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Histological and Histochemical Studies on the Effect of Single Dose of Cyclophosphamide on Migration of Primordial Germ Cells of Fetal Charles Foster Rat – A Preliminary Study

Year 2007, Volume: 12 Issue: 4, 246 - 250, 01.08.2007

Abstract

Objectives: Effect of single dose Cyclophosphamide (CP), a known teratogen on early ovarian development was studied to understand whether it affects the primordial germ cells (PGC) in migratory phase. Materials and Methods: Adult female pregnant Charles foster rats were divided into control and experimental groups. Experimental group was given intraperitoneal injection of CP at a dose of 2mg/kg of body weight on day 10 of gestation when PGCs were in migratory phase. 12 fetuses from each group were sacrificed on 16th day of gestation to process for histochemical studies (alkaline phosphatase staining), whereas the ovaries of 12 fetuses from each group were collected on 20th day of gestation for histological studies (haematoxyline and eosin staining). Results: Examination of sections of the fetuses of group I showed clusters of large circular PGCs showing blackish brown color in 16 day old fetus. In experimental group, only homogeneous light brown cells were present. Ovaries of 20 day old fetus showed ovarian follicles with large germ cells in the centre in controls. Germ cells were totally absent in treated group. Conclusion: It was concluded that the CP, inhibited migration of PGCs to gonadal ridge leading to absence in ovary. ©2007, Fırat University, Medical Faculty

References

  • Williams PL,Warwick R, Dyson M, Bannister LH, Eds. Grays anatomy of the human body. 38th edition. London: Churchill Livingstone.1995.
  • Merchant H. Rat gonadal and ovarian organogenesis with and without germ cells. An ultrastructural study. Developmental Biology 1975; 44: 1-21.
  • Kemper CH, Peters PWJ. Migration and proliferation of primordial germ cells in the rat. Teratology 1987; 36: 117-124.
  • Molyneaux KA, Stallock J, Schaible K, Wylie C. Time-lapse analysis of living mouse germ cell migration. Dev Biol 2001; 240: 488-498.
  • Tam PPL, Liu WK. Gonadal development and fertility of mice treated prenatally with cadmium during the early organogenesis stages. Teratology 1985; 32: 453-462.
  • Tam PPL, Snow MHL. Proliferation and migration of primordial germ cells during compensatory growth in mouse embryos. J Embryol Exp Morphol 1981; 64: 133-147.
  • Wide M. Lead exposure on critical days of fetal life affects fertility in the female mouse. Teratology 1985; 32: 375-380.
  • Wide M, Argy R. Effect of inorganic lead on the Primordial Germ Cells in the Mouse Embryo. Teratology 1986; 34: 207-212.
  • Foley GE, Friedman OM, Drolet BP. Studies on the mechanism of action of cytoxan  evidence of activation in vivo and in vitro. Cancer Res 1961; 21: 57-63.
  • Hales BF. Comparison of the mutagenicity and teratogenicity of cyclophosphamide and its active metabolites, 4hydroxycyclophosphamide, phosphoramide mustard and acrolein. Cancer Research 1982; 42: 3016-3021.
  • Mirkes PE, Greenway JC, Rogers JG, Brundrett RB. Role of Acrolein in Cyclophosphamide Teratogenicity in Rat Embryos in Vitro. Toxicol Appl Pharmacol 1984; 72: 281-291.
  • Mohanty C, Singh G, Das BK, Saxena AK. Teratogenicity of Acrolein in rats. J.Anat Soc India 2000; 49: 46-48.
  • Saxena AK, Singh G. Effect of cyclophosphamide on the proliferation of the cellular contents of seminiferous tubules in developing rat testis. In Vitro Cell. Dev. Biol. Animal. Wochenschr1999; 84 : 393;
  • Pearse AG E. Histochemistry. Theoretical and applied. London: Churchill 1968.
  • Jaglarz MK, Howard KR. The active migration of Drosophila primordial germ cells. Development 1995; 121: 3495-3503.
  • Cleine JH. Replacement of posterior by anterior endoderm reduces sterility in embryos from inverted eggs of Xenopus laevis. J Embryol Exp Morphol 1986; 94: 83-93.
  • Anderson R, Fassler R, Georges-Labouesse E, et al. Mouse primordial germ cells lacking beta1 integrins enter the germline but fail to migrate normally to the gonads. Development 1999; 126: 1655-1664.
  • Liu YX. Interaction and signal transduction between oocyte and samatic cells in the ovary. Front Biosci. 2007; 12: 2782-2796.
  • Gondos B. In: Butt WR, Crooke AC, Ryle M .Gonadotrophins and Ovarian Development. Edinburgh: Livingstone, 1970. 239248.
  • Wakayama T, Hamada K, Yamamoto M, Suda T, Iseki S. The expression of platelet endothelial cell adhesion molecule-1 in mouse primordial germ cells during their migration and early gonadal formation. Histochem Cell Biol. 2003; 119: 355-362.
  • Eddy EM, Clark JM. The Ultrastructure of Endocrine and Reproductive Organs. In: Hess M ed. Electron Microscopic Concepts of Secretion. New York: Wiley, 1975: 151-168.
  • Goel S, Sugimoto M, Minami N, Yamada M, Kume S, Imai H. Identification, isolation, and in vitro culture of porcine gonocytes. Biol Reprod. 2007; 77: 127-137.
  • Weidinger G, Wolke U, Köprunner M, Thisse C, Thisse B, Raz E. Regulation of zebrafish primordial germ cell migration by attraction towards an intermediate target. Development 2002; 129: 25-36.
  • Godin I, Wylie C, Heasman J. Genital ridges exert long-range effects on mouse primordial germ cell numbers and direction of migration in culture. Development 1990; 108: 357-363.
  • Kuwana T, Rogulska T. Migratory mechanisms of chick primordial germ cells toward gonadal anlage. Cell Mol Biol 1999; 45: 725-736.
  • Van Doren M, Broihier HT, Moore LA, Lehmann R . HMG-CoA reductase guides migrating primordial germ cells. Nature 1998; 396: 466-469.

