RN46A

Cat.No.: CSC-C9440J

Species: Rattus norvegicus (Rat)

Source: Embryo; Brain

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Cat.No.

CSC-C9440J

Description

RN46A , an immortalised serotonergic neuronal cell line, was cloned by serial dilution following infection of dissociated embryonic day 13 rat medullary raphe cells with a retrovirus encoding the temperature-sensitive mutant of SV40 large T-antigen (T-ag), RN46A cells are capable of differentiating at 39°C the non-permissive temperature. Under differentiation conditions, RN46A cells cease dividing, take on a neuronal morphology, and express enhanced levels of NSE and all three NF proteins. Differentiated RN46A cells express low-affinity nerve growth factor (NGF) receptor (p75NGFR) and are immunoreactive using an antibody that recognizes the carboxy-terminal 13 amino acids of all three trk proteins (pan-trk). Both immunoreactivities could be potentiated by treatment with brain-derived neurotrophic factor (BDNF), NGF, and adrenocorticotropic hormone, fragment 4-10 (ACTH4-10). Differentiated RN46A cells express low levels of tryptophan hydroxylase (TPH) immunoreactivity, which could be enhanced by treatment with ACTH4-10, BDNF, or NGF. Low levels of serotonin immunoreactivity are detected in differentiated RN46A cells, and this was potentiated by differentiating RN46A cells with BDNF for 8 d and 40 mM KCl for days 4-8. RN46A cells should prove useful to elucidate intracellular mechanisms that control neurofilament assembly and 5-HT expression in differentiating raphe neurons.

Species

Rattus norvegicus (Rat)

Source

Embryo; Brain

Recommended Medium

DMEM:Ham's F12 (1:1) + 2mM Glutamine + 10% Fetal Bovine Serum (FBS) + 0.25mg/ml Geneticin (G418). Alternatively CNS medium can be used.

Storage

Liquid Nitrogen (-180 °C).

Storage and Shipping

Creative Bioarray ships frozen cells on dry ice. On receipt, immediately transfer frozen cells to liquid nitrogen (-180 °C) until ready for experimental use. Never can cryopreserved cells be kept at -20 °C.

Synonyms

RN-46A

Citation Guidance

If you use this products in your scientific publication, it should be cited in the publication as: Creative Bioarray cat no. If your paper has been published, please click here to submit the PubMed ID of your paper to get a coupon.

RN46A cells, derived from the embryonic rat brain's raphe nucleus, are an immortalized line known for their abundance of serotonin-releasing neurons. RN46A Cells were immortalized with temperature-sensitive SV40 large T antigen and can be induced to proliferate or differentiate into serotonergic cells depending on the cell culture conditions. RN46A Cells therefore allow for an in vitro study of CNS serotonergic neuron development and function. Differentiated RN46A Cells behave similarly to neuronal cells in that they extend neurite-like processes and express neuronal markers. Furthermore, upon differentiation, RN46A cells display the expression of serotonergic neuron-specific markers, such as tryptophan hydroxylase (TPH), the serotonin transporter (SERT), and several neuronal cytoskeletal proteins.

Due to these characteristics RN46A Cells are used to study serotonin biosynthesis, transport, signaling, neuronal differentiation, neurodevelopment and synaptic regulation. RN46A cells are also a valuable tool for investigating neurotoxicity, neuroinflammation, and how drugs like antidepressants and other psychoactive substances impact serotonin-producing neurons.

Effects of MDMA and BDNF on Cell Viability by MTT Assay

MDMA ("Ecstasy") causes persistent serotonergic (5-HT) system alterations through unclear molecular mechanisms. Bavato et al. investigated whether BDNF pre-treatment prevents MDMA neurotoxicity and examined MDMA's effects on neurofilament light chain (NfL) in a 5-HT neuronal cell line.

5-HT neurons were differentiated from rat brain raphe nucleus RN46A cells, confirmed by MAP-2 immunocytochemistry (Supplementary Material 4). MDMA IC₅₀ was 1.75 mM at 24 h and 1.15 mM at 48 h; 1.3 mM was selected for subsequent experiments. BDNF (100 ng/ml, 1 h pre-treatment) partially counteracted MDMA-induced cell viability reduction at 24 h (F = 34.68, p < 0.001), but not at 48 h (F = 2.20, p = 0.160) (Fig. 1). Significant differences were found between MDMA and control, and BDNF + MDMA and control, at both time points (p < 0.001).

Effect of BDNF and MDMA treatment on RN46A serotonergic neurons viability. — Fig. 1. Effect of BDNF and MDMA treatment on RN46A serotonergic neurons viability (Bavato F, Stamatakos S, *et al*., 2022).

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