Neuroblastoma B104 cell line as a model for analysis of neurite outgrowth and neuronal aggregation induced by Reissner's fiber material

Abstract

The subcommissural organ (SCO) is a specialized ependymal structure of the brain that secretes glycoproteins into the cerebrospinal fluid (CSF), which condense to form a thread-like structure - Reissner's fiber (RF). The effects of soluble material released by RF were examined on neuroblastoma B104 cells grown in serum-free medium, using "low-density" and "high-density" culture systems. In the presence of soluble RF material, low-density cultures were suitable for analysis of the enhanced neurite outgrowth of B104 cells, while high-density cultures allowed the increased B104 cell aggregation to be examined. RF-induced neuronal aggregation and neuritic outgrowth were restricted to a perimeter around the RF. This standardized cell culture system reproduced in part the effects observed previously with primary cortical and spinal cord cell cultures and may serve the analysis of the mechanisms leading to aggregation and neurite outgrowth. In the present study, we analyzed variations in the rate of neural cell adhesion molecules, such as N-CAM and N-cadherin, induced by soluble RF material in high-density cultures.

Neuroprotection by the synthetic neurosteroid enantiomers ent-PREGS and ent-DHEAS against Aβ25–35 peptide-induced toxicity in vitro and in vivo in mice

“…Low-density cultures of B104 neuroblastoma cells are suitable for the analysis of the enhanced neurite outgrowth (El Bitar et al 1999). Three thousands cells in complete culture medium were seeded and allowed to attach for 24 h. They were incubated with ent -PREGS, PREGS, ent -DHEAS or DHEAS or no steroid (control), in the presence or absence of Aβ 25-35 in free serum culture medium for 3, 5 or 7 days.…”

Functional aspects of the subcommissural organ-Reissner's fiber complex with emphasis in the clearance of brain monoamines

“…Due to the finding that RF-glycoproteins may induce neuroblast differentiation (El Bitar et al, 1999), the possibility that in the adult SCO secretory Reproduced from E.M. Rodríguez et al (1992) with permission of the publisher).…”

Survival of mammalian B104 cells following neurite transection at different locations depends on somal Ca2+ concentration

“…Rat B104 cells, originally derived from a CNS neuroblastoma (Schubert et al, 1974;Bottenstein and Sato, 1979), generate action potentials (Gu and Waxman, 1996), release neurotransmitters [(e.g., GABA), (Tyndale et al, 1994)] and are often used as model systems to study cellular/molecular neuronal processes (Huber et al, 1985;El Bitar et al, 1999;Toda et al, 1999). B104 cells were grown in an incubator (humidified, regulated 5% CO 2 atmosphere at 37°C) in a "growth medium" consisting of a 1:1 mixture of Dulbecco's Modified Eagle Medium (DMEM) and Carlsbad,CA) supplemented for growth with 10% heat-inactivated fetal calf serum and 1% antibiotics (Sigma, St. Louis, MO; 10,000 units penicillin/ml and 10 mg streptomycin/ml).…”