Fetal Charles foster sıçanların primordial germ hücrelerinin migrayonu üzerine tek doz siklofosfamit'in etkilerinin histolojik ve histokimyasal olarak incelenmesi: Bir ön çalışma

Year 2007, Volume: 12 Issue: 4, 246 - 250, 01.08.2007

Abstract

Amaç: Erken ovaryan gelişimin üzerinde teratojen olduğu bilinen siklofosfamitin (CP), tek dozunun primordial germ hücrelerinin (PGC) migrasyon fazını etkileyip etkilemediği çalışıldı. Gereç ve Yöntem: Yetişkin gebe Charles foster sıçanları kontrol ve deney grubu olmak üzere iki gruba ayrıldı. Deney grubundaki hayvanlara, PGC'nin migrasyon fazında olduğu gestasyonun 10. gününde 2 mg/kg dozunda CP intraperitoneal olarak yapıldı. Her gruba ait 12 fetus gebeliğin 16. gününde histokimyasal çalışmalar (alkalin fosfataz boyama) için çıkarılırken, her gruba ait 12 fetüsün ovaryumları da gebeliğin 20. gününde histolojik çalışmalar (hematoksilen-eosin boyama) için toplandı. Bulgular: Grup I'e ait fetüslerin kesitlerinde geniş sirküler PGC kümeleri, 16 günlük fetüste siyahımsı kahverengi olarak göründü. Deney grubunda ise homojen açık kahverengi hücreler vardı. Kontrol grubuna ait 20 günlük fetüslerin overlerinde, merkezinde geniş germ hücrelerinin olduğu ovaryan foliküller gözlendi. Deney grubunda ise germ hücreleri tamamen yoktu. Sonuç: Siklofosfamit uygulaması primordial germ hücrelerinin gonadal kabartıya göçlerini inhibe etmekte ve overler içinde germ hücrelerinin olmamasına neden olmaktadır. ©2007, Fırat Üniversitesi, Tıp Fakültesi

