Hydrogen Sulfide in Proliferating and Differentiated Cells in Primary Cultures of Juvenile Brain of Masu Salmon Oncorhynchus masou
Affiliation(s)
1 Zhirmunsky Institute of Marine Biology, Far East Branch, Russian Academy of Sciences, Vladivostok, Russia. 2 Prof. Brien Holden Eye Research Centre, L.V. Prasad Eye Institute, Hyderabad, India.
1 Zhirmunsky Institute of Marine Biology, Far East Branch, Russian Academy of Sciences, Vladivostok, Russia. 2 Prof. Brien Holden Eye Research Centre, L.V. Prasad Eye Institute, Hyderabad, India.
ABSTRACT
Analysis of proliferative activity and the ability to neuron differentiation was performed in cultured cells of the brain and spinal cord of juvenile masu salmon Oncorhynchus masou. Proliferating cell nuclear antigen (PCNA) was used as a proliferative marker, while the markers of neuronal differentiation—a neuron protein HuCD, and a neuron-specific transcriptional factor with two DNA- binding sites Pax6—detected neurons. The results showed that cell proliferation occurred mainly in the suspension cell fraction. In monolayer, a few cells were only found to express PCNA. The results of morphological and immunohistochemical analysis allow us to conclude that proliferative activity in primary cultures from the O. masou brain is mainly connected with the suspension fraction of small cells. In contrast, a positive correlation between the cells expressing cystathionine β-synthase (CBS), a marker of H2S synthesis, and the cells expressing PCNA in the monolayer, indicates the participation of H2S in proliferative activity of neurons in primary cultures. The data obtained suggest that the hydrogen sulphide is also involved in the process of differentiation.
Analysis of proliferative activity and the ability to neuron differentiation was performed in cultured cells of the brain and spinal cord of juvenile masu salmon Oncorhynchus masou. Proliferating cell nuclear antigen (PCNA) was used as a proliferative marker, while the markers of neuronal differentiation—a neuron protein HuCD, and a neuron-specific transcriptional factor with two DNA- binding sites Pax6—detected neurons. The results showed that cell proliferation occurred mainly in the suspension cell fraction. In monolayer, a few cells were only found to express PCNA. The results of morphological and immunohistochemical analysis allow us to conclude that proliferative activity in primary cultures from the O. masou brain is mainly connected with the suspension fraction of small cells. In contrast, a positive correlation between the cells expressing cystathionine β-synthase (CBS), a marker of H2S synthesis, and the cells expressing PCNA in the monolayer, indicates the participation of H2S in proliferative activity of neurons in primary cultures. The data obtained suggest that the hydrogen sulphide is also involved in the process of differentiation.
Cite this paper
Pushchina, E. , Shukla, S. and Varaksin, A. (2015) Hydrogen Sulfide in Proliferating and Differentiated Cells in Primary Cultures of Juvenile Brain of Masu Salmon Oncorhynchus masou. Advances in Bioscience and Biotechnology, 6, 539-545. doi: 10.4236/abb.2015.68057.
Pushchina, E. , Shukla, S. and Varaksin, A. (2015) Hydrogen Sulfide in Proliferating and Differentiated Cells in Primary Cultures of Juvenile Brain of Masu Salmon Oncorhynchus masou. Advances in Bioscience and Biotechnology, 6, 539-545. doi: 10.4236/abb.2015.68057.
References
[1] Kizil, C., Kaslin, J., Kroehne, V. and Brand, M. (2012) Adult Neurogenesis and Brain Regeneration in Zebrafish. Developmental Neurobiology, 72, 429-461.
http://dx.doi.org/10.1002/dneu.20918 [2] Becker, T., Wullimann, M.F., Becker, C.G., et al. (1997) Axonal Regrowth after Spinal Cord Transection in Adult Zebrafish. Journal of Comparative Neurology, 377, 577-595.
http://dx.doi.org/10.1002/(SICI)1096-9861(19970127)377:4<577::AID-CNE8>3.0.CO;2-# [3] Candal, E., Anadon, R., DeGrip, W.J. and Rodriguez-Moldes, I. (2005) Patterns of Cell Proliferation and Cell Death in the Developing Retina and Optic Tectum of the Brown Trout. Developmental Brain Research, 154, 101-119.
http://dx.doi.org/10.1016/j.devbrainres.2004.10.008 [4] Sakowski, S.A., Lunn, J.S., Busta, A.S., et al. (2012) A Novel Approach to Study Motor Neurons from Zebrafish Embryos and Larvae in Culture. Journal of Neuroscience Methods, 205, 277-282.
http://dx.doi.org/10.1016/j.jneumeth.2012.01.007 [5] Arévalo, R.R., Alonso, J.R., Garcia-Ojeda, E., et al. (1995) NADPH-Diaphorase in the Central Nervous System of the Tench (Tinca tinca L., 1758). Journal of Comparative Neurology, 352, 398-420.
http://dx.doi.org/10.1002/cne.903520307 [6] Doe, C.Q., Fuerstenberg, S. and Peng, C.-Y. (1998) Neural Stem Cells: From Fly to Vertebrates. Journal of Neurobiology, 36, 111-127.
