Sithigorngul Paisarn, Panchan Nanthika, Chaivisuthangkura Parin, Longyant Siwaporn, Sithigorngul Weerawan, Petsom Amorn
Department of Biology, Faculty of Science, Srinakharinwirot University, Sukhumvit 23, Bangkok 10110, Thailand.
Peptides. 2002 Nov;23(11):1943-52. doi: 10.1016/s0196-9781(02)00180-8.
Mouse antiserum against C-terminal amide of Pem-CMG (a peptide in the family of CHH/MIH/GIH) penta-deca peptide (RPRQRNQYRAALQRLamide=CMG-15) was generated and used for localization of the peptide in tissue and extract of the eyestalk of Penaeus monodon by means of immunohistochemistry and dot-ELISA in comparison with anti-T+ antiserum (T+=YANAVQTVamide : the putative C-terminal amide of crustacean hyperglycemic hormone (CHH) of Macrobrachium rosenbergii). The anti-CMG-15 antiserum did not show cross-reactivity to T+ peptide by dot-ELISA and vice versa for anti-T+ antiserum. In dot-ELISA of eyestalk extract of P. monodon after one step separation by RP-HPLC, anti-CMG-15 antiserum recognized different peptide fractions (F38-39) from those recognized by anti-T+ antiserum (F19, 40-41 and 47-51). Most of the T+ immunoreactive fractions (except F19) show higher hyperglycemic activity than the CMG immunoreactive fractions. In immunohistochemical localization, anti-CMG antiserum recognized only 2-3 neurons in medulla terminalis X-organ complex (MTXO) with long processes terminated in the sinus gland. The CMG-immunoreactive neurons were clearly distinct from CHH containing neurons situated in the same area. This evidence confirms the existing of CMG peptide which may play distinct roles from CHHs in hormonal regulation in P. monodon.
制备了针对Pem - CMG(CHH/MIH/GIH家族中的一种肽)十五肽(RPRQRNQYRAALQRLamide = CMG - 15)C末端酰胺的小鼠抗血清,并通过免疫组织化学和斑点酶联免疫吸附测定(dot - ELISA)将该肽定位在斑节对虾眼柄组织和提取物中,同时与抗T +抗血清(T + = YANAVQTVamide:罗氏沼虾甲壳类高血糖激素(CHH)的假定C末端酰胺)进行比较。通过斑点酶联免疫吸附测定,抗CMG - 15抗血清与T +肽无交叉反应,抗T +抗血清反之亦然。在对斑节对虾眼柄提取物进行反相高效液相色谱(RP - HPLC)一步分离后的斑点酶联免疫吸附测定中,抗CMG - 15抗血清识别的肽段(F38 - 39)与抗T +抗血清识别的不同(F19、40 - 41和47 - 51)。大多数T +免疫反应性组分(除F19外)显示出比CMG免疫反应性组分更高的高血糖活性。在免疫组织化学定位中,抗CMG抗血清仅识别终髓X器官复合体(MTXO)中2 - 3个神经元,其长突起终止于窦腺。CMG免疫反应性神经元与位于同一区域的含CHH神经元明显不同。这一证据证实了CMG肽存在,其在斑节对虾的激素调节中可能发挥与CHHs不同的作用。