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Kolşisin ve Tripsin Uygulamalarının Yetişkin Dişi ve Erkek Nil Tilapya Dokularından ve Larvalarından Elde Edilen Metafaz Kromozom Yayılımı Üzerine Etkileri

Year 2024, Volume: 10 Issue: 3, 113 - 118, 28.12.2024
https://doi.org/10.58626/menba.1538957

Abstract

Birçok alanda olduğu gibi akuakültürde poliploidi, ginogenez ve androgenez gibi farklı biyoteknolojik yöntemlerin sonuçlarının değerlendirilmesi için sitogenetik tekniklere ihtiyaç duyulmaktadır. Düzenli dağılmış metafaz kromozom yayılımları elde etmek, sitogenetik teknikler için öncelikli bir gerekliliktir. Bu çalışmanın temel amacı kolşisin ve tripsin uygulamalarının, sayılabilir metafaz kromozom yayılımlarının verimi üzerindeki etkilerini değerlendirmektir. Bu amaçla, yetişkin dişi ve erkek Nil tilapyaları solungaç, dalak, böbrek ve kuyruk yüzgeci dokularından ve ayrıca larvalarının tüm vücudundan kromozom preparatları hazırlanmıştır. Önceki çalışmalara göz önüne alındığında, bu çalışma kolşisinin ve tripsinin metafaz kromozom yayılımları üzerindeki etkilerini cinsiyet farklılıkları ve larvalar açısından gösteren ilk sistematik ve kapsamlı çalışmadır. Kromozom preparatlarında “Tripsin (-) Kolşisin (-)”, “Tripsin (-) Kolşisin (+)”, “Tripsin (+) Kolşisin (-)” ve “Tripsin (+) Kolşisin (+)” olmak üzere dört uygulama belirlenmiştir. Kromozom preparatlarının hazırlanmasında, kolşisinin ve tripsinin kullanımları açısından etkileri, metafaz hücrelerinin sayıları belirlenerek değerlendirilmiştir. Bu amaçla, uygulamalar arasında tespit edilen metafaz hücrelerindeki farklılıkları ortaya çıkarmak için metafaz indeksi kullanılmıştır. Tüm dokularda en yüksek metafaz indeksi değerleri Tripsin (+) Kolşisin (+) uygulama grubunda bulunmuştur. Ayrıca, dalak ve böbrek dokularında bulunan metafaz indeksi değerleri, solungaç ve yüzgeç dokularında bulunanlardan daha yüksek olduğu saptamıştır. Bu çalışma ayrıca metafaz kromozom dağılımlarının kolşisin veya tripsin uygulamaları olmaksızın da elde edilebileceğini göstermiştir.

Ethical Statement

Bu çalışma Aksaray Üniversitesi Deney Hayvanları Uygulama Araştırma Merkezi 2021/1-3 sayılı kararı ile onaylanmıştır.

Supporting Institution

Bu çalışma Aksaray Üniversitesi Bilimsel Araştırma Projeleri Koordinasyon Birimi tarafından desteklenen 2021/021 numaralı proje kapsamında gerçekleştirilmiştir

Project Number

Aksaray University Coordinatorship of Scientific Research Projects (2021/021)

Thanks

Aksaray Üniversitesi'nden Prof. Dr. Mustafa Öz'e örneklemelerdeki yardımları için teşekkür ederim. Ayrıca yazar, Aksaray Üniversitesi Embriyo Transferi Eğitim Uygulama Araştırma Merkezi'nde floresan mikroskobu altında kesitleri inceleme fırsatı verdiği için Prof. Dr. Tahir Karaşahin'e özellikle teşekkürlerimi sunarım.

