Login G R, Yang J, Bryan K P, Digenis E C, McBride J, Elovic A, Quissell D O, Dvorak A M, Wong D T
Department of Oral Medicine and Diagnostic Sciences, Harvard School of Dental Medicine, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215, USA.
Am J Physiol. 1997 Mar;272(3 Pt 1):G553-62. doi: 10.1152/ajpgi.1997.272.3.G553.
Although the expression and biological role of transforming growth factor-alpha (TGF-alpha) have been explored in a variety of normal cells in mammalian species, little is known about the storage of TGF-alpha in secretory cells of exocrine organs. Parotid glands from four rats were homogenized for RNA isolation followed by reverse transcription-polymerase chain reaction to determine the presence of TGF-alpha message. In situ hybridization using a hamster-specific TGF-alpha riboprobe was done on paraffin sections. Parotid gland and isolated acinar cells were processed for transmission electron microscopy (TEM) and postembedding immunogold labeled for TGF-alpha. Gold particles were counted on approximately 200 granules in 10 acinar cells and in 10 intercalated duct cells. Labeling density was calculated as the number of gold particles per square micrometer +/- SD. Statistical significance was calculated using one-way analysis of variance. Using multiple technologies, we have established that rat parotid acinar and intercalated duct cells synthesize TGF-alpha and store the precursor form of this cytokine in their secretory granules.
尽管在哺乳动物的多种正常细胞中已经对转化生长因子α(TGF-α)的表达及生物学作用进行了探索,但对于TGF-α在外分泌器官分泌细胞中的储存情况却知之甚少。取4只大鼠的腮腺进行匀浆以分离RNA,随后进行逆转录-聚合酶链反应,以确定TGF-α信息的存在。使用仓鼠特异性TGF-α核糖探针在石蜡切片上进行原位杂交。对腮腺和分离出的腺泡细胞进行透射电子显微镜(TEM)处理,并在包埋后对TGF-α进行免疫金标记。在10个腺泡细胞和10个闰管细胞中,对大约200个颗粒上的金颗粒进行计数。标记密度计算为每平方微米金颗粒的数量±标准差。使用单因素方差分析计算统计学意义。通过多种技术,我们已证实大鼠腮腺腺泡细胞和闰管细胞可合成TGF-α,并将这种细胞因子的前体形式储存在其分泌颗粒中。