Lam S K, Harvey S, Hall T R
Gen Comp Endocrinol. 1986 Aug;63(2):178-85. doi: 10.1016/0016-6480(86)90154-1.
Basal and thyrotrophin (TSH)-stimulated release of iodothyronines (triiodothyronine, T3, and thyroxine, T4) from intact chicken thyroid glands was determined in vitro. In the absence of TSH, T3 and T4 were released in measurable amounts in the incubation media. The release of both iodothyronines was directly related to the media TSH concentrations and incubation period. Lineweaver-Burke analysis revealed that the Vmax for T3 was 99.4 pg/gland, with an apparent Km of 17.8 mU TSH, and that the Vmax for T4 was 323.35 ng/gland, with an apparent Km of 51.5 mU TSH, demonstrating that T4 is the major iodothyronine released by avian thyroid glands. The basal release of T4 was suppressed by the addition of a calcium chelator (ethyleneglycol-bis-(beta-aminoethylether)-N,N,N', N'-tetraacetic acid; EGTA), a calcium antagonist (cobalt chloride, CoCl2), or prostaglandin E1 (PGE1) to the incubation media. Basal T4 released was increased in the presence of a calcium agonist (lanthanum chloride, LaCl3), a calcium ionophore (A23187), dibutyryl cyclic adenosine 3'3'-monophosphate (dbcAMP), isobutylmethylxanthine (IBMX), indomethacin, magnesium chloride (MgCl2), and potassium iodide (KI). Thyrotrophin-stimulated T4 release was reduced by CoCl2, PGE1, and indomethacin but enhanced by LaCl3, MgCl2, and KI. These results demonstrate that it is possible to measure the release of thyroid hormones in an in vitro system in the chicken. Basal and stimulated iodothyronine release from the chicken thyroid gland appears to be mediated by calcium- and cAMP-dependent mechanisms.
在体外测定了完整鸡甲状腺中碘甲状腺原氨酸(三碘甲状腺原氨酸,T3,和甲状腺素,T4)的基础释放以及促甲状腺激素(TSH)刺激后的释放。在没有TSH的情况下,T3和T4在孵育培养基中以可测量的量释放。两种碘甲状腺原氨酸的释放都与培养基中TSH浓度和孵育时间直接相关。Lineweaver-Burke分析显示,T3的Vmax为99.4 pg/腺体,表观Km为17.8 mU TSH,T4的Vmax为323.35 ng/腺体,表观Km为51.5 mU TSH,表明T4是禽类甲状腺释放的主要碘甲状腺原氨酸。通过向孵育培养基中添加钙螯合剂(乙二醇双(β-氨基乙醚)-N,N,N',N'-四乙酸;EGTA)、钙拮抗剂(氯化钴,CoCl2)或前列腺素E1(PGE1),可抑制T4的基础释放。在存在钙激动剂(氯化镧,LaCl3)、钙离子载体(A23187)、二丁酰环腺苷3',5'-单磷酸(dbcAMP)、异丁基甲基黄嘌呤(IBMX)、吲哚美辛、氯化镁(MgCl2)和碘化钾(KI)的情况下,基础释放的T4增加。CoCl2、PGE1和吲哚美辛可降低TSH刺激的T4释放,但LaCl3、MgCl2和KI可增强该释放。这些结果表明,在鸡的体外系统中可以测量甲状腺激素的释放。鸡甲状腺中基础和刺激后的碘甲状腺原氨酸释放似乎由钙和cAMP依赖性机制介导。