References

  • Bertão, M.R., & Aguiar-Perecin, M.L.R. (2002). Maize somatic chromosome preparation: pretreatments and genotypes for obtention of high index of metaphase accumulation. Caryologia, 55(2): 115-119. https://doi.org/10.1080/00087114.2002.10589266
  • Blanco, D.R., L.A.C. Bertollo, R.L. Lui, M.R. Vicari, V.P. Margarido, R.F. Artoni & O. Moreira Filho. (2012). A new technique for obtaining mitotic chromosome spreads from fishes in the field. J. Fish Biol., 81:351-357. https://doi.org/10.1111/j.1095-8649.2012.03325.x
  • Cattin, P.M. & Ferreira, J.T. (1989). A rapid, nonsacrificial chromosome preparation technique for freshwater teleosts. South African Journal of Zoology, 24:76-78. https://doi.org/10.1080/02541858.1989.11448136
  • Chourrout, D., & Happe, A., (1986). Improved methods of direct chromosome preparation in rainbow trout, Salmo gairdneri. Aquaculture, 52: 255-261. https://doi.org/10.1016/0044-8486(86)90368-6
  • Demirok, N.K. & Ünlü, E., (2001). Karyotypes of Cyprinid fish Capoeta trutta and Capoeta capoeta umbla (Cyprinidae) from the Tigris River. Turkish Journal of Zoology, 25: 389-393.
  • Deng, W., Tsao, S.W., Lucas, J.N., Leung, C.S., & Cheung, A.L. (2003). A new method for improving metaphase chromosome spreading. Cytometry A, 51(1):46-51. https://doi.org/10.1002/cyto.a.10004
  • Foresti, F., Oliveira, C., & Foresti de Almeida-Toledo, L. (1993). A method for chromosome preparations from large fish specimens using in vitro short-term treatment with colchicine. Experientia, 49: 810-813. https://doi.org/10.1007/BF01923555
  • Harvey, S., Campos-Ramos, R., Kennedy, D.D., Ezaz, M.T., Bromage, N.R., Griffin, D.K., & Penman, D.J. (2002). Karyotype evolution in tilapia: mitotic and meiotic chromosome analysis of Oreochromis Karongae and O. Niloticus × O. Karongae Hybrids. Genetica 115:169-177. https://doi.org/10.1023/A:1020190918431
  • Henegariu, O., Heerema, N.A., Wright, L.L., Brayward, P., Ward, D.C., & Vance G.H. (2001). Improvements in cytogenetic slide preparation: Controlled chromosome spreading, chemical aging and gradual denaturing. Cytometry, 43: 101-109. https://doi.org/10.1002/1097-0320(20010201)43:2<101::AID-CYTO1024>3.0.CO;2-8
  • Howe, B., Umrigar, A., & Tsien, F. (2014). Chromosome preparation from cultured cells. J. Vis. Exp., 28(83):e50203. https://doi.org/10.3791/50203
  • Hussain, M.G., & McAndrew, B.J. (1994). An improved technique for chromosome karyotyping from embryonic and soft tissues of Tilapia and Salmonids. Asian Fisheries Science, 7: 187-190.
  • Moreira-Filho, O., & Bertollo, L.A.C. (1991). Extraction and use of the cephalic kidney for chromosome studies in small fi sh. Braz. J. Genet., 14:1085-1090.
  • Mukti, A.T., Carman, O., Alimuddin, & Zairin Jr, M. (2016). A rapid chromosome preparation technique without metaphase arrest for ploidy determination in Nile tilapia, Oreochromis niloticus, Caryologia, 69(2):175-180. https://doi.org/10.1080/00087114.2016.1152112
  • O'Connor, C. (2008). Karyotyping for chromosomal abnormalities. Nature Education 1(1):27.
  • Pope, L.H., Xiong, C., & Marko, J.F. (2006). Proteolysis of mitotic chromosomes induces gradual and anisotropic decondensation correlated with a reduction of elastic modulus and structural sensitivity to rarely cutting restriction enzymes. Mol. Biol. Cell., 17(1):104-113. https://doi.org/10.1091/mbc.e05-04-0321
  • Pradeep, P.J., Srijaya, T.C., Zain, R.B.M., Papini, A., & Chatterji, A.K. (2011). A simple technique for chromosome preparation from embryonic tissues of teleosts for ploidy verification. Caryologia, 64(2): 235-241. https://doi.org/10.1080/00087114.2002.10589788
  • Rieder, C.L., & Palazzo, R.E. (1992). Colcemid and the mitotic cycle. J. Cell Sci., 102(3):387-392. https://doi.org/10.1242/jcs.102.3.387
  • Schneider, C.A., Rasband, W.S., & Eliceiri, K.W. (2012). NIH Image to ImageJ: 25 years of image analysis. Nature Methods, 9(7):671-675. https://doi.org/10.1038/nmeth.2089
  • Supiwong, W., Tanomtong, A., Supanuam, P., Seetapan, K., Khakhong, S., & Sanoamuang, L. (2013). Chromosomal Characteristic of Nile Tilapia (Oreochromis niloticus) from Mitotic and Meiotic Cell Division by T-Lymphocyte Cell Culture. Cytologia, 78(1): 9-14. https://doi.org/10.1508/cytologia.78.9
  • Yamazaki, F., Onazato, H., & Arai, K. (1981). The chopping method for obtaining permanent chromosome preparation from embryos of teleost fishes. Bulletin of the Japanese Society of Scientific Fisheries, 47(7): 963. https://doi.org/10.2331/suisan.47.963

