Expression of the lipocalin-type prostaglandin D synthase gene in the reproductive tracts of Holstein bulls.

C M Rodríguez, J R Day, G J Killian
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引用次数: 11

Abstract

The aim of this study was to localize expression of the prostaglandin D synthase gene in the reproductive tracts of Holstein bulls using northern blotting and in situ hybridization. For northern blotting, a digoxigenin-labelled prostaglandin D synthase cDNA probe was used to probe blots containing RNA isolated from the testes, epididymides, vas deferens, ampullae, seminal vesicles, prostate and bulbourethral glands of bulls. The digoxigenin-labelled cDNA for the bovine homologue of prostaglandin D synthase hybridized to a single band (approximately 0.9 kb) to RNA samples from the caput, corpus and cauda epididymides, as well as RNA samples from the vas deferens and the ampulla. The probe also detected a single band in testis samples, although the transcript size was slightly larger (approximately 1.0 kb) than the transcript found in the other tissues. The highest expression of prostaglandin D synthase was observed in the testes and caput epididymides. Prostaglandin D synthase transcripts were not found in the seminal vesicles or the prostate or bulbourethral glands using northern blotting. For in situ hybridization, antisense and sense riboprobes were synthesized and used to hybridize to cryosections obtained from the reproductive tissues of bulls. In situ hybridization of bull testes showed that prostaglandin D synthase transcripts were present within the germ cells in the adluminal compartment of the seminiferous tubules containing round and elongated spermatids, indicating that expression varied with stage of development of the seminiferous tubules. Prostaglandin D synthase expression was observed in the epithelial cells of the epididymides with greatest expression occurring in the caput epididymidis. Some expression was also observed in the epithelial cells of the vas deferens and a few cells of some lobules in the prostate and bulbourethral glands. Expression of the prostaglandin D synthase gene was not detected in ampullae or seminal vesicles by in situ hybridization.

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脂钙素型前列腺素D合成酶基因在荷斯坦公牛生殖道中的表达。
本研究的目的是利用northern blotting和原位杂交技术定位前列腺素D合成酶基因在荷斯坦公牛生殖道中的表达。northern blotting采用地高辛标记的前列腺素D合成酶cDNA探针,对公牛睾丸、附睾、输精管、壶壶、精囊、前列腺和尿道球腺中分离的RNA进行印迹检测。用地高辛标记的牛前列腺素D合成酶同源物cDNA与来自附睾头、体和尾的RNA样本以及输精管和腹侧的RNA样本杂交成一条单带(约0.9 kb)。探针还在睾丸样本中检测到单个条带,尽管转录本的大小比在其他组织中发现的转录本稍大(约1.0 kb)。前列腺素D合酶在睾丸和附睾头的表达量最高。前列腺素D合成酶转录物未在精囊、前列腺和尿道球囊中发现。为了原位杂交,合成了反义和正核探针,并将其杂交到公牛生殖组织的冷冻切片上。原位杂交表明,含圆形和细长精子的精管腔室生殖细胞中存在前列腺素D合成酶转录本,表明其表达随精管发育阶段的不同而变化。前列腺素D合成酶在附睾上皮细胞中均有表达,其中以附睾头表达最多。在输精管上皮细胞和前列腺、尿道部分小叶细胞中也有表达。前列腺素D合成酶基因在壶腹和精囊中未见原位杂交表达。
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