Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
권호 표지

Changes in Serum IGF-I and Spermatogenesis Analysed by Flow Cytometry in Growing Male Rabbit

성장 중인 수토끼에서 혈청 IGF-I 수준과 Flow Cytometry 측정에 의한 정자 형성의 변화

  • Lee J. H. (Department of Animal Science & Technology, Chung-Ang University, BET Research Institute) ;
  • Kim C. K. (Department of Animal Science & Technology, Chung-Ang University, BET Research Institute) ;
  • Chang Y. M. (Department of Animal Science & Technology, Chung-Ang University, BET Research Institute) ;
  • Ryu J. W. (Department of Animal Science & Technology, Chung-Ang University, BET Research Institute) ;
  • Park M. Y. (Department of Animal Science & Technology, Chung-Ang University, BET Research Institute) ;
  • Chung Y. C. (Department of Animal Science & Technology, Chung-Ang University, BET Research Institute) ;
  • Pang M. G. (Department of Animal Science & Technology, Chung-Ang University, BET Research Institute)
  • 이주형 (중앙대학교 산업과학대학 동물자원과학과) ;
  • 김창근 (중앙대학교 산업과학대학 동물자원과학과) ;
  • 장유민 (중앙대학교 산업과학대학 동물자원과학과) ;
  • 류재원 (중앙대학교 산업과학대학 동물자원과학과) ;
  • 박민영 (중앙대학교 산업과학대학 동물자원과학과) ;
  • 정영채 (중앙대학교 산업과학대학 동물자원과학과) ;
  • 방명걸 (중앙대학교 산업과학대학 동물자원과학과)
  • Published : 2005.09.01
Download PDF
⟨ Previous Next ⟩

Abstract

The aim of this study was to investigate the changes in insulin-like growth factor-I (IGF-I) and growth hormone (GH) in serum, the quantitation of spermato-genesis and the comparable relationships among these measurements during pubertal period in New Zealand White male rabbits. To investigate the age-related testicular changes in DNA contents of spermatogenic cells, the fine-needle testicular biopsies from males aged 10 to 28 wks were evaluated by flow cytometry(FCM). Body weight increased significantly between the ages of 12 and 20 wks (P<0.05) and reached 3.4 kg at 28 wks of age. The highest serum IGF-I level (451.3ng/mL) was observed at 20wks of age (P<0.05) and thereafter remained stable at low levels. Serum GH level at 18 wks of age was 183.3 pg/mL which was significantly higher compared to the other ages (P<0.05), and the rising time in serum GH tend to be somewhat earlier than that of IGF-I. The relative percentage of It-cells in testicular cell compartments was $48.2\%$ at the age of 18 wks which significantly increased than those of 16-wk-old (P<0.05) and thereafter increased with the advance of age to $68\%$. The percentage of 2C-cells in testis was $26.8\%$ at 18 wks of age which was significantly lower than $54.3\%$ at 16 wks old (P<0.05). The percentage of 4C-cells was constantly maintained $2\~6\%$ except the $9.9\%$ at 18 wks of age. In conclusion, the results suggest that the puberty onset occurred at about the 18 wks of age and that the IGF-I and GH in serum during the pubertal period showed the age/growth-specific changes and these changes might be related to the spermatogenesis. The DNA FCM combined with fine-needle testicular biopsy could offer a very sensitive method to monitor the quantitative spermatogenic events related to the puberty onset.