Effects of Colchicine and Trypsin Treatments on Metaphase Chromosome Spreading from Larvae and Different Tissues of Adult Female and Male Nile Tilapia

Year 2024, Volume: 10 Issue: 3, 113 - 118, 28.12.2024
https://doi.org/10.58626/menba.1538957

Abstract

Assessment of results of different biotechnological methods in aquaculture such as ploidy, gynogenesis, and androgenesis are needed cytogenetical techniques. Achieving well-distributed metaphase chromosome spreads is a preeminent step for cytogenetical techniques. The aim of this study was to evaluate the effects of colchicine and trypsin treatments on the yield of countable metaphase chromosome spreads. With this aim, chromosome preparations from gill, spleen, kidney, and caudal fin tissues of adult females and males Nile tilapia, and also whole body of their larvae were carried out. Based on the previous studies reviewed so far, this is the first systematic and extensive study indicated that the effects of colchicine and trypsin on metaphase chromosome spreads regarding gender differences and larvae. In the chromosome preparations, it was designated four treatments as “Trypsin (-) Colchicine (-)”, “Trypsin (-) Colchicine (+)”, “Trypsin (+) Colchicine (-)”, and “Trypsin (+) Colchicine (+)”. Effects of colchicine and trypsin in terms of their usage in chromosome preparations were evaluated by determining the frequencies of metaphase cells. For this purpose, metaphase index were used to reveal differences in detected metaphase cells among the treatments. The highest metaphase index values were calculated in the Trypsin (+) Colchicine (+) treatment group of all tissues. Also, the scores of metaphase index found in spleen and kidney tissues were greater than those found in gill and fin tissues. The present study has also shown that metaphase chromosome spreads could be obtained without colchicine or trypsin treatments.

Ethical Statement

This study was conducted in Aksaray University Experimental Animal Application and Research Center (ASÜ-DEHAM) with the approval number 2021/1-3.

Supporting Institution

This study was financially supported by Aksaray University Coordinatorship of Scientific Research Projects with the project number of 2021/021.

Project Number

Aksaray University Coordinatorship of Scientific Research Projects (2021/021)

Thanks

The author would like to thank Prof. Dr. Mustafa Öz from Aksaray University for his assistance with the samplings. Also, the author is especially grateful to Prof. Dr. Tahir Karaşahin for the opportunity to examine the sections under the fluorescence microscope at Aksaray University Embryo Transfer Training Application Research Center.

