Targeting the annulus fibrosus of the intervertebral disc: Col1a2-Cre(ER)T mice show specific activity of Cre recombinase in the outer annulus fibrosus

Bedore, Jake; Quesnel, Katherine; Quinonez, Diana; Séguin, Cheryle A.; Leask, Andrew

doi:10.1007/s12079-016-0329-7

Cited by 14 publications

(15 citation statements)

References 37 publications

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“…TM inducible Col2‐creERT 42 and Col1‐creERT 43 mice have been shown to express Cre specifically in the IAF, GP, EP and NP, and OAF of the IVD, respectively. To further determine whether primary cilia involve in IVD development, maintenance, and degeneration, we deleted IFT80 in mouse IVD using Col2‐creERT and Col1‐creERT mice, respectively.…”

Section: Discussionmentioning

confidence: 99%

Ciliary IFT80 is essential for intervertebral disc development and maintenance

Yang

Han

et al. 2020

FASEB j.

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The intervertebral disc degeneration (IVDD)-related diseases occur in more than 90% of the population older than 50 years. Owing to the lack of understanding of the cellular mechanisms involved in IVDD formation effective treatment options are still unavailable. Primary cilia are microtubule-based organelles that play important roles in the organ development. Intraflagellar transport (IFT) proteins are essential for the assembly and bidirectional transport within the cilium. Role of cilia and IFT80 protein in intervertebral disc (IVD) development, maintenance, and degeneration are largely unknown. Using cilia-GFP mice, we found presence of cilia on growth plate (GP), cartilage endplate (EP) annulus fibrosus (AF), and nucleus pulposus (NP) with varying ciliary length. Cilia length in NP and AF during IVDD were significantly decreased. However, cilia numbers increased by 63% in AF during repair. Deletion of IFT80 in type II collagen-positive cells resulted in cilia loss in GP and EP, and disrupted IVD structure with disorganized and decreased GP, EP, and internal AF (IAF), and less compact and markedly decreased gel-like matrix in the NP. Deletion of IFT80 in type I collagen-positive cells led to a disorganized outer AF (OAF) with thinner, loosened, and disconnected fiber alignment. Mechanistic analyses showed that loss of IFT80 caused a significant increase in cell apoptosis in the IVD, and a marked decrease in expression of chondrogenic markers -type II collagen, sox9, aggrecan, and hedgehog (Hh) signaling components, including Gli1 and Patch1 in the IVD of IFT80 fl/fl ; Col2-creERT mice, and Gli1 and Patch1 expression in the OAF of IFT80 fl/fl ; Col1-creERT mice. Interestingly, Smoothened agonist-SAG rescued OAF cell proliferation and osteogenic differentiation. Our findings demonstrate that ciliary IFT80 is important for the maintenance of IVD cell organization and function through regulating the cell survival and Hh signaling. K E Y W O R D S annulus fibrosus, IFT80, intervertebral disc degeneration, nucleus pulposus, primary cilia 6742 | LI et aL.

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Section: Discussionmentioning

confidence: 99%

Ciliary IFT80 is essential for intervertebral disc development and maintenance

Yang

Han

et al. 2020

FASEB j.

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“…32 The majority of the rest target one of the three regions of the IVD. [33][34][35][36][37] The consequences of mutations in OSX on the IVD are unknown, but are associated with osteogenesis imperfect in global mutations 38…”

Section: Osterix In Healthy Ivdmentioning

confidence: 99%

Aging aggravates intervertebral disc degeneration by regulating transcription factors toward chondrogenesis

Silva¹,

Holguin

2019

FASEB j.

