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腦垂體pituitary巨噬細(xì)胞清除Macrophage depletion方案

更新時(shí)間:2024-01-07   點(diǎn)擊次數(shù):845次

垂體(pituitary)是下丘腦-垂體-腎上腺 (HPA) 軸的重要樞紐綠色化發展。垂體激素分泌細(xì)胞 (HPC) 釋放多種激素持續向好,以在正常和壓力條件下調(diào)節(jié)基本身體功能結構不合理。垂體內(nèi)分泌腺通過(guò)釋放促腎上腺皮質(zhì)激素 (ACTH) 來(lái)調(diào)節(jié)免疫系統(tǒng)有序推進,以響應(yīng)下丘腦的神經(jīng)元激活建設項目。然而自動化方案,目前尚不清楚全身炎癥如何調(diào)節(jié)垂體HPC的轉(zhuǎn)錄組學(xué)特征創新為先。在這里又進了一步,我們對(duì)小鼠垂體進(jìn)行了單細(xì)胞RNA測(cè)序(scRNA-seq)傳遞,發(fā)現(xiàn)在炎癥發(fā)生時(shí)融合,所有主要的垂體HPC都以細(xì)胞類型特異性方式產(chǎn)生強(qiáng)烈反應(yīng),其中皮質(zhì)促物表現(xiàn)出強(qiáng)烈的反應(yīng)相關性。全身炎癥還導(dǎo)致非典型生物活性分子的產(chǎn)生和釋放完成的事情,包括皮質(zhì)激素的 Nptx2,以調(diào)節(jié)免疫穩(wěn)態(tài)穩定。同時(shí)進入當下,HPCs上調(diào)了趨化因子的基因表達(dá),促進(jìn)了HPCs與免疫細(xì)胞之間的通訊效高化⌒麦w系?傊覀兊难芯拷沂玖舜贵w和免疫系統(tǒng)之間的廣泛相互作用創造,表明垂體在介導(dǎo)炎癥對(duì)身體生理學(xué)許多方面的影響方面發(fā)揮著多方面的作用不難發現。


Macrophage depletion and virus injection in the pituitary

Mice were anesthetized with pentobarbital (80 mg/kg, i.p.) before surgery and then placed in a mouse stereotaxic instrument. Injections were performed using a microsyringe pump and a Micro4 controller (World Precision Instruments). For macrophage depletion, liposome-PBS or liposome-Clodronate (Liposoma,CP-005-005) was stereotaxically microinjected into the anterior pituitary (2.5 mm posterior from Bregma, 0.4 mm lateral, 6 mm below pia). The liposomes were delivered to the target site at a rate of 60 nL/min for 500 nL per site. Mice received saline or 0.5 mg/kg LPS 18 h after liposome delivery and were killed 6 h after inflammation was established. For CCL2 expression and Nptx2-KO, AAV was delivered directly into the pituitary. The injection site, rate, and volume were the same as those used for the liposome injection. Subsequent experiments were performed at least 3 weeks after virus injection.


(C) Representative images showing Iba1 (the marker of macrophage) expression in the pituitary from mice that received liposome-PBS (left) or liposome-Clodronate (right) directly to the pituitary for 24 h. Scale bar, 100 μm.

(D) Serum concentrations of ACTH in saline- or LPS-treated (0.5 mg/kg LPS for 6 h) mice pretreated with liposome-PBS or liposome-Clodronate (n = 4–7 mice).

(E) Serum concentrations of corticosterone in LPS-treated (0.5 mg/kg LPS for 6 h) mice pretreated with liposome-PBS or liposome-Clodronate (n = 6–7 mice).




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