Ultrastructural factors affecting the quality of oocytes in aged mice

doi:10.16098/j.issn.0529-1356.2017.06.018

Abstract

Abstract:

Objective To investigate the morphology and distribution of cortical granules, mitochondria, microfilaments, spindles, and chromosomes in aged mice. Methods Kunming mice were divided into a young (4-8 weeks) and a old age group (48-52 weeks). Female mice were injected intraperitoneally with 10 IU prenant mare serum gonadotropin (PMSG), and followed 48 hours later by 10 IU of human chorionic gonadotrophin (hCG). MII oocytes were collected from the oviducts 14 hours after the hCG injection into M199 medium. The cumulus cells were removed by a brief incubation. Oocyte-corona-cumulus complex(OCCC) in fallopian tube were collected by superovulation to female mice. Anti-mouse α-tubulin monoclonal antibody was used to label the spindle microtubules, phalloidin labeled actin rhodamine to label microfilaments, lentil lectin combined with FITC to label cortical granules and Mito Tracker Green FM to label mitochondria. The specimen was examined under a laser scanning confocal microscope. Results Totally 372 oocytes were obtained, 179 oocytes from the young group and 193 oocytes from aged group. The abnormal rate of spindle of elderly mouse oocytes (71.78% ± 1.27%) was significantly higher than that of young mice (18.93% ± 1.27%) (P<0.01). The elderly mouse oocytes chromosomes arranged abnormal rate (65.16% ± 1.52%) was significantly higher than that of young mice (32.24% ± 1.10%) (P<0.01). Fluorescence intensity of aged mouse oocyte mitochondria (28.09 ± 6.62) was significantly lower than that of young mice (45.07 ± 5.90) (P<0.01). The aged mouse mature cortical granules (grade III cortical granules) rate (51% ± 1%) was significantly lower than that of young mice (94.04% ± 0.71%) (P<0.01). Conclusion The abnormal rate of aged mouse oocytes’ major organelles and cytoskeleton increased significantly than that of the younger mice, which may be the main reason for the low pregnancy rate in the elderly.

Key words: Oocytes, Mitochondria, Cortical granule, spindle, Microfilament, Chromosome, Laser scanning confocal microscopy, Mouse

LIU Mei-ju CHEN Xin-xia. Ultrastructural factors affecting the quality of oocytes in aged mice[J]. Acta Anatomica Sinica, 2017, 48(6): 740-745.

References

［1］Tan TY, Lau SK, Loh SF, et al.Female ageing and reproductive outcome in assisted reproduction cycles［J］. Singapore Med J, 2014, 55(6):305-309.

［2］Takahashi T, Igarashi H, Amita M, et al.Molecular mechanism of poor embryo development in postovulatory aged oocytes: mini review［J］. J Obstet Gynaecol Res, 2013, 39(10):1431-1439.

［3］Ge MX, Tao LL, Chen XP, et al. Clinical outcome of 409 cycles in vitro fertilization in elderly women［J］. Guangdong Medical Journal,2014,35(12):1907-1910. (in Chinese)

葛明晓，陶莉莉，陈小平，等.高龄妇女409个周期体外受精的临床结局［J］.广东医学, 2014, 35(12):1907-1910.

［4］Zhu ZY, Chen DY, Li JS, et al.Rotation of meiotic spindle is controlled by microfilaments in mouse oocytes［J］. Biol Reprod, 2003, 68(3):943-946.

［5］Shomper M, Lappa C, FitzHarris G.Kinetochore microtubule establishment is defective in oocytes from aged mice［J］. Cell Cycle, 2014, 13(7):1171-1179.

［6］Yang H, Liu F. The relationship between age, mitochondria and female reproduction［J］. Medical Journal of the Chinese People's Armed Police Forces,2008,19(3):272-274. (in Chinese)

杨华, 刘锋.年龄、线粒体与女性生殖能力的关系［J］.武警医学, 2008, 19(3):272-274.

