Bir yaşlanma modeli olarak Caenorhabditis elegans bireylerinde kalsinörin inhibisyonu ile ortaya çıkan değişiklikler
Abstract
Amaç: Kalsinörin inhibisyonunun Caenorhabditis elegans (C. elegans) bireylerinde yaşam süresini
uzattığı bilinmektedir. C. elegans
genlerinin önemli bir kısmı insan genlerine büyük benzerlik göstermektedir. Bu
çalışmada kalsinörin inhibisyonunun C.
elegans bireylerindeki yaşamsal fonksiyonlar üzerine olan etkilerinin
değerlendirilmesi amaçlandı.
Yöntem: Çalışma için bir
kalsinörin inhibitörü olan siklosporin A’nın 1 μM (1.doz), 0.1 μM (2.doz), 0.01
μM (3.doz) ve 0.001 μM (4.doz) dozunda çözeltileri hazırlanarak her bir
çözeltiden 1’er ml, içlerinde 10’ ar ml Nematod Growth Media (NGM) bulunduran
petrilere eklenmiştir. Kontrol grubu siklosporin eklenmemiş besiyerinde
beslenmiştir. Senkronizasyonu yapılmış C.
elegans’lardan siklosporinin belirtilen dozları ile hazırlanmış her bir
petriye 20’ şer adet aktarılmıştır. 6X–50X arasında büyütme sağlayan alttan
ışıklandırmalı stereo mikroskop ile C.
elegans’lar izlenmiştir. Bütün petrilerdeki C. elegans’lar ölene kadar her gün aynı saatte canlı hayvanlar
sayılmış ve kontrol grubuyla karşılaştırılmıştır.
Bulgular: Çalışma
sonucunda siklosporine maruz bırakılan C.
elegans’ların kontrol grubuna göre yaşam sürelerinde doz artışına bağlı
istatistiksel olarak anlamlı bir artış olduğu tespit edilmiştir (p<0,05).
Siklosporine maruz bırakılan C. elegans’ların
fiziksel gelişimlerinin doz artışına bağlı olarak anlamlı derecede azaldığı saptanmıştır
(p<0,05). Siklosporin dozundaki artış farinks pompalama sayısında
istatistiksel olarak anlamlı bir değişikliğe neden olmamıştır (p>0,05).
Sonuç: Kalsinörin
inhibisyonu, C. elegans’ların
gelişimsel sürecini yavaşlatır ve yaşam süresini uzatır. Kalsinörin
inhibitörleri, benzer mekanizmalarla insanlarda deri yaşlanması üzerine olumlu
etki yapabilir. Bu nedenle kalsinörin inhibisyonunun, deri yaşlanması üzerine
etkinliğini ve etki mekanizmasını açıklayacak ileri çalışmalar gereklidir.
References
- 1. Ünlü E. Deri yaşlanmasında korunma ve tedavi yöntemleri. Dermatoz 2010; 1: 23-31.
- 2. Ünal İ, Ertam İ. Factors Contributing to Skin Aging. Turkiye Klinikleri J Cosm Dermatol Special Topics 2008; 1: 1-7.
- 3. Hertweck M, Hoppe T, Baumeister R. C. elegans, a model for aging with highthroughput capacity. Exp Gerontol 2003; 38: 345-6.
- 4. Olsen A, Vantipalli MC, Lithgow GJ. Using Caenorhabditis elegans as a model for aging and age-related diseases. Ann N Y Acad Sci 2006; 1067: 120-8.
- 5. Smit NP, Van Rossum HH, Romijn FP, Sellar KJ, Breetveld M, Gibbs S et al. Calcineurin activity and inhibition in skin and (epi) dermal cell cultures. J Invest Dermatol 2008; 128:1686-90.
- 6. Fisher GJ, Duell EA, Nickoloff BJ, Annesley TM, Kowalke JK, Ellis CN et al. Levels of cyclosporin in epidermis of treated psoriasis patients differentially inhibit growth of keratinocytes cultured in serum free versus serum containing media. J Invest Dermatol 1988; 91: 142-6.
- 7. Nickoloff BJ, Fisher GJ, Mitra RS, Voorhees JJ. Direct cytopathic effects of cyclosporine A on rapidly proliferating cultured keratinocytes and dermal fibroblasts. Transplant Proc 1988; 20: 85-90.
