Harmouch A, Guerrero J M, Pozo D, Rafii-el-Idrissi M, Calvo J R, Reiter R J, Osuna C
Department of Medical Biochemistry and Molecular Biology, University of Seville School of Medicine, Spain.
J Endocrinol. 1997 Nov;155(2):305-12. doi: 10.1677/joe.0.1550305.
We have studied why rat pineal N-acetyltransferase (NAT) activity is relatively insensitive to isoproterenol in young rats when compared with adult rats. We report that activation by isoproterenol of pineal cyclic AMP production and NAT activity is higher in adult than in 2-week-old rats. However, the effect of dibutyryl cyclic AMP, which enters the pinealocyte and duplicates the effect of cyclic AMP, on NAT activity was similar at both ages. Moreover, we found that both alpha- and beta-adrenergic receptors are highly specific at both ages, since the binding of the specific radioligands used to their receptors could be displaced only by their corresponding agonists and antagonists. However, we observed differences between pineals from young and adult rats when several families of the alpha subunit of G-proteins were studied in cell membranes. ADP-ribosylation and immunoblot studies have shown clear differences in both 42 and 45 kDa forms of the Gs alpha Both forms exhibit low values in pineals from 2-week-old animals when compared with 6-week-old. We also show that the later appearance of both Gs alpha forms is roughly similar to the potent activation of cyclic AMP production and NAT activity in adult rats when compared with young rats. In conclusion, the results presented suggest that the relative lack of sensitivity of rat pineal gland to beta-adrenergic receptor agonists early in the postnatal development may be explained by the low levels of membrane Gs alpha, rather than postreceptor-mediated mechanisms or changes in the characteristics of the beta-adrenergic receptors on the pinealocyte membrane.
我们研究了与成年大鼠相比,幼鼠松果体N - 乙酰转移酶(NAT)活性为何对异丙肾上腺素相对不敏感。我们报告,异丙肾上腺素对松果体环磷酸腺苷(cAMP)生成和NAT活性的激活作用在成年大鼠中比在2周龄大鼠中更高。然而,二丁酰环磷酸腺苷(进入松果体细胞并复制cAMP作用)对NAT活性的影响在两个年龄段相似。此外,我们发现α和β肾上腺素能受体在两个年龄段都具有高度特异性,因为用于其受体的特异性放射性配体与其受体的结合仅能被其相应的激动剂和拮抗剂取代。然而,当研究细胞膜中G蛋白α亚基的几个家族时,我们观察到幼鼠和成年大鼠松果体之间存在差异。ADP - 核糖基化和免疫印迹研究表明,Gsα的42 kDa和45 kDa形式均存在明显差异。与6周龄动物相比,2周龄动物松果体中这两种形式的值均较低。我们还表明,与幼鼠相比,这两种Gsα形式在成年大鼠中出现较晚,这与成年大鼠中cAMP生成和NAT活性的有效激活大致相似。总之,所呈现的结果表明,出生后早期大鼠松果体对β肾上腺素能受体激动剂相对不敏感,可能是由于膜Gsα水平较低,而不是受体后介导机制或松果体细胞膜上β肾上腺素能受体特性的变化。