LeBaron M J, Ahonen T J, Nevalainen M T, Rui H
Thomas Jefferson University, Kimmel Cancer Center, Department of Cancer Biology, 233 South 10th Street, 330 BLSB, Philadelphia, PA 19107-5541, USA.
Endocrinology. 2007 Mar;148(3):989-1008. doi: 10.1210/en.2006-1219. Epub 2006 Nov 30.
To identify cell populations directly responsive to prolactin (PRL), GH, erythropoietin, or granulocyte-colony stimulating factor within the physiological setting of an intact mammal, we combined in situ detection of hormone-activated signal transducer and activator of transcription (Stat)-5 in rats with high-throughput tissue array analysis using cutting-edge matrix assembly (CEMA). Inducible activation of Stat5a/b, as judged by levels of nuclear-localized, phosphoTyr694/699-Stat5a/b, served as an immediate and sensitive in situ marker of receptor signaling in rat tissues after injection into male and female rats of a single, receptor-saturating dose of hormone for maximal receptor activation. CEMA tissue arrays facilitated analysis of most tissues, including architecturally complex, thin-walled, and stratified tissues such as gut and skin. In 40 tissues analyzed, 35 PRL-responsive and 32 GH-responsive cell types were detected, of which 22 cell types were responsive to both hormones. Interestingly, PRL but not GH activated Stat5 in nearly all of the endocrine glands. In mammary glands, PRL activated Stat5 in a majority of luminal epithelial cells but not myoepithelial cells, stromal fibroblasts, or adipocytes, whereas GH activated Stat5 in a significant fraction of myoepithelial cells, fibroblasts, and adipocytes but only in a minority of luminal cells. Finally, the organism-wide screening revealed a yet-to-be identified erythropoietin-responsive cell type in connective tissue. CEMA tissue arrays provide cost-effective in situ analysis of large numbers of tissues. Biomarker-based identification of cell populations responsive to individual hormones may shed new light on endocrine disease as well as improve understanding of effects and side effects of hormones and drugs.
为了在完整哺乳动物的生理环境中识别对催乳素（PRL）、生长激素（GH）、促红细胞生成素或粒细胞集落刺激因子直接产生反应的细胞群体，我们将大鼠体内激素激活的信号转导子和转录激活子（Stat）-5的原位检测与使用前沿基质组装（CEMA）的高通量组织阵列分析相结合。通过核定位的磷酸化Tyr694/699-Stat5a/b水平判断Stat5a/b的可诱导激活，作为向雄性和雌性大鼠注射单剂量受体饱和激素以实现最大受体激活后，大鼠组织中受体信号传导的即时且灵敏的原位标记。CEMA组织阵列有助于分析大多数组织，包括结构复杂、薄壁和分层的组织，如肠道和皮肤。在分析的40种组织中，检测到35种对PRL有反应的细胞类型和32种对GH有反应的细胞类型，其中22种细胞类型对两种激素都有反应。有趣的是，几乎所有内分泌腺中PRL激活了Stat5，而GH没有。在乳腺中，PRL在大多数腔上皮细胞中激活了Stat5，但在肌上皮细胞、基质成纤维细胞或脂肪细胞中未激活；而GH在相当一部分肌上皮细胞、成纤维细胞和脂肪细胞中激活了Stat5，但仅在少数腔细胞中激活。最后，全生物体范围的筛选揭示了结缔组织中一种尚未确定的对促红细胞生成素产生反应的细胞类型。CEMA组织阵列提供了对大量组织具有成本效益的原位分析。基于生物标志物识别对个体激素产生反应的细胞群体，可能为内分泌疾病带来新的见解，并增进对激素和药物的作用及副作用的理解。
