Isolation and characterization of cyp19a1a and cyp19a1b promoters in the protogynous hermaphrodite orange-spotted grouper (Epinephelus coioides)

Zhang, Weimin; Lu, Huijie; Jiang, Haiyan; Zhang, Shen; Liu, Qiongyou; Zhang, Lihong

doi:10.1016/j.ygcen.2011.12.005

Cited by 26 publications

(12 citation statements)

References 56 publications

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“…Similarly, most studies investigating a role for E2 and xenoestrogens in modulating brain function have focused on the estrogen induced up-regulation of the aromatase B gene ( cyp19a1b ) 68 – 70 which has been shown to be mediated through activation of nuclear estrogen receptors and binding to estrogen response elements in the cyp19a1b promoter 71 . While we did not identify any transcription factors known for modulating cyp19a1b gene expression in the CI list, we identified closely related proteins including regulatory factor X3 (RFX3) 72 , CAMP responsive element binding protein 3 (CREB3) 73 , CAMP responsive element binding protein 5 (CREB5) 73 , and POU class 3 homeobox 2 (POUF3) 72 in at least one sample. Results presented herein suggest that estrogen and progesterone receptor agonism, and potentially androgen receptor agonism, is capable of also activating rapid signaling mechanisms and may influence neurogenesis through phosphorylation of target proteins, potentially through activation of membrane-bound receptors.…”

Section: Discussionmentioning

confidence: 89%

Early phosphoproteomic changes for adverse outcome pathway development in the fathead minnow (Pimephales promelas) brain

Smith

Lavelle

Silva-Sanchez

et al. 2018

Sci Rep

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Adverse outcome pathways (AOPs) are conceptual frameworks that organize and link contaminant-induced mechanistic molecular changes to adverse biological responses at the individual and population level. AOPs leverage molecular and high content mechanistic information for regulatory decision-making, but most current AOPs for hormonally active agents (HAAs) focus on nuclear receptor-mediated effects only despite the overwhelming evidence that HAAs also activate membrane receptors. Activation of membrane receptors triggers non-genomic signaling cascades often transduced by protein phosphorylation leading to phenotypic changes. We utilized label-free LC-MS/MS to identify proteins differentially phosphorylated in the brain of fathead minnows (Pimephales promelas) aqueously exposed for 30 minutes to two HAAs, 17α-ethinylestradiol (EE2), a strong estrogenic substance, and levonorgestrel (LNG), a progestin, both components of the birth control pill. EE2 promoted differential phosphorylation of proteins involved in neuronal processes such as nervous system development, synaptic transmission, and neuroprotection, while LNG induced differential phosphorylation of proteins involved in axon cargo transport and calcium ion homeostasis. EE2 and LNG caused similar enrichment of synaptic plasticity and neurogenesis. This study is the first to identify molecular changes in vivo in fish after short-term exposure and highlights transduction of rapid signaling mechanisms as targets of HAAs, in addition to nuclear receptor-mediated pathways.

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Section: Discussionmentioning

confidence: 89%

Early phosphoproteomic changes for adverse outcome pathway development in the fathead minnow (Pimephales promelas) brain

Smith

Lavelle

Silva-Sanchez

et al. 2018

Sci Rep

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“…Then 1 lg total RNA was reverse transcribed with oligo (dT) 18 primers using the RevertAid H Minus First Strand cDNA Synthesis Kit (Fermentas) according to the manufacturer's instruction. The integrity of all the RNA samples was verified by the successful amplification of actb (actin, beta; AY510710).…”

Section: Rt-pcr Analysis Of Lrh-1 Mrna Tissue Patternsmentioning

confidence: 99%

“…The orange-spotted grouper cyp19a1a promoter reporter plasmids were constructed in our previous study [18]. The cDNA fragment encoding the orange-spotted grouper Lrh-1 was amplified with primers EcLRH1-F and EcLRH1a-R1, and inserted into pcDNA3.0 vector between EcoR I and Xho I restriction sites to generate the Lrh-1 expression construct.…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…All the cell culture media contained 1 mg/ml penicillin-streptomycin (Gibco), and cells were maintained at 378C in a humidified incubator under 5% CO 2 . TM4 cell line was chosen in the present study based on our previous report [18].…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…Our previous study showed that the orange-spotted grouper cyp19a1a promoter had two conserved Ftz-f1 binding sites, which could mediate the activation of cyp19a1a promoter by Ftz-f1 homologues [18]. However, a previously isolated Ftz-f1 homologue (Nr5a4) from the orangespotted grouper [19] could not activate cyp19a1a transcription in vitro [18]. In the present study, a Lrh-1 homologue is isolated from the orange-spotted grouper, and its possible involvement in the regulation of cyp19a1a in the ovary is explored.…”

