Developmental Increase in Ecto-5-Nucleotidase Activity Overlaps with Appearance of Two Immunologically Distinct Enzyme Isoforms in Rat Hippocampal Synaptic Plasma Membranes
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2014
Authors
Grković, IvanaBjelobaba, Ivana
Nedeljković, Nadežda
Mitrović, Nataša Lj.
Drakulić, Dunja R.
Stanojlović, Miloš R.
Horvat, Anica
Article (Published version)
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Ecto-5-nucleotidase (e-5NT), a glycosylphosphatidylinositol-linked membrane protein, catalyzes a conversion of AMP to adenosine, which influences nearly every aspect of brain physiology, including embryonic and postnatal brain development. The present study aimed to investigate a pattern of expression, activity and kinetic properties of e-5NT in the hippocampal formation and synaptic plasma membrane (SPM) preparations in rats at postnatal days (PDs) 7, 15, 20, 30 and 90. By combining gene expression analysis and enzyme histochemistry, we observed that e-5NT mRNA reached the adult level at PD20, while the enzyme activity continued to increase beyond this age. Further analysis revealed that hippocampal layers rich in synapses expressed the highest levels of e-5NT activity, while in layers populated with neuronal cell bodies, the enzyme activity was weak or absent. Therefore, activity and expression of e-5NT were analyzed in SPM preparations isolated from rats at different ages. The prese...nce of two protein bands of about 65 and 68 kDa was determined by immunoblot analysis. The 65-kDa band was present at all ages, and its abundance increased from PD7 to PD20. The 68-kDa band appeared at PD15 and increased until PD30, coinciding with the increase of e-5NT activity, substrate affinity and enzymatic efficiency. Since distinct e-5NT isoforms may derive from different patterns of the enzyme protein N-glycosylation, we speculate that long-term regulation of e-5NT activity in adulthood may be effectuated at posttranslational level and without overall change in the gene and protein expression.
Keywords:
Ecto-5 -nucleotidase / Adenosine / Synaptic plasma membranes / Hippocampus / Development / PlasticitySource:
Journal of Molecular Neuroscience, 2014, 54, 1, 109-118Funding / projects:
- Molecular mechanisms of cellular responses on pathological changes in central neuronal system and peripheral organs of mammals (RS-MESTD-Basic Research (BR or ON)-173044)
- Cellular and molecular basis of neuroinflamation: potential targets for translational medicine and therapy (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-41014)
DOI: 10.1007/s12031-014-0256-0
ISSN: 0895-8696; 1559-1166
PubMed: 24563227
WoS: 000340474200012
Scopus: 2-s2.0-84906345111
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VinčaTY - JOUR AU - Grković, Ivana AU - Bjelobaba, Ivana AU - Nedeljković, Nadežda AU - Mitrović, Nataša Lj. AU - Drakulić, Dunja R. AU - Stanojlović, Miloš R. AU - Horvat, Anica PY - 2014 UR - https://vinar.vin.bg.ac.rs/handle/123456789/64 AB - Ecto-5-nucleotidase (e-5NT), a glycosylphosphatidylinositol-linked membrane protein, catalyzes a conversion of AMP to adenosine, which influences nearly every aspect of brain physiology, including embryonic and postnatal brain development. The present study aimed to investigate a pattern of expression, activity and kinetic properties of e-5NT in the hippocampal formation and synaptic plasma membrane (SPM) preparations in rats at postnatal days (PDs) 7, 15, 20, 30 and 90. By combining gene expression analysis and enzyme histochemistry, we observed that e-5NT mRNA reached the adult level at PD20, while the enzyme activity continued to increase beyond this age. Further analysis revealed that hippocampal layers rich in synapses expressed the highest levels of e-5NT activity, while in layers populated with neuronal cell bodies, the enzyme activity was weak or absent. Therefore, activity and expression of e-5NT were analyzed in SPM preparations isolated from rats at different ages. The presence of two protein bands of about 65 and 68 kDa was determined by immunoblot analysis. The 