Long-term organ culture of rat anterior pituitary glands.
Long-term organ culture of rat anterior pituitary glands.
复制标题
大鼠垂体前叶的长期器官培养。
作者：
J. E. Martin;D. Klein
一种用于新生大鼠垂体前腺的器官培养系统，其细胞完整性和对LH释放激素（LHRH）的反应性可维持至少4天。取Sprague-Dawley大鼠垂体前腺，在95% O2-5% CO2的环境中，37℃单独培养，BGJb培养基，Fitton-Jackson改性。5日龄大鼠垂体在对照培养基中培养96 h，组织学检查未见组织坏死。相比之下，在相同条件下，40日龄大鼠垂体前腺仅在孵育24小时后就出现了中央坏死。用合成LHRH处理新生儿腺体24 h，用双抗体放射免疫法测定中、垂体LH FSH浓度。在培养的第一天，LHRH引起LH的剂量相关释放，最小有效剂量在10(-10)~ 10(-9)M之间。10(-6)M LHRH似乎达到了接近最大的释放，这导致中LH水平比对照组提高了12倍。10(-9)M LHRH刺激后3小时内释放明显。垂体在培养0、24和72小时时对该剂量有反应，lhrh刺激和对照LH水平的比值分别为3、13和24，逐渐增加。虽然在培养的第一天没有观察到LHRH释放FSH，但在第二天LHRH引起了该激素的剂量相关分泌。在添加LHRH和促甲状腺激素释放激素（TRH）后，垂体反应对LH和TSH释放的特异性得到证实。这种新生大鼠垂体前叶器官培养系统的简单、灵敏、可重复性和长期生存能力使其成为研究促性腺激素分泌的一种有价值的技术。
An organ culture system for neonatal rat anterior pituitary glands has been developed in which cellular integrity and responsiveness to LH releasing hormone (LHRH) are maintained for at least 4 days. Anterior pituitary glands from Sprague-Dawley rats were cultured individually at 37 C in an atmosphere of 95% O2-5% CO2 in BGJb culture medium, Fitton-Jackson modification. Histological examination of 5-day-old rat pituitary glands cultured for 96 h in control medium revealed no evidence of tissue necrosis. By contrast, 40-day-old-rat anterior pituitary glands were centrally necrotic after only 24 h of incubation under the same conditions. The neonatal glands were treated for 24 h with synthetic LHRH, and medium and pituitary LH FSH concentrations were determined by double antibody radioimmunoassay. On the first day of culture, LHRH caused a dose-related release of LH with the minimal effective dose between 10(-10) and 10(-9)M. Near-maximal release appeared to be attained with 10(-6)M LHRH, which induced a 12-fold elevation in medium LH over controls. Release was evident within 3 h following stimulation with 10(-9)M LHRH. Pituitary glands were responsive to this dose at 0,24 and 72 h of culture with progressively increasing ratios between LHRH-stimulated and control LH levels of 3,13, and 24, respectively. Although FSH release by LHRH was not observed on the first day of culture, on the second day LHRH caused a dose-related secretion of this hormone. Specificity of the pituitary response was demonstrated for LH and TSH release following addition of LHRH and thyrotropin releasing hormone (TRH). The simplicity, sensitivity, reproducibility, and long-term viability of this neonatal rat anterior pituitary organ culture system make it a valuable technique for the study of gonadotropin secretion.