Distribution of Secretory Pathway Ca 2+ ATPase (SPCA1) in Neuronal and Glial Cell Cultures
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1. Secretory pathway Ca2+ ATPase type 1 (SPCA1) is a newly recognized Ca2+/Mn2+-transporting pump localized in membranes of the Golgi apparatus.
2. The expression level of SPCA1 in brain tissue is relatively high in comparison with other tissues.
3. With the aim to determine the expression of SPCA1 within the different types of neural cells, we investigated the distribution of SPCA1 in neuronal, astroglial, oligodendroglial, ependymal, and microglial cell cultures derived from rat brains.
4. Western Blot analysis with rabbit anti-SPCA1 antibodies revealed the presence of SPCA1 in homogenates derived from neuronal, astroglial, ependymal, and oligodendroglial, but not from microglial cells.
5. Cell cultures that gave rise to positive signal in the immunoblot analysis were also examined immunocytochemically.
6. Immunocytochemical double-labeling experiments with anti-SPCA1 serum in combination with antibodies against cell-type specific proteins showed a localization of the SPCA1signal within cells stained positively also for GFAP, α-tubulin or MBP.
7. These results definitely established the expression of SPCA1 in astroglial, ependymal, and oligodendroglial cells.
8. In addition, the evaluation of neuronal cultures for the presence of SPCA1 revealed an SPCA1-specific immunofluorescence signal in cells identified as neurons.
KEY WORDS:Golgi apparatus secretory pathway Ca2+ ATPase SPCA1 neuron glial cell immunocytochemistry
The authors would like to thank Barbara Birk for her expert technical help in the preparation of the ependymal cell cultures. Part of this work was presented as an abstract on the 5th International Symposium on Experimental and Clinical Neurobiology, Tatranska Lomnica-Stara Lesna, Slovak Republic, September 2005. This study was supported by research grants: VEGA No. 1/0034/03, VEGA No. 3380/06, APVT No. 51-127404, and MVTS 39.
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