Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes

Katsuhiro Hitomi

doi:doi:10.11648/j.cb.20140204.11

| Peer-Reviewed

Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes

Katsuhiro Hitomi

Published in Cell Biology (Volume 2, Issue 4)

Received: 20 August 2014 Accepted: 6 September 2014 Published: 20 September 2014

Views: Downloads:

Download PDF

Share This Article

Twitter
Linked In
Facebook

Abstract

Telomeres are the physical ends of chromosomes, and are cleaved with each cell division. Telomere loss results from not only incomplete DNA replication (“end replication problem”), followed by the processing of chromosome, but also exposure to various DNA damaging agents. Human epidermis is constantly replaced through replication at the basal layer, followed by upward movement and terminal differentiation at the suprabasal layer. The objective of this study was to assess differences in telomere length between basal and suprabasal keratinocytes, and variations of telomere lengths among basal cells. We investigated terminal restriction fragment (TRF) length in fractions of epidermal cells by Southern blot analysis. Time-dependent trypsin treatment to epidermis divided into basal cell rich and suprabasal cell rich fractions. In the five skin samples from various sites of the body, the mean TRF length were 5,932 ± 470 (range 5,182-6,630) bp in basal cell rich fraction showing 21.5% ± 5.8% (range 12.6-28.6%) of keratin 10 (K10)-positivity, which is marker of keratinizing suprabasal cell, and 5,320 ± 640 (range 4,495-6,212) bp in suprabasal cell rich fraction showing 78.8% ± 4.0% (range 72.8-83.0%) of K10-positivity. It was calculated that K10 positive cells have 1,091 ± 963 (range 248-2,650) bp shorter telomeres compared to K10 negative cells. In addition, fluorescence in situ hybridization (FISH) study showed that columnar groups of keratinocytes with statistically longer telomere in each nucleus than the other epidermal cells in the vicinity occupied occasionally between the tip of dermal papilla and the bottom of rete ridge. The present study showed that the keratinizing suprabasal cells have shorter telomeres compared to basal cells. FISH study showed that some groups of epidermal cells having different replicative histories among the groups of epidermal cells in the vicinity.

Published in	Cell Biology (Volume 2, Issue 4)
DOI	10.11648/j.cb.20140204.11
Page(s)	28-35
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Human Skin, Keratinocyte, Fluorescence in Situ Hybridization, Telomere, Terminal Restriction Fragment