［7] Mahrous E, Yang Q, Clarke HJ. Regulation of mitochondrial DNA accumulation during oocyte growth and meiotic maturation in the mouse［J］.Reproduction, 2012, 144(2):177-185.［8］Cotterill M, Harris SE, Collado Fernandez E, et al. The activity and copy number of mitochondrial DNA in ovine oocytes throughout oogenesis in vivo and during oocyte maturation in vitro［J］. Mol Hum Reprod, 2013, 19(7):444-450.

［9］Santos TA, El Shourbagy S, St John JC. Mitochondrial content reflects oocyte variability and fertilization outcome［J］. Fertil Steril, 2006, 85(3):584-591.

［10］May-Panloup P, Boucret L, Chao de la Barca JM, et al.Ovarian ageing: the role of mitochondria in oocytes and follicles［J］. Hum Reprod Update, 2016, 22(6):725-743.

［11］Iwata H, Goto H, Tanaka H, et al. Effect of maternal age on mitochondrial DNA copy number,ATP content and IVF outcome of bovine oocytes［J］. Reprod Fertil Dev, 2011, 23(3):424-432.

［12］Zhao H, Li Y, Gao X, et al. Timing of meiotic progression and morphological changes in human oocytes［J］. Acta Anatomica Sinica,2006,37(4):479-483. (in Chinese)

赵涵, 李媛, 高选, 等.人类卵母细胞减数分裂进程及形态学研究［J］. 解剖学报，2006，37（4）：479-483.

［13］Carneiro GF, Liu IK, Hyde D, et al. Quantification and distribution of equine oocyte cortical granules during meiotic maturation and after activation［J］. Mol Reprod Dev, 2002, 63(4):451-458.

［14］Katie Howe, Greg FitzHarris. Recent insights into spindle function in mammalian oocytes and early embryos［J］. Biol Reprod, 2013, 89(3):71-79.

［15］Nagaoka SI, Hassold TJ, Hunt PA.Human aneuploidy: mechanisms and new insights into an age-old problem［J］. Nat Rev Genet, 2012, 13(7):493-504.

［16］Battaglia DE, Goodwin P, Klein NA, et al. Influence of maternal age on meiotic spindle assembly in oocytes from naturally cycling women［J］. Hum Reprod, 1996, 11(10):2217-2222.

［17］Pellestor F, Andréo B, Arnal F, et al. Maternal aging and chromosomal abnormalities: new data drawn from in vitro unfertilized human oocytes［J］. Hum Genet, 2003, 112(2):195-203.

［18] Ma R, Zhang Y, Zhang L, et al.Sirt1 protects pig oocyte against in vitro aging［J］. Anim Sci J, 2015, 86(9):826-832.

［19］Jin YX, Cui XS, Yu XF, et al.Alterations of spindle and microfilament assembly in aged cat oocytes［J］. Reprod Domest Anim, 2010, 45(5):865-871.

