Shoemaker Justin M, Riley Larry G, Hirano Tetsuya, Grau E Gordon, Rubin David A
Department of Biological Sciences, Illinois State University, Normal, IL 61790, USA.
Gen Comp Endocrinol. 2006 Apr;146(2):186-94. doi: 10.1016/j.ygcen.2005.10.009. Epub 2005 Dec 20.
A new parathyroid hormone (PTH)-like endocrine system has been identified in mammals and fishes consisting of the PTH type-2 receptor (PTH2R) and tuberoinfundibular peptide 39 (TIP39). Although the mammalian PTH2R-TIP39 system is involved in nociception and pituitary regulation, the function(s) of this system in fishes is undetermined. Using degenerate primers based on conserved zebrafish and fugu TIP39 nucleotide sequences, 3'-RACE reactions isolated the coding region of a putative TIP38 cDNA in the Nile tilapia (Oreochromis niloticus). Tilapia-specific primers were subsequently used in 5'-RACE reactions to isolate the remaining portion of the transcript. The cDNA encoding O. niloticus TIP (OnTIP38) was determined to yield a 38 amino acid secreted hormone. A second tilapia TIP38 cDNA was isolated from the euryhaline Mozambique tilapia (OmTIP38). Except for the 39th residue, both tilapia cDNA sequences showed significant identity to human, bovine, murine, fugu, and zebrafish TIP39. To determine the tissue-specific expression of OnTIP38 and OmTIP38, real-time quantitative RT-PCR (rQRT-PCR) was performed on skin, gill, kidney, testis, heart, and brain. In freshwater (FW)-acclimated Nile tilapia, OnTIP38 showed highest levels of expression in kidney and lowest levels in skin and gill. In Mozambique tilapia tissues, expression of OmTIP38 and G6Pase (glucose-6 phosphatase) were higher in salt water (SW)-acclimated fish than in FW-acclimated fish. G6Pase expression, and not OmTIP38, showed significant differences among various tissues in FW- and SW-acclimated fish. Results of the present study clearly indicate that the TIP38/39-PTH2R system shows considerable conservation in sequence identity and tissue-specific expression in mammals and fishes.
在哺乳动物和鱼类中已鉴定出一种新的甲状旁腺激素(PTH)样内分泌系统,该系统由2型PTH受体(PTH2R)和结节漏斗肽39(TIP39)组成。尽管哺乳动物的PTH2R - TIP39系统参与痛觉感受和垂体调节,但该系统在鱼类中的功能尚未确定。基于斑马鱼和河豚TIP39保守核苷酸序列设计简并引物,通过3'-RACE反应分离出尼罗罗非鱼(Oreochromis niloticus)中一个假定的TIP38 cDNA的编码区。随后使用罗非鱼特异性引物进行5'-RACE反应,以分离转录本的其余部分。编码尼罗罗非鱼TIP(OnTIP38)的cDNA被确定产生一种由38个氨基酸组成的分泌激素。从广盐性莫桑比克罗非鱼中分离出第二个罗非鱼TIP38 cDNA(OmTIP38)。除第39位残基外,两种罗非鱼的cDNA序列与人类、牛、小鼠、河豚和斑马鱼的TIP39均有显著同源性。为了确定OnTIP38和OmTIP38的组织特异性表达,对皮肤、鳃、肾脏、睾丸、心脏和大脑进行了实时定量RT-PCR(rQRT-PCR)。在淡水(FW)适应的尼罗罗非鱼中,OnTIP38在肾脏中的表达水平最高,在皮肤和鳃中的表达水平最低。在莫桑比克罗非鱼组织中,盐适应(SW)的鱼中OmTIP38和葡萄糖-6磷酸酶(G6Pase)的表达高于淡水适应的鱼。在淡水和盐适应的鱼中,G6Pase的表达在不同组织间存在显著差异,而OmTIP38的表达无显著差异。本研究结果清楚地表明,TIP38/39 - PTH2R系统在哺乳动物和鱼类的序列同源性和组织特异性表达方面具有相当程度的保守性。