References

  • Williams PL,Warwick R, Dyson M, Bannister LH, Eds. Grays anatomy of the human body. 38th edition. London: Churchill Livingstone.1995.
  • Merchant H. Rat gonadal and ovarian organogenesis with and without germ cells. An ultrastructural study. Developmental Biology 1975; 44: 1-21.
  • Kemper CH, Peters PWJ. Migration and proliferation of primordial germ cells in the rat. Teratology 1987; 36: 117-124.
  • Molyneaux KA, Stallock J, Schaible K, Wylie C. Time-lapse analysis of living mouse germ cell migration. Dev Biol 2001; 240: 488-498.
  • Tam PPL, Liu WK. Gonadal development and fertility of mice treated prenatally with cadmium during the early organogenesis stages. Teratology 1985; 32: 453-462.
  • Tam PPL, Snow MHL. Proliferation and migration of primordial germ cells during compensatory growth in mouse embryos. J Embryol Exp Morphol 1981; 64: 133-147.
  • Wide M. Lead exposure on critical days of fetal life affects fertility in the female mouse. Teratology 1985; 32: 375-380.
  • Wide M, Argy R. Effect of inorganic lead on the Primordial Germ Cells in the Mouse Embryo. Teratology 1986; 34: 207-212.
  • Foley GE, Friedman OM, Drolet BP. Studies on the mechanism of action of cytoxan  evidence of activation in vivo and in vitro. Cancer Res 1961; 21: 57-63.
  • Hales BF. Comparison of the mutagenicity and teratogenicity of cyclophosphamide and its active metabolites, 4hydroxycyclophosphamide, phosphoramide mustard and acrolein. Cancer Research 1982; 42: 3016-3021.
  • Mirkes PE, Greenway JC, Rogers JG, Brundrett RB. Role of Acrolein in Cyclophosphamide Teratogenicity in Rat Embryos in Vitro. Toxicol Appl Pharmacol 1984; 72: 281-291.
  • Mohanty C, Singh G, Das BK, Saxena AK. Teratogenicity of Acrolein in rats. J.Anat Soc India 2000; 49: 46-48.
  • Saxena AK, Singh G. Effect of cyclophosphamide on the proliferation of the cellular contents of seminiferous tubules in developing rat testis. In Vitro Cell. Dev. Biol. Animal. Wochenschr1999; 84 : 393;
  • Pearse AG E. Histochemistry. Theoretical and applied. London: Churchill 1968.
  • Jaglarz MK, Howard KR. The active migration of Drosophila primordial germ cells. Development 1995; 121: 3495-3503.
  • Cleine JH. Replacement of posterior by anterior endoderm reduces sterility in embryos from inverted eggs of Xenopus laevis. J Embryol Exp Morphol 1986; 94: 83-93.
  • Anderson R, Fassler R, Georges-Labouesse E, et al. Mouse primordial germ cells lacking beta1 integrins enter the germline but fail to migrate normally to the gonads. Development 1999; 126: 1655-1664.
  • Liu YX. Interaction and signal transduction between oocyte and samatic cells in the ovary. Front Biosci. 2007; 12: 2782-2796.
  • Gondos B. In: Butt WR, Crooke AC, Ryle M .Gonadotrophins and Ovarian Development. Edinburgh: Livingstone, 1970. 239248.
  • Wakayama T, Hamada K, Yamamoto M, Suda T, Iseki S. The expression of platelet endothelial cell adhesion molecule-1 in mouse primordial germ cells during their migration and early gonadal formation. Histochem Cell Biol. 2003; 119: 355-362.
  • Eddy EM, Clark JM. The Ultrastructure of Endocrine and Reproductive Organs. In: Hess M ed. Electron Microscopic Concepts of Secretion. New York: Wiley, 1975: 151-168.
  • Goel S, Sugimoto M, Minami N, Yamada M, Kume S, Imai H. Identification, isolation, and in vitro culture of porcine gonocytes. Biol Reprod. 2007; 77: 127-137.
  • Weidinger G, Wolke U, Köprunner M, Thisse C, Thisse B, Raz E. Regulation of zebrafish primordial germ cell migration by attraction towards an intermediate target. Development 2002; 129: 25-36.
  • Godin I, Wylie C, Heasman J. Genital ridges exert long-range effects on mouse primordial germ cell numbers and direction of migration in culture. Development 1990; 108: 357-363.
  • Kuwana T, Rogulska T. Migratory mechanisms of chick primordial germ cells toward gonadal anlage. Cell Mol Biol 1999; 45: 725-736.
  • Van Doren M, Broihier HT, Moore LA, Lehmann R . HMG-CoA reductase guides migrating primordial germ cells. Nature 1998; 396: 466-469.
There are 26 citations in total.