본 연구는 New Zealand White 수토끼에서 춘기발동 기간 동안에 혈청내 IGF-I(insulin-like growth factor-I)과 GH(growth hormone) 수준의 변화, 정자 형성에 따른 정소내 세포 구성 변화 및 이들 측정치들 간에 관계를 조사하기 위하여 실시되었다. 주령과 관련된 정소내 세포의 DNA 함량 변화 조사를 위하여 $10\~28$주령 수토끼 정소 조직의 fine-needle biopsy를 flow cytometry(FCM)로 분석하였다. 생체중은 $12\~20$주령 때 크게 증가되었고(P<0.05), 28주령 체중은 3.4kg이었다. 혈청 IGF-I 수준(451.3ng/mL)은 20주령에서 가장 높았으며(P<0.05), 그 후 낮은 수준으로 유지되었다. 혈청 GH 수준은 183.3pg/mL으로 다른 주령 때보다 현저히 높았으며(P<0.05), 상승시기가 IGF-I보다는 다소 빨랐다. 정소 조직세포 중 1C-세포의 상대적 비율은 18주령 때 $48.2\%$로 16주령보다 크게 상승되었고(P<0.05), 주령 증가와 더불어 $68\%$로 증가되었다. 2C-세포 비율은 18주령 때 $26.8\%$로 16주령의 $54.3\%$보다 현저히 낮았다(P<0.05). 4C-세포 비율은 18주령 때 $9.9\%$를 제외하고 $2\~6\%$를 유지하였다. 이상 결과에서 토끼는 춘기 발동 개시가 약 18주령에 일어나고 이 기간 중 IGF-I과 GH 수준의 변화가 나이 또는 체성장과 관계가 있었으며 그 영향이 정자 형성과 관련이 있음을 알 수 있었다. Fine-needle biopsy와 연계된 FCM은 춘기 발동 개시와 관련된 정자의 형성 과정을 평가하는데 매우 정확한 방법임을 확인할 수 있었다. 평균 CK-MB치는 $17.4\pm29.7\;IU/L$였다. 전체적인 이식편의 개통률은 $99.1\%\;(214/216)$(좌내유동맥 $100\%$, 요골동맥. $98.4\%$, 우내유동맥: $100\%$)를 보였다. 결론: 다중 복합 Y 동맥 이식편을 이용한 완전 동맥 무인공 심폐 바이패스 관상동맥우회술을 기술적인 문제점 없이 시행할 수 있었으며, 우수한 초기 임상 결과와 개통률을 보였다. 저자들은 특히 심장의 둔각 모서리 부위의 우회술시에 기존의 연쇄 문합에 비해 기술적으로 더 용이하며 복잡한 관상동맥우회술이 필요한 환자에게도 도움이 될 것으로 생각한다.보존하는 일에 등한시하여 왔다. 이 때문에 우리 민족 고유의 뿌리를 점차 잃어가고 있다. 이러한 시점에서 도시노점상을 정리하기 위한 목적으로 정부에서 도시 5일장을 개장한 것은 역사의 아이러니라 아니할 수 없다. 이렇게 정부주도로 개장된 5일장이 운영되어 온 지 2년이 되어가고 있다. 개장 초기에는 시에서의 지원도 적극적이고 소비자들의 호응도 좋았으나, 최근에 들어 활성화의 속도가 둔화되면서 도시 5일장의 개념을 재정립할 필요성이 제기되고 있다. 정부의 주도로 시작된 5일장이므로 정부의 적극적인 추진하에 풍물시장 번영회와 활성화 방안을 모색해야 한다. 쌍다리 풍물시장의 5일장을 활성화하기 위하여 도시 5일장의 개념을 ${\ulcorner}$국민들의 생활수준이 향상되고 여가를 즐길 수 있는 여건이 형성되고 있으므로 전통문화(향토문화)를 유지하고 시민들의 정서함양에 기여할 수 있는 여가공간 조성${\lrcorner}$으로 규정해야 한다. 이러한 개념 하에 5일장의 주체를 명확히 하기 위해 시청 지역경제과를 중심으로 지자제의 실시에 발맞춰 지역특색에 맞게 5일장을 활성화하기 위한 전학을 수립해야 한다

