Tomás-Zapico Cristina, Antonio Boga José, Caballero Beatriz, Vega-Naredo Ignacio, Sierra Verónica, Alvarez-García Oscar, Tolivia Delio, Josefa Rodríguez-Colunga María, Coto-Montes Ana
Departamento de Morfología y Biología Celular, Universidad de Oviedo, Oviedo, Spain.
J Pineal Res. 2005 Aug;39(1):21-6. doi: 10.1111/j.1600-079X.2005.00210.x.
Melatonin acts through several specific receptors, including membrane receptors (MT(1) and MT(2)) and members of the RZR/ROR nuclear receptors family, which have been identified in a large variety of mammalian and nonmammalian cells types. Both membrane and nuclear melatonin receptors have been partially characterized in Harderian gland of the Syrian hamster. Nevertheless, the identities of these receptors were unknown until this study, where the coexistence of MT(1) and RORalpha(1) in this gland was determined by nested RT-PCR followed by amplicon sequencing and Western-blot. Furthermore, the cellular localization of both receptors was determined by immunohistochemistry. Thus, MT(1) receptor was localized exclusively at the basal side of the cell acini, supporting the hypothesis that this receptor is activated by the pineal-synthesized melatonin. On the contrary, although a RORalpha(1)-immunoreactivity was observed in nuclei of epithelial cells of both sexes, an extranuclear specific staining, which was more frequently among those cells of males, was also seen. The implication of this possible nuclear exclusion of RORalpha(1) on the role of this indoleamine against oxidative stress is discussed.
褪黑素通过多种特定受体发挥作用,包括膜受体(MT(1)和MT(2))以及RZR/ROR核受体家族成员,这些受体已在多种哺乳动物和非哺乳动物细胞类型中被鉴定出来。膜性和核性褪黑素受体在叙利亚仓鼠的哈德氏腺中已得到部分表征。然而,在本研究之前这些受体的身份尚不清楚,在本研究中,通过巢式逆转录聚合酶链反应(RT-PCR),随后进行扩增子测序和蛋白质免疫印迹法,确定了MT(1)和RORα(1)在该腺体中的共存情况。此外,通过免疫组织化学确定了这两种受体的细胞定位。因此,MT(1)受体仅定位于腺泡细胞的基底侧,这支持了该受体由松果体合成的褪黑素激活的假说。相反,尽管在两性上皮细胞的细胞核中均观察到RORα(1)免疫反应性,但在细胞核外也可见特异性染色,且在雄性细胞中更为常见。本文讨论了RORα(1)这种可能的核外排对这种吲哚胺抗氧化应激作用的影响。