References

  • Bertão, M.R., & Aguiar-Perecin, M.L.R. (2002). Maize somatic chromosome preparation: pretreatments and genotypes for obtention of high index of metaphase accumulation. Caryologia, 55(2): 115-119. https://doi.org/10.1080/00087114.2002.10589266
  • Blanco, D.R., L.A.C. Bertollo, R.L. Lui, M.R. Vicari, V.P. Margarido, R.F. Artoni & O. Moreira Filho. (2012). A new technique for obtaining mitotic chromosome spreads from fishes in the field. J. Fish Biol., 81:351-357. https://doi.org/10.1111/j.1095-8649.2012.03325.x
  • Cattin, P.M. & Ferreira, J.T. (1989). A rapid, nonsacrificial chromosome preparation technique for freshwater teleosts. South African Journal of Zoology, 24:76-78. https://doi.org/10.1080/02541858.1989.11448136
  • Chourrout, D., & Happe, A., (1986). Improved methods of direct chromosome preparation in rainbow trout, Salmo gairdneri. Aquaculture, 52: 255-261. https://doi.org/10.1016/0044-8486(86)90368-6
  • Demirok, N.K. & Ünlü, E., (2001). Karyotypes of Cyprinid fish Capoeta trutta and Capoeta capoeta umbla (Cyprinidae) from the Tigris River. Turkish Journal of Zoology, 25: 389-393.
  • Deng, W., Tsao, S.W., Lucas, J.N., Leung, C.S., & Cheung, A.L. (2003). A new method for improving metaphase chromosome spreading. Cytometry A, 51(1):46-51. https://doi.org/10.1002/cyto.a.10004
  • Foresti, F., Oliveira, C., & Foresti de Almeida-Toledo, L. (1993). A method for chromosome preparations from large fish specimens using in vitro short-term treatment with colchicine. Experientia, 49: 810-813. https://doi.org/10.1007/BF01923555
  • Harvey, S., Campos-Ramos, R., Kennedy, D.D., Ezaz, M.T., Bromage, N.R., Griffin, D.K., & Penman, D.J. (2002). Karyotype evolution in tilapia: mitotic and meiotic chromosome analysis of Oreochromis Karongae and O. Niloticus × O. Karongae Hybrids. Genetica 115:169-177. https://doi.org/10.1023/A:1020190918431
  • Henegariu, O., Heerema, N.A., Wright, L.L., Brayward, P., Ward, D.C., & Vance G.H. (2001). Improvements in cytogenetic slide preparation: Controlled chromosome spreading, chemical aging and gradual denaturing. Cytometry, 43: 101-109. https://doi.org/10.1002/1097-0320(20010201)43:2<101::AID-CYTO1024>3.0.CO;2-8
  • Howe, B., Umrigar, A., & Tsien, F. (2014). Chromosome preparation from cultured cells. J. Vis. Exp., 28(83):e50203. https://doi.org/10.3791/50203
  • Hussain, M.G., & McAndrew, B.J. (1994). An improved technique for chromosome karyotyping from embryonic and soft tissues of Tilapia and Salmonids. Asian Fisheries Science, 7: 187-190.
  • Moreira-Filho, O., & Bertollo, L.A.C. (1991). Extraction and use of the cephalic kidney for chromosome studies in small fi sh. Braz. J. Genet., 14:1085-1090.
  • Mukti, A.T., Carman, O., Alimuddin, & Zairin Jr, M. (2016). A rapid chromosome preparation technique without metaphase arrest for ploidy determination in Nile tilapia, Oreochromis niloticus, Caryologia, 69(2):175-180. https://doi.org/10.1080/00087114.2016.1152112
  • O'Connor, C. (2008). Karyotyping for chromosomal abnormalities. Nature Education 1(1):27.
  • Pope, L.H., Xiong, C., & Marko, J.F. (2006). Proteolysis of mitotic chromosomes induces gradual and anisotropic decondensation correlated with a reduction of elastic modulus and structural sensitivity to rarely cutting restriction enzymes. Mol. Biol. Cell., 17(1):104-113. https://doi.org/10.1091/mbc.e05-04-0321
  • Pradeep, P.J., Srijaya, T.C., Zain, R.B.M., Papini, A., & Chatterji, A.K. (2011). A simple technique for chromosome preparation from embryonic tissues of teleosts for ploidy verification. Caryologia, 64(2): 235-241. https://doi.org/10.1080/00087114.2002.10589788
  • Rieder, C.L., & Palazzo, R.E. (1992). Colcemid and the mitotic cycle. J. Cell Sci., 102(3):387-392. https://doi.org/10.1242/jcs.102.3.387
  • Schneider, C.A., Rasband, W.S., & Eliceiri, K.W. (2012). NIH Image to ImageJ: 25 years of image analysis. Nature Methods, 9(7):671-675. https://doi.org/10.1038/nmeth.2089
  • Supiwong, W., Tanomtong, A., Supanuam, P., Seetapan, K., Khakhong, S., & Sanoamuang, L. (2013). Chromosomal Characteristic of Nile Tilapia (Oreochromis niloticus) from Mitotic and Meiotic Cell Division by T-Lymphocyte Cell Culture. Cytologia, 78(1): 9-14. https://doi.org/10.1508/cytologia.78.9
  • Yamazaki, F., Onazato, H., & Arai, K. (1981). The chopping method for obtaining permanent chromosome preparation from embryos of teleost fishes. Bulletin of the Japanese Society of Scientific Fisheries, 47(7): 963. https://doi.org/10.2331/suisan.47.963
There are 20 citations in total.

