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A Novel Gonadotropin-Releasing Hormone 1 (Gnrh1) Enhancer-Derived Noncoding RNA Regulates Gnrh1 Gene Expression in GnRH Neuronal Cell Models.

PloS one | 2016

Gonadotropin-releasing hormone (GnRH), a neuropeptide released from a small population of neurons in the hypothalamus, is the central mediator of the hypothalamic-pituitary-gonadal axis, and is required for normal reproductive development and function. Evolutionarily conserved regulatory elements in the mouse, rat, and human Gnrh1 gene include three enhancers and the proximal promoter, which confer Gnrh1 gene expression specifically in GnRH neurons. In immortalized mouse hypothalamic GnRH (GT1-7) neurons, which show pulsatile GnRH release in culture, RNA sequencing and RT-qPCR revealed that expression of a novel long noncoding RNA at Gnrh1 enhancer 1 correlates with high levels of GnRH mRNA expression. In GT1-7 neurons, which contain a transgene carrying 3 kb of the rat Gnrh1 regulatory region, both the mouse and rat Gnrh1 enhancer-derived noncoding RNAs (GnRH-E1 RNAs) are expressed. We investigated the characteristics and function of the endogenous mouse GnRH-E1 RNA. Strand-specific RT-PCR analysis of GnRH-E1 RNA in GT1-7 cells revealed GnRH-E1 RNAs that are transcribed in the sense and antisense directions from distinct 5' start sites, are 3' polyadenylated, and are over 2 kb in length. These RNAs are localized in the nucleus and have a half-life of over 8 hours. In GT1-7 neurons, siRNA knockdown of mouse GnRH-E1 RNA resulted in a significant decrease in the expression of the Gnrh1 primary transcript and Gnrh1 mRNA. Over-expression of either the sense or antisense mouse GnRH-E1 RNA in immature, migratory GnRH (GN11) neurons, which do not express either GnRH-E1 RNA or GnRH mRNA, induced the transcriptional activity of co-transfected rat Gnrh1 gene regulatory elements, where the induction requires the presence of the rat Gnrh1 promoter. Together, these data indicate that GnRH-E1 RNA is an inducer of Gnrh1 gene expression. GnRH-E1 RNA may play an important role in the development and maturation of GnRH neurons.

Pubmed ID: 27389022 RIS Download

Research resources used in this publication

None found

Antibodies used in this publication

None found

Associated grants

  • Agency: NICHD NIH HHS, United States
    Id: T32 HD007203
  • Agency: NIEHS NIH HHS, United States
    Id: P42 ES010337
  • Agency: NCI NIH HHS, United States
    Id: U54 CA155435
  • Agency: NICHD NIH HHS, United States
    Id: R01 HD020377
  • Agency: NIDDK NIH HHS, United States
    Id: P30 DK063491
  • Agency: NIDDK NIH HHS, United States
    Id: T32 DK007541
  • Agency: BLRD VA, United States
    Id: I01 BX002709
  • Agency: NCI NIH HHS, United States
    Id: P30 CA023100
  • Agency: NIDDK NIH HHS, United States
    Id: T32 DK007494
  • Agency: NICHD NIH HHS, United States
    Id: F32 HD058427
  • Agency: NICHD NIH HHS, United States
    Id: U54 HD012303
  • Agency: NICHD NIH HHS, United States
    Id: R01 HD072754
  • Agency: NICHD NIH HHS, United States
    Id: R01 HD082567
  • Agency: NICHD NIH HHS, United States
    Id: P50 HD012303
  • Agency: NCI NIH HHS, United States
    Id: R01 CA196853

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Ambion Inc. (tool)

RRID:SCR_008406

A division of Applied Biosystems selling products for the isolation, detection, quantification, amplification, and characterization of RNA.

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FastQC (tool)

RRID:SCR_014583

Quality control software that perform checks on raw sequence data coming from high throughput sequencing pipelines. This software also provides a modular set of analyses which can give a quick impression of the quality of the data prior to further analysis.

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Cufflinks (tool)

RRID:SCR_014597

Software tool for transcriptome assembly and differential expression analysis for RNA-Seq. Includes script called cuffmerge that can be used to merge together several Cufflinks assemblies. It also handles running Cuffcompare as well as automatically filtering a number of transfrags that are likely to be artifacts. If the researcher has a reference GTF file, the researcher can provide it to the script to more effectively merge novel isoforms and maximize overall assembly quality.

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GT1-7 (tool)

RRID:CVCL_0281

Cell line GT1-7 is a Transformed cell line with a species of origin Mus musculus (Mouse)

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NIH 3T3 (tool)

RRID:CVCL_0594

Cell line NIH 3T3 is a Spontaneously immortalized cell line with a species of origin Mus musculus

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