Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, 362-763, Republic of Korea.
Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, 362-763, Republic of Korea.
Brain Res Bull. 2019 Oct;152:225-235. doi: 10.1016/j.brainresbull.2019.07.028. Epub 2019 Jul 26.
Calbindin-D (CaBP-9k), one of the major calcium-binding and calcium-buffering proteins, is important in the physiological functioning of organs. The neuroanatomical localization of CaBP-9k in the rodent brain has not been reported; thus, this study investigated the neuroanatomical distribution of CaBP-9k and the regulation of CaBP-9k expression on steroid hormones in the immature rat brain. To confirm the influence of steroid hormones on CaBP-9k expression, immature female rats were injected for 5 days with estrogen (E2), progesterone (P4), dexamethasone (DEX), and their antagonists (ICI 182, 780 and RU 486). The localization and expression of the CaBP-9k protein in brain regions were identified by immunofluorescence and western blot assays, respectively. We observed that CaBP-9k expression was especially strong in hypothalamus, cerebellum, and brain stem. In addition, CaBP-9k was colocalized with mature-, GABAergic, dopaminergic, and oxytocinergic neurons. We also observed that the CaBP-9k protein level was significantly increased by P4 and reversed by antagonist RU 486 treatment in immature rat brain. In summary, CaBP-9k positive cells have a wide distribution in the immature rat brain, and CaBP-9k expression is regulated by P4. We suggest that CaBP-9k expression regulated by steroid hormone may serve as an important regulator of cytosolic calcium concentration in the brain.
钙结合蛋白-9k(CaBP-9k)是一种主要的钙结合和钙缓冲蛋白,对器官的生理功能很重要。CaBP-9k 在啮齿动物大脑中的神经解剖定位尚未报道;因此,本研究调查了 CaBP-9k 在未成熟大鼠大脑中的神经解剖分布以及类固醇激素对 CaBP-9k 表达的调节。为了确认类固醇激素对 CaBP-9k 表达的影响,对未成熟雌性大鼠连续 5 天注射雌激素(E2)、孕酮(P4)、地塞米松(DEX)及其拮抗剂(ICI 182、780 和 RU 486)。通过免疫荧光和 Western blot 检测分别鉴定脑区 CaBP-9k 蛋白的定位和表达。我们观察到 CaBP-9k 在下丘脑、小脑和脑干中的表达特别强烈。此外,CaBP-9k 与成熟、GABA 能、多巴胺能和催产素能神经元共定位。我们还观察到 P4 可显著增加 CaBP-9k 蛋白水平,而 P4 的拮抗剂 RU 486 处理则逆转这一作用。综上所述,CaBP-9k 阳性细胞在未成熟大鼠脑中分布广泛,且 CaBP-9k 的表达受 P4 调节。我们认为,类固醇激素调节的 CaBP-9k 表达可能作为脑细胞溶质钙离子浓度的重要调节剂。