in the endometrium in relation to the number of implantation sites after RU486 treatment. The results indicated that RU486 could significantly inhibit embryo implantation and change the expression of MMP-2 and TIMP-1,-3 in a dose-dependent pattern. When the mice were treated with 12 mg/kg RU486, there were a few embryos implanted as compared with the control. The expression of matrix metalloproteinase MMP-2 was low during the period of "implantation window", while the tissue inhibitor of metalloproteinase in the endometrial cells was high, suggesting that the activity of the proteolytic enzyme was strictly controlled by its inhibitors. After RU486 treatment, the generation of TIMP-1,3 was decreased while the MMP-2 was significantly increased, indicating that the normal balance between the activators and their inhibitors in the tissue was broken and the extracellular matrix was excessively degraded, subsequently the embryo implantation was inhibited. Therefore, it is suggested that the anti-implantation effect of RU486 may be mediated by MMPs and their inhibitors TIMPs.
    Science in China Series C Life Sciences 09/2002; 45(4):406-11. · 1.61 Impact Factor
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    ABSTRACT: To investigate the effects of mifepristone on apoptosis and expression of relative genes in early pregnant chorionic villi and decidua. The specimen of early pregnant chorionic villi and decidua obtained from 10 cases of requesting termination of pregnancy by curettage, 20 cases of mifepriston contragestation. The paraffin sections were used to determine apoptotic cells by TdT-mediated dUTP-biotin nick end labeling method, to identify bcl-2, bax, fas, fasL and proliferating cell nuclear antigen (PCNA) by immunohistochemistry, to demonstrate fas and fasL mRNA by in situ hybridization. In normal early pregnant specimens, apoptotic cells were mainly observed in syncytiotrophoblast, but not in cytotrophoblast cells, occasionally seen in decidua cells. The antigen of bax, fas, fasL were present in syncytiotrophoblast cells and decidua with lower amount. While bcl-2 antigen staining was strong in cytotrophoblastic cells and in decidua. PCNA protein was present in cytotrophoblastic and decidual cells only. In the specimens treated with mifepristone, apoptotic cells were increased in syncytiotrophoblastic cells of villi and visualized in decidua cells. The expression of fas, fasL and bax was also higher than that of normal. Mifepristone increased apoptosis in syncytiotrophoblastic and decidua cells, but had no effect on the expression of bcl-2 and PCNA.
    Zhonghua fu chan ke za zhi 05/2002; 37(4):220-2.
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    ABSTRACT: Human placental tissues from the first and second trimesters of gestation have been investigated using riboprobein situ hybridisation of mRNA sequences coding for membrane type metalloproteinase (MT-1-MMP) and tissue inhibitors of metalloproteinase-1 (TIMP-1). Results show that (i) both mRNAs express at a relatively high level in the chorion laeve trophoblast cells and the adjacent decidual cells of fetal membrane; (ii) the most abundant expression of the two mRNAs was found in the extravillous trophoblast between Rohrs and Nitabuch striae of basal plate, trophoblast shell and gland cells of the decidua; (iii) isolated or small groups of cytotrophoblast cells in the chorionic villi and in the cells lining arterioles in decidua and stem villi also expressed both MT-1-MMP and TIMP-1 at defferent extents. The data suggest that the coordinated expression of the MT-MMP and its inhibitor TIMP in defferent cells of the placental tissue may play an essential role in trophoblast invasion and angiogenesis related to placentation in the first two trimesters of gestation. They may also have an ability to effect separation of fetal from material tissue at a favorable junctional site during parturition.
    Chinese Science Bulletin 07/2000; 45(16):1484-1489. · 1.37 Impact Factor
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    ABSTRACT: Expression and cellular localization of orphan receptor TR2 mRNA in relation to germ cell apoptosis in cryptorchid testes of rat and rhesus monkey have been studied by usingin situ hybridization andin situ 3′-end labeling of DNA fragments (TUNEL). The results show that: (i) TR2 mRNA is specifically expressed in the germ cells, mainly in the spermatocytes, round and elongated spermatids. The expression level of TR2 mRNA varies with the seminiferous cycle, (ii) In the rat cryptorchid testes on days 3 and 5 after the surgery, the germ cells began to undergo apoptosis with no evident decrease in TR2 mRNA level. On day 7.5, however, most germ cells underwent apoptosis, while the expression level of TR2 mRNA declined markedly, and TR2 mRNA was rarely expressed on day 10 thereafter, (iii) On days 15 and 20 of the cryptorchid testes of rhesus monkey, TR2 mRNA was only expressed in a few of primary spermatocytes and the mRNA was almost undetectable on days 30, 45, 60. These results suggest that TR2 mRNA probably plays an important role in spermatogenesis and germ cell apoptosis.
    Chinese Science Bulletin 03/2000; 45(8):720-725. · 1.37 Impact Factor
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    ABSTRACT: By using DNA 3′-end labeling, immunocytochemistry and mRNA insitu hybridization detection techniques, the expression of inhibin subunits and LH receptor in the granulosa cells and tissue-type plasminogen activator (tPA) in the oocytes has been studied in relation to follicular development and atresia. The results demonstrated that: (i) tPA activity in the oocytes of normal developing follicles is undetectable, and increases significantly in the follicle undergoing atresia; (ii) the production of inhibin subunits in granulosa cells is negatively correlated with the expression of oocyte tPA activity, indicating that they may be an important regulator of oocyte tPA production and follicular development; (iii) in atretic follicles, granulosa cells do not express LH receptor and inhibin subunits. It is therefore suggested that tPA may play a role in oocyte self-destruction and clearance in some of atretic follicles, and inhibin of granulosa-origin might be an inhibitory factor for the translation of tPA in the oocyte.
    Science in China Series C Life Sciences 12/1999; 42(6):583-590. · 1.61 Impact Factor

Publication Stats

97 Citations
46.97 Total Impact Points


  • 2004–2014
    • Chinese Academy of Sciences
      • State Key Laboratory of Reproductive Biology
      Peping, Beijing, China
  • 2002–2013
    • Northeast Institute of Geography and Agroecology
      • State Key Laboratory of Reproductive Biology
      Beijing, Beijing Shi, China