Division of Reproductive Endocrinology and Infertility, University Hospitals Cleveland Medical Center, Cleveland, Ohio.
Department of Reproductive Biology, Case Western Reserve University School of Medicine, Cleveland, Ohio.
F S Sci. 2021 May;2(2):198-206. doi: 10.1016/j.xfss.2021.03.003. Epub 2021 Mar 20.
To investigate the effect of superovulation with human chorionic gonadotropin (hCG) or gonadotropin-releasing hormone agonist (GnRHa) trigger on leukocyte density and expression of leukocyte-specific genes in the peri-implantation period in the mouse uterus.
Laboratory research.
University laboratory facility.
Female mice were mated to fertile male mice in one of three protocols: (1) natural mating or mating following injection with pregnant mare serum gonadotropin followed by trigger with (2) GnRHa or (3) hCG. Female mice were killed prior to implantation, 3 days after ovulation (E3.5), and the ovaries and uterine tissue were collected. Total RNA was isolated and assayed using quantitative reverse transcription polymerase chain reaction, and the uterine tissue was stained for histologic analysis of immune cell markers.
Endometrial leukocyte (CD45) and vessel density (CD31) by immunohistochemical staining; expression of leukocyte markers CD11b, CD335, and CD22, by quantitative reverse transcription polymerase chain reaction in the whole uterine tissue.
Superovulation decreased (compared with controls) the endometrial leukocyte density, based on the number of cells staining for CD45, and endometrial vessel density, based on the number of cells staining for CD31. Leukocyte density was additionally decreased in the GnRHa trigger group compared with that in the hCG trigger group. Superovulation with hCG and GnRHa triggers decreased the uterine expression of the B-cell marker CD22 compared with controls. The expression of the natural killer cell marker CD11b was decreased by the hCG trigger but not by the GnRHa. Abundance of mRNA encoding the CD335 natural killer cell marker was not affected by superovulation or trigger agent.
In mice, superovulation with the GnRHa trigger compared with that with the hCG trigger differentially alters key immunologic factors in the uterine peri-implantation. These altered immunologic factors have roles in angiogenesis that may assist in elucidating the effects of assisted reproductive technologies on implantation efficiency and fetal growth and development.
研究人绒毛膜促性腺激素（hCG）或促性腺激素释放激素激动剂（GnRHa）扳机对小鼠植入期子宫内膜白细胞密度和白细胞特异性基因表达的影响。
实验室研究。
大学实验室设施。
雌性小鼠与生育能力强的雄性小鼠交配，分为以下三种方案之一：（1）自然交配或注射孕马血清促性腺激素后交配，然后用（2）GnRHa 或（3）hCG 触发。在植入前、排卵后 3 天（E3.5）处死雌性小鼠，并收集卵巢和子宫组织。分离总 RNA，采用定量逆转录聚合酶链反应进行检测，并用免疫组织化学染色法检测子宫组织中免疫细胞标志物。
子宫内膜白细胞（CD45）和血管密度（CD31）的免疫组织化学染色；整个子宫组织中白细胞标志物 CD11b、CD335 和 CD22 的表达，采用定量逆转录聚合酶链反应进行检测。
与对照组相比，超排卵减少了（CD45 染色的）子宫内膜白细胞密度和（CD31 染色的）子宫内膜血管密度。GnRHa 触发组的白细胞密度也比 hCG 触发组低。hCG 和 GnRHa 触发的超排卵导致与对照组相比，子宫中 B 细胞标志物 CD22 的表达减少。hCG 触发降低了自然杀伤细胞标志物 CD11b 的表达，但 GnRHa 触发没有。超排卵或触发剂对编码 CD335 天然杀伤细胞标志物的 mRNA 丰度没有影响。
在小鼠中，与 hCG 触发相比，GnRHa 触发的超排卵会对子宫植入期的关键免疫因素产生不同的影响。这些改变的免疫因素在血管生成中起作用，这可能有助于阐明辅助生殖技术对植入效率和胎儿生长发育的影响。