Keywords

References

  1. Amann RP, Lambiase, JTJr (1967): The male rabbit. I. change in semen characteristics and sperm output between puberty and one year of age. J Reprod Fertil 14:329-332 https://doi.org/10.1530/jrf.0.0140329
  2. Cailleau J, Vermeire S, Verhoeven G (1990): Independent control of the production of insulin-like growth factor I and its binding protein by cultured testicular cells. Mol Cell Endocrinol 69:79-89 https://doi.org/10.1016/0303-7207(90)90091-L
  3. Chadrashekar V, Bartke A (1993): Induction of exogenous insulin-like growth factor-I secretion alters the hypothalamic-pituitary-testicular function in growth hormone-deficient adult dwarf mice. Biol Reprod 47:544-551 https://doi.org/10.1095/​biolreprod48.3.544
  4. Chandrashekar V, Bartke A, Awoniyi CA, Tsai-Morris CH, Dafau ML, Russell LD, Kopchick JJ (2001): Testicular endocrine function in GH receptor gene disrupted mice. Endocrinology 142(8):3443-3450 https://doi.org/10.1210/en.142.8.3443
  5. Chandrashekar V, Zaczek D, Bartke A (2004): The consequences of altered somatotropic system on reproduction. Biol Reprod 71:17-27 https://doi.org/10.1095/biolreprod.103.027060
  6. Clark PA, Rogol AD (1996): Growth hormones and sex steroid interactions at puberty. Endocrinol Metab Clin North Am 25:665-681 https://doi.org/10.1016/S0889-8529(05)70346-7
  7. Clausen OPF, Purvis K, Hansson V (1977): Application of microflow fluorometry to studies of meiosis in the male rat. Biol Reprod 17:555-560 https://doi.org/10.1095/biolreprod17.4.555
  8. Codner E, Cassorla E (2002): Growth hormone and reproductive function. Mol Cell Endocrinol 186:133-136 https://doi.org/10.1016/S0303-7207(01)00653-0
  9. Copeland KC, Kuehl TJ. Castracane VD (1982): Pubertal endocrinology of the baboon : elevated somatomedin-c/insulin-like growth factor I at puberty. J Clin Endocrinol Metab 55:1198-1201 https://doi.org/10.1210/jcem-55-6-1198
  10. Daughaday W, Rotwein, P (1989): Insulin-like growth factors I and II : peptide, messenger ribonucleic acid and gene structure, serum, and tissue concentrations. Endocr Rev 10:68-91 https://doi.org/10.1210/edrv-10-1-68
  11. Davis SR, Hodgkinson SC, Prosser CG, Gluckman PD, Bcconomo FC, Collier RJ (1992): Effect of growth hormone treatment on the distribution of insulinlike growth factor-I between plasma and lymph of lactating sheep. J Endocrinol 132:339-344 https://doi.org/10.1677/joe.0.1320339
  12. Dey P, Mondal A, Singh SK, Vohra H (2000): Quantitation of spermatogenesis by DNA flow cytometry from fine-needle aspiration cytology material. Diagn Cytopathol 6:386-387
  13. Foote RH, Carney EW (2000): The rabbit as a model for reproductive and developmental toxicity studies. Reprod Toxicol 14:477-493 https://doi.org/10.1016/S0890-6238(00)00101-5
  14. Frame SR, Hurtt ME, Green JW (1994): Testicular maturation in prepubertal New Zealand rabbits. Vet Pathol 31:341-345
  15. Glass AR, Dahms WT, Swerdloff RS (1979): Body fat at puberty in rats: alteration by changes in diet. Pediatr Res 12:263-267
  16. Gnessi L, Fabbri A, Spera G (1997): Gonadal peptides as mediators of development and functional control of the testis : an integrated system with hormones and local environment. Endocr Rev 18: 541-608 https://doi.org/10.1210/er.18.4.541
  17. Hacker U, Schumann J, Gohde W, Muller K (1981): Mammalian spermatogenesis as a biologic dosimeter for radiation. Acta Radiol Oncol Radiat Phys Biol 20:279-282
  18. Handelsman DJ, Spaliviero JA, Scott CD, Baxter RC (1987): Normal regulation of the peripubertal surge of insulin-like growth factor-I in the rat. Endocrinology 120:491-496 https://doi.org/10.1210/endo-120-2-491
