Involvement and expression of growth hormone/insulin-like growth factor member mRNAs in the ovarian development of turbot (Scophthalmus maximus)

Jia, Yudong; Jing, Qiqi; Gao, Yunhong; Huang, Bin

doi:10.1007/s10695-018-0604-z

Cited by 12 publications

(8 citation statements)

References 45 publications

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“…In our previous studies, we cloned and identified the functional properties of hypothalamus-pituitary-ovary-liver (HPOL) axis-related genes, gonadotrophin receptors and estrogen receptors (ers) during the ovarian development of turbot [22][23][24] . The 18s, actb and ctsd are the most stable housekeeping gene used for turbot luteinizing hormone receptor, ers, growth hormone and insulin-like growth factor expression analysis 22,24,25 . Thus, no single reference gene is universally applicable in turbot HPOL axis related genes expression analysis.…”

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confidence: 99%

Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus)

Gao

Huang

et al. 2020

Sci Rep

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Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is a powerful and sensitive method used in gene expression analysis. Suitable reference genes, which are stable under all experimental circumstances and tissues significantly improve the accuracy of qRT-PCR data. In this study, the stability of six genes, namely, 18S ribosomal RNA (18s), beta-actin (actb), elongation factor 1-alpha (ef1α), glyceraldehyde-3-phosphate-dehydrogenase (gapdh), cathepsin D (ctsd), and beta-2-microglobulin (b2m) were evaluated as potential references for qRT-PCR analysis. The genes were examined in the hypothalamus-pituitary-ovary-liver (HPOL) axis throughout turbot ovarian development via using the geNorm, NormFinder and BestKeeper algorithms. Results showed that the most stable reference genes were ef1α, actb, and ctsd in the hypothalamus, pituitary, ovary and liver, respectively. The best-suited gene combinations for normalization were 18s, ef1α, and ctsd in the hypothalamus; actb, ctsd, and 18s in the pituitary; actb, and ctsd in the ovary; gapdh and ctsd in the liver. Moreover, the expression profile of estrogen receptor α (erα) manifested no significant difference normalization to the aforementioned best-suited gene during turbot ovarian development. However, no single gene or pair of genes is suitable as an internal control and account for the amplification differences among the four tissues during ovarian development. In summary, these results provide a basic data for the optimal reference gene selection and obtain highly accurate normalization of qRT-PCR data in HPOL axis-related gene expression analysis during turbot ovarian development. Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is widely used in gene expression analysis because of the advantages of its sensitivity and the accurate detection of mRNA at extremely low transcription levels 1. The successful application of qRT-PCR depends on accurate transcript normalization via the selection of suitable reference genes. The reference genes minimize errors attributed to the use of biological samples, such as experimental operations, and RNA and cDNA qualities 2. The ideal reference gene for qRT-PCR exhibits a stable expression level in all target tissues/cells and should be no affected by various experimental conditions or treatments. In general, the commonly used reference genes mainly include glyceraldehyde-3-phosphate-dehydrogenase (gapdh), 18S ribosomal RNA (18s), beta actin (actb), elongation factor 1-alpha (ef1α), cathepsin D (ctsd), and beta-2-microglobulin-like (b2m) in turbot 3,4 and other fish species 5-8. However, these classical reference genes vary at the transcription level in different tissues, at different developmental stages and experimental conditions or treatments 9-11. Inappropriate reference genes that serve as internal controls can affect the accuracy of qRT-PCR results and cause the significantly different conclusion 5,12,13. Actually, no single reference gene is universally applicable in ...

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confidence: 99%

Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus)

Gao

Huang

et al. 2020

Sci Rep

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“…At a phenotypical level, the size of liver in teleosts tends to increase during vitellogenesis, as it has been shown in several fish species [43,44]. By contrast, the hepatosomatic index tends to decrease during lack of food/fasting, when vitellogenesis is not involved [45,46].…”

Section: Plos Onementioning

confidence: 84%

Unravelling the changes during induced vitellogenesis in female European eel through RNA-Seq: What happens to the liver?

Bertolini

Jørgensen²,

Henkel³

et al. 2020

PLoS ONE

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The life cycle of European eel (Anguilla anguilla), a catadromous species, is complex and enigmatic. In nature, during the silvering process prior to their long spawning migration, reproductive development is arrested, and they cease feeding. In studies of reproduction using hormonal induction, eels are equivalently not feed. Therefore, in female eels that undergo vitellogenesis, the liver plays different, essential roles being involved both in vitellogenins synthesis and in reallocating resources for the maintenance of vital functions, performing the transoceanic reproductive migration and completing reproductive development. The present work aimed at unravelling the major transcriptomic changes that occur in the liver during induced vitellogenesis in female eels. mRNA-Seq data from 16 animals (eight before induced vitellogenesis and eight after nine weeks of hormonal treatment) were generated and differential expression analysis was performed comparing the two groups. This analysis detected 1,328 upregulated and 1,490 downregulated transcripts. Overrepresentation analysis of the upregulated genes included biological processes related to biosynthesis, response to estrogens, mitochondrial activity and localization, while downregulated genes were enriched in processes related to morphogenesis and development of several organs and tissues, including liver and immune system. Among key genes, the upregulated ones included vitellogenin genes (VTG1 and VTG2) that are central in vitellogenesis, together with ESR1 and two novel genes not previously investigated in European eel (LMAN1 and NUPR1), which have been linked with reproduction in other species. Moreover, several upregulated genes, such as CYC1, ELOVL5, KARS and ACSS1, are involved in the management of the effect of fasting and NOTCH, VEGFA and NCOR are linked with development, autophagy and liver maintenance in other species. These results increase the understanding of the molecular changes that occur in the liver during vitellogenesis in this complex and distinctive fish species, bringing new insights on European eel reproduction and broodstock management.

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“…GH is produced by the pituitary gland, and its regulation is governed by the hypothalamic neuropeptides (Bolander, 2004). It also regulates nutrient metabolism along with growth, development, reproduction and some other physiological processes (Abdolahnejad et al., 2015; Jia et al., 2019; Mir et al., 2019; Pérez‐Sanchez et al., 1991; Sakamoto & Hirano, 1991).…”

Section: Discussionmentioning

confidence: 99%

mRNA expression of growth hormone (GH) and Insulin during the initial ontogeny of tropical gar larvae (Atractosteus tropicus)

et al. 2020

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Involvement and expression of growth hormone/insulin-like growth factor member mRNAs in the ovarian development of turbot (Scophthalmus maximus)

Cited by 12 publications

References 45 publications

Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus)

Reference gene validation for quantification of gene expression during ovarian development of turbot (Scophthalmus maximus)

Unravelling the changes during induced vitellogenesis in female European eel through RNA-Seq: What happens to the liver?

mRNA expression of growth hormone (GH) and Insulin during the initial ontogeny of tropical gar larvae (Atractosteus tropicus)

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