Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Detection of BrdU-label retaining cells in the lacrimal gland: implications for tissue repair

  • 183 Accesses

  • 15 Citations

Abstract

The purpose of the present study was to determine if the lacrimal gland contains 5-bromo-2′-deoxyuridine (BrdU)-label retaining cells and if they are involved in tissue repair. Animals were pulsed daily with BrdU injections for 7 consecutive days. After a chase period of 2, 4, or 12 weeks, the animals were sacrificed and the lacrimal glands were removed and processed for BrdU immunostaining. In another series of experiments, the lacrimal glands of 12-week chased animals were either left untreated or were injected with interleukin 1 (IL-1) to induce injury. Two and half days post-injection, the lacrimal glands were removed and processed for BrdU immunostaining. After 2 and 4 weeks of chase period, a substantial number of lacrimal gland cells were BrdU+ (11.98 ± 1.84 and 7.95 ± 1.83 BrdU+ cells/mm2, respectively). After 12 weeks of chase, there was a 97% decline in the number of BrdU+ cells (0.38 ± 0.06 BrdU+ cells/mm2), suggesting that these BrdU-label retaining cells may represent slow-cycling adult stem/progenitor cells. In support of this hypothesis, the number of BrdU labeled cells increased over 7-fold during repair of the lacrimal gland (control: 0.41 ± 0.09 BrdU+ cells/mm2; injured: 2.91 ± 0.62 BrdU+ cells/mm2). Furthermore, during repair, among BrdU+ cells 58.2 ± 3.6 % were acinar cells, 26.4 ± 4.1% were myoepithelial cells, 0.4 ± 0.4% were ductal cells and 15.0 ± 3.0% were stromal cells. We conclude that the murine lacrimal gland contains BrdU-label retaining cells that are mobilized following injury to generate acinar, myoepithelial and ductal cells.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. Alison MR, Golding M, Sarraf CE (1997) Liver stem cells: when the going gets tough they get going. Int J Exp Pathol 78:365–381

  2. Bjerknes M, Cheng H (2002) Multipotential stem cells in adult mouse gastric epithelium. Am J Physiol Gastrointest Liver Physiol 283:G767–777

  3. Blanpain C, Horsley V, Fuchs E (2007) Epithelial stem cells: turning over new leaves. Cell 128:445–458

  4. Bron AJ, Tiffany JM, Gouveia SM, Yokoi N, Voon LW (2004) Functional aspects of the tear film lipid layer. Exp Eye Res 78:347–360

  5. Calissendorff B, el Azazi M, Lonnqvist B (1989) Dry eye syndrome in long-term follow-up of bone marrow transplanted patients. Bone Marrow Transpl 4:675–678

  6. Chan RW, Gargett CE (2006) Identification of label-retaining cells in mouse endometrium. Stem Cells 24:1529–1538

  7. Cotsarelis G, Cheng SZ, Dong G, Sun TT, Lavker RM (1989) Existence of slow-cycling limbal epithelial basal cells that can be preferentially stimulated to proliferate: implications on epithelial stem cells. Cell 57:201–209

  8. Cotsarelis G, Sun TT, Lavker RM (1990) Label-retaining cells reside in the bulge area of pilosebaceous unit: implications for follicular stem cells, hair cycle, and skin carcinogenesis. Cell 61:1329–1337

  9. Dartt DA (2009) Neural regulation of lacrimal gland secretory processes: relevance in dry eye diseases. Prog Retin Eye Res 28:155–177

  10. De Vita S, Damato R, De Marchi G, Sacco S, Ferraccioli G (2002) True primary Sjögren’s syndrome in a subset of patients with hepatitis C infection: a model linking chronic infection to chronic sialadenitis. Isr Med Assoc J 4:1101–1105

  11. DeCarlo DK, Penner SL, Schamerloh RJ, Fullard RJ (1995) Dry eye among males infected with the human immunodeficiency virus. J Am Optom Assoc 66:533–538

  12. Drosos AA, Constantopoulos SH, Psychos D, Stefanou D, Papadimitriou CS, Moutsopoulos HM (1989) The forgotten cause of sicca complex; sarcoidosis. J Rheumatol 16:1548–1551

  13. Duvillie B, Attali M, Aiello V, Quemeneur E, Scharfmann R (2003) Label-retaining cells in the rat pancreas: location and differentiation potential in vitro. Diabetes 52:2035–2042

  14. Forbes SJ, Vig P, Poulsom R, Wright NA, Alison MR (2002) Adult stem cell plasticity: new pathways of tissue regeneration become visible. Clin Sci (Lond) 103:355–369

  15. Gipson IK, Argueso P (2003) Role of mucins in the function of the corneal and conjunctival epithelia. Int Rev Cytol 231:1–49

  16. Gomperts BN, Strieter RM (2007) Stem cells and chronic lung disease. Annu Rev Med 58:285–298

  17. Hisatomi Y, Okumura K, Nakamura K, Matsumoto S, Satoh A, Nagano K, Yamamoto T, Endo F (2004) Flow cytometric isolation of endodermal progenitors from mouse salivary gland differentiate into hepatic and pancreatic lineages. Hepatology 39:667–675

  18. Hodges RR, Dartt DA (2003) Regulatory pathways in lacrimal gland epithelium. Int Rev Cytol 231:129–196

  19. Kim YJ, Kwon HJ, Shinozaki N, Hashimoto S, Shimono M, Cho SW, Jung HS (2008) Comparative analysis of ABCG2-expressing and label-retaining cells in mouse submandibular gland. Cell Tissue Res 334:47–53