Section: Introductionmentioning

confidence: 99%

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Cytoplasmic Localization of Lrh-1 Down-Regulates Ovarian Follicular cyp19a1a Expression in a Teleost, the Orange-Spotted Grouper Epinephelus coioides1

Zhang

Liu

et al. 2014

Biology of Reproduction

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Liver receptor homolog-1 (LRH-1) is a conserved member of the NR5A subfamily in vertebrates and a potential regulator of estrogen synthesis in the ovarian granulosa cells. An Lrh-1 homologue was obtained from the orange-spotted grouper Epinephelus coioides that contains the conserved structural features of NR5A and is phylogenetically closely related to NR5A2. The expression of the orange-spotted grouper Lrh-1 is tissue-specific with relatively higher levels in the liver and ovary. The immunoreactive signals for Lrh-1 and Cyp19a1a were present in the ovarian follicular cells and germ cells. In the ovarian follicular cells, Lrh-1 was present both in the nucleus and cytoplasm, and colocalized with Cyp19a1a. The expression levels of both increased during vitellogenesis whereas only Cyp19a1a dramatically decreased toward maturation when Lrh-1 was localized almost exclusively to the cytoplasm of the follicular cells. The orange-spotted grouper Lrh-1 could upregulate cyp19a1a transcription in vitro via the two conserved Ftz-f1 sites in cyp19a1a promoter. Chromatin immunoprecipitation analysis showed that the orange-spotted grouper Lrh-1 could bind cyp19a1a promoter in vivo with a higher abundance in the vitellogenic ovary, whereas the binding was dramatically decreased in the mature ovary. Taken together, the results of present study demonstrate that Lrh-1 plays an important role in up-regulating cyp19a1a gene in the ovarian follicular cells during vitellogenesis, and the sequestration of Lrh-1 to the cytoplasm may down-regulate cyp19a1a expression in the mature ovary. This mechanism for modifying transcriptional roles of the orange-spotted grouper Lrh-1 may shed new light on the regulation of Cyp19a1 expression in other vertebrates as well.

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Diethylstilbestrol impaired oogenesis of yellow catfish juveniles through disrupting hypothalamic–pituitary–gonadal axis and germ cell development

Wang

Chen

et al. 2017

J of Applied Toxicology

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Diethylstilbestrol (DES), a non-steroidal estrogen, has been found to cause altered germ cell development and disordered ovarian development in fish females. However, the mechanisms that might be involved are poorly understood. In this study, female juveniles of yellow catfish (Pelteobagrus fulvidraco) (120 days post-hatching) were exposed to two doses (10 and 100 ng l ) of DES for 28 days. After the endpoint of exposure, decreased ovary weight and gonadosomatic index, as well as various ovarian impairments were observed in response to DES. Besides, DES elevated the mRNA levels of vitellogenin 1 (vtg 1) and estrogen receptor 1 (esr 1) in liver and decreased 17β-estradiol level in plasma. Correspondingly, suppressed mRNA levels of the key genes in the hypothalamic-pituitary-gonadal axis (such as cyp19a1b, gnrh-II, fshβ and lhβ in brain and fshr, lhr and cyp19a1a in ovary) after DES exposure were also observed. The declined level of plasma 17β-estradiol and altered gene expressions of genes in the hypothalamic-pituitary-gonadal axis were thus supposed to be closely related to the disrupted oogenesis in DES-treated fish. Analyses further demonstrated that, higher concentration of DES elevated the expression ratio of bax/bcl-2, indicating the enhanced apoptosis occurred in ovary. Moreover, DES upregulated the expressions of genes involved in proliferation (cyclin d1 and pcna), meiotic entry (cyp26a1 and scp3) and meiotic maintenance (dmc1), resulting in arrested oogenesis in catfish. The present study greatly extended our understanding on the mechanisms underlying of reproductive toxicity of DES on fish oogenesis.

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Isolation and characterization of cyp19a1a and cyp19a1b promoters in the protogynous hermaphrodite orange-spotted grouper (Epinephelus coioides)

Cited by 26 publications

References 56 publications

Early phosphoproteomic changes for adverse outcome pathway development in the fathead minnow (Pimephales promelas) brain

Early phosphoproteomic changes for adverse outcome pathway development in the fathead minnow (Pimephales promelas) brain

Cytoplasmic Localization of Lrh-1 Down-Regulates Ovarian Follicular cyp19a1a Expression in a Teleost, the Orange-Spotted Grouper Epinephelus coioides1

Diethylstilbestrol impaired oogenesis of yellow catfish juveniles through disrupting hypothalamic–pituitary–gonadal axis and germ cell development

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