Dataset Information

Regulation and Function of C-Type Natriuretic Peptide (CNP) in Gonadotrope-Derived Cell Lines.

ABSTRACT: C-type natriuretic peptide (CNP) is the most conserved member of the mammalian natriuretic peptide family, and is implicated in the endocrine regulation of growth, metabolism and reproduction. CNP is expressed throughout the body, but is particularly abundant in the central nervous system and anterior pituitary gland. Pituitary gonadotropes are regulated by pulsatile release of gonadotropin releasing hormone (GnRH) from the hypothalamus, to control reproductive function. GnRH and CNP reciprocally regulate their respective signalling pathways in ?T3-1 gonadotrope cells, but effects of pulsatile GnRH stimulation on CNP expression has not been explored. Here, we examine the sensitivity of the natriuretic peptide system in L?T2 and ?T3-1 gonadotrope cell lines to continuous and pulsatile GnRH stimulation, and investigate putative CNP target genes in gonadotropes. Multiplex RT-qPCR assays confirmed that primary mouse pituitary tissue express Nppc, Npr2 (encoding CNP and guanylyl cyclase B (GC-B), respectively) and Furin (a CNP processing enzyme), but failed to express transcripts for Nppa or Nppb (encoding ANP and BNP, respectively). Pulsatile, but not continuous, GnRH stimulation of L?T2 cells caused significant increases in Nppc and Npr2 expression within 4 h, but failed to alter natriuretic peptide gene expression in ?T3-1 cells. CNP enhanced expression of cJun, Egr1, Nr5a1 and Nr0b1, within 8 h in L?T2 cells, but inhibited Nr5a1 expression in ?T3-1 cells. Collectively, these data show the gonadotrope natriuretic peptide system is sensitive to pulsatile GnRH signalling, and gonadotrope transcription factors are putative CNP-target genes. Such findings represent additional mechanisms by which CNP may regulate reproductive function.

SUBMITTER: Mirczuk SM

PROVIDER: S-EPMC6769446 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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Regulation and Function of C-Type Natriuretic Peptide (CNP) in Gonadotrope-Derived Cell Lines.

Mirczuk Samantha M SM Lessey Andrew J AJ Catterick Alice R AR Perrett Rebecca M RM Scudder Christopher J CJ Read Jordan E JE Lipscomb Victoria J VJ Niessen Stijn J SJ Childs Andrew J AJ McArdle Craig A CA McGonnell Imelda M IM Fowkes Robert C RC

Cells 20190914 9

C-type natriuretic peptide (CNP) is the most conserved member of the mammalian natriuretic peptide family, and is implicated in the endocrine regulation of growth, metabolism and reproduction. CNP is expressed throughout the body, but is particularly abundant in the central nervous system and anterior pituitary gland. Pituitary gonadotropes are regulated by pulsatile release of gonadotropin releasing hormone (GnRH) from the hypothalamus, to control reproductive function. GnRH and CNP reciprocall ...[more]

PMID: 31540096

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Project description:ObjectiveThis study sought to correlate the presence of pleural-based B-lines seen by emergency department ultrasound performed with the linear transducer with B-type natriuretic peptide (BNP) level in patients with suspected congestive heart failure.MethodsThe study was a prospective convenience sample on adult patients in an academic, urban emergency department with over 100,000 annual patient visits. Adult patients with a BNP level ordered by the treating physician were prospectively enrolled by one of four physicians, blinded to the BNP level. The enrolling physicians included an emergency ultrasound director, two emergency ultrasound fellows, and a senior emergency medicine resident. Bedside ultrasound was performed using a 3-12 MHz linear broadband transducer in four lung fields. The serum BNP level was correlated with bilateral B-lines, defined as three or more comet-tail artifacts arising from the pleural line extending to the far field without a decrease in intensity on the right and left thorax.ResultsSixty three patients were consented and enrolled during a four-month period. Fifteen patients had the presence of bilateral B-lines. The median BNP in patients with bilateral B-lines was 1560 pg/mL (95% confidence interval (CI) 1141-3706 pg/mL), compared with 538 pg/mL (95% confidence interval 310-1917 pg/mL) in patients without B-lines. The distributions in the two groups differed significantly (p=0.0006). Based on the threshold level of BNP 500 pg/mL, the sensitivity of finding bilateral B-lines on ultrasound was 33.3% (95% CI: 0.19-0.50), and the specificity was 91.7% (95% CI: 0.73-0.99). In addition, bilateral B-lines were absent in all patients with a BNP<100 pg/mL.ConclusionThe presence of bilateral B-lines identified with the linear probe is associated with significantly higher BNP levels than patients without B-lines. In our patient population, the presence of B-lines was specific but not sensitive for BNP>500. Further research may show that it can be applied to quickly assess patients with undifferentiated dyspnea.

Project description:C-type natriuretic peptide (CNP) has been recently identified as an important anabolic regulator of endochondral bone growth, but the molecular mechanism mediating these effects are not completely understood. Here we demonstrate that CNP activates the p38 MAP kinase pathway in chondrocytes and that pharmacological inhibition of p38 blocks the anabolic effects of CNP in a tibia organ culture system. We further show that CNP stimulates endochondral bone growth largely through expansion of the hypertrophic zone of the growth plate, while delaying mineralization. Both effects are reversed by p38 inhibition. We performed Affymetrix microarray analyses to identify CNP target genes in the organ culture system. These studies confirmed that hypertrophic chondrocytes are the main targets of CNP signaling in the growth plate, potentially because cGMP-dependent kinases I and II, important transducers of CNP signaling and are expressed at much higher levels in these cells than in other areas of the tibia. One of the genes most strongly induced by CNP was the Ptgs2 gene, encoding Cox2. Real-time PCR confirmed that Cox2 expression was induced by CNP in hypertrophic chondrocytes, but surprisingly in a p38-independent manner. Moreover, Cox2 inhibition â in contrast to p38 inhibition - did not block the anabolic effects of CNP. In summary, our data identify novel target genes of CNP and demonstrate that the p38 pathway is a novel, essential mediator of CNP effects on endochondral ossification, with potential implications for numerous skeletal diseases. Experiment Overall Design: Tibiae from E15.5 day old embryonic mice were isolated and cultured in minimal media in the presence of vehicle, BSA/HCl (1mM), or C-type natriuretic peptide, CNP (10-6M). On the sixth day of treatment cultured tibias were micro-dissected into the resting/proliferating, hypertrophic, and mineralized areas. Distinct zones from approximately 24 bones were pooled together, from which RNA was isolated using the Qiagen RNeasy Lipid Extraction Kit. Once the quality of total RNA from three independent trials was determined using the Agilent 2100 bioanalyzer, microarray analyses were performed at the London Regional Genomics Centre using MOE430_2.0 Affymetrix arrays. Results were analyzed using GeneSpring 7.2 software.

Dataset Information

Regulation and Function of C-Type Natriuretic Peptide (CNP) in Gonadotrope-Derived Cell Lines.

Publications

Regulation and Function of C-Type Natriuretic Peptide (CNP) in Gonadotrope-Derived Cell Lines.

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