Details

Primary Language English
Subjects Aquaculture and Fisheries (Other)
Journal Section Araştırmalar
Authors

Burak Evren İnanan 0000-0002-2888-8457

Project Number Aksaray University Coordinatorship of Scientific Research Projects (2021/021)
Publication Date December 28, 2024
Submission Date August 26, 2024
Acceptance Date November 9, 2024
Published in Issue Year 2024 Volume: 10 Issue: 3

Cite

APA İnanan, B. E. (2024). Effects of Colchicine and Trypsin Treatments on Metaphase Chromosome Spreading from Larvae and Different Tissues of Adult Female and Male Nile Tilapia. Menba Kastamonu Üniversitesi Su Ürünleri Fakültesi Dergisi, 10(3), 113-118. https://doi.org/10.58626/menba.1538957
AMA İnanan BE. Effects of Colchicine and Trypsin Treatments on Metaphase Chromosome Spreading from Larvae and Different Tissues of Adult Female and Male Nile Tilapia. Menba Kastamonu Üniversitesi Su Ürünleri Fakültesi Dergisi. December 2024;10(3):113-118. doi:10.58626/menba.1538957
Chicago İnanan, Burak Evren. “Effects of Colchicine and Trypsin Treatments on Metaphase Chromosome Spreading from Larvae and Different Tissues of Adult Female and Male Nile Tilapia”. Menba Kastamonu Üniversitesi Su Ürünleri Fakültesi Dergisi 10, no. 3 (December 2024): 113-18. https://doi.org/10.58626/menba.1538957.
EndNote İnanan BE (December 1, 2024) Effects of Colchicine and Trypsin Treatments on Metaphase Chromosome Spreading from Larvae and Different Tissues of Adult Female and Male Nile Tilapia. Menba Kastamonu Üniversitesi Su Ürünleri Fakültesi Dergisi 10 3 113–118.
IEEE B. E. İnanan, “Effects of Colchicine and Trypsin Treatments on Metaphase Chromosome Spreading from Larvae and Different Tissues of Adult Female and Male Nile Tilapia”, Menba Kastamonu Üniversitesi Su Ürünleri Fakültesi Dergisi, vol. 10, no. 3, pp. 113–118, 2024, doi: 10.58626/menba.1538957.
ISNAD İnanan, Burak Evren. “Effects of Colchicine and Trypsin Treatments on Metaphase Chromosome Spreading from Larvae and Different Tissues of Adult Female and Male Nile Tilapia”. Menba Kastamonu Üniversitesi Su Ürünleri Fakültesi Dergisi 10/3 (December 2024), 113-118. https://doi.org/10.58626/menba.1538957.
JAMA İnanan BE. Effects of Colchicine and Trypsin Treatments on Metaphase Chromosome Spreading from Larvae and Different Tissues of Adult Female and Male Nile Tilapia. Menba Kastamonu Üniversitesi Su Ürünleri Fakültesi Dergisi. 2024;10:113–118.
MLA İnanan, Burak Evren. “Effects of Colchicine and Trypsin Treatments on Metaphase Chromosome Spreading from Larvae and Different Tissues of Adult Female and Male Nile Tilapia”. Menba Kastamonu Üniversitesi Su Ürünleri Fakültesi Dergisi, vol. 10, no. 3, 2024, pp. 113-8, doi:10.58626/menba.1538957.
Vancouver İnanan BE. Effects of Colchicine and Trypsin Treatments on Metaphase Chromosome Spreading from Larvae and Different Tissues of Adult Female and Male Nile Tilapia. Menba Kastamonu Üniversitesi Su Ürünleri Fakültesi Dergisi. 2024;10(3):113-8.