Volume 31 Issue 3
May  2010
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LIN Peng-Fei, HAO Yuan-Bing, GUO Hui-Li, LIU Hong-Lin, RUI Rong. Role of Fas/FasL on Apoptosis of Porcine Follicular Granulose Cells Derived from Isolated Follicles During Culture in vitro. Zoological Research, 2010, 31(3): 268-274. doi: 10.3724/SP.J.1141.2010.03268
Citation: LIN Peng-Fei, HAO Yuan-Bing, GUO Hui-Li, LIU Hong-Lin, RUI Rong. Role of Fas/FasL on Apoptosis of Porcine Follicular Granulose Cells Derived from Isolated Follicles During Culture in vitro. Zoological Research, 2010, 31(3): 268-274. doi: 10.3724/SP.J.1141.2010.03268

Role of Fas/FasL on Apoptosis of Porcine Follicular Granulose Cells Derived from Isolated Follicles During Culture in vitro

doi: 10.3724/SP.J.1141.2010.03268
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  • Corresponding author: RUI Rong
  • Received Date: 2009-06-22
  • Rev Recd Date: 2009-12-29
  • Publish Date: 2010-06-22
  • The whole antral follicles were isolated from porcine ovaries and classified as follows: healthy follicles (HF), early atretic follicles (EF) and progressed atretic follicles (PF). The isolated porcine follicles were used for routine histological section and HE staining after examination by eyesight. Morphological research shows that the accuracy rate of eyesight examination for HF is 92%. Healthy follicles were chosen for further experiment and divided into 3 groups: large follicle (Æ>5 mm), medium follicles (3−5 mm) and small follicles (≤3 mm). All follicles were cultured for 8, 16 and 24 h, respectively and the apoptosis of of their granulose cells were examined by Annexin V-FITC/PI double-labeling. It showed that the total apoptotic rate of granulose cells derived from cultured follicles could reach over 70% at 8 h after culture and be 81.1%-94.6% at 24 h after culture. Granulosa cells from groups were collected at 0, 8, 16, 24, 48 and 72 h after culture without serum and used for the examination of expression of FasL and Fas mRNA with real time PCR SYBRgreen method. The expression level of FasL mRNA of granulose cells from different size of follicles increased with culture time and reached the highest level at 24 h after culture (P<0.05). Expression level of FasL mRNA of granulose cells from small follicles was higher than those from large and medium follicles. There exists no difference for expression level of Fas mRNA of granulose cells among groups before culture but significantly increased at 8 h after culture and reached the highest level at 48 h after culture. It showed in the present experiment that the follicular culture system without serum used could effectively induce the apoptosis of follicular granulose cells. Cell apoptosis is the main cause of follicular atresia, the degree of which varied with the size of follicles. Small follicles seemed to be easier atretic than medium and large follicles.
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  • [1]
    Alonso-Pozos I, Rosales-Torres AM, Ávalos-Rodrígueza A, Vergara-Onofre M, Rosado-García A. 2003. Mechanism of granulosa cell death during follicular atresia depends on follicular size [J]. Theriogenology, 60: 1071-1081.
    [2]
    Bridgham JT, Johnson AL. 2001. Expression and regulation of Fas antigen and tumor necrosis factor receptor type I in hen granulosa cells [J]. Biol Reprod, 65: 733-739.
    [3]
    Canipari R. 2000. Oocyte--granulosa cell interactions[J]. Hum Reprod Update, 6(3): 279-289.
    [4]
