Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
JCR Impact Factor (IF) – 2.1 (5-Year IF – 2.0)
Journal Citation Indicator (JCI) (2023) – 0.4
Scopus CiteScore – 3.7 (CiteScore Tracker 3.3)
Index Copernicus  – 161.11; MNiSW – 70 pts

ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
Periodicity – monthly

Download original text (EN)

Advances in Clinical and Experimental Medicine

2013, vol. 22, nr 3, May-June, p. 327–335

Publication type: original article

Language: English

The Detrimental Effects of Diabetes on Pluripotency Determined by KLF4, SOX2, C-MYC and OCT4 Immunoreactivity in Rat Testes

Szkodliwy wpływ cukrzycy na omnipotencjalność określoną przez immunoreaktywność Klf4, Sox2, c-myc i Oct4 w jądrze szczura

Huseyin Aktug1,A,B,C,E, Aysegul Uysal1,C,D,E,F, Altug Yavasoglu1,A,B,C,D, Fatih Oltulu1,B,C,D, Saadet O. Akarca1,B,C,D, Ozlem Yilmaz-Dilsiz1,D,E,F, Gulperi Oktem1,A,B,C,E

1 Ege University Faculty of Medicine, Department of Histology and Embryology, Bornova, Izmir, Turkey

Abstract

Background. Diabetes mellitus (DM) is a multisystem disorder. Type 1 DM can be experimentally induced in rats with streptozotocin (STZ). Diabetic conditions result in testicular oxidative stress and suppressed male reproductive activity as well as decreases in both testicular organ weights and subject weights
Objectives. The purpose of this study was to investigate immunohistochemical differences in testicular tissue due to STZ induced diabetes regarding pluripotency via transcription factors like Klf4, Sox2, c-Myc and Oct4, and to determine weight changes in both the subjects and the testes during the experiment.
Material and Methods. Diabetes was induced in male adult rats for this study. A healthy control group and a diabetic group were observed for one month. Blood glucose levels over 250 mg/dL were considered diabetic.
Results. On days 0, 3, 15 and 30, the subjects’ weights and testicular organ weights were determined and analyzed. The results revealed statistically significant decreases (p < 0.05 and p < 0.001, respectively). Semiquantitative immunohistochemical analyses of Klf4, Sox2, c-Myc and Oct4 were studied in testes paraffin sections via light microscopy. Decreased immunoreactivity of Klf4 was observed in the diabetic group in comparison to the controls. Spermatogonial cells and Sertoli cells showed increased immunostaining for Sox2 and c-Myc, while decreased immunoreactivity of Oct4 was noted for both spermatogenic and Sertoli cells compared to the control group.
Conclusion. This study clearly demonstrated that Klf4, Sox2 and Oct4 immunopositive cells in adult male rat testes manifested sustainable pluripotency and that diabetes has dramatically detrimental effects on this trait.

Streszczenie

Wprowadzenie.Cukrzyca (DM) jest wielonarządową chorobą. Typ 1 DM można doświadczalnie wywołać u szczurów za pomocą streptozotocyny (STZ). Warunki, jakie wywołuje cukrzyca, powodują stres oksydacyjny jąder i hamują aktywność reprodukcyjną osobników płci męskiej, a także zmniejsza się masa jąder i samych osobników.
Cel pracy. Zbadanie immunohistochemicznych różnic tkanki jąder na skutek cukrzycy wywołanej przez STZ dotyczących omnipotencjalności za pomocą czynników transkrypcyjnych, takich jak Klf4, Sox2, c-myc i Oct4, oraz ustalenie zmian masy ciała u badanych osobników i ich jąder podczas badania doświadczalnego.
Materiał i metody. W tym badaniu cukrzycę wywołano u dorosłych samców szczurów. Grupę kontrolną i badaną obserwowano przez miesiąc. Jeśli stężenie glukozy we krwi było powyżej 250 mg/dl, osobnika uznawano za chorego na cukrzycę.
Wyniki. W dniach 0, 3, 15 i 30 zmierzono masę ciała osobników i ich jąder. Wyniki wykazały jej statystycznie istotne zmniejszenie (p < 0,05 i p < 0,001). Wykonano półilościową immunohistochemiczną analizę Klf4, Sox2, c-myc i Oct4 parafinowych wycinków tkanki jąder za pomocą mikroskopu optycznego. Wykryto zmniejszoną immunoreaktywność Klf4 w grupie chorych na cukrzycę, w porównaniu do grupy kontrolnej. Spermatogonia i komórki Sertoliego wykazały zwiększone barwienie immunologiczne dla Sox2 i c-myc, a zmniejszoną immunoreaktywność Oct4 zaobserwowano zarówno dla spermatogoniów, jak i komórek Sertoliego w porównaniu z grupą kontrolną.
Wnioski. Badanie to wyraźnie pokazało, że immunopozytywne komórki Klf4, Sox2 i Oct4 w jądrach dorosłych szczurów wykazują zrównoważoną omnipotencjalność i że cukrzyca ma znacząco niekorzystny wpływ na tę cechę.