- 8. Bandyopadhyay J, Lee J, Lee JI, Yu JR, Jee C, Cho JH, et al. Calcineurin, a calcium/calmodulin-dependent protein phosphatase, is involved in movement, fertility, egg laying, and growth in Caenorhabditis elegans. Mol Biol Cell 2002; 13: 3281-93.
- 9. Kuhara A, Inada H, Katsura I, Mori I. Negative regulation and gain control of sensory neurons by the C. elegans calcineurin TAX-6. Neuron 2002; 33: 751-63.
- 10. Lee J, Jee C, Song HO, Bandyopadhyay J, Lee JI, Yu JR, et al. Opposing functions of calcineurin and CaMKII regulate g-protein signaling in egg-laying behavior of C. elegans. J Mol Biol 2004; 344: 585-95.
- 11. Sutphin GL, Kaeberlein M. Measuring Caenorhabditis elegans lifespan on solid media. J Vis Exp 2009; 12: 1152.
- 12. Hekimi S and Guarente L. Genetics and the specificity of the aging process. Science 2003; 299:1351-54.
- 13. Luo Y. Long-lived worms and aging. Redox Rep 2004; 9: 65-9.
- 14. Corsi AK. A Biochemist guide to C. elegans. Anal Biochem 2006; 359: 1-17.
- 15. Guarente L, Kenyon C. Genetic pathways that regulate aging in model organisms. Nature 2000; 408: 255-62.
- 16. Klass MR. A method for the isolation of longevity mutants in the nematode caenorhabditis elegans and initial results. Mech Age Dev 1983; 22: 279-86.
- 17. Crabtree GR. Generic signals and specific minireview outcomes: signaling through Ca2+, calcineurin, and NF-at. Cell 1999; 96: 611-4.
- 18. Kingsbury TJ and Cunningham KW. A conserved family of calcineurin regulators. Genes Dev 2000; 14: 1595-604.
- 19. Lee JI, Mukherjee S, Yoon KH, Dwivedi M, Bandyopadhyay J. The multiple faces of calcineurin signaling in Caenorhabditis elegans: development, behaviour and aging. J Biosci 2013; 38:417-31.
- 20. Saygılı E, Rana O. R, Meyer C, et all. The angiotensin–calcineurin–NFAT pathway mediates stretch-induced up-regulation of matrix metalloproteinases-2/-9 in atrial myocytes. Basic Res Cardiol 2009; 104: 435-48.
- 21. Lee YM, Kim YK, Chung JH. Increased expression of TRPV1 channel in intrinsically aged and photoaged human skin in vivo. Exp Dermatol 2008; 18: 431-6.
- 22. Vincenti MP, Brinckerhoff CE. Transcriptional regulation of collagenase (MMP-1, MMP-13) genes in arthritis: integration of complex signaling pathways for the recruitment of gene specific transcription factors. Arthritis research 2002; 4: 157-64.
- 23. Fisher GJ, Wang ZQ, Datta SC, Varani J, Kang S, Voorhees JJ. Pathophysiology of premature skin aging induced by ultraviolet light. N Engl J Med 1997; 337: 1419-28.
- 24. Varani J, Warner RL, Gharaee-Kermani M et al. Vitamin A antagonizes decreased cell growth and elevated collagen-degrading matrix metalloproteinases and stimulates collagen accumulation in naturally aged human skin. J Invest Dermatol 2000; 114: 480-6.
- 25. Varani J, Hattori Y, Chi Y, Schmidt T, Perone P, Zeigler ME, et al. Elaboration of collagenolytic and gelatinolytic matrix metalloproteinases and their inhibitors by basal cell carcinomas of skin: comparison with normal skin. Br J Cancer 2000; 82: 657-65.
- 26. Hwang YP, Kim HG, Han EH, et al. N-Acetylglucosamine suppress collagenases activation in ultraviolet B-irradiated human dermal fibroblasts: Involvement of calcium ions and mitogen-activated protein kinases. J Dermatol Sci 2011; 63: 93-103.
- 27. Clapham DE. Calcium signaling. Cell 1995; 80: 259-68.
- 28. Hunter T. Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signaling. Cell 1995; 80: 225-36.
