Immunocytochemical analysis of prolactin production by monolayer cultures of GH3 rat anterior pituitary tumor cells: II. Variation in prolactin content of individual cell colonies, and dynamics of stimulation with thyrotropin-releasing hormone (TRH).
Immunocytochemical analysis of prolactin production by monolayer cultures of GH3 rat anterior pituitary tumor cells: II. Variation in prolactin content of individual cell colonies, and dynamics of stimulation with thyrotropin-releasing hormone (TRH).
复制标题
GH3大鼠垂体前叶肿瘤细胞单层培养物产生催乳素的免疫细胞化学分析：II。
DOI：
10.1002/ar.1091970206
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发表时间：
1980
期刊：
影响因子：
--
通讯作者：
TashjianJr,AH
中科院分区：
文献类型：
--
作者：
HoytJr,RF;TashjianJr,AH
前述报告（Hoyt 和 Tashjian，'80）将免疫细胞化学定位与 TRH 最大刺激的 GH3 单层中的平均催乳素浓度相关联；目前在 TRH 治疗期间这样做。低密度接种产生大量离散的GH3细胞集落。施用 TRH (50 ng/ml) 或盐水（对照）后 1/2、4、12、24、72 和 144 小时收获培养物。至少 10 个显微视野/单层、1 个细胞集落/视野中的所有细胞（总共 42,658 个）被分类为催乳素未染色、重度 (H)、中度 (M) 或弱 (W) 染色。在对照中，集落每 100 个群体包含 51-91 个催乳素阳性细胞。阳性细胞较少的集落的 W 细胞比 M 细胞多得多，而阳性细胞较多的集落则相反。在所有菌落中，TRH 的作用都是双相的。催乳素的初始（0-4 小时）释放与 3-4 小时开始的细胞内激素逐渐增加重叠。 144小时后，处理后的培养物的催乳素含量增加至对照的190%，催乳素阳性细胞数量更多（对照的114%）。 TRH 处理 48 小时后，这些增加低于前一篇论文中报道的水平，当时细胞内催乳素等于对照的 450%，阳性细胞等于对照的 129%。这些不一致反映了催乳素产生的控制水平的差异，而不是 TRH 的绝对效果的差异，在连续的实验中，TRH 的绝对效果实际上是相同的。我们的结论是：（1）TRH 的作用是改变已经产生催乳素的细胞中激素的产生； (2) TRH 一定程度上增加了含催乳素细胞的数量； (3) 这种“新”细胞对激素输出增加的相对贡献取决于催乳素产生的基础水平，该水平在各个 GH3 细胞集落之间存在差异，并且随着培养时间的推移而变化。这种多样性并没有削弱 GH3 细胞作为激素生物合成的生化模型的有用性。它确实阻碍了它们有效的形态学评估，显然必须像生化实验一样仔细控制，并且应该包括免疫细胞化学定位，至少对于所讨论的激素而言。
The preceeding report (Hoyt and Tashjian, '80) correlates immunocytochemical localizations and mean prolactin concentrations in GH3monolayers maximally stimulated with TRH; the present does so over the duration of TRH treatment. Low density seeding produced numerous discrete GH3cell colonies. Cultures were harvested ½, 4, 12, 24, 72, and 144 hr after administration of TRH (50 ng/ml) or saline (control). All cells (42,658 total) in at least 10 microscopic fields/monolayer, 1 cell colony/field, were classed as unstained, heavily (H), moderately (M), or weakly (W) stained for prolactin. In controls, colonies contained 51‐91 prolactin‐positive cells/100 of population. Colonies with few positive cells had many more W than M cells, and the reverse was true in those with many positive cells. In all colonies, the effect of TRH was biphasic. Initial (0‐4 hr) release of prolactin was overlapped, beginning at 3‐4 hr, by a progressive increase of intracellular hormone. After 144 hr, the prolactin content of treated cultures had increased to 190% of control, and prolactin‐positive cells were more numerous (114% of control). These increases were lower than those reported in the preceeding paper after 48 hr of TRH treatment, when intracellular prolactin equalled 450% of control and positive cells equalled 129% of control. These inconsistencies reflect differences in the control level of prolactin production rather than in the absolute effects of TRH, which were virtually identical in the successive experiments. We conclude that: (1) TRH acts to alter hormone production in cells already making prolactin; (2) TRH increases somewhat the number of prolactin‐containing cells; (3) the relative contribution of such “new” cells to increased hormone output depends on the basal level of prolactin production, which differs among individual GH3cell colonies and varies over time in culture. This diversity does not diminish the usefulness of GH3cells as biochemical models of hormone biosynthesis. It does hinder their valid morphological evaluation, which apparently must be controlled as carefully as biochemical experiments and should include immunocytochemical localizations, at least for the hormone at issue.