Key words

Klf4 protein, SOX2, c-Myc transcription factor, Oct4 protein, rat, testes, diabetes mellitus, experimental.

Słowa kluczowe

białko Klf4, Sox2, czynnik transkrypcyjny c-Myc, białko Oct4, szczur, jądra, cukrzyca, badanie doświadczalne.

References (31)

  1. Amaral S, Oliveira PJ, Ramalho-Santos J: Diabetes and the impairment of reproductive function: possible role of mitochondria and reactive oxygen species. Curr Diabetes rev 2008, 4(1), 46–54.
  2. Masuzawa K, Goto K, Jesmin S, Maeda S, Miyauchi T, Kaji Y et al.: An endothelin type A receptor antagonist reverses upregulated VEGF and ICAM-1 levels in streptozotocin-induced diabetic rat retina. Curr Eye res 2006, 31(1), 79–89.
  3. Agarwal A, Saleh RA, Bedaiwy MA: role of reactive oxygen species in the pathophysiology of human reproduction. Fertil Steril 2003, 79(4), 829–843.
  4. Navarro-Casado L, Juncos-Tobarra MA, Cháfer-Rudilla M, de Onzoño LÍ, Blázquez-Cabrera JA, MirallesGarcía JM: Effect of experimental diabetes and STZ on male fertility capacity. Study in rats. J Androl 2010, 31(6), 584–592.
  5. Ratajczak MZ, Machalinski B, Wojakowski W, Ratajczak J, Kucia M: A hypothesis for an embryonic origin of pluripotent Oct4 (+) stem cells in adult bone marrow and other tissues. Leukemia 2007, 21(5), 860–867.
  6. Sun C, Liu YK: Induced pluripotent cancer cells: progress and application. J Cancer res Clin Oncol 2011 Jan, 137(1), 1–8. doi: 10.1007/s00432-010-0955-z. Epub 2010 Nov 26.
  7. Takahashi K, Yamanaka S: Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006, 126(4), 663–676.
  8. Watanabe S, Hirai H, Asakura Y, Tastad C, Verma M, Keller C et al.: MyoD gene suppression by oct4 is required for reprogramming in myoblasts to produce induced pluripotent stem cells. Stem Cells 2011, 29(3), 505–516.
  9. Godmann M, Kosan C, Behr R: Kruppel-like factor 4 is widely expressed in the mouse male and female reproductive tract and responds as an immediate early gene to activation of the protein kinase A in TM4 Sertoli cells. reproduction 2010, 139(4),771–782.
  10. Wei Z, Yang Y, Zhang P, Andrianakos R, Hasegawa K, Lyu J et al.: Klf4 interacts directly with Oct4 and Sox2 to promote reprogramming. Stem Cells 2009, 27(12), 2969–2978.
  11. Rosner MH, Vigano MA, Ozato K, Timmons PM, Poirier F, Rigby PW et al.: A pOU-domain transcription factor in early stem cells and germ cells of the mammalian embryo. Nature 1990, 345(6277), 686–692.
  12. Schöler HR, Ruppert S, Suzuki N, Chowdhury K, Gruss P: New type of pOU domain in germ line-specific protein Oct-4. Nature 1990, 344(6265), 435–439.
  13. Niwa H: How is pluripotency determined and maintained? Development 2007, 134(4), 635–646.
  14. Pelengaris S, Khan M, Evan G: c-Myc: more than just a matter of life and death. Nature rev Cancer 2002, 2(10), 764–776.
  15. Lengner CJ: ipS cell technology in regenerative medicine. Ann NY Acad Sci 2010, 1192, 38–44.
  16. Ekwall H, Jansson A, Sjöberg P, Plöen L: Differentiation of the rat testis between 20 and 120 days of age. Arch Androl 1984, 13(1), 27–36.