[1]	HONG Xiao-min LI San-qiang CUI Qin-yi ZHENG Run-yue YANG Meng-li LUO Ren-li LI Qian-hui. Nuclear factor-κB signaling pathway and gender differences in alcoholic liver fibrosis [J]. Acta Anatomica Sinica, 2024, 55(1): 55-61.
[2]	YUAN Lei YANG Zhi-wei YANG Hui LIU Zheng-hai HE Jie WAN Wei. Panax notoginseng saponin relieving the inflammatory pain caused by complete Freund’s adjuvant by inhibiting the activation of astrocytes in mice [J]. Acta Anatomica Sinica, 2024, 55(1): 25-31.
[3]	MA Ting-ting CHEN Jian-hong LIU Ai-cui LI Hai-ning. Role of inhibiting lncRNA TUG1 to down-regulate nucleotide binding oligomerization domain like receptor protein 1 inflammasome in delaying the progression of Alzheimer’s disease [J]. Acta Anatomica Sinica, 2024, 55(1): 32-42.
[4]	ZOU Cheng-gong FENG Hao CHEN Bing TANG Hui SHAO Chuan SUN Mou YANG Rong HE Jia-quan . Research on the dynamic changes of neurological dysfunction and cognitive function impairment in traumatic brain injury [J]. Acta Anatomica Sinica, 2024, 55(1): 43-48.
[5]	LÜ Hai-yan SHEN Xi-ya ZHAO Fu-sheng ZHU Mei . Effects of tricholoma matsutake polysaccharides on 1-methy-4-pehnyl-pyridine ion-induced PC12 cell damage [J]. Acta Anatomica Sinica, 2024, 55(1): 49-54.
[6]	ZHANG Qin YANG Jun-xia JIANG Qing-song. Effect of fosinopril on angiotensin converting enzyme 2 and vascular endothelial growth factor in diabetic retinopathy mice [J]. Acta Anatomica Sinica, 2023, 54(6): 628-634.
[7]	CHEN Zi-xuan SI Li-na SHU Wei-han ZHANG Xin WEI Chen-yang CHENG Lu-yang YANG Song-he QIAO Yue-bing. Mechanism of melatonin regulation of PI3K/Akt/mTOR signaling in hypothalamus delaying the initiation of puberty in female mice mice [J]. Acta Anatomica Sinica, 2023, 54(6): 644-651.
[8]	LIU Zhe-chuan LI Kun MA Shuai-nan MENG Jia-qi WANG Yan-qin. Effects of liraglutide on inflammation and mitochondrial fusion/division in Parkinson’s disease model of mice induced by paraquat [J]. Acta Anatomica Sinica, 2023, 54(6): 676-681.
[9]	LIU Ye CHU Ya-nan XU Ce-lu HE Jia-cheng SU Bing-yin TAI Hao-ran. Roscovitine rescuing neuronal loss and neuroinflammation in brain regions associated with Parkinson’s disease mice [J]. Acta Anatomica Sinica, 2023, 54(6): 635-643.
[10]	WEI Chun-chun WANG Ping LIN Fang-xing MA Xian-hua ZHANG Wei-ping XIE Zhi-fang. An improved fixation method for preparing mouse brown adipose tissue for transmission electron microscopy [J]. Acta Anatomica Sinica, 2023, 54(6): 738-743.
[11]	YAN Kun WU Xiao-lin LIU Ying-juan GE Ke-li1 REN Lei-ming LI Hong-yun . Effect of cerebrotein hydrolysate-Ⅰon intestinal microflora regulation of mice with Parkinson’s disease [J]. Acta Anatomica Sinica, 2023, 54(5): 497-504.
[12]	Ling GU ZHANG Ping SONG KeXin Na Li Hong-yu Chen Zhang Jie CONG Jing ZHAI XiaoYue GU Ling ZHANG Ping SONG Ke-xin LI Na CHEN Hong-yu ZHANG Jie CONG -Jing ZHAI Xiao-yue . Morphometry of the ureteric bud branching in the developing mouse kidney [J]. Acta Anatomica Sinica, 2023, 54(5): 593-598.
[13]	HE Ke-ke LI Yuan-di WEN Ting-hao ZHU Jie GAO Jie HU Rong HAN Feng SU Min . Erythrocyte membrane-associated protein inhibiting Th17 cell differentiation via IL-6/STAT3/ROR-γt signaling pathway in experimental autoimmune encephalomyelitis mice encephalomyelitis mice [J]. Acta Anatomica Sinica, 2023, 54(5): 538-545.
[14]	FAN Wen-juan CHEN Xu-dong CHEN Yong-fang YANG Xu-guang JIN Shao-ju ZHAO Zhi-jun DENG Jin-bo. Developmental comparison between cerebral organoids in vitro and body’s cortices in vivo [J]. Acta Anatomica Sinica, 2023, 54(4): 383-391.
[15]	ZHANG Yi-zhou LI Sha MI Shi-xiong ZUO Hong-chun ZHANG Qian-qian LI Han-lin LEI Zi-han ZHANG Dong-ze CUI Hui-xian. Effect of estradiol regulating sortilin-related receptor A expression on hippocampal spine density and synaptic protein expression via estrogen receptor of mouse [J]. Acta Anatomica Sinica, 2023, 54(3): 261-268.