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发情周期、早期妊娠期间及性类固醇处理后猪子宫中雄激素受体的分布及含量变化

Distribution and changes in amounts of the androgen receptor in the pig uterus during the estrous cycle, early pregnancy and after treatment with sex steroids.

作者信息

Cárdenas H, Pope W F

机构信息

Department of Animal Sciences, The Ohio State University, 2027 Coffey Road, Columbus, Ohio 43210, USA.

出版信息

J Endocrinol. 2003 Jun;177(3):461-9. doi: 10.1677/joe.0.1770461.

Abstract

Two experiments were performed to examine the expression of the androgen receptor (AR) gene in the pig uterus. In experiment 1, immunohistochemistry (IHC) was used to determine the distribution of the AR in uterine tIssue of pigs when collected at the first day of estrus (day 0) and the mid-luteal phase (day 12) of the estrous cycle, or early pregnancy (day 12, n=4 gilts per group). In experiment 2, AR immunostaining and AR mRNA in uterine tIssue were compared among ovariectomized gilts (n=4 per group) following treatment for 4 days with daily injections of: (1) progesterone (2 mg/kg bodyweight (BW)), (2) estradiol-17beta (E(2,) 2 micro g/kg BW), (3) E(2) plus progesterone (same dosages as 1 and 2 combined), (4) 5alpha-dihydrotestosterone (DHT, 7 micro g/kg BW), or (5) vehicle (corn oil). Data were analyzed using ANOVA. In experiment 1, nuclear staining for AR in luminal and glandular epithelia was strong and did not differ in intensity between the two locations. Immunostaining of AR in the myometrium was less (P<0.001) intense than in the luminal and glandular epithelia. Nuclei of stromal cells contained AR immunostaining that varied in intensity from strong (mainly in subepithelial stroma) to weak or no staining. Stages of the estrous cycle or early pregnancy did not influence AR immunostaining in the endometrial epithelia and myometrium. In experiment 2, immunostaining of AR in glandular and luminal epithelia and myometrium of ovariectomized gilts treated with vehicle or DHT was less (P<0.05) than in gilts treated with E(2), progesterone, or E(2) plus progesterone. Immunostaining of AR did not differ between ovariectomized gilts treated with vehicle or DHT, or between gilts treated with E(2), progesterone, or E(2) plus progesterone. In both experiments, intensity of AR immunostaining was greater in glandular epithelium located at the adluminal region compared with glandular epithelium located at the basal region of the endometrium. Competitive reverse-transcription PCR (RT-PCR) indicated a stimulatory effect (P<0.01) of E(2) on amounts of AR mRNA in whole endometrium. This increase in AR mRNA after E(2) treatment was not detected when E(2) was combined with progesterone. Endometrial AR mRNA was not influenced by DHT or progesterone relative to vehicle-treated gilts. In conclusion, immunoreactive AR is mainly present in luminal and glandular epithelia of the pig uterus and to a lesser extent in the myometrium, and does not change significantly during the estrous cycle or early pregnancy. Expression of the AR gene in the pig endometrium and myometrium appears to be regulated by E(2) and progesterone.

摘要

进行了两项实验以检测雄激素受体(AR)基因在猪子宫中的表达。在实验1中,采用免疫组织化学(IHC)方法确定发情周期第1天(第0天)、发情周期的黄体中期(第12天)或早期妊娠(第12天,每组4头后备母猪)时猪子宫组织中AR的分布。在实验2中,比较了卵巢切除的后备母猪(每组4头)在每日注射以下物质处理4天后子宫组织中的AR免疫染色和AR mRNA:(1)孕酮(2 mg/kg体重(BW)),(2)雌二醇-17β(E₂,2 μg/kg BW),(3)E₂加孕酮(与1和2联合使用的相同剂量),(4)5α-双氢睾酮(DHT,7 μg/kg BW),或(5)赋形剂(玉米油)。数据采用方差分析进行分析。在实验1中,管腔和腺上皮中AR的核染色较强,且两个部位的染色强度无差异。子宫肌层中AR的免疫染色强度低于管腔和腺上皮(P<0.001)。基质细胞的细胞核含有AR免疫染色,强度从强(主要在上皮下基质)到弱或无染色不等。发情周期阶段或早期妊娠不影响子宫内膜上皮和子宫肌层中的AR免疫染色。在实验2中,用赋形剂或DHT处理的卵巢切除后备母猪的腺上皮、管腔上皮和子宫肌层中的AR免疫染色低于用E₂、孕酮或E₂加孕酮处理的后备母猪(P<0.05)。用赋形剂或DHT处理的卵巢切除后备母猪之间,或用E₂、孕酮或E₂加孕酮处理的后备母猪之间,AR免疫染色无差异。在两项实验中,与位于子宫内膜基部区域的腺上皮相比,位于近腔区域的腺上皮中AR免疫染色强度更大。竞争性逆转录PCR(RT-PCR)表明E₂对整个子宫内膜中AR mRNA的量有刺激作用(P<0.01)。当E₂与孕酮联合使用时,未检测到E₂处理后AR mRNA的这种增加。相对于用赋形剂处理的后备母猪,子宫内膜AR mRNA不受DHT或孕酮的影响。总之,免疫反应性AR主要存在于猪子宫的管腔和腺上皮中,在子宫肌层中含量较少,在发情周期或早期妊娠期间无显著变化。猪子宫内膜和子宫肌层中AR基因的表达似乎受E₂和孕酮的调节。

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