Functional expression of corticotropin-releasing hormone (CRH) receptor 1 in cultured rat microglia

被引:39
作者
Wang, W
Ji, P
Riopelle, RJ
Dow, KE
机构
[1] Kingston Gen Hosp, Dept Pediat, Kingston, ON K7L 2V7, Canada
[2] Queens Univ, Dept Anat & Cell Biol, Kingston, ON, Canada
[3] Queens Univ, Dept Med, Kingston, ON K7L 3N6, Canada
关键词
brain; cAMP; glia; ligand binding;
D O I
10.1046/j.0022-3042.2001.00687.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Corticotropin-releasing hormone (CRH), known as a key regulator of the hypothalamic-pituitary-ad renal axis response to stress, elicits its biological effects by binding to two membrane receptors (CRH-R1 and CRH-R2). The present studies examined the presence of functional expression of CRH receptors in cultured microglia of rat. CRH-R1 mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR), western blotting and receptor chemical cross-linking assay in cultured microglia. CRH-R2 mRNA was undectable by RT-PCR. The radioligand binding analysis using [I-125]Tyr-rat/human CRH revealed a high affinity binding site (K-d of 1.2 nm and B-max of 84 fmol/mg of protein). Competition studies using CRH and related peptides indicated kinetic and pharmacological characteristics consistent with the CRH-R1 receptor subtype. Receptor chemical cross-linking assay demonstrated a single band of CRH receptor with a molecular weight of similar to77 kDa, which was inhibited in the presence of excess unlabeled rat/human CRH in a dose-dependent manner and inhibited by a CRH receptor antagonist astressin. Functional coupled cAMP production in cultured microglia was stimulated by exogenous addition of CRH and related peptides in a dose-dependent manner and blocked by astressin. Our findings suggest the functional expression of CRH-R1 receptor in rat microglia, indicating an important mechanism of interaction between immune and neuroendocrine systems in brain physiological and pathological conditions.
引用
收藏
页码:287 / 294
页数:8
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