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儿茶酚胺对鲶鱼(胡鲶)视前区和卵巢中九肽前体mRNA表达的差异调节:一项体外研究

Catecholamines modulate differentially nonapeptide precursor mRNA expression in the preoptic area and ovary of the catfish Heteropneustes fossilis: An in vitro study.

作者信息

Banerjee Putul, Chaube Radha, Joy Keerikkattil Paily

机构信息

Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi 221005, India.

Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi 221005, India.

出版信息

Gen Comp Endocrinol. 2023 Mar 1;333:114211. doi: 10.1016/j.ygcen.2023.114211. Epub 2023 Jan 13.

Abstract

In the catfish Heteropneustes fossilis, three nonapeptide hormone genes were identified in the brain preoptic area (POA) and ovary: a pro-vasotocin (pro-vt) and two isotocin gene paralogs viz., a novel pro-ita and conventional pro-itb. In the present study, the regulatory role of catecholamines [CA: dopamine (DA), noradrenaline (NA), adrenaline (AD)] on the expression of these genes were investigated in vitro. DA (1, 10, and 100 ng/mL) inhibited significantly the mRNA expression in both the POA and ovary. NA upregulated the POA mRNA expression in a biphasic manner, the lower concentrations (1 ng and 10 ng) scaled up and the higher concentration (100 ng) scaled down the expression of pro-vt and pro-itb, while only the 1 ng NA scaled up the pro-ita expression. In the ovary, NA upregulated the mRNA expressions at all concentrations; the pro-vt expression was stimulated only at 10 and 100 ng. AD stimulated pro-vt and pro-ita expression in the POA at all concentrations but the pro-itb expression was inhibited at 1 and 10 ng, and stimulated at 100 ng concentrations. In the ovary, AD elicited varied effects; no significant change in pro-vt, a stimulation of pro-ita, and an inhibition of pro-itb at 1 ng, and stimulation of pro-itb at the 10 and 100 ng. The incubation of the POA and ovary with α-methylparatyrosine (MPT, 250 µg/mL, a tyrosine hydroxylase inhibitor) for 8 h downregulated the mRNA expression in the POA but unaltered the expression in the ovary. Pre-incubation with MPT for 4 h, followed by co-incubation with DA, NA or AD for 4 h elicited varied effects. In the POA, the co-incubations with the CAs rescued the inhibition due to MPT. The MPT + DA and MPT + AD treatments reduced the magnitude of the inhibition of pro-vt and pro-itb by MPT. But the pro-ita expression was modestly stimulated in the MPT + AD group. On the other hand, the MPT + NA treatment rescued the MPT effect and elicited 10-folds increase in the expression levels. In the ovary, the changes were: an inhibition in the MPT + DA group, no significant alteration in the MPT + NA group, and a mild stimulation in the MPT + AD group. The results suggest that CAs modulate brain and ovarian nonapeptide gene expression differentially, which is important in the neuroendocrine/endocrine integration of reproduction in the catfish.

摘要

在鲶鱼(Heteropneustes fossilis)中,在脑视前区(POA)和卵巢中鉴定出三种九肽激素基因:一种血管加压素原(pro-vt)和两种异催产素基因旁系同源物,即一种新型的pro-ita和传统的pro-itb。在本研究中,体外研究了儿茶酚胺[CA:多巴胺(DA)、去甲肾上腺素(NA)、肾上腺素(AD)]对这些基因表达的调节作用。DA(1、10和100 ng/mL)显著抑制POA和卵巢中的mRNA表达。NA以双相方式上调POA中的mRNA表达,较低浓度(1 ng和10 ng)上调,较高浓度(100 ng)下调pro-vt和pro-itb的表达,而只有1 ng NA上调pro-ita的表达。在卵巢中,NA在所有浓度下均上调mRNA表达;pro-vt表达仅在10和100 ng时受到刺激。AD在所有浓度下均刺激POA中的pro-vt和pro-ita表达,但pro-itb表达在1和10 ng时受到抑制,在100 ng浓度时受到刺激。在卵巢中,AD产生不同的影响;pro-vt无显著变化,pro-ita在1 ng时受到刺激,pro-itb在1 ng时受到抑制,在10和100 ng时受到刺激。将POA和卵巢与α-甲基对酪氨酸(MPT,250 μg/mL,一种酪氨酸羟化酶抑制剂)孵育8小时下调了POA中的mRNA表达,但未改变卵巢中的表达。用MPT预孵育4小时,然后与DA、NA或AD共孵育4小时产生了不同的影响。在POA中,与儿茶酚胺的共孵育挽救了由于MPT引起的抑制作用。MPT + DA和MPT + AD处理降低了MPT对pro-vt和pro-itb的抑制程度。但在MPT + AD组中,pro-ita表达受到适度刺激。另一方面,MPT + NA处理挽救了MPT的作用,并使表达水平增加了10倍。在卵巢中,变化如下:MPT + DA组受到抑制,MPT + NA组无显著变化,MPT + AD组受到轻度刺激。结果表明,儿茶酚胺对脑和卵巢九肽基因表达的调节存在差异,这在鲶鱼繁殖的神经内分泌/内分泌整合中很重要。

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