http://dx.doi.org/10.1002/(SICI)1097-4695(199808)36:2<111::AID-NEU2>3.0.CO;2-4 [7] Zupanc, G.K. and Sîrbulescu, R.F. (2013) Teleost Fish as a Model System to Study Successful Regeneration of the Central Nervous System. Current Topics in Microbiology and Immunology, 367, 193-233.
http://dx.doi.org/10.1007/82_2012_297 [8] Pushchina, E.V., Obukhov, D.K. and Varaksin, A.A. (2013) Features of Adult Neurogenesis and Neurochemical Signaling in the Cherry Salmon Oncorhynchus masou Brain. Neural Regeneration Research, 8, 13-23. [9] Wang, R. (2012) Physiological Implication of Hydrogen Sulfide: A Whiff Exploration That Blossomed. Physiological Reviews, 92, 791-896.
http://dx.doi.org/10.1152/physrev.00017.2011 [10] Waseem, N.H. and Lane, D.P. (1990) Monoclonal Antibody Analysis of the Proliferating Cell Nuclear Antigen (PCNA). Structural Conservation and the Detection of a Nucleolar Form. Journal of Cell Science, 96, 121-129. [11] Vriz, S., Lemaitre, J.M., Leibovici, M., et al. (1992) Comparative Analysis of the Intracellular Localization of c-Myc, c-Fos, and Replicative Proteins during Cell Cycle Progression. Molecular and Cellular Biology, 12, 3548-3555. [12] Blackmore, M.G., Moore, D.L., Smith, R.P., et al. (2010) High Content Screening of Cortical Neurons Identifies Novel Regulators of Axon Growth. Molecular and Cellular Neuroscience, 44, 43-54.
http://dx.doi.org/10.1016/j.mcn.2010.02.002
[1] Kizil, C., Kaslin, J., Kroehne, V. and Brand, M. (2012) Adult Neurogenesis and Brain Regeneration in Zebrafish. Developmental Neurobiology, 72, 429-461.
http://dx.doi.org/10.1002/dneu.20918 [2] Becker, T., Wullimann, M.F., Becker, C.G., et al. (1997) Axonal Regrowth after Spinal Cord Transection in Adult Zebrafish. Journal of Comparative Neurology, 377, 577-595.
http://dx.doi.org/10.1002/(SICI)1096-9861(19970127)377:4<577::AID-CNE8>3.0.CO;2-# [3] Candal, E., Anadon, R., DeGrip, W.J. and Rodriguez-Moldes, I. (2005) Patterns of Cell Proliferation and Cell Death in the Developing Retina and Optic Tectum of the Brown Trout. Developmental Brain Research, 154, 101-119.
http://dx.doi.org/10.1016/j.devbrainres.2004.10.008 [4] Sakowski, S.A., Lunn, J.S., Busta, A.S., et al. (2012) A Novel Approach to Study Motor Neurons from Zebrafish Embryos and Larvae in Culture. Journal of Neuroscience Methods, 205, 277-282.
http://dx.doi.org/10.1016/j.jneumeth.2012.01.007 [5] Arévalo, R.R., Alonso, J.R., Garcia-Ojeda, E., et al. (1995) NADPH-Diaphorase in the Central Nervous System of the Tench (Tinca tinca L., 1758). Journal of Comparative Neurology, 352, 398-420.
http://dx.doi.org/10.1002/cne.903520307 [6] Doe, C.Q., Fuerstenberg, S. and Peng, C.-Y. (1998) Neural Stem Cells: From Fly to Vertebrates. Journal of Neurobiology, 36, 111-127.
http://dx.doi.org/10.1002/(SICI)1097-4695(199808)36:2<111::AID-NEU2>3.0.CO;2-4 [7] Zupanc, G.K. and Sîrbulescu, R.F. (2013) Teleost Fish as a Model System to Study Successful Regeneration of the Central Nervous System. Current Topics in Microbiology and Immunology, 367, 193-233.
http://dx.doi.org/10.1007/82_2012_297 [8] Pushchina, E.V., Obukhov, D.K. and Varaksin, A.A. (2013) Features of Adult Neurogenesis and Neurochemical Signaling in the Cherry Salmon Oncorhynchus masou Brain. Neural Regeneration Research, 8, 13-23. [9] Wang, R. (2012) Physiological Implication of Hydrogen Sulfide: A Whiff Exploration That Blossomed. Physiological Reviews, 92, 791-896.
http://dx.doi.org/10.1152/physrev.00017.2011 [10] Waseem, N.H. and Lane, D.P. (1990) Monoclonal Antibody Analysis of the Proliferating Cell Nuclear Antigen (PCNA). Structural Conservation and the Detection of a Nucleolar Form. Journal of Cell Science, 96, 121-129. [11] Vriz, S., Lemaitre, J.M., Leibovici, M., et al. (1992) Comparative Analysis of the Intracellular Localization of c-Myc, c-Fos, and Replicative Proteins during Cell Cycle Progression. Molecular and Cellular Biology, 12, 3548-3555. [12] Blackmore, M.G., Moore, D.L., Smith, R.P., et al. (2010) High Content Screening of Cortical Neurons Identifies Novel Regulators of Axon Growth. Molecular and Cellular Neuroscience, 44, 43-54.
http://dx.doi.org/10.1016/j.mcn.2010.02.002