title: Human fetal brain cells in aggregate culture: a model system to study regulatory processes of the developing human neuropeptide Y (NPY)-producing neuron.
abstract: Neuropeptide Y (NPY) is one of the most abundant neuropeptides in the brain and it has been implicated in a wide range of brain functions, including mentation. The aim of this study was to establish a culture system of human fetal brain cells expressing NPY in a regulated manner. The NPY production in response to forskolin and phorbol 12-myristate 13-acetate (PMA) was taken as a criterion for regulated expression of NPY. Aggregates were formed from dissociated cells derived from the cerebral hemispheres of human fetuses (12.5-19 weeks' gestation) by constant rotation and were maintained in serum-free medium. A 24 hr exposure to 10 microM forskolin + 20 nM PMA led to a 2-6-fold increase in NPY content of the cultures, most of which (80-90%) was secreted into the medium. The latter consisted of two substances differing in size: one corresponding in size to proNPY and the other to NPY. Thus, forskolin + PMA led to an increased production of NPY. Exposure to PMA alone led to an increase in NPY production comparable to that seen after forskolin + PMA and this effect of PMA was dose-dependent. In contrast, forskolin alone did not induce NPY production. Conditioned medium, derived from monolayer cultures enriched with human astrocytes, enhanced NPY production in response to forskolin+PMA in an age-dependent manner. The NPY production by aggregates derived from a 12.5 week-, 14-week- and 18-week-old fetus was enhanced 3-3.6-fold, 1.6-2-fold and 1.1-fold, respectively. Thus, expression of the NPY neurons in this culture system is a regulated process. The NPY production is enhanced markedly by activation of the protein kinase C pathway and by an astrocyte-derived soluble substance(s). Based on these results, we propose that this culture system can serve as a model for the study of regulatory processes of the human developing NPY neuron.
authors.affiliation: Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center at Dallas 75235-9032, USA.
authors.forename: N
authors.initials: N
authors.lastname: Aguila-Mansilla
authors.forename: A
authors.initials: A
authors.lastname: Barnea
mesh_terms: D001253:Astrocytes / Q000378:metabolism / Q000502:physiology; D002273:Carcinogens / Q000494:pharmacology; D002478:Cells, Cultured / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology; D002540:Cerebral Cortex / Q000166:cytology*; D005576:Colforsin / Q000494:pharmacology; D015925:Cryopreservation; D017077:Culture Media, Conditioned / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004357:Drug Synergism; D005333:Fetus / Q000166:cytology; D006801:Humans; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D009478:Neuropeptide Y / Q000096:biosynthesis* / Q000378:metabolism; D010703:Phorbol Esters / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D013997:Time Factors
chemical_list: D002273:Carcinogens; D017077:Culture Media, Conditioned; D009478:Neuropeptide Y; D010703:Phorbol Esters; D005576:Colforsin; D000242:Cyclic AMP; D011493:Protein Kinase C