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Gene expression and the physiological role of transforming growth factor-alpha in the mouse pituitary

机译：小鼠垂体中的基因表达和转化生长因子-α的生理作用

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Transforming growth factor-alpha (TGF-alpha), a member of the epidermal growth factor (EGF) family, is produced within the mouse anterior pituitaries. However, the cell types of TGF-alpha-expressing cells and the physiological roles of TGF-a within mouse pituitary glands remain unclear. The aim of the present study was to localize TGF-alpha mRNA-expressing cells, and to clarify the involvement of TGF-alpha in estrogen-induced DNA replication in mouse anterior pituitary cells. Northern blot analysis demonstrated TGF-alpha mRNA expression in adult male and female mouse anterior pituitaries. In situ hybridization analysis of the pituitaries in these mice showed that TGF-alpha mRNA-expressing cells in the anterior pituitary are round, oval, and medium-sized. TGF-alpha mRNA was colocalized in most of the growth hormone (GH) mRNA-expressing cells, while only some of the prolactin (PRL) mRNA-expressing cells. DNA replication in the anterior pituitary cells was detected by monitoring the cellular uptake of a thymidine analogue, bromodeoxyuridine (BrdU) in a primary serum-free culture system. Estradiol-17beta (E2) and TGF-alpha treatment increased the number of BrdU-labelled mammotrophs, indicating that E2 and TGF-alpha treatment stimulates the DNA replication in mammotrophs. Immunoneutralization of TGF-alpha with anti-TGF-alpha-antibodies nullified the E2-induced increase in DNA replication. RT-PCR analysis of TGF-alpha mRNA expression in ovariectomized female mice revealed that E2 increases TGF-alpha mRNA levels. These results indicate that the TGF-alpha produced primarily in the somatotrophs mediates the stimulatory effects of estrogen on the DNA replication of pituitary cells in a paracrine or autocrine manner.
机译：转化生长因子-α（TGF-alpha）是表皮生长因子（EGF）家族的成员，在小鼠前垂体内产生。但是，尚不清楚小鼠垂体中表达TGF-α的细胞的细胞类型和TGF-α的生理作用。本研究的目的是定位表达TGF-αmRNA的细胞，并阐明TGF-α与雌激素诱导的小鼠垂体前叶DNA复制有关。 Northern印迹分析表明成年雄性和雌性小鼠前垂体中TGF-αmRNA的表达。对这些小鼠的垂体进行原位杂交分析表明，垂体前叶中表达TGF-αmRNA的细胞为圆形，椭圆形和中等大小。 TGF-αmRNA在大多数表达生长激素（GH）的细胞中共定位，而只有一些催乳激素（PRL）表达的细胞。通过监测初级无血清培养系统中胸苷类似物溴脱氧尿苷（BrdU）的细胞摄取来检测垂体前叶细胞中的DNA复制。雌二醇17beta（E2）和TGF-α处理可增加BrdU标记的乳腺营养菌的数量，表明E2和TGF-α处理可刺激乳腺营养菌中的DNA复制。用抗TGF-α抗体对TGF-α进行免疫原化处理可消除E2诱导的DNA复制增加。切除卵巢的雌性小鼠中TGF-αmRNA表达的RT-PCR分析显示，E2可提高TGF-αmRNA水平。这些结果表明主要在体养动物中产生的TGF-α以旁分泌或自分泌方式介导雌激素对垂体细胞DNA复制的刺激作用。

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