References

[1]	Blackburn EH. Structure and function of telomeres. Nature 1991; 350: 569-73.
[2]	Zakian VA. Telomeres: beginning to understand the end. Science 1995; 270: 1601-7.
[3]	Moyzis RK, Buckingham JM, Cram LS, Dani M, Deaven LL, Jones MD, Meyne J, Ratliff RL, Wu JR. A highly conserved repetitive DNA sequence, (TTAGGG)n, present at the telomeres of human chromosomes. Proc Natl Acad Sci U S A. 1988; 85: 6622-6.
[4]	Harley CB. “Telomeres and aging.” in Telomeres, Blackburn EH, Greider CW, Eds. Cold Spring Harbor Laboratory Press, 1995, pp. 247-63.
[5]	Aubert G, Lansdorp PM. Telomeres and aging. Physiol Rev 2008; 88: 557-79.
[6]	Blanpain C, Fuchs E. Epidermal homeostasis: a balancing act of stem cells in the skin. Nat Rev Mol Cell Biol. 2009; 10: 207-17.
[7]	Clayton E, Doupé DP, Klein AM, Winton DJ, Simons BD, Jones PH. A single type of progenitor cell maintains normal epidermis. Nature 2007; 446: 185-9.
[8]	Mascré G, Dekoninck S, Drogat B, Youssef KK, Broheé S, Sotiropoulou PA, Simons BD, Blanpain C. Distinct contribution of stem and progenitor cells to epidermal maintenance. Nature 2012; 489: 257-62.
[9]	de Rosa L, de Luca M. Cell biology: Dormant and restless skin stem cells. Nature 2012; 489: 215-7.
[10]	Lindsey J, McGill NI, Lindsey LA, Green DK, Cooke HJ. In vivo loss of telomeric repeats with age in humans. Mutat Res. 1991; 256: 45-8.
[11]	Friedrich U, Griese E, Schwab M, Fritz P, Thon K, Klotz U. Telomere length in different tissues of elderly patients. Mech Ageing Dev. 2000; 119: 89-99.
[12]	Nakamura K, Izumiyama-Shimomura N, Sawabe M, Arai T, Aoyagi Y, Fujiwara M, Tsuchiya E, Kobayashi Y, Kato M, Oshimura M, Sasajima K, Nakachi K, Takubo K. Comparative analysis of telomere lengths and erosion with age in human epidermis and lingual epithelium. J Invest Dermatol. 2002; 119: 1014-9.
[13]	Sugimoto M, Yamashita R, Ueda M. Telomere length of the skin in association with chronological aging and photoaging. J Dermatol Sci. 2006; 43: 43-7.
[14]	Krunic D, Moshir S, Greulich-Bode KM, Figueroa R, Cerezo A, Stammer H, Stark HJ, Gray SG, Nielsen KV, Hartschuh W, Boukamp P. Tissue context-activated telomerase in human epidermis correlates with little age-dependent telomere loss. Biochim Biophys Acta. 2009; 1792: 297-308.
[15]	Ishikawa N, Nakamura K, Izumiyama-Shimomura N, Aida J, Ishii A, Goto M, Ishikawa Y, Asaka R, Matsuura M, Hatamochi A, Kuroiwa M, Takubo K. Accelerated in vivo epidermal telomere loss in Werner syndrome. Aging (Albany NY). 2011; 3: 417-29.
[16]	Härle-Bachor C, Boukamp P. Telomerase activity in the regenerative basal layer of the epidermis in human skin and in immortal and carcinoma-derived skin keratinocytes. Proc Natl Acad Sci U S A. 1996; 93: 6476-81.
[17]	Bryant JE, Hutchings KG, Moyzis RK, Griffith JK. Measurement of telomeric DNA content in human tissues. Biotechniques. 1997; 23: 476-8.
[18]	de Lange T, Shiue L, Myers RM, Cox DR, Naylor SL, Killery AM, Varmus HE. Structure and variability of human chromosome ends. Mol Cell Biol. 1990; 10: 518-27.
[19]	Aida J, Izumiyama-Shimomura N, Nakamura K, Ishikawa N, Poon SS, Kammori M, Sawabe M, Arai T, Matsuura M, Fujiwara M, Kishimoto H, Takubo K. Basal cells have longest telomeres measured by tissue Q-FISH method in lingual epithelium. Exp Gerontol. 2008; 43: 833-9.
[20]	Takubo K, Fujita M, Izumiyama N, Nakamura K, Ishikawa N, Poon SS, Fujiwara M, Sawabe M, Matsuura M, Grabsch H, Arai T, Aida J. Q-FISH analysis of telomere and chromosome instability in the oesophagus with and without squamous cell carcinoma in situ. J Pathol. 2010; 221: 201-9.
[21]	Montpetit AJ, Alhareeri AA, Montpetit M, Starkweather AR, Elmore LW, Filler K, Mohanraj L, Burton CW, Menzies VS, Lyon DE, Jackson-Cook CK. Telomere length: a review of methods for measurement. Nurs Res. 2014; 63: 289-99.
[22]	Miyata Y, Okada K, Fujimoto A, Hata K, Kagami H, Tomita Y, Ueda M. The effect of the long-term cultivation on telomere length and morphology of cultured epidermis. J Dermatol Sci. 2004; 34: 221-30.
[23]	Buckingham EM, Klingelhutz AJ. The role of telomeres in the ageing of human skin. Exp Dermatol. 2011; 20: 297-302.
[24]	Ikeda H, Aida J, Hatamochi A, Hamasaki Y, Izumiyama-Shimomura N, Nakamura K, Ishikawa N, Poon SS, Fujiwara M, Tomita K, Hiraishi N, Kuroiwa M, Matsuura M, Sanada Y, Kawano Y, Arai T, Takubo K. Quantitative fluorescence in situ hybridization measurement of telomere length in skin with/without sun exposure or actinic keratosis. Hum Pathol. 2014; 45: 473-80.
[25]	Jones PH, Harper S, Watt FM. Stem cell patterning and fate in human epidermis. Cell. 1995; 80: 83-93.
[26]	Wellinger RJ. In the end, what's the problem? Mol Cell. 2014; 53: 855-6.

Cite This Article

Plain Text BibTeX RIS

APA Style

Katsuhiro Hitomi. (2014). Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes. Cell Biology, 2(4), 28-35. https://doi.org/10.11648/j.cb.20140204.11

Copy | Download

ACS Style

Katsuhiro Hitomi. Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes. Cell Biol. 2014, 2(4), 28-35. doi: 10.11648/j.cb.20140204.11

Copy | Download

AMA Style

Katsuhiro Hitomi. Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes. Cell Biol. 2014;2(4):28-35. doi: 10.11648/j.cb.20140204.11

