Differential function of the alpha2A-adrenoceptor and Phosphodiesterase-3B in human adipocytes of different origin.
OBJECTIVE
Human adipocytes can be obtained in vitro by differentiation of human preadipocytes or mesenchymal stem cells (hMSC). Although functionally similar to freshly isolated cells, no detailed comparison of the different cell types has been performed. The antilipolytic alpha2A-adrenoceptor (AR) and the cAMP-degrading enzyme Phosphodiesterase-3B (PDE3B) have been implicated in the fine-tuning of lipolysis but little is known regarding their role in human adipocytes nor whether their expression and/or function differs in fat cells from different precursors.


METHODS
The effects of alpha2A-AR and PDE3B inhibition in mature adipocytes was determined and compared to that in differentiated preadipocytes and hMSC-derived fat cells. Gene expression was determined by real-time PCR and protein expression by Western blot.


RESULTS
Noradrenaline (NA) stimulated lipolysis in preadipocytes and mature adipocytes but markedly reduced lipolysis in differentiated hMSC derived-adipocytes. This was due to a potent stimulation of alpha2A-AR since co-incubation with NA and the alpha2-AR-inhibitor yohimbine restored NA-induced lipolysis. The order of Yohimbine response was hMSC>preadipocytes>mature adipocytes. Although alpha2-AR mRNA expression was highest in mature adipocytes there was no difference in alpha2A-AR protein levels between the cell types. In contrast, Galphai2 mRNA and protein expression was significantly higher in MSC-derived adipocytes, suggesting that differences in the response to alpha2A-AR inhibition reside at the postreceptor level. Incubation with the cAMP-analog 8-bromo(8b) cAMP increased lipolysis in hMSC-derived fat cells while co-incubation with the PDE3-specific inhibitor OPC3911 did not alter the lipolytic effect. In contrast, OPC3911 increased 8bcAMP-induced lipolysis significantly in preadipocytes and mature adipocytes. The response to PDE3B inhibition was; mature adipocytes>preadipocytes>hMSC a finding that correlated significantly with both PDE3B mRNA expression and enzymatic activity.


CONCLUSIONS
Although differentiated adipocytes of different origins display similar functional characteristics there are important differences in the regulation of lipolysis with a marked alpha2A-AR and less pronounced PDE3B effect in fat cells from MSCs.
Dicker
A
A
Department of Medicine, Karolinska University Hospital, Stockholm, Sweden.
Kaaman
M
M
van Harmelen
V
V
Aström
G
G
Blanc
K L
KL
Rydén
M
M
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000378:metabolism*; D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000328:Adult; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology*; D000242:Cyclic AMP / Q000378:metabolism; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3; D005260:Female; D006801:Humans; D008066:Lipolysis / Q000187:drug effects*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D015363:Quinolones / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D018341:Receptors, Adrenergic, alpha-2 / Q000378:metabolism*; D015016:Yohimbine / Q000494:pharmacology
D015363:Quinolones; D012333:RNA, Messenger; D018341:Receptors, Adrenergic, alpha-2; D015124:8-Bromo Cyclic Adenosine Monophosphate; D015016:Yohimbine; C060853:OPC 3911; D000242:Cyclic AMP; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3; C517291:PDE3B protein, human