Mice with an N-Ethyl-N-Nitrosourea (ENU) Induced Tyr209Asn Mutation in Natriuretic Peptide Receptor 3 (NPR3) Provide a Model for Kyphosis Associated with Activation of the MAPK Signaling Pathway

Christopher T. Esapa, Sian E. Piret, M. Andrew Nesbit, Nellie Y. Loh, Gethin Thomas, Peter I. Croucher, Matthew A. Brown, Steve D. M. Brown, Roger D. Cox, Rajesh V. Thakker

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Non-syndromic kyphosis is a common disorder that is associated with significant morbidityand has a strong genetic involvement; however, the causative genes remain to be identified,as such studies are hampered by genetic heterogeneity, small families and various modesof inheritance. To overcome these limitations, we investigated 12 week old progeny of micetreated with the chemical mutagen N-ethyl-N-nitrosourea (ENU) using phenotypic assessmentsincluding dysmorphology, radiography, and dual-energy X-ray absorptiometry. Thisidentified a mouse with autosomal recessive kyphosis (KYLB). KYLB mice, when comparedto unaffected littermates, had: thoraco-lumbar kyphosis, larger vertebrae, and increasedbody length and increased bone area. In addition, female KYLB mice had increases in bonemineral content and plasma alkaline phosphatase activity. Recombination mapping localizedthe Kylb locus to a 5.5Mb region on chromosome 15A1, which contained 51 genes,including the natriuretic peptide receptor 3 (Npr3) gene. DNA sequence analysis of Npr3identified a missense mutation, Tyr209Asn, which introduced an N-linked glycosylation consensus sequence. Expression of wild-type NPR3 and the KYLB-associated Tyr209AsnNPR3 mutant in COS-7 cells demonstrated the mutant to be associated with abnormal Nlinkedglycosylation and retention in the endoplasmic reticulum that resulted in its absencefrom the plasma membrane. NPR3 is a decoy receptor for C-type natriuretic peptide (CNP),which also binds to NPR2 and stimulates mitogen-activated protein kinase (MAPK) signaling,thereby increasing the number and size of hypertrophic chondrocytes. Histomorphometricanalysis of KYLB vertebrae and tibiae showed delayed endochondral ossification
LanguageEnglish
Pagese0167916
JournalPLoS ONE
Volume11
Issue number12
DOIs
Publication statusPublished - 13 Dec 2016

Fingerprint

N-ethyl-N-nitrosourea
Ethylnitrosourea
Natriuretic Peptides
Kyphosis
Peptide Receptors
Mitogen-Activated Protein Kinases
mitogen-activated protein kinase
mutagenesis
Genes
Chemical activation
vertebrae
Mutation
mice
Spine
C-Type Natriuretic Peptide
Glycosylation
natriuretic peptides
mutants
missense mutation
dual-energy X-ray absorptiometry