    Chen Q, Yano T, Matsumi H, Osuga Y, Yano N, Xu J, Wada O, Koga K, Fujiwara T, Kugu K, Taketani Y. 2005. Cross-talk between Fas/Fas ligand system and nitric oxide in the pathway subserving granulosa cell apoptosis: a possible regulatory mechanism for ovarian follicle atresia [J]. Endocrinology, 146: 808-815.
    [5]
    Dharma SJ, Kelkar RL, Nandedkar TD. 2003. Fas and Fas ligand protein and mRNA in normal and atretic mouse ovarian follicles [J]. Reproduction, 126: 783-789.
    [6]
    Diaz FJ, Wigglesworth K, Eppig JJ. 2007. Oocytes determine cumulus cell lineage in mouse ovarian follicles [J]. J Cell Sci, 120: 1330-1340.
    [7]
    Guthrie HD, Garrett WM, Cooper BS. 1998. Follicle-stimulating hormone and insulin-like growth factor-I attenuate apoptosis in cultured porcine granulosa cells [J]. Biol Reprod, 58: 390-396.
    [8]
    Inoue N, Maeda A, Matsuda-minehata F, Fukuta K, Manabe N. 2006. Expression and localization of Fas ligand and Fas during atresia in porcine ovarian follicles[J]. J Reprod Dev, 52(6): 723-730.
    [9]
    Inoue N, Manabe N, Matsui T, Maeda A,Nakagawa S, Wada S, Miyamoto H. 2003. Roles of tumor necrosis factor-related apoptosis-inducing ligand signaling pathway in granulosa cell apoptosis during atresia in pig ovaries [J]. J Reprod Dev, 49: 313-321.
    [10]
    Jolly PD, Smith PR, Heath DA, Hudson NL, Lun S, Still LA, Watts CH, McNatty KP. 1997. Morphological evidence of apoptosis and the prevalence of apoptotic versus mitotic cells in the membrana granulosa of ovarian follicles during spontaneous and induced atresia in ewes [J]. Biol Reprod, 56: 837-846.
    [11]
    Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△Ct method [J]. Methods, 25: 402-408.
    [12]
    Maillet G, Benhaïm A, Mittre H, Féral C. 2003. Involvement of theca cells and steroids in the regulation of granulosa cell apoptosis in rabbit preovulatory follicles [J]. Reproduction, 125(5): 709-716.
    [13]
    Manabe N, Goto Y, Matsuda-Minehata F, Inoue N, Maeda A, Sakamaki K, Miyano T. 2004. Regulation mechanism of selective atresia in porcine follicles: regulation of granulosa cell apoptosis during atresia [J]. J Reprod Dev, 50(5): 493-514.
    [14]
    Matsuda-minehata F, Inoue N, Goto Y, Manabe N. 2006. The regulation of ovarian granulosa cell death by pro- and anti-apoptotic molecules [J]. J Reprod Dev, 52(6): 695-705.
    [15]
    Moor RM, Dai Y, Lee C, Fulka J Jr. 1998. Oocyte maturation and embryonic failure [J]. Hum Reprod Update, 4(3): 223-236.
    [16]
    Nakayama M, Manabe N, Inoue N, Matsui T, Miyamoto H. 2003. Changes in the expression of tumor necrosis factor (TNF)?, TNF ɑ receptor (TNFR) 2 and TNFR-associated factor 2 in granulosa cells during atresia in pig ovaries [J]. Biol Reprod, 68: 530-535.
    [17]
    Nakayama M, Manabe N, Nishihara S, Miyamoto H. 2000. Species specific differences in apoptotic cell localization in granulosa and theca interna cells during follicular atresia in porcine and bovine ovaries [J]. J Reprod Dev, 46: 147-156.
    [18]
    Parborell F, Irusta G, Vitale A, Gonzalez O, Pecci A, Tesone M. 2005. Gonadotropin-releasing hormone antagonist antide inhibits apoptosis of preovulatory follicle cells in rat ovary [J]. Biol Reprod, 72: 659-666.
    [19]
    Porter DA, Vickers SL, Cowan RG, Huber SC, Quirk SM. 2000. Expression and function of Fas antigen vary in bovine granulosa and theca cells during ovarian follicular development and atresia [J]. Biol Reprod, 62: 62-66.
    [20]
    Quirk SM, Cowan RG, Joshi SG, Henrikson KP. 1995. Fas antigen-mediated apoptosis in human granulosa/luteal cells[J]. Biol Reprod, 52: 279-287.
    [21]
     
    [22]
     
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