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GATA2 protein is expressed in the TRH neurons in the rat hypothalamic PVN. (A) The specificity of anti-GATA2 antibody. CV1 cells were transfected with empty vector (a and b) or mouse GATA2-expression plasmid (c and d). These cells were stained with 0.1% anti-GATA2 antibody (B9922A, Perseus Proteomics, Japan) (a and c) or DAPI (b and d). Several GATA2-positive cells were detected in (c) but not in (a), while the numbers of nuclei stained with DAPI were comparable between (b) and (d). Transfection efficiency was approximately 5% (compare c and d). (B) (a, b, and c) Immunohistochemical staining of rat PVN with 0.2% anti-TRH antibody. Magnification: ×10 (a), ×40 (b), and ×100 (c). The cytoplasms were stained brown (open arrowhead). The boxed areas in (a) and (b) were magnified in (b) and (c), respectively. (d, e and f) Immunohistochemical staining of the rat PVN with anti-mouse 0.1% GATA2 antibody. Magnification: ×10 (d), ×40 (e) and ×100 (f). The nuclei were strongly stained blue (solid arrowhead). The boxed areas in (d) and (e) were magnified in (e) and (f), respectively. (C) Immunohistochemical double staining of the rat PVN with 0.2% anti-TRH antibody and 0.1% anti-mouse GATA2 antibody (a, b and c). Magnification: ×10 (a), ×40 (b), and ×100 (c). The cytoplasms of TRH neuron were stained brown with anti-TRH antibody (open arrowhead) and nuclei were stained blue with anti-mouse GATA2 antibody (solid arrowhead). The boxed areas in (a) and (b) were magnified in (b) and (c), respectively. https://doi.org/10.1371/journal.pone.0242380.g001
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Thyroid hormone (T3) inhibits thyrotropin-releasing hormone (TRH) synthesis in the hypothalamic paraventricular nucleus (PVN). Although the T3 receptor (TR) β2 is known to mediate the negative regulation of the prepro-TRH gene, its molecular mechanism remains unknown. Our previous studies on the T3-dependent negative regulation of the thyrotropin β...
Contexts in source publication
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... examine GATA2 protein expression in the TRH neurons in rat PVN, we employed an anti-GATA2 antibody [10,29,32]. As shown in Fig 1A-a and 1A-c, the presence of GATA2 protein was detected by this antibody in the nuclei of CV1 cells transfected with GATA2 expression plasmid but not those with empty vector. The numbers of the cell nuclei were comparable between these dishes (Fig 1A-b and 1A-d). ...
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... shown in Fig 1A-a and 1A-c, the presence of GATA2 protein was detected by this antibody in the nuclei of CV1 cells transfected with GATA2 expression plasmid but not those with empty vector. The numbers of the cell nuclei were comparable between these dishes (Fig 1A-b and 1A-d). As shown in Fig 1B, frozen brain sections of adult male rats were immunohistochemically stained with an anti-TRH primary antibody followed by an HRP-conjugated secondary antibody. ...
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... numbers of the cell nuclei were comparable between these dishes (Fig 1A-b and 1A-d). As shown in Fig 1B, frozen brain sections of adult male rats were immunohistochemically stained with an anti-TRH primary antibody followed by an HRP-conjugated secondary antibody. This primary antibody is known to be able to detect the TRH expression in PVN [33,34] and median eminence, where TRH is transported [35]. ...
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... primary antibody is known to be able to detect the TRH expression in PVN [33,34] and median eminence, where TRH is transported [35]. As shown in Fig 1B-a, 1B-b and 1B-c, we detected TRH-positive cells (brown in cytoplasms) in regions corresponding to PVN. Using sections of the PVN, we also observed numerous cells, nuclei of which were stained blue with the anti-GATA2 primary antibody and an ALP-conjugated secondary antibody (Fig 1B-d, 1B-e and 1B-f). ...
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... shown in Fig 1B-a, 1B-b and 1B-c, we detected TRH-positive cells (brown in cytoplasms) in regions corresponding to PVN. Using sections of the PVN, we also observed numerous cells, nuclei of which were stained blue with the anti-GATA2 primary antibody and an ALP-conjugated secondary antibody (Fig 1B-d, 1B-e and 1B-f). Finally, double immunostain using anti-TRH and anti-GATA2 antibodies revealed that the nuclei of the TRH-positive neurons (brown in cytosol) were also GATA2-positive (blue in nuclei), suggesting the expression of GATA2 protein in TRH neurons (Fig 1C-a, 1C-b and 1C- c). ...
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... sections of the PVN, we also observed numerous cells, nuclei of which were stained blue with the anti-GATA2 primary antibody and an ALP-conjugated secondary antibody (Fig 1B-d, 1B-e and 1B-f). Finally, double immunostain using anti-TRH and anti-GATA2 antibodies revealed that the nuclei of the TRH-positive neurons (brown in cytosol) were also GATA2-positive (blue in nuclei), suggesting the expression of GATA2 protein in TRH neurons (Fig 1C-a, 1C-b and 1C- c). ...