Keywords

  • kyphosis
  • mutation
  • mouse model
  • signalling

Cite this

Esapa, Christopher T. ; Piret, Sian E. ; Nesbit, M. Andrew ; Loh, Nellie Y. ; Thomas, Gethin ; Croucher, Peter I. ; Brown, Matthew A. ; Brown, Steve D. M. ; Cox, Roger D. ; Thakker, Rajesh V. / Mice with an N-Ethyl-N-Nitrosourea (ENU) Induced Tyr209Asn Mutation in Natriuretic Peptide Receptor 3 (NPR3) Provide a Model for Kyphosis Associated with Activation of the MAPK Signaling Pathway. In: PLoS ONE. 2016 ; Vol. 11, No. 12. pp. e0167916.
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abstract = "Non-syndromic kyphosis is a common disorder that is associated with significant morbidityand has a strong genetic involvement; however, the causative genes remain to be identified,as such studies are hampered by genetic heterogeneity, small families and various modesof inheritance. To overcome these limitations, we investigated 12 week old progeny of micetreated with the chemical mutagen N-ethyl-N-nitrosourea (ENU) using phenotypic assessmentsincluding dysmorphology, radiography, and dual-energy X-ray absorptiometry. Thisidentified a mouse with autosomal recessive kyphosis (KYLB). KYLB mice, when comparedto unaffected littermates, had: thoraco-lumbar kyphosis, larger vertebrae, and increasedbody length and increased bone area. In addition, female KYLB mice had increases in bonemineral content and plasma alkaline phosphatase activity. Recombination mapping localizedthe Kylb locus to a 5.5Mb region on chromosome 15A1, which contained 51 genes,including the natriuretic peptide receptor 3 (Npr3) gene. DNA sequence analysis of Npr3identified a missense mutation, Tyr209Asn, which introduced an N-linked glycosylation consensus sequence. Expression of wild-type NPR3 and the KYLB-associated Tyr209AsnNPR3 mutant in COS-7 cells demonstrated the mutant to be associated with abnormal Nlinkedglycosylation and retention in the endoplasmic reticulum that resulted in its absencefrom the plasma membrane. NPR3 is a decoy receptor for C-type natriuretic peptide (CNP),which also binds to NPR2 and stimulates mitogen-activated protein kinase (MAPK) signaling,thereby increasing the number and size of hypertrophic chondrocytes. Histomorphometricanalysis of KYLB vertebrae and tibiae showed delayed endochondral ossification",
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Mice with an N-Ethyl-N-Nitrosourea (ENU) Induced Tyr209Asn Mutation in Natriuretic Peptide Receptor 3 (NPR3) Provide a Model for Kyphosis Associated with Activation of the MAPK Signaling Pathway. / Esapa, Christopher T.; Piret, Sian E.; Nesbit, M. Andrew; Loh, Nellie Y.; Thomas, Gethin; Croucher, Peter I.; Brown, Matthew A.; Brown, Steve D. M.; Cox, Roger D.; Thakker, Rajesh V.

In: PLoS ONE, Vol. 11, No. 12, 13.12.2016, p. e0167916.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Mice with an N-Ethyl-N-Nitrosourea (ENU) Induced Tyr209Asn Mutation in Natriuretic Peptide Receptor 3 (NPR3) Provide a Model for Kyphosis Associated with Activation of the MAPK Signaling Pathway

AU - Esapa, Christopher T.

AU - Piret, Sian E.

AU - Nesbit, M. Andrew

AU - Loh, Nellie Y.

AU - Thomas, Gethin

AU - Croucher, Peter I.

AU - Brown, Matthew A.

AU - Brown, Steve D. M.

AU - Cox, Roger D.

AU - Thakker, Rajesh V.

PY - 2016/12/13

Y1 - 2016/12/13

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AB - Non-syndromic kyphosis is a common disorder that is associated with significant morbidityand has a strong genetic involvement; however, the causative genes remain to be identified,as such studies are hampered by genetic heterogeneity, small families and various modesof inheritance. To overcome these limitations, we investigated 12 week old progeny of micetreated with the chemical mutagen N-ethyl-N-nitrosourea (ENU) using phenotypic assessmentsincluding dysmorphology, radiography, and dual-energy X-ray absorptiometry. Thisidentified a mouse with autosomal recessive kyphosis (KYLB). KYLB mice, when comparedto unaffected littermates, had: thoraco-lumbar kyphosis, larger vertebrae, and increasedbody length and increased bone area. In addition, female KYLB mice had increases in bonemineral content and plasma alkaline phosphatase activity. Recombination mapping localizedthe Kylb locus to a 5.5Mb region on chromosome 15A1, which contained 51 genes,including the natriuretic peptide receptor 3 (Npr3) gene. DNA sequence analysis of Npr3identified a missense mutation, Tyr209Asn, which introduced an N-linked glycosylation consensus sequence. Expression of wild-type NPR3 and the KYLB-associated Tyr209AsnNPR3 mutant in COS-7 cells demonstrated the mutant to be associated with abnormal Nlinkedglycosylation and retention in the endoplasmic reticulum that resulted in its absencefrom the plasma membrane. NPR3 is a decoy receptor for C-type natriuretic peptide (CNP),which also binds to NPR2 and stimulates mitogen-activated protein kinase (MAPK) signaling,thereby increasing the number and size of hypertrophic chondrocytes. Histomorphometricanalysis of KYLB vertebrae and tibiae showed delayed endochondral ossification

KW - kyphosis

KW - mutation

KW - mouse model

KW - signalling

U2 - 10.1371/journal.pone.0167916

DO - 10.1371/journal.pone.0167916

M3 - Article

VL - 11

SP - e0167916

JO - PLoS ONE

T2 - PLoS ONE

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