Mishra Surabhi, Chaube Radha
Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi 221005, India.
Department of Zoology, Institute of Science, Banaras Hindu University, Varanasi 221005, India.
Gen Comp Endocrinol. 2017 Jan 15;241:80-88. doi: 10.1016/j.ygcen.2016.04.031. Epub 2016 May 6.
In vertebrates, steroids are synthesized de novo in the central and peripheral nervous system, independent of peripheral steroidogenic glands, such as the adrenal, gonads and placenta. 3β-Hydroxysteroid dehydrogenase/Δ5-Δ4-isomerase (3β-HSD) is a key steroidogenic enzyme in vertebrate gonads, placenta and adrenal. It mediates the oxidation and isomerization reactions of progesterone from pregnenolone, 17-hydroxyprogesterone from 17-hydroxypregnenolone and androstenedione from dehydroepiandrosterone. In the present study, we examined the expression of 3β-HSD cDNA by real time-PCR and localization of the mRNA by in situ hybridization in the brain and its regions during the different phases of the reproductive cycle of the catfish Heteropneustes fossilis. Further, 3β-HSD activity was assayed biochemically to show seasonal variations. We showed significant seasonal and sexual dimorphic changes in the levels of transcript abundance in the whole brain and its regions. In whole brain, level was the highest in post-spawning phase and lowest in spawning phase in males. In females, there was a progressive increase through resting phase to pre-spawning phase, a decline in the spawning phase and increase in the post-spawning phase. In the preparatory phase, the highest transcript level was seen in medulla oblongata and the lowest in pituitary in males. In females, the level was the highest in the hypothalamus and lowest in olfactory bulb and pituitary. However, in the pre-spawning phase, in males it was the highest in telencephalon and hypothalamus and lowest in pituitary. In females, the highest transcript level was in olfactory bulb and lowest in pituitary. 3β-HSD enzyme activity showed significant seasonal variation in the brain, the highest in the resting phase and lowest in the preparatory and spawning phases. In situ hybridization showed the presence of 3β-HSD transcript was especially high in the cerebellum region. The presence of 3β-HSD in the brain may indicate steroidogenesis in the catfish brain.
在脊椎动物中,类固醇在中枢神经系统和外周神经系统中从头合成,独立于外周类固醇生成腺,如肾上腺、性腺和胎盘。3β-羟基类固醇脱氢酶/Δ5-Δ4-异构酶(3β-HSD)是脊椎动物性腺、胎盘和肾上腺中的一种关键类固醇生成酶。它介导孕烯醇酮转化为孕酮、17-羟孕烯醇酮转化为17-羟孕酮以及脱氢表雄酮转化为雄烯二酮的氧化和异构化反应。在本研究中,我们通过实时PCR检测了鲶鱼(Heteropneustes fossilis)生殖周期不同阶段大脑及其区域中3β-HSD cDNA的表达,并通过原位杂交确定了mRNA的定位。此外,通过生化方法测定了3β-HSD活性以显示季节性变化。我们发现全脑及其区域的转录本丰度水平存在显著的季节性和两性差异变化。在全脑中,雄性在产后阶段水平最高,在产卵阶段最低。在雌性中,从静止期到产卵前期逐渐升高,在产卵期下降,在产后阶段升高。在准备阶段,雄性延髓中的转录本水平最高,垂体中的最低。在雌性中,下丘脑的水平最高,嗅球和垂体中的最低。然而,在产卵前期,雄性端脑和下丘脑中的水平最高,垂体中的最低。在雌性中,转录本水平最高的是嗅球,最低的是垂体。3β-HSD酶活性在大脑中显示出显著的季节性变化,在静止期最高,在准备期和产卵期最低。原位杂交显示小脑区域3β-HSD转录本的存在特别高。大脑中3β-HSD的存在可能表明鲶鱼大脑中存在类固醇生成。