RESUMO
In fetal liver, bipotential hepatoblasts differentiate into hepatocytes and bile duct cells (cholangiocytes). The persistence of such progenitor cells in adult mouse liver is still debated. In damaged liver of adult murine animals, when hepatocyte proliferation is compromised, bipotential oval cells emerge, probably from bile ducts, proliferate, and differentiate to regenerate the liver. However, treatment to elicit oval cell proliferation is not necessary to obtain bipotential stem cells from adult mouse liver. Here, we have isolated bipotential clonal cell lines from healthy liver of 8-10-week-old C57BL/6 mice. Primary cultures established from hepatocyte-enriched suspensions were characterized by time-lapse image acquisition, immunocytology, and RNA transcript analysis. Although hepatocytes dedifferentiated with loss of apical polarity and other hepatocyte markers, they rapidly activated expression of bile duct/oval cell markers. Reversibility of these processes was achieved in part by culture under dilute Matrigel or by aging of confluent cultures. Cell lines were obtained at high frequency from mass cultures, from isolated colonies, and by primary cloning of the hepatocyte-enriched suspension. Cells of the clonal cell lines do not grow in soft agar and are nontumorigenic, and they express cytokeratin 19, A6 antigen, and alpha6 integrin, as well as a large panel of hepatocyte functions. Furthermore, they can participate in liver regeneration in albumin-urokinase-type plasminogen activator/severe combined immune-deficient mice, where they differentiate in clusters of hepatocytes and occasionally bile ducts. These results demonstrate the existence, in normal adult mouse liver, of a significant pool of clonogenic cells that are (or can become) bipotential.
Assuntos
Diferenciação Celular , Hepatócitos/citologia , Fígado/citologia , Animais , Ductos Biliares/citologia , Evolução Biológica , Biomarcadores/metabolismo , Linhagem Celular , Células Cultivadas , Cromossomos de Mamíferos/genética , Células Clonais , Feminino , Cariotipagem , Regeneração Hepática , Camundongos , Camundongos Endogâmicos C57BL , Camundongos SCID , MitoseRESUMO
The specificity of cyclic adenosine monophosphate (cAMP)-mediated signaling events is achieved by the composition and biochemical properties of the different cAMP-dependent protein kinase holoenzymes (PKAI and II) and by compartmentalization of PKA to discrete subcellular locations. Intracellular localization is mediated by interaction with A-kinase anchoring proteins (AKAPs) that recruit PKAII close to its substrates and to sites where it can respond optimally to local changes in intracellular cAMP concentration, thereby directing and amplifying the effects of cAMP. This review presents recent evidence that indicates that specific AKAPs mediate PKAI anchoring through interaction with its regulatory subunit RI alpha, notably at the neuromuscular junction of skeletal muscle.