Nerve growth factor affects Ca2+ currents via the p75 receptor to enhance prolactin mRNA levels in GH3 rat pituitary cells.
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 approximately 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 approximately 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 (>or= 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.
López-Domínguez
Adriana M
AM
Department of Physiology, Biophysics and Neurosciences, Cinvestav-IPN, AP 14-740, Mexico, DF 07000, Mexico.
Espinosa
Juan Luis
JL
Navarrete
Araceli
A
Avila
Guillermo
G
Cota
Gabriel
G
D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020746:Calcium Channels, L-Type / Q000187:drug effects / Q000502:physiology; D020747:Calcium Channels, T-Type / Q000187:drug effects / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000502:physiology; D020932:Nerve Growth Factor / Q000494:pharmacology*; D018408:Patch-Clamp Techniques; D010902:Pituitary Gland / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011388:Prolactin / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D020800:Receptor, Nerve Growth Factor / Q000502:physiology*; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology
D020746:Calcium Channels, L-Type; D020747:Calcium Channels, T-Type; D012333:RNA, Messenger; D020800:Receptor, Nerve Growth Factor; D013973:Thyrotropin-Releasing Hormone; D004815:Epidermal Growth Factor; D011388:Prolactin; D020932:Nerve Growth Factor; D002118:Calcium