Differentiated rabbit prostatic stromal cells in primary culture display functional alpha1A-adrenoceptors.
OBJECTIVE
BPH is characterized by uncontrolled proliferation and increased contractility of prostatic smooth muscle cells. The activation of alpha1-adrenoceptors (alpha1-AR) seems involved in the latter event, but the lack of in vitro models expressing these receptors has hampered a more specific characterization of their role. In order to do so, we attempted to develop a new model of rabbit cultured prostatic stromal cells (PSC) in a non-proliferative and differentiated state.


METHODS
The expression of cytoskeletal and stromal markers was confirmed by immunohistochemistry on primary cultured PSC. Alpha1-AR subtype expression was assessed by RT-PCR, while receptor coupling to the ERK1/ERK2 and calcium pathways was studied by Western Blot and Fura-2 calcium imaging, respectively.


RESULTS
Cells grown under non-proliferative conditions displayed a differentiated phenotype, with expression of contractile cytoskeletal and stromal proteins. Furthermore, the alpha1A-AR was shown to activate ERK1/ERK2 as well as calcium signaling.


CONCLUSIONS
These results emphasize the interest of this model for the characterization of PSC adrenergic regulation, in particular through the little-known alpha1A-AR.
Boulbès
Delphine
D
EA 2995, Faculté de Pharmacie, Montpellier, France.
Choquet
Armelle
A
Barrère-Lemaire
Stéphanie
S
Costa
Pierre
P
Soustelle
Laurent
L
Nargeot
Joël
J
Bali
Jean-Pierre
JP
Hollande
Frédéric
F
Magous
Richard
R
D000818:Animals; D015153:Blotting, Western; D001973:Bromodeoxyuridine / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D002469:Cell Separation; D048430:Cell Shape; D002478:Cells, Cultured; D003285:Contractile Proteins / Q000502:physiology; D003599:Cytoskeleton / Q000502:physiology; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D048049:Extracellular Signal-Regulated MAP Kinases / Q000502:physiology; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008297:Male; D010641:Phenotype; D011467:Prostate / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits; D018340:Receptors, Adrenergic, alpha-1 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity; D017154:Stromal Cells / Q000378:metabolism*
C547713:ADRA1A protein, human; D003285:Contractile Proteins; D012333:RNA, Messenger; D018340:Receptors, Adrenergic, alpha-1; D004247:DNA; D048049:Extracellular Signal-Regulated MAP Kinases; D001973:Bromodeoxyuridine; D002118:Calcium