Receptors to steroid hormones and aromatase are expressed by cultured motoneurons but not by glial cells derived from rat embryo spinal cord.
The aim of this study was to examine the expression of aromatase and receptors to steroid hormones in cultured motoneurons (MNs). We first developed an original method for obtaining rat MN cultures. Dissociated E15 rat spinal cords were purified using metrizamide and bovine serum albumin density gradients, and cells were then seeded on the culture substratum. We optimized the culture parameters and found that simple addition of rat muscle extract (ME) and conditioned culture medium (CM) from glial cell lines (GCL) derived from spinal cord were sufficient to obtain almost pure MN cultures. MNs were characterized by the presence of specific MN markers and electrophysiology. MNs could be kept alive for 2 weeks. We demonstrate that ME and CM are essential for MN development and survival respectively. Immunocytochemistry and aromatase activity assay indicated the presence of androgen and estrogen receptors as well as aromatase in MNs but not in GCL. This is the first report demonstrating the presence of both female and male sex hormone receptors and a key enzyme in steroid hormone metabolism in MNs and its absence in GCL, at least in our culture conditions. This in vitro model appears to be valuable for elucidating the impact of the sex hormone circuit in neuronal maturation. The relevance of this model for the comprehension of neurodegenerative diseases is discussed.
Rakotoarivelo
Clovis
C
Unité 583, INSERM, Instituts des Neurosciences de Montpellier, Hôpital Saint-Eloi, 80, rue Augustin-Fliche, FR-34091 Montpellier, France.
Petite
Didier
D
Lambard
Sophie
S
Fabre
Christine
C
Rouleau
Caroline
C
Lumbroso
Serge
S
de Weille
Jan
J
Privat
Alain
A
Carreau
Serge
S
Mersel
Marcel
M
D000818:Animals; D001141:Aromatase / Q000378:metabolism*; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D004591:Electrophoresis, Polyacrylamide Gel; D004622:Embryo, Mammalian; D007150:Immunohistochemistry; D008564:Membrane Potentials / Q000502:physiology; D009046:Motor Neurons / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism; D009457:Neuroglia / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D011987:Receptors, Steroid / Q000378:metabolism*; D013116:Spinal Cord / Q000166:cytology*
D017077:Culture Media, Conditioned; D011987:Receptors, Steroid; D001141:Aromatase