Details

Primary Language Turkish
Journal Section Articles
Authors

Biswabina Ray This is me

Bhagath Kumar Potu This is me

Publication Date August 1, 2007
Published in Issue Year 2007 Volume: 12 Issue: 4

Cite

APA Ray, B., & Potu, B. K. (2007). Fetal Charles foster sıçanların primordial germ hücrelerinin migrayonu üzerine tek doz siklofosfamit'in etkilerinin histolojik ve histokimyasal olarak incelenmesi: Bir ön çalışma. Fırat Tıp Dergisi, 12(4), 246-250.
AMA Ray B, Potu BK. Fetal Charles foster sıçanların primordial germ hücrelerinin migrayonu üzerine tek doz siklofosfamit'in etkilerinin histolojik ve histokimyasal olarak incelenmesi: Bir ön çalışma. Fırat Tıp Dergisi. August 2007;12(4):246-250.
Chicago Ray, Biswabina, and Bhagath Kumar Potu. “Fetal Charles Foster sıçanların Primordial Germ hücrelerinin Migrayonu üzerine Tek Doz siklofosfamit'In Etkilerinin Histolojik Ve Histokimyasal Olarak Incelenmesi: Bir ön çalışma”. Fırat Tıp Dergisi 12, no. 4 (August 2007): 246-50.
EndNote Ray B, Potu BK (August 1, 2007) Fetal Charles foster sıçanların primordial germ hücrelerinin migrayonu üzerine tek doz siklofosfamit'in etkilerinin histolojik ve histokimyasal olarak incelenmesi: Bir ön çalışma. Fırat Tıp Dergisi 12 4 246–250.
IEEE B. Ray and B. K. Potu, “Fetal Charles foster sıçanların primordial germ hücrelerinin migrayonu üzerine tek doz siklofosfamit'in etkilerinin histolojik ve histokimyasal olarak incelenmesi: Bir ön çalışma”, Fırat Tıp Dergisi, vol. 12, no. 4, pp. 246–250, 2007.
ISNAD Ray, Biswabina - Potu, Bhagath Kumar. “Fetal Charles Foster sıçanların Primordial Germ hücrelerinin Migrayonu üzerine Tek Doz siklofosfamit'In Etkilerinin Histolojik Ve Histokimyasal Olarak Incelenmesi: Bir ön çalışma”. Fırat Tıp Dergisi 12/4 (August 2007), 246-250.
JAMA Ray B, Potu BK. Fetal Charles foster sıçanların primordial germ hücrelerinin migrayonu üzerine tek doz siklofosfamit'in etkilerinin histolojik ve histokimyasal olarak incelenmesi: Bir ön çalışma. Fırat Tıp Dergisi. 2007;12:246–250.
MLA Ray, Biswabina and Bhagath Kumar Potu. “Fetal Charles Foster sıçanların Primordial Germ hücrelerinin Migrayonu üzerine Tek Doz siklofosfamit'In Etkilerinin Histolojik Ve Histokimyasal Olarak Incelenmesi: Bir ön çalışma”. Fırat Tıp Dergisi, vol. 12, no. 4, 2007, pp. 246-50.
Vancouver Ray B, Potu BK. Fetal Charles foster sıçanların primordial germ hücrelerinin migrayonu üzerine tek doz siklofosfamit'in etkilerinin histolojik ve histokimyasal olarak incelenmesi: Bir ön çalışma. Fırat Tıp Dergisi. 2007;12(4):246-50.