A sensitive method to analyse the effect of putative regulatory ligands on the release of glycoprotein from primary cultures of dispersed bovine subcommissural organ cells

The subcommissural organ (SCO) releases into the cerebrospinal fluid (CSF) large glycoproteins that polymerize forming the Reissner's fibre (RF), which is involved in CSF circulation and homeostasis. We obtained high purity primary cultures of bovine secretory SCO cells and measured glycoprotei...

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Bibliographic Details
Published in:Journal of neuroscience methods 2010-08, Vol.191 (2), p.239-243
Main Authors: Bermúdez-Silva, F.J., Pérez, J., Cifuentes, M., Pérez-Martín, M., Grondona, J.M., López-Ávalos, M.D., Estivill-Torrús, G., Fernández-Llebrez, P.
Format: Article
Language:English
Subjects:
5-Hydroxytriptamine
Acetylcholine
Adenosine Triphosphate - pharmacology
Animals
ATP
Carbachol
Carbachol - pharmacology
Cattle
Cell culture
Cell Culture Techniques - methods
Cells, Cultured
Cerebrospinal Fluid - metabolism
Dose-Response Relationship, Drug
ELISA
Enzyme-Linked Immunosorbent Assay - methods
Ependyma
Ependyma - drug effects
Ependyma - metabolism
Ependyma - secretion
Exocytosis
Glycoproteins - antagonists & inhibitors
Glycoproteins - biosynthesis
Glycoproteins - secretion
Ligands
Potassium - pharmacology
Serotonin
Serotonin - pharmacology
Subcommissural Organ - drug effects
Subcommissural Organ - metabolism
Subcommissural Organ - secretion
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Summary:The subcommissural organ (SCO) releases into the cerebrospinal fluid (CSF) large glycoproteins that polymerize forming the Reissner's fibre (RF), which is involved in CSF circulation and homeostasis. We obtained high purity primary cultures of bovine secretory SCO cells and measured glycoprotein release by a reliable and sensitive ELISA method. We also analysed the effect of regulatory ligands known to control the secretory activity of the SCO. Cells cultured for short time (4 h) released a high amount of glycoproteins that decreased with time. In young cultures, ATP increased and serotonin inhibited secretion rate. By contrast the acetylcholine agonist carbachol and high potassium did not evoke any detectable change in SCO glycoprotein release. These results support not only the suitability of the methodological approach but an important role of both ATP and serotonin in regulating SCO secretory activity as well.
ISSN:0165-0270
1872-678X
DOI:10.1016/j.jneumeth.2010.07.002