Comparative and Molecular Endocrinology Group, CCMAR, CIMAR Laboratório Associado, University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal.
Gen Comp Endocrinol. 2013 Sep 15;191:102-12. doi: 10.1016/j.ygcen.2013.05.014. Epub 2013 Jun 7.
Endocrine factors play an essential role in the formation and turnover of the skeleton in vertebrates. In the present study sea bream vertebral bone transcripts for PTH1R and PTH3R were identified and the action of intermittent administration of parathyroid hormone related protein (PTHrP) on the proteome of vertebral bone was analysed. Treatment of immature sea bream (Sparus auratus, n=6) for 5days with homologous recombinant PTHrP(1-125; 150ng/g body weight) modified bone metabolism and caused a significant (p<0.05) reduction in both tartrate resistant acid phosphatase (TRACP) and alkaline phosphatase (ALP) in relation to control fish. However, the ratio of TRACP: ALP in PTHrP treated fish (1.3 to 2.2 cf. control) suggested it had an anabolic response. A sea bream vertebral bone proteome of 157 protein spots was generated and putative identity assigned to 118 (75.2%) proteins of which 72% had homology to proteins/transcripts from teleosts many of which have not previously been reported in teleost bone. Classification of bone proteins using gene ontology revealed those with protein or metal/ion (e.g., calcium, magnesium, zinc) binding (∼53%) activities were most abundant. The expression of eight proteins was significantly (p<0.05) modified in the vertebra of PTHrP treated compared to control fish; three were up-regulated, betainehomocystein S-methyltransferase, glial fibrillary acidic protein, parvalbumin beta and five were down-regulated, annexin A5, apolipoprotein A1, myosin light chain 2, fast skeletal myosin light chain 3, troponin C. In conclusion, intermittent administration of PTHrP to sea bream is associated with an anabolic response in vertebral bone metabolism and modifies calcium binding proteins in the proteome.
内分泌因素在脊椎动物骨骼的形成和更新中起着至关重要的作用。在本研究中，鉴定了牙鲆椎骨的甲状旁腺素 1 型受体（PTH1R）和甲状旁腺素 3 型受体（PTH3R）的转录本，并分析了间歇性给予甲状旁腺素相关蛋白（PTHrP）对椎骨蛋白质组的作用。用同源重组的 PTHrP（1-125；150ng/g 体重）处理未成熟的牙鲆（Sparus auratus，n=6）5 天，改变了骨代谢，使抗酒石酸酸性磷酸酶（TRACP）和碱性磷酸酶（ALP）与对照鱼相比显著降低（p<0.05）。然而，PTHrP 处理鱼的 TRACP：ALP 比值（1.3 至 2.2 与对照相比）表明其具有合成代谢反应。生成了 157 个牙鲆椎骨蛋白质斑点的蛋白质组，并将假定的身份分配给了 118 个（75.2%）蛋白质，其中 72%与硬骨鱼类的蛋白质/转录本具有同源性，其中许多蛋白质以前在硬骨鱼类的骨骼中没有报道过。使用基因本体论对骨骼蛋白进行分类，发现具有蛋白质或金属/离子（如钙、镁、锌）结合（约 53%）活性的蛋白质最为丰富。与对照组相比，PTHrP 处理的牙鲆椎骨中 8 种蛋白质的表达显著（p<0.05）改变；有 3 种上调，甜菜碱同型半胱氨酸 S-甲基转移酶、神经胶质纤维酸性蛋白、副肌球蛋白 beta，5 种下调，载脂蛋白 A1、肌球蛋白轻链 2、快速骨骼肌肌球蛋白轻链 3、肌钙蛋白 C。总之，间歇性给予 PTHrP 与牙鲆椎骨代谢的合成代谢反应有关，并改变蛋白质组中的钙结合蛋白。