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Osterix is a critical transcription factor of mesenchymal stem cell fate, where its loss or loss of Wnt signaling diverts differentiation to a chondrocytic lineage. Intervertebral disc (IVD) degeneration activates the differentiation of prehypertrophic chondrocyte‐like cells and inactivates Wnt signaling, but its interactive role with osterix is unclear. First, compared to young‐adult (5 mo), mechanical compression of old (18 mo) IVD induced greater IVD degeneration. Aging (5 vs 12 mo) and/or compression reduced the transcription of osterix and notochordal marker T by 40‐75%. Compression elevated the transcription of hypertrophic chondrocyte marker MMP13 and pre‐osterix transcription factor RUNX2, but less so in 12 mo IVD. Next, using an Ai9/td reporter and immunohistochemical staining, annulus fibrosus and nucleus pulposus cells of young‐adult IVD expressed osterix, but aging and compression reduced its expression. Lastly, in vivo LRP5‐deficiency in osterix‐expressing cells inactivated Wnt signaling in the nucleus pulposus by 95%, degenerated the IVD to levels similar to aging and compression, reduced the biomechanical properties by 45‐70%, and reduced the transcription of osterix, notochordal markers and chondrocytic markers by 60‐80%. Overall, these data indicate that age‐related inactivation of Wnt signaling in osterix‐expressing cells may limit regeneration by depleting the progenitors and attenuating the expansion of chondrocyte‐like cells.

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“…Each of these components contains distinct cell types that are believed to express specific matrix components, as well as signal molecules and growth factors to delicately regulate IVD development and homeostasis. Disruption of these regulations may cause IVD degeneration (Bedore, Quesnel, Quinonez, Seguin, & Leask, ). However, efforts to identify the contribution of the specific cell type to the disease development are hindered by the limited tools that can target specific cell type populations within the IVDs.…”

Section: Introductionmentioning

confidence: 99%

Characterization of Cre recombinase mouse lines enabling cell type‐specific targeting of postnatal intervertebral discs

Zheng

Liu

et al. 2019

Journal Cellular Physiology

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Cre/loxP technology is an important tool for studying cell type‐specific gene functions. Cre recombinase mouse lines, including Agc1‐CreERT2, Col2a1‐Cre; Col2a1‐CreERT2, Shh‐Cre, Shh‐CreERT2, and Osx‐Cre, have been proven to be valuable tools to elucidate the biology of long bones, yet the information for their activity in postnatal intervertebral disc (IVD) tissues was very limited. In this study, we used R26‐mTmG fluorescent reporter to systematically analyze cell specificity and targeting efficiency of these six mouse lines in IVD tissues at postnatal growing and adult stages. We found that Agc1‐CreERT2 is effective to direct recombination in all components of IVDs, including annulus fibrosus (AF), nucleus pulposus (NP), and cartilaginous endplate (CEP), upon tamoxifen induction at either 2 weeks or 2 months of ages. Moreover, Col2a1‐Cre targets most of the cells in IVDs, except for some cells in the outer AF (OAF) and NP. In contrast, the activity of Col2a1‐CreERT2 is mainly limited to the IAF of IVD tissues at either stage of tamoxifen injection. Similarly, Shh‐Cre directs recombination specifically in all NP cells, whereas Shh‐CreERT2 is active only in a few NP cells when tamoxifen is administered at either stage. Finally, Osx‐Cre targets cells in the CEP, but not in the NP or AF of IVDs tissues at these two stages. Thus, our data demonstrated that all these Cre lines can direct recombination in IVD tissues at postnatal stages with different cell type specificity and/or targeting efficiency, and can, therefore, serve as valuable tools to dissect cell type‐specific gene functions in IVD development and homeostasis.

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Targeting the annulus fibrosus of the intervertebral disc: Col1a2-Cre(ER)T mice show specific activity of Cre recombinase in the outer annulus fibrosus

Cited by 14 publications

References 37 publications

Ciliary IFT80 is essential for intervertebral disc development and maintenance

Ciliary IFT80 is essential for intervertebral disc development and maintenance

Aging aggravates intervertebral disc degeneration by regulating transcription factors toward chondrogenesis

Characterization of Cre recombinase mouse lines enabling cell type‐specific targeting of postnatal intervertebral discs

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