Adverse neurodevelopmental effects of dexamethasone modeled in PC12 cells: identifying the critical stages and concentration thresholds for the targeting of cell acquisition, differentiation and viability.
The use of dexamethasone (DEX) to prevent respiratory distress in preterm infants is suspected to produce neurobehavioral deficits. We used PC12 cells to model the effects of DEX on different stages of neuronal development, utilizing exposures from 24 h up to 11 days and concentrations from 0.01 to 10 microM, simulating subtherapeutic, therapeutic, and high-dose regimens. In undifferentiated cells, even at the lowest concentration, DEX inhibited DNA synthesis and produced a progressive deficit in the number of cells as evaluated by DNA content, whereas cell growth (evaluated by the total protein to DNA ratio) and cell viability (Trypan blue exclusion) were promoted. When cell differentiation was initiated with nerve growth factor, the simultaneous inclusion of DEX still produced a progressive deficit in cell numbers and promoted cell growth and viability while retarding the development of neuritic projections as monitored by the membrane/total protein ratio. Again, even 0.01 microM DEX was effective. We next assessed effects at mid-differentiation by introducing nerve growth factor for 4 days followed by coexposure to DEX. Although effects on cell number, growth, and neurite extension were still detectable, the outcomes were generally less notable. DEX also shifted the fate of PC12 cells away from the cholinergic phenotype and toward the adrenergic phenotype, with the maximum effect achieved at the outset of differentiation. Our results indicate that DEX directly disrupts neuronal cell replication, differentiation, and phenotype at concentrations below those required for the therapy of preterm infants, providing a mechanistic link between glucocorticoid use and neurodevelopmental sequelae.
Jameson
Ruth R
RR
Department of Pharmacology & Cancer Biology, Duke University Medical Center, Durham, NC 27710, USA.
Seidler
Frederic J
FJ
Qiao
Dan
D
Slotkin
Theodore A
TA
D000818:Animals; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D048429:Cell Size / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects* / Q000502:physiology; D003907:Dexamethasone / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D001523:Mental Disorders / Q000139:chemically induced / Q000473:pathology; D016716:PC12 Cells; D051381:Rats; D013997:Time Factors
D003907:Dexamethasone