- 29. Akyol M, Özpınar N, Hayta SB, Özçelik S. Deri yaşlanması için bir model olarak Caenorhabditis elegans bireylerinde stronsiyum klorid heksahidrat’ın etkileri. XXV. Ulusal Dermatoloji Kongresi, 21-25 Ekim 2014, Antalya.
- 30. Donohoe DR, Jarvis RA, Weeks K, Aamodt EJ and Dwyer DS. Behavioral adaptation in C. elegans produced by antipsychotic drugs requires serotonin and is associated with calcium signaling and calcineurin inhibition. Neurosci Res 2009; 64: 280-89.
Abstract
Objective: It is known that
calcineurin inhibition prolongs life in individuals with Caenorhabditis elegans (C.
elegans). An important part of C.
elegans genes shows great similarity to human genes. This study aims to
evaluate the effects of a calcineurin inhibitor on vital functions in C. elegans individuals.
Method: In this study,
solutions of cyclosporine A which is a calcineurin inhibitör 1 μM (1.dose), 0.1
μM (2.dose), 0.01 μM (3.dose) and 0.001 μM (4.dose) were prepared and 1 ml from
each solution was taken and added to petri dishes, containing 10 ml Nematode
Growth Media (NGM). The control group was fed the medium which was not added
cyclosporine. The indicated doses of cyclosporine whose synchronization has
been completed were transferred to petri dishes in order that each would have
20 pieces. C. elegans were observed
using bottom light stereo microscop that provided 6X-50X magnification. Live
animals at the same time every day were counted and compared with the control
group until all C. elegans in all
petri dishes died.
Results: The results have
showed that there was a statistically significant increase in life span periods
of C. elegans exposed to cyclosporin
depending on dose increase (p<0,05).
It has been found out that physical development of C. elegans exposed to increased doses of cyclosporin decreased
significantly (p<0,05). The increase in doses of cyclosporin have not caused
statitically significant change in the number of pharynx pumping (p>0,05).
Conclusions: Calcineurin
inhibition slows down the developmental process in C. elegans and prolongs survival, and may produce positive effects
on aging.
References
- 1. Ünlü E. Deri yaşlanmasında korunma ve tedavi yöntemleri. Dermatoz 2010; 1: 23-31.
- 2. Ünal İ, Ertam İ. Factors Contributing to Skin Aging. Turkiye Klinikleri J Cosm Dermatol Special Topics 2008; 1: 1-7.
- 3. Hertweck M, Hoppe T, Baumeister R. C. elegans, a model for aging with highthroughput capacity. Exp Gerontol 2003; 38: 345-6.
- 4. Olsen A, Vantipalli MC, Lithgow GJ. Using Caenorhabditis elegans as a model for aging and age-related diseases. Ann N Y Acad Sci 2006; 1067: 120-8.
- 5. Smit NP, Van Rossum HH, Romijn FP, Sellar KJ, Breetveld M, Gibbs S et al. Calcineurin activity and inhibition in skin and (epi) dermal cell cultures. J Invest Dermatol 2008; 128:1686-90.
- 6. Fisher GJ, Duell EA, Nickoloff BJ, Annesley TM, Kowalke JK, Ellis CN et al. Levels of cyclosporin in epidermis of treated psoriasis patients differentially inhibit growth of keratinocytes cultured in serum free versus serum containing media. J Invest Dermatol 1988; 91: 142-6.
- 7. Nickoloff BJ, Fisher GJ, Mitra RS, Voorhees JJ. Direct cytopathic effects of cyclosporine A on rapidly proliferating cultured keratinocytes and dermal fibroblasts. Transplant Proc 1988; 20: 85-90.
- 8. Bandyopadhyay J, Lee J, Lee JI, Yu JR, Jee C, Cho JH, et al. Calcineurin, a calcium/calmodulin-dependent protein phosphatase, is involved in movement, fertility, egg laying, and growth in Caenorhabditis elegans. Mol Biol Cell 2002; 13: 3281-93.
- 9. Kuhara A, Inada H, Katsura I, Mori I. Negative regulation and gain control of sensory neurons by the C. elegans calcineurin TAX-6. Neuron 2002; 33: 751-63.
- 10. Lee J, Jee C, Song HO, Bandyopadhyay J, Lee JI, Yu JR, et al. Opposing functions of calcineurin and CaMKII regulate g-protein signaling in egg-laying behavior of C. elegans. J Mol Biol 2004; 344: 585-95.