65-kDa band was present at all ages, and its abundance increased from PD7 to PD20. The 68-kDa band appeared at PD15 and increased until PD30, coinciding with the increase of e-5NT activity, substrate affinity and enzymatic efficiency. Since distinct e-5NT isoforms may derive from different patterns of the enzyme protein N-glycosylation, we speculate that long-term regulation of e-5NT activity in adulthood may be effectuated at posttranslational level and without overall change in the gene and protein expression. T2 - Journal of Molecular Neuroscience T1 - Developmental Increase in Ecto-5-Nucleotidase Activity Overlaps with Appearance of Two Immunologically Distinct Enzyme Isoforms in Rat Hippocampal Synaptic Plasma Membranes VL - 54 IS - 1 SP - 109 EP - 118 DO - 10.1007/s12031-014-0256-0 ER -
@article{ author = "Grković, Ivana and Bjelobaba, Ivana and Nedeljković, Nadežda and Mitrović, Nataša Lj. and Drakulić, Dunja R. and Stanojlović, Miloš R. and Horvat, Anica", year = "2014", abstract = "Ecto-5-nucleotidase (e-5NT), a glycosylphosphatidylinositol-linked membrane protein, catalyzes a conversion of AMP to adenosine, which influences nearly every aspect of brain physiology, including embryonic and postnatal brain development. The present study aimed to investigate a pattern of expression, activity and kinetic properties of e-5NT in the hippocampal formation and synaptic plasma membrane (SPM) preparations in rats at postnatal days (PDs) 7, 15, 20, 30 and 90. By combining gene expression analysis and enzyme histochemistry, we observed that e-5NT mRNA reached the adult level at PD20, while the enzyme activity continued to increase beyond this age. Further analysis revealed that hippocampal layers rich in synapses expressed the highest levels of e-5NT activity, while in layers populated with neuronal cell bodies, the enzyme activity was weak or absent. Therefore, activity and expression of e-5NT were analyzed in SPM preparations isolated from rats at different ages. The presence of two protein bands of about 65 and 68 kDa was determined by immunoblot analysis. The 65-kDa band was present at all ages, and its abundance increased from PD7 to PD20. The 68-kDa band appeared at PD15 and increased until PD30, coinciding with the increase of e-5NT activity, substrate affinity and enzymatic efficiency. Since distinct e-5NT isoforms may derive from different patterns of the enzyme protein N-glycosylation, we speculate that long-term regulation of e-5NT activity in adulthood may be effectuated at posttranslational level and without overall change in the gene and protein expression.", journal = "Journal of Molecular Neuroscience", title = "Developmental Increase in Ecto-5-Nucleotidase Activity Overlaps with Appearance of Two Immunologically Distinct Enzyme Isoforms in Rat Hippocampal Synaptic Plasma Membranes", volume = "54", number = "1", pages = "109-118", doi = "10.1007/s12031-014-0256-0" }
Grković, I., Bjelobaba, I., Nedeljković, N., Mitrović, N. Lj., Drakulić, D. R., Stanojlović, M. R.,& Horvat, A.. (2014). Developmental Increase in Ecto-5-Nucleotidase Activity Overlaps with Appearance of Two Immunologically Distinct Enzyme Isoforms in Rat Hippocampal Synaptic Plasma Membranes. in Journal of Molecular Neuroscience, 54(1), 109-118. https://doi.org/10.1007/s12031-014-0256-0
Grković I, Bjelobaba I, Nedeljković N, Mitrović NL, Drakulić DR, Stanojlović MR, Horvat A. Developmental Increase in Ecto-5-Nucleotidase Activity Overlaps with Appearance of Two Immunologically Distinct Enzyme Isoforms in Rat Hippocampal Synaptic Plasma Membranes. in Journal of Molecular Neuroscience. 2014;54(1):109-118. doi:10.1007/s12031-014-0256-0 .
Grković, Ivana, Bjelobaba, Ivana, Nedeljković, Nadežda, Mitrović, Nataša Lj., Drakulić, Dunja R., Stanojlović, Miloš R., Horvat, Anica, "Developmental Increase in Ecto-5-Nucleotidase Activity Overlaps with Appearance of Two Immunologically Distinct Enzyme Isoforms in Rat Hippocampal Synaptic Plasma Membranes" in Journal of Molecular Neuroscience, 54, no. 1 (2014):109-118, https://doi.org/10.1007/s12031-014-0256-0 . .