Nerve growth factor affects Ca 2+ currents via the p75 receptor to enhance prolactin mRNA levels in GH 3 rat pituitary cells

• Published in: The Journal of physiology Vol. 574; no. 2; pp. 349 - 365
• Main Authors: López‐Domínguez, Adriana M., Espinosa, Juan Luis, Navarrete, Araceli, Avila, Guillermo, Cota, Gabriel
• Format: Journal Article
• Language: English
• Published: 15.07.2006
• ISSN: 0022-3751; 1469-7793
• DOI: 10.1113/jphysiol.2006.110791

In clonal pituitary GH 3 cells, spontaneous action potentials drive the opening of Ca v 1 (L‐type) channels, leading to Ca 2+ transients that are coupled to prolactin gene transcription. Nerve growth factor (NGF) has been shown to stimulate prolactin synthesis by GH 3 cells, but the underlying mechanisms are unknown. Here we studied whether NGF influences prolactin gene expression and Ca 2+ currents. By using RT‐PCR, NGF (50 ng ml −1 ) was found to augment prolactin mRNA levels by ∼80% when applied to GH 3 cells for 3 days. A parallel change in the prolactin content was detected by Western blotting. Both NGF‐induced responses were mimicked by an agonist (Bay K 8644) and prevented by a blocker (nimodipine) of L‐type channels. In whole‐cell patch‐clamp experiments, NGF enhanced the L‐type Ca 2+ current by ∼2‐fold within 60 min. This effect reversed quickly upon growth factor withdrawal, but was maintained for days in the continued presence of NGF. In addition, chronic treatment (≥ 24 h) with NGF amplified the T‐type current, which flows through Ca v 3 channels and is thought to support pacemaking activity. Thus, NGF probably increases the amount of Ca 2+ that enters per action potential and may also induce a late increase in spike frequency. MC192, a specific antibody for the p75 neurotrophin receptor, but not tyrosine kinase inhibitors (K252a and lavendustin A), blocked the effects of NGF on Ca 2+ currents. Overall, the results indicate that NGF activates the p75 receptor to cause a prolonged increase in Ca 2+ influx through L‐type channels, which in turn up‐regulates the prolactin mRNA.