Characterization of Quiescent Epithelial Cells in Mouse Meibomian Glands and Hair Follicle/Sebaceous Glands by Immunofluorescence Tomography

Parfitt, Geraint J.; Geyfman, Mikhail; Xie, Yong; Jester, James V.

doi:10.1038/jid.2014.484

Cited by 16 publications

(17 citation statements)

References 11 publications

(11 reference statements)

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“…This is supported by a previous study that documented the LRCs in the eyelid and meibomian gland using immunofluorescence tomography. 37 The localization of the quiescent epithelial cells to the fornix further supports the hypothesis that the fornix is the region of progenitor or adult stem cells in the conjunctiva. 9,24,38 In conclusion, the quiescent epithelial cell population of the cornea occupies almost the entire peripheral limbus of the mouse cornea at 28 days chase; however, only 3% of limbal epithelial cells are LRCs at 56 days chase.…”

Section: Discussionsupporting

confidence: 54%

“…Recently, we used immunofluorescence tomography to characterize and quantify the meibomian gland adult epithelial stem cells 26 using the H2B-GFP/K5tTA bi-genic mouse. 27 In this study, immunofluorescence tomography was used to 3-dimensionally (3-D) reconstruct the limbus of the H2B-GFP/ K5tTA mouse to colocalize putative stem cell immunomarkers, such as sox9, lrig1, and blimp1, with LRCs.…”

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confidence: 99%

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Immunofluorescence Tomography of Mouse Ocular Surface Epithelial Stem Cells and Their Niche Microenvironment

Parfitt

Kavianpour

et al. 2015

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. Currently, there are no definitive immunomarkers for epithelial stem cells (corneal and conjunctival) or their poorly understood niche microenvironment. The H2B-GFP/K5tTA mouse enables visualization of label-retaining cells (LRCs), which exhibit the functional marker of stem cell quiescence. We used immunofluorescence tomography to evaluate putative stem cell markers and LRCs of the mouse ocular surface.METHODS. H2B-GFP/K5tTA mice were pulsed for 56 days and then chased with doxycycline to label LRCs. Limbus and eyelid tissue was 3-dimensionally (3-D) reconstructed using immunofluorescence tomography to identify and characterize LRCs using the putative stem cell markers sox9, keratin 19, lrig1, blimp1, and abcb5.RESULTS. After 28 days of chase, LRCs were localized to the entire limbus epithelium and, infrequently, the anterior limbal stroma. Label-retaining cells comprised 3% of limbal epithelial cells after 56 days of chase. Conjunctival LRCs were localized to the fornix and comprised 4% of the total fornix epithelial cells. No stem cell immunomarker was specific for ocular surface LRCs; however, blimp1 enriched for limbal basal epithelial cells and 100% of green fluorescent protein-positive (GFP þ ) cells at the limbus and fornix were found to be lrig1-positive.CONCLUSIONS. Label-retaining cells represent a larger population of the mouse limbus than previously thought. They decrease in number with increased doxycycline chase, suggesting that LRC populations with different cell cycle lengths exist at the limbus. We conclude that current immunomarkers are unable to colocalize with the functional marker of epithelial stem cell quiescence; however, blimp1 may enrich for limbal epithelial basal cells.

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Section: Discussionsupporting

confidence: 54%

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confidence: 99%

Immunofluorescence Tomography of Mouse Ocular Surface Epithelial Stem Cells and Their Niche Microenvironment

Parfitt

Kavianpour

et al. 2015

Invest. Ophthalmol. Vis. Sci.

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“…To address this question, we used the H2B-GFP/K5tTA ‘tet-off’ transgenic mouse to initially label proliferating cells with nuclear GFP and then follow labeled cells after tetracycline treatment to turn-off nuclear GFP expression (Parfitt et al, 2015). IT was then used to identify, localize and quantify the label retaining cells (LRCs) at different intervals after tetracycline treatment.…”

Section: Meibomian Gland Stem Cellsmentioning

confidence: 99%

Meibocyte differentiation and renewal: Insights into novel mechanisms of meibomian gland dysfunction (MGD)

Hwang

Parfitt

Brown

et al. 2017

Experimental Eye Research

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This paper reviews our current understanding of age-related meibomian gland dysfunction (MGD) and the role of the nuclear receptor, peroxisome proliferator-activated receptor gamma (PPARγ), in the regulation of meibomian gland function, meibocyte differentiation and lipid synthesis. The studies suggest that PPARγ is a master regulator of meibocyte differentiation and function, whose expression and nuclear signaling coupled with meibocyte renewal is altered during aging, potentially leading to atrophy of the meibomian gland as seen in clinical MGD. Study of meibomian gland stem cells also suggest that there is a limited number of precursor meibocytes that provide progeny to the acini, that may be susceptible to exhaustion as occurs during aging and other environmental factors. Further study of pathways regulating PPARγ expression and function as well as meibocyte stem cell maintenance may provide clues to establishing cellular and molecular mechanisms underlying MGD and the development of novel therapeutic strategies to treating this disease.

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“…Meibum is synthesized by enlarged, terminally differentiated meibocytes that form the acini of the meibomian gland, and is delivered through the central duct to the mucocutaneous junction of the eyelid margin. The mechanism that explains how the acinar and ductal epithelia renew throughout life has not been well described, and there is only limited information about a putative meibomian gland stem cell population ( Knop et al., 2011 , Olami et al., 2001 , Parfitt et al., 2015 ).…”

Section: Introductionmentioning

confidence: 99%

Renewal of the Holocrine Meibomian Glands by Label-Retaining, Unipotent Epithelial Progenitors

et al. 2016

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SummaryThe meibomian and sebaceous glands secrete lipids to prevent desiccation of the ocular surface and skin, respectively. Precisely how these holocrine tissues regenerate is not well understood. To address this, we characterized keratin 5+ (K5) label-retaining cells (LRCs) and the lineage tracing of keratin 14 (K14) progenitors in mouse meibomian glands. Using the tet-off H2B-GFP/K5tTA mouse, H2B-GFP fluorescence dilutes 2-fold with every division in K5+ cell nuclei after doxycycline administration. In 3D reconstructions generated over a >28-day doxycycline chase, we observed LRCs at the acinus entrance where K6+ ductal epithelium terminates. For lineage tracing, K14CreERT2-Confetti mice were injected intraperitoneally with tamoxifen and euthanized at 23 and 59 weeks later. Meibomian gland acini in these mice were either monochromatic or dual-colored, whereas the duct exhibited multiple colors. In conclusion, LRCs are likely to direct meibomian gland turnover and may exist as two distinct unipotent progenitors that renew ductal and acinar tissue separately.

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Characterization of Quiescent Epithelial Cells in Mouse Meibomian Glands and Hair Follicle/Sebaceous Glands by Immunofluorescence Tomography

Cited by 16 publications

References 11 publications

Immunofluorescence Tomography of Mouse Ocular Surface Epithelial Stem Cells and Their Niche Microenvironment

Immunofluorescence Tomography of Mouse Ocular Surface Epithelial Stem Cells and Their Niche Microenvironment

Meibocyte differentiation and renewal: Insights into novel mechanisms of meibomian gland dysfunction (MGD)

Renewal of the Holocrine Meibomian Glands by Label-Retaining, Unipotent Epithelial Progenitors

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