Citations
... Notably, these transcription factors are essential for the differentiation of the paraventricular nucleus in the hypothalamus [9]. Our recent studies further revealed that GATA2 is a critical activator of the preproTRH gene [10]. ...
... In 2011, germline heterozygous mutations in GATA2 were identified to cause four previously described hematological syndromes: MonoMAC syndrome, dendritic cell, monocyte, and lymphoid deficiency (DCML), familial myelodysplastic/acute myeloid leukemia syndromes, and Emberger syndrome [16]. Although GATA2 plays an essential role in regulating both the preproTRH and the TSHβ genes [7,10,17,18], little is known about the effect of naturally occuring GATA2 mutations on the HPT axis. ...
... We previously reported that the expression of GATA2 is important for basal activation of rTRH(547)-CAT in CV1 cells that lack endogenous GATA2 expression [10]. We first compared GATA2 mutants with wild-type GATA2 for their ability to transactivate the preproTRH promoter. ...
The transcription factor GATA2 regulates gene expression in several cells and tissues, including hematopoietic tissues and the central nervous system. Recent studies revealed that loss-of-function mutations in GATA2 are associated with hematological disorders. Our earlier in vitro studies showed that GATA2 plays an essential role in the hypothalamus–pituitary–thyroid axis (HPT axis) by regulating the genes encoding prepro-thyrotropin-releasing hormone (preproTRH) and thyroid-stimulating hormone β (TSHβ). However, the effect of GATA2 mutants on the transcriptional activity of their promoters remains unelucidated. In this study, we created five human GATA2 mutations (R308P, T354M, R396Q, R398W, and S447R) that were reported to be associated with hematological disorders and analyzed their functional properties, including transactivation potential and DNA-binding capacity toward the preproTRH and the TSHβ promoters. Three mutations (T354M, R396Q, and R398W) within the C-terminal zinc-finger domain reduced the basal GATA2 transcriptional activity on both the preproTRH and the TSHβ promoters with a significant loss of DNA binding affinity. Interestingly, only the R398W mutation reduced the GATA2 protein expression. Subsequent analysis demonstrated that the R398W mutation possibly facilitated the GATA2 degradation process. R308P and S447R mutants exhibited decreased transcriptional activity under protein kinase C compared to the wild-type protein. In conclusion, we demonstrated that naturally occurring GATA2 mutations impair the HPT axis through differential functional mechanisms in vitro.
The hippocampus is considered the center for learning and memory in the brain, and its development and function is greatly affected by the thyroid and stress axes. Thyroid hormone (TH) and glucocorticoids (GC) are known to have a synergistic effect on developmental programs across several vertebrate species, and their effects on hippocampal structure and function are well-documented. However, there are few studies that focus on the processes and genes that are cooperatively regulated by the two hormone axes. Cross-regulation of the thyroid and stress axes in the hippocampus occurs on multiple levels such that TH can regulate the expression of the GC receptor (GR) while GC can modulate tissue sensitivity to TH by controlling the expression of TH receptor (TR) and enzymes involved in TH biosynthesis. Thyroid hormone and GC are also known to synergistically regulate the transcription of genes associated with neuronal function and development. Synergistic gene regulation by TH and GC may occur through the direct, cooperative action of TR and GR on common target genes, or by indirect mechanisms involving gene regulatory cascades activated by TR and GR. In this chapter, we describe the known physiological effects and underlying molecular mechanisms of TH and GC synergistic gene regulation in the hippocampus.
Hypophysiotropic neurons of the paraventricular nucleus of the hypothalamus
that express thyrotropin-releasing hormone (TRH) control the synthesis and
release of thyrotropin, the pituitary hormone that regulates the synthesis and
release of thyroid hormones. Thyroid hormones are pleiotropic hormones with
multiple functions involved in growth, development, and energy homeostasis.
The TRH neuroendocrine cells receive neuronal inputs from different parts of the
brain, as well as local and hormonal signals, and integrate and transduce the
information as a hormone (TRH) output. They are glutamatergic neurons; most
co-express cocaine- and amphetamine-regulated transcripts; however, their
transcriptomic characterization is still in its infancy but suggests functional
diversity. Transcription of the Trh gene is rapidly but transiently increased by
multiple signals, some of which also cause the release of TRH. This review
summarizes the basic mechanisms involved in the generation of TRH in
hypophysiotropic neurons and turnover in median eminence, and recapitulates
the multiple factors that regulate Trh synthesis and the amount of TRH that
reaches thyrotropes, the physiological conditions and environmental stressors
that alter TRH neurons and thyroid axis status during development and in adult
animals, as well as critical sex differences
