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Characterization of Krt19 (CreERT) allele for targeting the nucleus pulposus cells in the postnatal mouse intervertebral disc

Intervertebral disc degeneration and associated back pain are relatively common but sparsely understood conditions, affecting over 70% of the population during some point of life. Disc degeneration is often associated with a loss of nucleus pulposus (NP) cells. Genetic mouse models offer convenient...

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Autores principales: Mohanty, Sarthak, Pinelli, Robert, Dahia, Chitra Lekha
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6778700/
https://www.ncbi.nlm.nih.gov/pubmed/31187500
http://dx.doi.org/10.1002/jcp.28952
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author Mohanty, Sarthak
Pinelli, Robert
Dahia, Chitra Lekha
author_facet Mohanty, Sarthak
Pinelli, Robert
Dahia, Chitra Lekha
author_sort Mohanty, Sarthak
collection PubMed
description Intervertebral disc degeneration and associated back pain are relatively common but sparsely understood conditions, affecting over 70% of the population during some point of life. Disc degeneration is often associated with a loss of nucleus pulposus (NP) cells. Genetic mouse models offer convenient avenues to understand the cellular and molecular regulation of the disc during its formation, growth, maintenance, and aging. However, due to the lack of inducible driver lines to precisely target NP cells in the postnatal mouse disc, progress in this area of research has been moderate. NP cells are known to express cytokeratin 19 (Krt19), and tamoxifen (Tam)‐inducible Krt19 (CreERT) allele is available. The current study describes the characterization of Krt19 (CreERT) allele to specifically and efficiently target NP cells in neonatal, skeletally mature, middle‐aged, and aged mice using two independent fluorescent reporter lines. The efficiency of recombination at all ages was validated by immunostaining for KRT19. Results show that following Tam induction, Krt19 (CreERT) specifically drives recombination of NP cells in the spine of neonatal and aged mice, while no recombination was detected in the surrounding tissues. Knee joints from skeletally mature Tam‐treated Krt19 (CreERT/+); R26 (tdTOM) mouse show the absence of recombination in all tissues and cells of the knee joint. Thus, this study provides evidence for the use of Krt19 (CreERT) allele for genetic characterization of NP cells at different stages of the mouse life.
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spelling pubmed-67787002019-12-19 Characterization of Krt19 (CreERT) allele for targeting the nucleus pulposus cells in the postnatal mouse intervertebral disc Mohanty, Sarthak Pinelli, Robert Dahia, Chitra Lekha J Cell Physiol Original Research Articles Intervertebral disc degeneration and associated back pain are relatively common but sparsely understood conditions, affecting over 70% of the population during some point of life. Disc degeneration is often associated with a loss of nucleus pulposus (NP) cells. Genetic mouse models offer convenient avenues to understand the cellular and molecular regulation of the disc during its formation, growth, maintenance, and aging. However, due to the lack of inducible driver lines to precisely target NP cells in the postnatal mouse disc, progress in this area of research has been moderate. NP cells are known to express cytokeratin 19 (Krt19), and tamoxifen (Tam)‐inducible Krt19 (CreERT) allele is available. The current study describes the characterization of Krt19 (CreERT) allele to specifically and efficiently target NP cells in neonatal, skeletally mature, middle‐aged, and aged mice using two independent fluorescent reporter lines. The efficiency of recombination at all ages was validated by immunostaining for KRT19. Results show that following Tam induction, Krt19 (CreERT) specifically drives recombination of NP cells in the spine of neonatal and aged mice, while no recombination was detected in the surrounding tissues. Knee joints from skeletally mature Tam‐treated Krt19 (CreERT/+); R26 (tdTOM) mouse show the absence of recombination in all tissues and cells of the knee joint. Thus, this study provides evidence for the use of Krt19 (CreERT) allele for genetic characterization of NP cells at different stages of the mouse life. John Wiley and Sons Inc. 2019-06-11 2020-01 /pmc/articles/PMC6778700/ /pubmed/31187500 http://dx.doi.org/10.1002/jcp.28952 Text en © 2019 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Research Articles
Mohanty, Sarthak
Pinelli, Robert
Dahia, Chitra Lekha
Characterization of Krt19 (CreERT) allele for targeting the nucleus pulposus cells in the postnatal mouse intervertebral disc
title Characterization of Krt19 (CreERT) allele for targeting the nucleus pulposus cells in the postnatal mouse intervertebral disc
title_full Characterization of Krt19 (CreERT) allele for targeting the nucleus pulposus cells in the postnatal mouse intervertebral disc
title_fullStr Characterization of Krt19 (CreERT) allele for targeting the nucleus pulposus cells in the postnatal mouse intervertebral disc
title_full_unstemmed Characterization of Krt19 (CreERT) allele for targeting the nucleus pulposus cells in the postnatal mouse intervertebral disc
title_short Characterization of Krt19 (CreERT) allele for targeting the nucleus pulposus cells in the postnatal mouse intervertebral disc
title_sort characterization of krt19 (creert) allele for targeting the nucleus pulposus cells in the postnatal mouse intervertebral disc
topic Original Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6778700/
https://www.ncbi.nlm.nih.gov/pubmed/31187500
http://dx.doi.org/10.1002/jcp.28952
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