- 11. Sutphin GL, Kaeberlein M. Measuring Caenorhabditis elegans lifespan on solid media. J Vis Exp 2009; 12: 1152.
- 12. Hekimi S and Guarente L. Genetics and the specificity of the aging process. Science 2003; 299:1351-54.
- 13. Luo Y. Long-lived worms and aging. Redox Rep 2004; 9: 65-9.
- 14. Corsi AK. A Biochemist guide to C. elegans. Anal Biochem 2006; 359: 1-17.
- 15. Guarente L, Kenyon C. Genetic pathways that regulate aging in model organisms. Nature 2000; 408: 255-62.
- 16. Klass MR. A method for the isolation of longevity mutants in the nematode caenorhabditis elegans and initial results. Mech Age Dev 1983; 22: 279-86.
- 17. Crabtree GR. Generic signals and specific minireview outcomes: signaling through Ca2+, calcineurin, and NF-at. Cell 1999; 96: 611-4.
- 18. Kingsbury TJ and Cunningham KW. A conserved family of calcineurin regulators. Genes Dev 2000; 14: 1595-604.
- 19. Lee JI, Mukherjee S, Yoon KH, Dwivedi M, Bandyopadhyay J. The multiple faces of calcineurin signaling in Caenorhabditis elegans: development, behaviour and aging. J Biosci 2013; 38:417-31.
- 20. Saygılı E, Rana O. R, Meyer C, et all. The angiotensin–calcineurin–NFAT pathway mediates stretch-induced up-regulation of matrix metalloproteinases-2/-9 in atrial myocytes. Basic Res Cardiol 2009; 104: 435-48.
- 21. Lee YM, Kim YK, Chung JH. Increased expression of TRPV1 channel in intrinsically aged and photoaged human skin in vivo. Exp Dermatol 2008; 18: 431-6.
- 22. Vincenti MP, Brinckerhoff CE. Transcriptional regulation of collagenase (MMP-1, MMP-13) genes in arthritis: integration of complex signaling pathways for the recruitment of gene specific transcription factors. Arthritis research 2002; 4: 157-64.
- 23. Fisher GJ, Wang ZQ, Datta SC, Varani J, Kang S, Voorhees JJ. Pathophysiology of premature skin aging induced by ultraviolet light. N Engl J Med 1997; 337: 1419-28.
- 24. Varani J, Warner RL, Gharaee-Kermani M et al. Vitamin A antagonizes decreased cell growth and elevated collagen-degrading matrix metalloproteinases and stimulates collagen accumulation in naturally aged human skin. J Invest Dermatol 2000; 114: 480-6.
- 25. Varani J, Hattori Y, Chi Y, Schmidt T, Perone P, Zeigler ME, et al. Elaboration of collagenolytic and gelatinolytic matrix metalloproteinases and their inhibitors by basal cell carcinomas of skin: comparison with normal skin. Br J Cancer 2000; 82: 657-65.
- 26. Hwang YP, Kim HG, Han EH, et al. N-Acetylglucosamine suppress collagenases activation in ultraviolet B-irradiated human dermal fibroblasts: Involvement of calcium ions and mitogen-activated protein kinases. J Dermatol Sci 2011; 63: 93-103.
- 27. Clapham DE. Calcium signaling. Cell 1995; 80: 259-68.
- 28. Hunter T. Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signaling. Cell 1995; 80: 225-36.
- 29. Akyol M, Özpınar N, Hayta SB, Özçelik S. Deri yaşlanması için bir model olarak Caenorhabditis elegans bireylerinde stronsiyum klorid heksahidrat’ın etkileri. XXV. Ulusal Dermatoloji Kongresi, 21-25 Ekim 2014, Antalya.
- 30. Donohoe DR, Jarvis RA, Weeks K, Aamodt EJ and Dwyer DS. Behavioral adaptation in C. elegans produced by antipsychotic drugs requires serotonin and is associated with calcium signaling and calcineurin inhibition. Neurosci Res 2009; 64: 280-89.
Details
| Subjects | Health Care Administration |
|---|---|
| Journal Section | Basic Science Research Articles |
| Authors | |
| Publication Date | September 19, 2017 |
| Acceptance Date | September 6, 2017 |
| Published in Issue | Year 2017 |