  17. Taşkara E, Gör A, Kutlu O, Karagüzel E, Cobanoğlu U, Topbaş M et al.: Does propofol prevent testicular ischemia-reperfusion injury due to torsion in the long term? pediatr Surg Int 2011, Apr 6. [Epub ahead of print]
  18. Shalaby MA, Hamowieh AR: Safety and efficacy of Zingiber officinale roots on fertility of male diabetic rats. Food Chem Toxicol 2010, 48(10), 2920–2924.
  19. Godmann M, Kromberg I, Mayer J, Behr R: The mouse Kruppel-like factor 4 (Klf4) gene: four functional polyadenylation sites which are used in a cell-specific manner as revealed by testicular transcript analysis and multiple processed pseudogenes. Gene 2005, 361, 149–156.
  20. Godmann M, Gashaw I, Katz JP, Nagy A, Kaestner KH, Behr R: Kruppel-like factor 4, a “pluripotency transcription factor” highly expressed in male postmeiotic germ cells, is dispensable for spermatogenesis in the mouse. Mech Dev 2009, 126(8–9), 650–664.
  21. Behr R, Deller C, Godmann M, Muller T, Bergmann M, Ivell R et al.: Kruppel-like factor 4 expression in normal and pathological human testis. Mol Hum reprod 2007, 13(11), 815–820.
  22. McLean DJ, Friel PJ, Pouchnik D, Griswold MD: Oligonucleotide microarray analysis of gene expression in follicle-stimulating hormone treated rat Sertoli cells. Mol Endocrinol 2002, 16(12), 2780–2792.
  23. Zindy F, Eischen CM, Randle DH, Kamijo T, Cleveland JL, Sherr CJ et al.: Myc signaling via the ArF tumor suppressor regulates p53-dependent apoptosis and immortalization. Genes Dev 1998, 12(15), 2424–2433.
  24. Zhang W, Geiman DE, Shields JM, Dang DT, Mahatan CS, Kaestner KH et al.: The gut-enriched Kruppel-like factor (Kruppel-like factor 4) mediates the transactivating effect of p53 on the p21WAF1/Cip1 promoter. J Biol Chem 2000, 275(24), 18391–18398.
  25. Seoane J, Le HV, Massague J: Myc suppression of the p21(Cip1) Cdk inhibitor influences the outcome of the p53 response to DNA damage. Nature 2002, 419(6908), 729–734.
  26. Sánchez-Sánchez AV, Camp E, García-España A, Leal-Tassias A, Mullor JL: Medaka Oct4 is expressed during early embryo development, and in primordial germ cells and adult gonads. Dev Dyn 2010, 239(2), 672–679.
  27. Scaldaferri ML, Fera S, Grisanti L, Sanchez M, Stefanini M, De Felici M et al.: Identification of side population cells in mouse primordial germ cells and prenatal testis. Int J Dev Biol 2011, 55, 209–214.
  28. Gubbay J, Collignon J, Koopman P, Capel B, Economou A, Münsterberg A et al.: A gene mapping to the sexdetermining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes. Nature 1990, 346(6281), 245–250.
  29. Xiao YM, Chen L, Liu J, Liu WB, Chen HG, Zou LJ et al.: Contrast expression patterns of JNK1 during sex reversal of the rice-field eel. J Exp Zool B Mol Dev Evol 2010, 314(3), 242–256.
  30. Fernandez PC, Frank SR, Wang L, Schroeder M, Liu S, Greene J et al.: Genomic targets of the human c-Myc protein. Genes Dev 2003, 17(9), 1115–1129.
  31. Cheung L, Zervou S, Mattsson G, Abouna S, Zhou L, Ifandi V et al.: c-Myc directly induces both impaired insulin secretion and loss of β-cell mass, independently of hyperglycemia in vivo. Islets 2010, 2(1), 37–45.
© Wroclaw Medical University