Limits...
Ontogeny of ATP hydrolysis and isoform expression of the plasma membrane Ca(2+)-ATPase in mouse brain.

Marcos D, Sepulveda MR, Berrocal M, Mata AM - BMC Neurosci (2009)

Bottom Line: Western blots revealed the presence of the four isoforms in all regions, with similar increase in their expression patterns as those seen for the activity profile.Immunohistochemistry assays in cortex and hippocampus showed co-expression of all isoforms in the neuropil associated with synapses and in the plasma membrane of pyramidal cells soma, while cerebellum showed a more isoform-specific distribution pattern in Purkinje cells.Overall, our findings support a close relationship between the ontogeny of PMCA isoforms and specific requirements of Ca2+ during development of different brain areas.

View Article: PubMed Central - HTML - PubMed

Affiliation: Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Ciencias, Universidad de Extremadura, 06071 Badajoz, Spain. danielmc@unex.es

ABSTRACT

Background: Plasma membrane Ca2+-ATPases (PMCAs) are high affinity Ca2+ transporters actively involved in intracellular Ca2+ homeostasis. Considering the critical role of Ca2+ signalling in neuronal development and plasticity, we have analyzed PMCA-mediated Ca2+-ATPase activity and PMCA-isoform content in membranes from mouse cortex, hippocampus and cerebellum during postnatal development.

Results: PMCA activity was detected from birth, with a faster evolution in cortex than in hippocampus and cerebellum. Western blots revealed the presence of the four isoforms in all regions, with similar increase in their expression patterns as those seen for the activity profile. Immunohistochemistry assays in cortex and hippocampus showed co-expression of all isoforms in the neuropil associated with synapses and in the plasma membrane of pyramidal cells soma, while cerebellum showed a more isoform-specific distribution pattern in Purkinje cells.

Conclusion: These results show an upregulation of PMCA activity and PMCA isoforms expression during brain development in mouse, with specific localizations mainly in cerebellum. Overall, our findings support a close relationship between the ontogeny of PMCA isoforms and specific requirements of Ca2+ during development of different brain areas.

Show MeSH
PMCA activity in developing cortex, hippocampus and cerebellum. Twenty μg of MV obtained from the indicated brain regions at different developmental stages were used to measure PMCA activity, as described in the Experimental Procedures. Data are the mean ± SE values obtained from three experiments performed in triplicate, and from two preparations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2749858&req=5

Figure 1: PMCA activity in developing cortex, hippocampus and cerebellum. Twenty μg of MV obtained from the indicated brain regions at different developmental stages were used to measure PMCA activity, as described in the Experimental Procedures. Data are the mean ± SE values obtained from three experiments performed in triplicate, and from two preparations.

Mentions: The PMCA protein showed the capacity to catalyze ATP-hydrolysis in a Ca2+-dependent manner from the first stage in all areas (Fig. 1). Besides, the PMCA activity during postnatal development followed two patterns, reaching the Vmax at P8 in the cortex and at P15 stage in hippocampus and cerebellum. Although the values of Vmax were similar in all regions (0.237 ± 0.017 μmol·min-1·mg-1) the increase of activity was more pronounced (around 4-fold) in cortex and hippocampus than in cerebellum (around 2.3-fold).


Ontogeny of ATP hydrolysis and isoform expression of the plasma membrane Ca(2+)-ATPase in mouse brain.

Marcos D, Sepulveda MR, Berrocal M, Mata AM - BMC Neurosci (2009)

PMCA activity in developing cortex, hippocampus and cerebellum. Twenty μg of MV obtained from the indicated brain regions at different developmental stages were used to measure PMCA activity, as described in the Experimental Procedures. Data are the mean ± SE values obtained from three experiments performed in triplicate, and from two preparations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2749858&req=5

Figure 1: PMCA activity in developing cortex, hippocampus and cerebellum. Twenty μg of MV obtained from the indicated brain regions at different developmental stages were used to measure PMCA activity, as described in the Experimental Procedures. Data are the mean ± SE values obtained from three experiments performed in triplicate, and from two preparations.
Mentions: The PMCA protein showed the capacity to catalyze ATP-hydrolysis in a Ca2+-dependent manner from the first stage in all areas (Fig. 1). Besides, the PMCA activity during postnatal development followed two patterns, reaching the Vmax at P8 in the cortex and at P15 stage in hippocampus and cerebellum. Although the values of Vmax were similar in all regions (0.237 ± 0.017 μmol·min-1·mg-1) the increase of activity was more pronounced (around 4-fold) in cortex and hippocampus than in cerebellum (around 2.3-fold).

Bottom Line: Western blots revealed the presence of the four isoforms in all regions, with similar increase in their expression patterns as those seen for the activity profile.Immunohistochemistry assays in cortex and hippocampus showed co-expression of all isoforms in the neuropil associated with synapses and in the plasma membrane of pyramidal cells soma, while cerebellum showed a more isoform-specific distribution pattern in Purkinje cells.Overall, our findings support a close relationship between the ontogeny of PMCA isoforms and specific requirements of Ca2+ during development of different brain areas.

View Article: PubMed Central - HTML - PubMed

Affiliation: Departamento de Bioquímica y Biología Molecular y Genética, Facultad de Ciencias, Universidad de Extremadura, 06071 Badajoz, Spain. danielmc@unex.es

ABSTRACT

Background: Plasma membrane Ca2+-ATPases (PMCAs) are high affinity Ca2+ transporters actively involved in intracellular Ca2+ homeostasis. Considering the critical role of Ca2+ signalling in neuronal development and plasticity, we have analyzed PMCA-mediated Ca2+-ATPase activity and PMCA-isoform content in membranes from mouse cortex, hippocampus and cerebellum during postnatal development.

Results: PMCA activity was detected from birth, with a faster evolution in cortex than in hippocampus and cerebellum. Western blots revealed the presence of the four isoforms in all regions, with similar increase in their expression patterns as those seen for the activity profile. Immunohistochemistry assays in cortex and hippocampus showed co-expression of all isoforms in the neuropil associated with synapses and in the plasma membrane of pyramidal cells soma, while cerebellum showed a more isoform-specific distribution pattern in Purkinje cells.

Conclusion: These results show an upregulation of PMCA activity and PMCA isoforms expression during brain development in mouse, with specific localizations mainly in cerebellum. Overall, our findings support a close relationship between the ontogeny of PMCA isoforms and specific requirements of Ca2+ during development of different brain areas.

Show MeSH