  19. Janca FC, Jost LK, Evenson DP (1986): Mouse testicular and sperm cell development characterized from birth to adulthood by dual parameter flow cytometry. Biol Reprod 34:613-623 https://doi.org/10.1095/biolreprod34.4.613
  20. Jones JL, Clemmons DR (1995): Insulin-like growth factor and their binding proteins : biological actions. Endocr Rev 16:3-34
  21. Katongole CB, Naftolin F, Short RV (1971): Relationship between blood levels of luteinizing hormone and testosterone in bulls and the effect of sexual stimulation. J Endocrinol 50:457-466 https://doi.org/10.1677/joe.0.0500457
  22. Keene DE, Suescun MO, Bostwick MG, Chandrashekar V, Bartke A, Kopchick, JJ (2002): Puberty is delayed in male growth hormone receptor gene-disrupted mice. J Androl 23:661-668
  23. Laron Z (2002): Growth hormone insensitivity (Laron syndrome). Rev Endocr Metab Disord 3:347-355 https://doi.org/10.1023/A:1020905725012
  24. Lee PDK, Rosenfeld RG (1987): Clinical utility of insulin-like growth factor assays. Pediatrician 14:154-161
  25. Lin T, Wang D, Calkins JH, Guo H, Chi R, Housley PR (1990): Regulation of insulin-like growth factor-I messenger ribonucleic acid expression in Leydig cells. Mol Cell Endocrinol 73:147-152 https://doi.org/10.1016/0303-7207(90)90127-T
  26. McCarthy MS, Hafs HD, Convey EM (1979): Serum hormone patterns associated with growth and sexual developments in bulls. J Anim Sci 49:1012-1020 https://doi.org/10.2527/jas1979.4941012x
  27. McFee AF (1963): Changes in testicular morphology and reproductive capacity of male rabbits following pre-pubertal exposures to gamma radiation. PhD thesis Cornell Univ
  28. Nakae J, Kido Y, Acelili D (2001): Distinct and overlaping functions of insulin and IGF-I receptors. Endocr Rev 22:818-835 https://doi.org/10.1210/er.22.6.818
  29. Ohyama K, Ohta M, Nakagomi Y, Yamori T, Sano T, Shimura Y, Sato K, Nakazawa S (1995): Effects of growth hormone and insulin-like growth factor I on testosterone secretion in premature male rats. Endocr J 42:817-820 https://doi.org/10.1507/endocrj.42.817
  30. Ojeda SR, Urbanski HF (1988): Puberty in the rat. In: Knobil, E. Neill, J. The physiology of reproduction. Raven press. New York, USA, pp 1699
  31. Pell JM, Bates PC (1992): Differential actions of growth hormone and insulin-like growth factor-I on tissue protein metabolism in dwarf mice. Endocrinology 130:1942-1950 https://doi.org/10.1210/en.130.4.1942
  32. Rawlings NC, Hafs HD, Swanson LV (1972): Testicular and blood serum androgens in Holstein bulls from birth through puberty. J Anim Sci 34: 435-440 https://doi.org/10.2527/jas1972.343435x
  33. Santos RL, Silva, CM, Ribeiro AFC, Vasconcelos AC, Pesquero JL, Coelho SG, Serakides R, Reis SR (1999): Effects of growth hormone and induced IGF-I release on germ cell population and apoptosis in the bovine testis. Theriogenology 51:975-984 https://doi.org/10.1016/S0093-691X(99)00043-6
  34. Skinner JD, Rowson A (1968): Puberty in suffolk and creoss-bred rams. J Reprod Fert 16:479-488 https://doi.org/10.1530/jrf.0.0160479
  35. Wang GM, O'Shaughnessy PJ, Chubb C, Robaire B, Hardy MP (2003): Effects of insulin-like growth factor I on steroidogenic enzyme expression levels in mouse Leydig cells. Endocrinology 144:5058-5064 https://doi.org/10.1210/en.2003-0563
  36. Yoon CY, Hong CM, Cho YY, Song JY, Hong IJ, Cho DH, Lee BJ, Song HI, Kim CK (2001): Flow cytometric evaluation on the age-dependent change of testicular DNA contents in rats. J Vet Sci 2:43-46