Three-dimensional imaging of hormone-secreting cells and their microvessel environment in estrogen-induced prolactinoma of the rat pituitary gland by confocal laser scanning microscopy
Three-dimensional imaging of hormone-secreting cells and their microvessel environment in estrogen-induced prolactinoma of the rat pituitary gland by confocal laser scanning microscopy
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DOI：
10.1097/00022744-200112000-00013
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发表时间：
2001-12-01
影响因子：
1.6
通讯作者：
Osamura, RY
中科院分区：
文献类型：
--
作者：
Itoh, J;Kawai, T;Osamura, RY
本研究的重点是雌激素诱导的大鼠垂体催乳素瘤中激素分泌细胞及其微血管环境的三维成像。成年雌性Wistar-Imamichi大鼠注射二丙酸雌二醇，7周后处死。一些给予雌激素7周的大鼠在处死前也注射了溴隐亭。为了获得详细的三维图像的微血管，透析的异硫氰酸荧光素（FITC）结合的明胶注射到左心室的大鼠心脏。灌注后，切除垂体并进行免疫组织化学（IHC）。为了评价雌激素和溴隐亭的作用，用抗催乳素（PRL）、促肾上腺皮质激素（ACTH）和生长激素（GH）的抗体进行IHC。与组合，微血管和细胞含有PRL，ACTH和GH可以清楚地确定通过共聚焦激光扫描显微镜（CLSM）。PRL细胞数量增加，并在长期暴露于雌激素后变得肥大。然而，雌激素治疗后给予溴隐亭，PRL细胞数量减少，并变得萎缩。目前的研究表明，雌激素和溴隐亭对PRL分泌和微血管环境有显着影响。因此，这种技术（FITC注射和IHC）与CLSM是适合于在各种条件下的细胞分泌机制的三维成像。
This study focused on the three-dimensional imaging of hormone-secreting cells and their microvascular environment in estrogen-induced prolactinoma of the rat pituitary gland. Adult female Wistar-Imamichi rats were injected with estradiol dipropionate and killed 7 weeks later. Some rats given estrogen for 7 weeks also were injected with bromocriptine before killing. To obtain a detailed three-dimensional image of microvessels, dialyzed fluorescein isothiocyanate (FITC)-conjugated gelatin was injected into the left ventricle of the rat heart. After the perfusion, the pituitary glands were resected and subjected to immunohistochemistry (IHC). To evaluate the effects of estro-en and bromocriptine, IHC was performed with antibodies against prolactin (PRL), adrenocorticotropic hormone (ACTH), and growth hormone (GH). With the combination, microvessels and cells containing PRL, ACTH, and GH could be clearly identified by confocal laser scanning microscopy (CLSM). The PRL cells increased in number and became hypertrophic after prolonged exposure to estrogen. With bromocriptine administration after estrogen treatment, however, PRL cells decreased in number and became atrophic. The current study revealed that estrogen and bromocriptine had significant effects on PRL secretion and the microvascular environment. Therefore, this technique (FITC injection and IHC) with CLSM is suitable for the three-dimensional imaging of hormone-secreting mechanisms under various conditions.