Copy | Download

@article{10.11648/j.cb.20140204.11,
  author = {Katsuhiro Hitomi},
  title = {Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes},
  journal = {Cell Biology},
  volume = {2},
  number = {4},
  pages = {28-35},
  doi = {10.11648/j.cb.20140204.11},
  url = {https://doi.org/10.11648/j.cb.20140204.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.cb.20140204.11},
  abstract = {Telomeres are the physical ends of chromosomes, and are cleaved with each cell division. Telomere loss results from not only incomplete DNA replication (“end replication problem”), followed by the processing of chromosome, but also exposure to various DNA damaging agents. Human epidermis is constantly replaced through replication at the basal layer, followed by upward movement and terminal differentiation at the suprabasal layer. The objective of this study was to assess differences in telomere length between basal and suprabasal keratinocytes, and variations of telomere lengths among basal cells. We investigated terminal restriction fragment (TRF) length in fractions of epidermal cells by Southern blot analysis. Time-dependent trypsin treatment to epidermis divided into basal cell rich and suprabasal cell rich fractions. In the five skin samples from various sites of the body, the mean TRF length were 5,932 ± 470 (range 5,182-6,630) bp in basal cell rich fraction showing 21.5% ± 5.8% (range 12.6-28.6%) of keratin 10 (K10)-positivity, which is marker of keratinizing suprabasal cell, and 5,320 ± 640 (range 4,495-6,212) bp in suprabasal cell rich fraction showing 78.8% ± 4.0% (range 72.8-83.0%) of K10-positivity. It was calculated that K10 positive cells have 1,091 ± 963 (range 248-2,650) bp shorter telomeres compared to K10 negative cells. In addition, fluorescence in situ hybridization (FISH) study showed that columnar groups of keratinocytes with statistically longer telomere in each nucleus than the other epidermal cells in the vicinity occupied occasionally between the tip of dermal papilla and the bottom of rete ridge. The present study showed that the keratinizing suprabasal cells have shorter telomeres compared to basal cells. FISH study showed that some groups of epidermal cells having different replicative histories among the groups of epidermal cells in the vicinity.},
 year = {2014}
}

Copy | Download

TY  - JOUR
T1  - Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes
AU  - Katsuhiro Hitomi
Y1  - 2014/09/20
PY  - 2014
N1  - https://doi.org/10.11648/j.cb.20140204.11
DO  - 10.11648/j.cb.20140204.11
T2  - Cell Biology
JF  - Cell Biology
JO  - Cell Biology
SP  - 28
EP  - 35
PB  - Science Publishing Group
SN  - 2330-0183
UR  - https://doi.org/10.11648/j.cb.20140204.11
AB  - Telomeres are the physical ends of chromosomes, and are cleaved with each cell division. Telomere loss results from not only incomplete DNA replication (“end replication problem”), followed by the processing of chromosome, but also exposure to various DNA damaging agents. Human epidermis is constantly replaced through replication at the basal layer, followed by upward movement and terminal differentiation at the suprabasal layer. The objective of this study was to assess differences in telomere length between basal and suprabasal keratinocytes, and variations of telomere lengths among basal cells. We investigated terminal restriction fragment (TRF) length in fractions of epidermal cells by Southern blot analysis. Time-dependent trypsin treatment to epidermis divided into basal cell rich and suprabasal cell rich fractions. In the five skin samples from various sites of the body, the mean TRF length were 5,932 ± 470 (range 5,182-6,630) bp in basal cell rich fraction showing 21.5% ± 5.8% (range 12.6-28.6%) of keratin 10 (K10)-positivity, which is marker of keratinizing suprabasal cell, and 5,320 ± 640 (range 4,495-6,212) bp in suprabasal cell rich fraction showing 78.8% ± 4.0% (range 72.8-83.0%) of K10-positivity. It was calculated that K10 positive cells have 1,091 ± 963 (range 248-2,650) bp shorter telomeres compared to K10 negative cells. In addition, fluorescence in situ hybridization (FISH) study showed that columnar groups of keratinocytes with statistically longer telomere in each nucleus than the other epidermal cells in the vicinity occupied occasionally between the tip of dermal papilla and the bottom of rete ridge. The present study showed that the keratinizing suprabasal cells have shorter telomeres compared to basal cells. FISH study showed that some groups of epidermal cells having different replicative histories among the groups of epidermal cells in the vicinity.
VL  - 2
IS  - 4
ER  -

Copy | Download

Author Information

Katsuhiro Hitomi

Department of Dermatology, Yamagata University School of Medicine, Yamagata, Japan; Present Affiliation: Department of Dermatology, Saitama Medical Center, Saitama Medical University, Kawagoe, Japan

Download PDF

Sections

Plain Text BibTeX RIS

APA Style

Katsuhiro Hitomi. (2014). Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes. Cell Biology, 2(4), 28-35. https://doi.org/10.11648/j.cb.20140204.11

Copy | Download

ACS Style

Katsuhiro Hitomi. Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes. Cell Biol. 2014, 2(4), 28-35. doi: 10.11648/j.cb.20140204.11

