Kodithuwakku Suranga P, Miyamoto Akio, Wijayagunawardane Missaka P B
Department of Animal Science, University of Peradeniya, Peradeniya 20400, Sri Lanka.
Reproduction. 2007 Jun;133(6):1087-94. doi: 10.1530/REP-06-0201.
The dynamic action of oviductal secretory compounds on spermatozoa function is well documented. In contrast, the effect of spermatozoa on oviductal function remains poorly characterized. Previously, our lab and others have shown that prostaglandin (PG), together with other vasoactive peptides, plays major roles in oviductal contractions and sperm transport. We therefore examined the effect of spermatozoa on the production of PG by cow oviductal epithelial cells (OEC). A bovine spermatozoa-OEC co-culture system was utilized for this purpose. OECs in the second passage were co-cultured for 0, 1, 3, 6, 12, and 24 h with six doses of motile, killed, or truncated spermatozoa heads (control; without spermatozoa, 10(2)-10(6) spermatozoa/ml medium). The levels of PGE(2) and PGF(2alpha) in the medium were measured using enzyme immunoassays. Messenger RNA expression of cyclooxygenase-2, PGF synthase (PGFS), and PGE synthase (PGES) was investigated using real-time RT-PCR. The results indicated that motile spermatozoa increased the secretion of PG by OEC as well as cellular expression of mRNA for cyclooxygenase, PGES, and PGFS in a dose- and time-dependent manner. A maximum three- to fivefold increased secretion of PG was observed with a dose of 10(5) spermatozoa/ml after a 12-h co-incubation. Neither killed spermatozoa nor truncated spermatozoa heads stimulated oviductal biosynthesis and secretion of PG at any dose or time point observed. The results provide the first evidence that live spermatozoa in the oviduct up-regulate the local PG system, and thereby, enhance oviductal contractions. Thus, spermatozoa may bear a role in accelerating their own transport into the fertilization site.
输卵管分泌化合物对精子功能的动态作用已有充分记录。相比之下,精子对输卵管功能的影响仍鲜为人知。此前,我们实验室和其他研究表明,前列腺素(PG)与其他血管活性肽一起,在输卵管收缩和精子运输中起主要作用。因此,我们研究了精子对牛输卵管上皮细胞(OEC)产生PG的影响。为此采用了牛精子-OEC共培养系统。将第二代OEC与六剂活动、灭活或截断的精子头部(对照;无精子,10² - 10⁶个精子/毫升培养基)共培养0、1、3、6、12和24小时。使用酶免疫测定法测量培养基中PGE₂和PGF₂α的水平。使用实时RT-PCR研究环氧合酶-2、PGF合酶(PGFS)和PGE合酶(PGES)的信使RNA表达。结果表明,活动精子以剂量和时间依赖性方式增加了OEC分泌的PG以及环氧合酶、PGES和PGFS的mRNA细胞表达。共孵育12小时后,在10⁵个精子/毫升的剂量下观察到PG分泌最多增加三到五倍。在所观察的任何剂量或时间点,灭活精子和截断的精子头部均未刺激输卵管PG的生物合成和分泌。这些结果首次证明输卵管中的活精子上调局部PG系统,从而增强输卵管收缩。因此,精子可能在加速自身向受精部位的运输中发挥作用。