Laboratory of Freshwater Fishery Germplasm Resources, Ministry of Agriculture, Shanghai Ocean University, Shanghai 201306, China.
Fish Physiol Biochem. 2013 Aug;39(4):881-93. doi: 10.1007/s10695-012-9748-4. Epub 2012 Nov 27.
A fully developed stomach, characterized by the secretion of pepsinogens and chlorhydric acid, is vital for digestion and survival of fish larvae. To further understand the functional development of the stomach of mandarin fish (Siniperca chuatsi) during early ontogeny, the temporal and spatial expression of pepsinogens (PG A1, A2 and C), as well as proton pump genes were analyzed in the stomach from 0 to 40 days post-hatch (dph) by reverse transcription polymerase chain reaction (RT-PCR) and in situ hybridization (ISH) techniques. Pepsinogen C cDNA was firstly cloned with a full length of 1,557 bp, which contained a 37-bp 5'-untranslated region (UTR), an open reading frame of 1,164 bp encoding a polypeptide of 387 amino acids (aa) residues and a 356-bp 3'-UTR. RT-PCR analysis revealed a sequential expression mode of three pepsinogens (PG A1, A2 and C) along the ontogeny of the stomach in mandarin fish. Pepsinogen A1 was firstly detected at 4 dph (84 degree-days, dd) ahead of the appearance of gastric glands; pepsinogen A2 appeared at 12 dph (252 dd) and became the predominant form in the stomach after 19 dph (399 dd); pepsinogen C was the latest expressed gene at 14 dph (294 dd). Expression of proton pump at 12 dph (252 dd), coinciding with the expression time of pepsinogen A2 showed an excellent coordinated transcription mode between proton pump and pepsinogens. ISH analysis located the expression of three pepsinogens and α subunit of proton pump in the same gastric gland cells, which confirmed that they belonged to oxynticopeptic cells. In addition, oxynticopeptic cells developed and increased gradually from 14 to 40 dph. No transcripts of pepsinogens or proton pump were detected in surface mucous cells and mucous neck cells of the gastric mucosa. Our results implied the functional development of stomach in mandarin fish was closely related to pepsinogens expression.
一个充分发育的胃，其特征是胃蛋白酶原和盐酸的分泌，对于鱼类幼体的消化和生存至关重要。为了进一步了解鳜鱼（Siniperca chuatsi）早期胚胎发生过程中胃的功能发育，通过逆转录聚合酶链反应（RT-PCR）和原位杂交（ISH）技术，分析了胃蛋白酶原（PG A1、A2 和 C）和质子泵基因在孵化后 0 至 40 天（dph）期间的时空表达。首次克隆了全长为 1557bp 的胃蛋白酶原 C cDNA，其中包含 37bp 的 5'-非翻译区（UTR）、1164bp 的开放阅读框，编码 387 个氨基酸（aa）残基的多肽和 356bp 的 3'-UTR。RT-PCR 分析显示，三种胃蛋白酶原（PG A1、A2 和 C）在鳜鱼胃的发育过程中呈现出一种连续表达模式。胃蛋白酶原 A1 最早在 4 dph（84 度日，dd）被检测到，早于胃腺的出现；胃蛋白酶原 A2 出现在 12 dph（252dd），并在 19 dph（399dd）后成为胃中的主要形式；胃蛋白酶原 C 是最晚表达的基因，出现在 14 dph（294dd）。质子泵的表达在 12 dph（252dd），与胃蛋白酶原 A2 的表达时间一致，显示出质子泵和胃蛋白酶原之间极好的转录协调模式。ISH 分析将三种胃蛋白酶原和质子泵的α亚基的表达定位在相同的胃腺细胞中，这证实它们属于泌酸细胞。此外，泌酸细胞从 14 日龄到 40 日龄逐渐发育和增加。在胃黏膜的表面黏液细胞和黏液颈细胞中未检测到胃蛋白酶原或质子泵的转录物。我们的结果表明，鳜鱼胃的功能发育与胃蛋白酶原的表达密切相关。
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