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基于在体活体连续活检的马黄体中的时相基因表达。

Temporal gene expression in equine corpora lutea based on serial biopsies in vivo.

机构信息

Colorado State University, Animal Reproduction and Biotechnology Laboratory, Fort Collins 80523, USA.

出版信息

J Anim Sci. 2011 Feb;89(2):389-96. doi: 10.2527/jas.2010-3247. Epub 2010 Sep 17.

Abstract

A biopsy procedure was developed to enable repeated sampling of a single equine corpus luteum (CL) over the course of an estrous cycle. The tissue collected was utilized in characterizing mRNA abundance for genes involved in luteal formation, function, and regression in the cyclic mare. Serial biopsies of CL in cyclic mares (2.7 to 27.5 mg per biopsy) were collected using an ultrasound-guided transvaginal technique. Biopsies were collected from each mare on d 2 and 5 (d 0 = ovulation) of the estrous cycle, and every other day from d 12 through luteolysis. Samples were obtained from 4 mares with normal estrous cycles and 1 mare with a retained CL. The biopsy procedure did not adversely affect luteal size or function, as measured by luteal area and serum concentrations of progesterone. Real-time reverse-transcription PCR was used to quantify steady state mRNA concentrations in each tissue sample obtained. Mean abundance of steroidogenic acute regulatory protein (StAR) mRNA was not different (P = 0.102 to 0.964) on any of the sampling dates, but a trend for mRNA encoding StAR to decrease between d 12 and 14 (P = 0.10) was observed. Values for mRNA encoding StAR were positively correlated to serum concentrations of progesterone on d 5 (R = 0.95; P = 0.05) and 14 (R > 0.99; P < 0.01). Steady-state abundance of mRNA for 3β-hydroxysteroid dehydrogenase, Δ 5-Δ 4 isomerase (3β-HSD) declined between d 12 and 14 (P = 0.15). There were positive correlations between mRNA for 3β-HSD and concentrations of progesterone on d 5 (R = 0.94; P = 0.06) and 12 (R > 0.99; P = 0.05). No difference was detected in abundance of mRNA encoding cyclooxygenase-2 (cox-2; P = 0.340 to 0.840) or caspase-3 (P = 0.517 to 0.882) between any of the sampling dates. A successful luteal biopsy procedure was developed that did not negatively affect luteal function, and abundance of mRNA encoding StAR, 3β-HSD, cox-2, and caspase-3 was characterized in luteal biopsy tissue collected on d 2, 5, 12, and 14 of the estrous cycle in the mare.

摘要

建立了一种活检程序,以实现对发情周期中单匹马的黄体(CL)进行多次采样。所收集的组织用于表征与循环母马黄体形成、功能和退化相关的基因的 mRNA 丰度。使用超声引导的经阴道技术对发情周期中的马 CL 进行连续活检(每次活检 2.7 至 27.5 毫克)。在发情周期的第 2 天(d0=排卵)和第 5 天(d0=排卵)以及第 12 天至黄体溶解期间每隔一天从每匹马中采集活检。从 4 匹正常发情周期的母马和 1 匹保留 CL 的母马中获得样本。活检程序不会影响黄体的大小或功能,因为黄体面积和血清孕酮浓度没有变化。使用实时逆转录 PCR 定量每个组织样本中稳定状态的 mRNA 浓度。在任何采样日期,类固醇急性调节蛋白(StAR)mRNA 的丰度没有差异(P=0.102 至 0.964),但在第 12 天和第 14 天之间观察到编码 StAR 的 mRNA 减少的趋势(P=0.10)。第 5 天(R=0.95;P=0.05)和第 14 天(R>0.99;P<0.01)时,编码 StAR 的 mRNA 值与血清孕酮浓度呈正相关。第 12 天和第 14 天之间(P=0.15),3β-羟甾脱氢酶、Δ5-Δ4 异构酶(3β-HSD)的 mRNA 丰度呈正相关。第 5 天(R=0.94;P=0.06)和第 12 天(R>0.99;P=0.05)时,编码 3β-HSD 的 mRNA 与孕酮浓度呈正相关。在任何采样日期之间,环加氧酶-2(cox-2;P=0.340 至 0.840)或半胱氨酸蛋白酶-3(P=0.517 至 0.882)编码 mRNA 的丰度均无差异。建立了一种成功的黄体活检程序,该程序不会对黄体功能产生负面影响,并在发情周期的第 2、5、12 和 14 天对马黄体活检组织中编码 StAR、3β-HSD、cox-2 和半胱氨酸蛋白酶-3 的 mRNA 丰度进行了特征描述。

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