Das Shubash Chandra, Isobe Naoki, Yoshimura Yukinori
Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan.
Reproduction. 2009 Mar;137(3):527-36. doi: 10.1530/REP-08-0175. Epub 2008 Jun 6.
The aim of this study was to determine the physiological significance of interleukin-1beta (IL1B) and lipopolysaccharide-induced TNF factor (LITAF) in the fate of sperm in the oviduct of laying hens after artificial insemination (AI). Laying hens were inseminated with fresh semen, PBS or seminal plasma and tissues from different oviductal segments were collected to observe the general histology, changes in the mRNA expression of IL1B and LITAF and the localization of positive cells expressing immunoreactive IL1B (irIL1B). Semi-quantitative RT-PCR was used to observe the changes in mRNA expression of these molecules in the infundibulum, uterus, utero-vaginal junction (UVJ), and vagina after insemination. Intact sperm in the lumen and between the primary or secondary folds of the vagina were found until 6 h after insemination but were degraded at 12 h. The mRNA expression of IL1B and LITAF was significantly increased in the vagina until 6 h after AI but remained unchanged in the other oviductal segments. In the tissue of the vagina and UVJ, irIL1B was localized in the mucosal stroma. The number of irIL1B-positive cells was increased in the vagina but almost unchanged in UVJ after insemination with semen. Significant changes were not observed in the mRNA expression and irIL1B-positive cells in the vagina after PBS or seminal plasma insemination. The increase of IL1B and LITAF in the vagina may lead to sperm degradation and elimination by cilia of surface epithelium, whereas their lower levels in UVJ may permit sperm to survive in sperm storage tubules.
本研究的目的是确定白细胞介素 - 1β(IL1B)和脂多糖诱导的肿瘤坏死因子(LITAF)在人工授精(AI)后蛋鸡输卵管中精子命运方面的生理意义。给蛋鸡分别输入新鲜精液、磷酸盐缓冲液(PBS)或精浆,收集不同输卵管节段的组织,观察其一般组织学、IL1B和LITAF的mRNA表达变化以及表达免疫反应性IL1B(irIL1B)的阳性细胞定位。采用半定量逆转录 - 聚合酶链反应(RT - PCR)观察授精后漏斗部、子宫、子宫 - 阴道连接处(UVJ)和阴道中这些分子的mRNA表达变化。授精后6小时内,在阴道腔以及阴道初级或次级皱襞之间可发现完整精子,但在12小时时精子发生降解。AI后6小时内,阴道中IL1B和LITAF的mRNA表达显著增加,但在其他输卵管节段保持不变。在阴道和UVJ组织中,irIL1B定位于黏膜基质。精液授精后,阴道中irIL1B阳性细胞数量增加,但UVJ中几乎不变。PBS或精浆授精后,阴道中mRNA表达和irIL1B阳性细胞未观察到显著变化。阴道中IL1B和LITAF的增加可能导致精子被表面上皮纤毛降解和清除,而它们在UVJ中较低的水平可能使精子在精子储存小管中存活。