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 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 (4h) 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.
Bermúdez-Silva
F J
FJ
Laboratorio de Medicina Regenerativa, Fundación IMABIS, Avenida Carlos Haya 82, 29010 Málaga, Spain.
Pérez
J
J
Cifuentes
M
M
Pérez-Martín
M
M
Grondona
J M
JM
López-Avalos
M D
MD
Estivill-Torrús
G
G
Fernández-Llebrez
P
P
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D002555:Cerebrospinal Fluid / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004805:Ependyma / Q000187:drug effects* / Q000378:metabolism; D006023:Glycoproteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*; D008024:Ligands; D011188:Potassium / Q000494:pharmacology; D012701:Serotonin / Q000494:pharmacology; D013351:Subcommissural Organ / Q000187:drug effects* / Q000378:metabolism
D006023:Glycoproteins; D008024:Ligands; D012701:Serotonin; D000255:Adenosine Triphosphate; D002217:Carbachol; D011188:Potassium