Department of Biology, East China Normal University, Shanghai, China.
PLoS One. 2010 Jun 17;5(6):e11175. doi: 10.1371/journal.pone.0011175.
Leptin is an adipocyte-derived hormone with multiple functions that regulates energy homeostasis and reproductive functions. Increased knowledge of leptin receptor function will enhance our understanding of the physiological roles of leptin in animals.
METHODOLOGY/PRINCIPAL FINDINGS: In the present study, a full-length leptin receptor (lepr) cDNA, consisting of 1,353 nucleotides, was cloned from Chinese mitten crab (Eriocheir sinensis) using rapid amplification of cDNA ends (RACE) following the identification of a single expressed sequence tag (EST) clone in a cDNA library. The lepr cDNA consisted of a 22-nucleotide 5'-untranslated region (5' UTR), a 402-nucleotide open reading frame (ORF) and a 929-nucleotide 3' UTR. Multiple sequence alignments revealed that Chinese mitten crab lepr shared a conserved vacuolar protein sorting 55 (Vps55) domain with other species. Chinese mitten crab lepr expression was determined in various tissues and at three different reproductive stages using quantitative real-time RT-PCR. Lepr expression was highest in the intestine, thoracic ganglia, gonad, and accessory gonad, moderate in hepatopancreas and cranial ganglia, and low in muscle, gill, heart, haemocytes, and stomach. Furthermore, lepr expression was significantly higher in the intestine, gonad and thoracic ganglia in immature crabs relative to precocious and mature crabs. In contrast, lepr expression was significantly lower in the hepatopancreas of immature crabs relative to mature crabs.
CONCLUSIONS/SIGNIFICANCE: We are the first to identify the lepr gene and to determine its gene expression patterns in various tissues and at three different reproductive stages in Chinese mitten crab. Taken together, our results suggest that lepr may be involved in the nutritional regulation of metabolism and reproduction in Chinese mitten crabs.
瘦素是一种由脂肪细胞分泌的激素，具有多种功能，可调节能量平衡和生殖功能。对瘦素受体功能的深入了解将有助于我们理解瘦素在动物中的生理作用。
方法/主要发现：本研究采用 cDNA 末端快速扩增技术（RACE），根据 cDNA 文库中一个单一表达序列标签（EST）克隆，从中华绒螯蟹（Eriocheir sinensis）中克隆出全长瘦素受体（lepr）cDNA，由 22 个核苷酸的 5'非翻译区（5'UTR）、402 个核苷酸的开放阅读框（ORF）和 929 个核苷酸的 3'UTR 组成。序列比对显示，中华绒螯蟹 lepr 与其他物种共享一个保守的液泡分选 55（Vps55）结构域。采用实时定量 RT-PCR 技术，检测了不同组织和三个不同繁殖阶段中华绒螯蟹 lepr 的表达情况。lepr 在肠、胸神经节、性腺和副性腺中表达最高，在肝胰腺和脑神经节中表达中等，在肌肉、鳃、心脏、血细胞和胃中表达较低。此外，与早熟和成熟蟹相比，未成熟蟹的肠、性腺和胸神经节中 lepr 的表达显著较高。相反，与成熟蟹相比，未成熟蟹的肝胰腺中 lepr 的表达显著较低。
结论/意义：我们首次鉴定了中华绒螯蟹 lepr 基因，并在不同组织和三个不同繁殖阶段检测了其基因表达模式。综上所述，我们的研究结果表明，lepr 可能参与中华绒螯蟹的代谢和生殖营养调控。
