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

Autor: Sarthak Mohanty, Robert Pinelli, Chitra Lekha Dahia
Rok vydání: 2019
Předmět:
Zdroj: Journal of Cellular Physiology
ISSN: 1097-4652
0021-9541
DOI: 10.1002/jcp.28952
Popis: 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.
An inducible driver line to precisely target nucleus pulposus (NP) cells during the postnatal is required to understand the role of these cells during development and aging. The current study describes the characterization of Krt19CreERT allele to efficiently target NP cells in neonatal, skeletally mature, middle‐aged, and aged mice using two independent fluorescent reporter lines. Moreover, this allele did not target the surrounding cells in the spine or any cell in the knee joint, validating its specificity to target NP cells in the postnatal mouse.
Databáze: OpenAIRE
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