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Immortalized hippocampal cell line H19‐7 express endothelin system genes
Author(s) -
Kanyicska Bela,
Liu Huiling
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.1044.4
Subject(s) - endothelins , receptor , endothelin receptor , hippocampal formation , endothelin 1 , cell culture , cytosol , immortalised cell line , biology , microbiology and biotechnology , endocrinology , medicine , chemistry , biochemistry , enzyme , genetics
Previous studies indicate that endothelins (ETs) may play a neuromodulatory role in the central nervous system. However, there is a paucity of data regarding ETs’ action on specific subsets of neurons. We evaluated the hippocampal cell line H19‐7 (conditionally immortalized by a temperature‐sensitive SV40 large T antigen) as an in vitro model to investigate the role of ETs in neural activity. The relative abundance of endothelin system genes mRNA was assessed by real‐time PCR method using specific primers for endothelins (ET‐1 and ET‐3), endothelin receptors (ETA and ETB), and endothelin converting enzyme‐1 (ECE‐1). Under non‐differentiating conditions (34°C), we found a robust expression of ECE1, ETA and ET‐1, while ETB and ET‐3 mRNA levels were undetectable. The expression patterns of ET system genes were essentially similar when the cells were placed under differentiating condition (39°C) for 24–36 hours. When changes in cytosolic free Ca2+ levels were assessed by the Fura‐2 method, undifferentiated cells were unresponsive to ET1. However, 20 nM ET‐1 elicited a robust bi‐phasic increase of cytosolic free Ca2+ (a hallmark effect of ETA receptor activation) on differentiated cells, and markedly enhanced capacitative Ca2+ entry as well. Thus, differentiated H19‐7 cells seem suitable for mechanistic studies of ETs’ action on neuronal targets.