Identification, tissue distribution and functional characterization of the ghrelin receptor in West African lungfish, Protopterus annectens

被引:8
|
作者
Kaiya, Hiroyuki [1 ]
Konno, Norifumi [2 ]
Kangawa, Kenji [1 ]
Uchiyama, Minoru [2 ]
Miyazato, Mikiya [1 ]
机构
[1] Natl Cardiovasc Ctr, Res Inst, Dept Biochem, Suita, Osaka 5658565, Japan
[2] Toyama Univ, Grad Sch Sci & Engn, Lab Regulatory Biol, Gofuku 3190, Toyama 9308555, Japan
基金
日本学术振兴会;
关键词
cDNA cloning; Functional characterization; Ghrelin receptor; GHS-R1a; GHS-R1b; Lungfish; HORMONE SECRETAGOGUE RECEPTOR; 2ND EXTRACELLULAR LOOP; COMPLEMENTARY DEOXYRIBONUCLEIC-ACID; MESSENGER-RNA EXPRESSION; GROWTH-HORMONE; RAINBOW-TROUT; MOLECULAR CHARACTERIZATION; GENOME DUPLICATION; GASTROINTESTINAL-TRACT; (GHS-R)-LIKE RECEPTOR;
D O I
10.1016/j.ygcen.2014.07.021
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
We identified two ghrelin receptor isoforms, the ghrelin receptor type-la (GHS-R1a) and its alternative splice form (GHS-R1b) for West African lungfish, Protopterus annectens. Lungfish GHS-R1a and 1b comprised 361 and 281 amino acids, respectively. Lungfish GHS-R1a showed the highest identity to coelacanth GHS-R1a (80.4%). The highest expression of GHS-R1a mRNAs was seen in the brain, liver, ovary, heart, intestine, and gills. GHS-R1b mRNAs were also detected in the same tissues with GHS-R1a, but their expression level was 1/20 that of GHS-R1a. In human embryonic kidney 293 cells transiently expressing lungfish GHS-R1a, rat and bullfrog ghrelin, and two GHS-R1a agonists, GHRP-6 and hexarelin, increased intracellular Ca2+ concentrations. The intensity of the Ca2+ increases induced by GHS-R1a agonists was twice when compared to that induced by ghrelin, although the median effective doses (ED50) were similar, suggesting a long-lasting effect of GHS-R1 a agonists with similar affinity. We also examined changes in the GHS-R gene expression during an eight-week estivation. Body weight was slightly lowered, but plasma sodium and glucose concentrations decreased; plasma urea concentration increased significantly 4 weeks after the start of estivation. Overall, expression of GHS-Rla mRNA decreased, but changes in GHS-Rlb mRNA expression were inconsistent with those of GHS-R1a during estivation, suggesting an involvement of GHS-R in energy homeostasis, as seen in mammals. Our results suggest that the ghrelinGHS-Rla system is present in this lungfish although ghrelin has not yet been found. The structure of GHSRla is closer to that of tetrapods than Actinopterygian fish, indicating a process of evolution that follows the Crossopterygii such as coelacanth. (C) 2014 Elsevier Inc. All rights reserved.
引用
收藏
页码:106 / 117
页数:12
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