Copy | Download

AMA Style

Katsuhiro Hitomi. Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes. Cell Biol. 2014;2(4):28-35. doi: 10.11648/j.cb.20140204.11

Copy | Download

@article{10.11648/j.cb.20140204.11,
  author = {Katsuhiro Hitomi},
  title = {Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes},
  journal = {Cell Biology},
  volume = {2},
  number = {4},
  pages = {28-35},
  doi = {10.11648/j.cb.20140204.11},
  url = {https://doi.org/10.11648/j.cb.20140204.11},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.cb.20140204.11},
  abstract = {Telomeres are the physical ends of chromosomes, and are cleaved with each cell division. Telomere loss results from not only incomplete DNA replication (“end replication problem”), followed by the processing of chromosome, but also exposure to various DNA damaging agents. Human epidermis is constantly replaced through replication at the basal layer, followed by upward movement and terminal differentiation at the suprabasal layer. The objective of this study was to assess differences in telomere length between basal and suprabasal keratinocytes, and variations of telomere lengths among basal cells. We investigated terminal restriction fragment (TRF) length in fractions of epidermal cells by Southern blot analysis. Time-dependent trypsin treatment to epidermis divided into basal cell rich and suprabasal cell rich fractions. In the five skin samples from various sites of the body, the mean TRF length were 5,932 ± 470 (range 5,182-6,630) bp in basal cell rich fraction showing 21.5% ± 5.8% (range 12.6-28.6%) of keratin 10 (K10)-positivity, which is marker of keratinizing suprabasal cell, and 5,320 ± 640 (range 4,495-6,212) bp in suprabasal cell rich fraction showing 78.8% ± 4.0% (range 72.8-83.0%) of K10-positivity. It was calculated that K10 positive cells have 1,091 ± 963 (range 248-2,650) bp shorter telomeres compared to K10 negative cells. In addition, fluorescence in situ hybridization (FISH) study showed that columnar groups of keratinocytes with statistically longer telomere in each nucleus than the other epidermal cells in the vicinity occupied occasionally between the tip of dermal papilla and the bottom of rete ridge. The present study showed that the keratinizing suprabasal cells have shorter telomeres compared to basal cells. FISH study showed that some groups of epidermal cells having different replicative histories among the groups of epidermal cells in the vicinity.},
 year = {2014}
}

Copy | Download

TY  - JOUR
T1  - Telomere Length Differences upon Keratinization and its Variations in Normal Human Epidermal Keratinocytes
AU  - Katsuhiro Hitomi
Y1  - 2014/09/20
PY  - 2014
N1  - https://doi.org/10.11648/j.cb.20140204.11
DO  - 10.11648/j.cb.20140204.11
T2  - Cell Biology
JF  - Cell Biology
JO  - Cell Biology
SP  - 28
EP  - 35
PB  - Science Publishing Group
SN  - 2330-0183
UR  - https://doi.org/10.11648/j.cb.20140204.11
AB  - Telomeres are the physical ends of chromosomes, and are cleaved with each cell division. Telomere loss results from not only incomplete DNA replication (“end replication problem”), followed by the processing of chromosome, but also exposure to various DNA damaging agents. Human epidermis is constantly replaced through replication at the basal layer, followed by upward movement and terminal differentiation at the suprabasal layer. The objective of this study was to assess differences in telomere length between basal and suprabasal keratinocytes, and variations of telomere lengths among basal cells. We investigated terminal restriction fragment (TRF) length in fractions of epidermal cells by Southern blot analysis. Time-dependent trypsin treatment to epidermis divided into basal cell rich and suprabasal cell rich fractions. In the five skin samples from various sites of the body, the mean TRF length were 5,932 ± 470 (range 5,182-6,630) bp in basal cell rich fraction showing 21.5% ± 5.8% (range 12.6-28.6%) of keratin 10 (K10)-positivity, which is marker of keratinizing suprabasal cell, and 5,320 ± 640 (range 4,495-6,212) bp in suprabasal cell rich fraction showing 78.8% ± 4.0% (range 72.8-83.0%) of K10-positivity. It was calculated that K10 positive cells have 1,091 ± 963 (range 248-2,650) bp shorter telomeres compared to K10 negative cells. In addition, fluorescence in situ hybridization (FISH) study showed that columnar groups of keratinocytes with statistically longer telomere in each nucleus than the other epidermal cells in the vicinity occupied occasionally between the tip of dermal papilla and the bottom of rete ridge. The present study showed that the keratinizing suprabasal cells have shorter telomeres compared to basal cells. FISH study showed that some groups of epidermal cells having different replicative histories among the groups of epidermal cells in the vicinity.
VL  - 2
IS  - 4
ER  -

Copy | Download