  20. Kimoto M, Yura Y, Kishino M, Toyosawa S, Ogawa Y (2008) Label-retaining cells in the rat submandibular gland. J Histochem Cytochem 56:15–24

  21. Kishi T, Takao T, Fujita K, Taniguchi H (2006) Clonal proliferation of multipotent stem/progenitor cells in the neonatal and adult salivary glands. Biochem Biophys Res Commun 340:544–552

  22. Kume S (2005) Stem-cell-based approaches for regenerative medicine. Dev Growth Differ 47:393–402

  23. Lemullois M, Rossignol B, Mauduit P (1996) Immunolocalization of myoepithelial cells in isolated acini of rat exorbital lacrimal gland: cellular distribution of muscarinic receptors. Biol Cell 86:175–181

  24. Maeshima A, Yamashita S, Nojima Y (2003) Identification of renal progenitor-like tubular cells that participate in the regeneration processes of the kidney. J Am Soc Nephrol 14:3138–3146

  25. Meinhardt A, Spicher A, Roehrich ME, Glauche I, Vogt P, Vassalli G (2011) Immunohistochemical and flow cytometric analysis of long-term label-retaining cells in the adult heart. Stem Cells Dev 20:211–222

  26. Ogawa Y, Kuwana M (2003) Dry eye as a major complication associated with chronic graft-versus-host disease after hematopoietic stem cell transplantation. Cornea 22:S19–27

  27. Okumura K, Nakamura K, Hisatomi Y, Nagano K, Tanaka Y, Terada K, Sugiyama T, Umeyama K, Matsumoto K, Yamamoto T, Endo F (2003) Salivary gland progenitor cells induced by duct ligation differentiate into hepatic and pancreatic lineages. Hepatology 38:104–113

  28. Pflugfelder SC (2004) Antiinflammatory therapy for dry eye. Am J Ophthalmol 137:337–342

  29. Potten CS, Hume WJ, Reid P, Cairns J (1978) The segregation of DNA in epithelial stem cells. Cell 15:899–906

  30. Poulsom R, Alison MR, Forbes SJ, Wright NA (2002) Adult stem cell plasticity. J Pathol 197:441–456

  31. Schaumberg DA, Sullivan DA, Buring JE, Dana MR (2003) Prevalence of dry eye syndrome among US women. Am J Ophthalmol 136:318–326

  32. Schaumberg DA, Dana R, Buring JE, Sullivan DA (2009) Prevalence of dry eye disease among US men: estimates from the Physicians’ Health Studies. Arch Ophthalmol 127:763–768

  33. Smith GH (2005) Label-retaining epithelial cells in mouse mammary gland divide asymmetrically and retain their template DNA strands. Development 132:681–687

  34. Stern ME, Beuerman RW, Fox RI, Gao J, Mircheff AK, Pflugfelder SC (1998) The pathology of dry eye: the interaction between the ocular surface and lacrimal glands. Cornea 17:584–589

  35. Tiffany JM (2008) The normal tear film. Dev Ophthalmol 41:1–20

  36. Wei Z, Cotsarelis G, Sun T, Lavker R (1995) Label-retaining cells are preferentially located in fornical epithelium: implications on conjunctival epithelial homeostasis. Invest Ophthalmol Vis Sci 36:236–246

  37. You S, Kublin CL, Avidan O, Miyasaki D, Zoukhri D (2011) Isolation and propagation of mesenchymal stem cells from the lacrimal gland. Invest Ophthalmol Vis Sci 52:2087–2094

  38. You S, Avidan O, Tariq A, Ahluwalia, I, Stark, P, Kublin CL, Zoukhri D (2011) Role of epithelial-mesenchymal transition in repair of the lacrimal gland following experimentally induced injury. Invest Ophthalmol Vis Sci (in press)

  39. Zhang YQ, Kritzik M, Sarvetnick N (2005) Identification and expansion of pancreatic stem/progenitor cells. J Cell Mol Med 9:331–344

  40. Zoukhri D (2006) Effect of inflammation on lacrimal gland function. Exp Eye Res 82:885–898

  41. Zoukhri D (2010) Mechanisms involved in injury and repair of the murine lacrimal gland: role of programmed cell death and mesenchymal stem cells. Ocul Surf 8:60–69

  42. Zoukhri D, Macari E, Kublin CL (2007) A single injection of interleukin-1 induces reversible aqueous-tear deficiency, lacrimal gland inflammation, and acinar and ductal cell proliferation. Exp Eye Res 84:894–904

  43. Zoukhri D, Fix A, Alroy J, Kublin CL (2008) Mechanisms of murine lacrimal gland repair after experimentally induced inflammation. Invest Ophthalmol Vis Sci 49:4399–4406

Download references

Acknowledgements

The authors gratefully acknowledge the BRB Preclinical Repository of the NCI for the generous gift of recombinant human cytokines, the assistance of Tufts Center for Neuroscience Research P30 NS047243 (Jackson), Ms. Robin Hodges and Dr. Darlene Dartt for their critical reading of the manuscript and Dr. Fara Sourie for her invaluable contribution to this work.

Author information

Correspondence to Driss Zoukhri.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

You, S., Tariq, A., Kublin, C.L. et al. Detection of BrdU-label retaining cells in the lacrimal gland: implications for tissue repair. Cell Tissue Res 346, 317–326 (2011). https://doi.org/10.1007/s00441-011-1271-x

Download citation

Keywords

  • Progenitor cells
  • BrdU-label retaining cells
  • Tissue repair
  • Lacrimal gland