母体年龄相关的卵母细胞老化对生育力的影响机制:斑马鱼模型的转录组学测序分析  

Mechanisms of the Effect of Maternal Age-Related Oocyte Aging on Fertility:Transcriptomic Sequencing Analysis of a Zebrafish Model

作　　者：竹琳 林子媛 刘燕燕[3] 孙华钦 孙春堂 陈凤 ZHU Lin;LIN Ziyuan;LIU Yanyan;SUN Huaqin;SUN Chuntang;CHEN Feng(Hemodialysis Room,Department of Nephrology,West China Hospital,Sichuan University/West China School of Nursing,Sichuan university,Chengdu 610041,China;West China Institutes for Women and Children’s Health,West China Second University Hospital,SCU-CUHK Joint Laboratory for Reproductive Medicine,and Key Laboratory of Birth Defects and Related Diseases of Women of the Ministry of Education,Sichuan University,Chengdu 610041,China;Medical Genetics Department,West China Second University Hospital,Sichuan University,Chengdu 610041,China;Department of Tumor Radiotherapy and Chemotherapy,West China Second University Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院肾脏内科血液透析室/四川大学华西护理学院,成都610041 [2]四川大学华西第二医院西部妇幼医学研究院,四川大学-香港中文大学生殖医学联合实验室,出生缺陷与相关妇儿疾病教育部重点实验室,成都610041 [3]四川大学华西第二医院医学遗传科,成都610041 [4]四川大学华西第二医院肿瘤放化疗科,成都610041

出　　处：《四川大学学报（医学版）》2024年第3期588-595,共8页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.82271692);四川省自然科学基金(No.2022NSFSC0782、No.2022NSFSC0797)资助。

摘　　要：目的 以斑马鱼为模型,研究母体年龄相关的卵母细胞老化对生育力的影响,寻找随年龄增长的生育力下降的潜在分子机制。方法 随机选取6月龄、12月龄、18月龄雌性斑马鱼各8条,均与6月龄雄性斑马鱼交配产卵。对各月龄组的结局指标进行比较。主要结局指标为胚胎受精率,次要结局指标为雌性斑马鱼单次产卵数量、胚胎死亡率和胚胎畸形率。收集不同月龄雌性斑马鱼的卵母细胞以及后代囊胚期胚胎进行转录组学测序分析。结果 18月龄组受精率(86.8±5.5)%低于6月龄组(94.9±3.6)%及12月龄组(92.3±4.2)%,差异有统计学意义(P均<0.05)。与6月龄组相比,12月龄与18月龄组雌性斑马鱼胚胎死亡率均升高(P均<0.05)。各组单次产卵数量和胚胎畸形率之间的差异无统计学意义。母体年龄相关的囊胚期胚胎差异性表达基因主要富集于MAPK信号通路以及脂肪酸降解等通路途径。母体年龄相关的卵母细胞差异性表达基因主要富集于细胞黏附分子,蛋白质消化吸收等通路途径。结论 母体年龄可能是卵母细胞受精能力降低及早期胚胎死亡率升高的影响因素,母体年龄相关的卵母细胞老化影响生育力及后代胚胎的发育。Objective Female fertility gradually decreases with the increase in women’s age.The underlying reasons include the decline in the quantity and quality of oocytes.Oocyte aging is an important manifestation of the decline in oocyte quality,including in vivo oocyte aging before ovulation and in vitro oocyte aging after ovulation.Currently,few studies have been done to examine oocyte aging,and the relevant molecular mechanisms are not fully understood.Therefore,we used zebrafish as a model to investigate oocyte aging.Three different age ranges of female zebrafish were selected to mate with male zebrafish of the best breeding age.In this way,we studied the effects of maternal age-related oocyte aging on fertility and investigated the potential molecular mechanisms behind maternal age-related fertility decline.Methods Eight female zebrafish aged between 158 and 195 d were randomly selected for the 6-month age group(180±12)d,8 female zebrafish aged between 330 and 395 d were randomly selected for the 12-month age group(360±22)d,and 8 female zebrafish aged between 502 and 583 d were randomly selected for the 18-month age group(540±26)d.Male zebrafish of(180±29)d were randomly selected from zebrafish aged between 158 and 195 d and mated with female zebrafish in each group.Each mating experiment included 1 female zebrafish and 1 male zebrafish.Zebrafish embryos produced by the mating experiments were collected and counted.The embryos at 4 hours post-fertilization were observed under the microscope,the total number of embryos and the number of unfertilized embryos were counted,and the fertilization rate was calculated accordingly.The numbers of malformed embryos and dead embryos were counted 24 hours after fertilization,and the rates of embryo malformation and mortality were calculated accordingly.The primary outcome measure was the embryo fertilization rate,and the secondary outcome measures were the number of embryos per spawn(the total number of embryos laid within 1.5 hours after the beginning of mating and reprod

关 键 词：斑马鱼 母亲年龄 受精 生育力 

分 类 号：R714[医药卫生—妇产科学] R-332[医药卫生—临床医学]