Activació de les fosfolipases C i D per neurotransmissors

Els fosfolípids de les membranes cel·lulars compleixen un paper fonamental en el manteniment de 1'estructura i la compartició cel·lular, gràcies a la seva naturalesa molecular amfipàtica, i cada cop més són observats des de la perspectiva de la transducció de senyals com a precursors de missatgers intracel·lulars. Diverses fosfolipases, sensibles a l'activació per neurotransmissors i altres estímuls extracel·lulars, catalitzen la seva hidròlisi trencant enllaços ester o fosfoester, i l'acció, com a segons missatgers, dels productes que en poden derivar permet dibuixar una xarxa d'esdeveniments que se succeeixen i s'entrellacen configurant una part important del complex comandament del metabolisme i de les funcions cel·lulars. En aquest capítol tractarem alguns aspectes relacionats amb la intervenció de les fosfolipases C de fosfoinosítids (PLC) i de les fosfolipases D (PLD) en sistemes de transducció

Early impairment of lung mechanics in a murine model of Marfan syndrome

Early morbidity and mortality in patients with Marfan syndrome (MFS) -a connective tissue disease caused by mutations in fibrillin-1 gene- are mainly caused by aorta aneurysm and rupture. However, the increase in the life expectancy of MFS patients recently achieved by reparatory surgery promotes clinical manifestations in other organs. Although some studies have reported respiratory alterations in MFS, our knowledge of how this connective tissue disease modifies lung mechanics is scarce. Hence, we assessed whether the stiffness of the whole lung and of its extracellular matrix (ECM) is affected in a well-characterized MFS mouse model (FBN1 C1039G/+ ). The stiffness of the whole lung and of its ECM were mea- sured by conventional mechanical ventilation and atomic force microscopy, respectively. We studied 5-week and 9-month old mice, whose ages are representative of early and late stages of the disease. At both ages, the lungs of MFS mice were significantly more compli- ant than in wild type (WT) mice. By contrast, no significant differences were found in local lung ECM stiffness. Moreover, histopathological lung evaluation showed a clear emphyse- matous-like pattern in MFS mice since alveolar space enlargement was significantly increased compared with WT mice. These data suggest that the mechanism explaining the increased lung compliance in MFS is not a direct consequence of reduced ECM stiffness, but an emphysema-like alteration in the 3D structural organization of the lung. Since lung alterations in MFS are almost fully manifested at an early age, it is suggested that respira- tory monitoring could provide early biomarkers for diagnosis and/or follow-up of patients with the Marfan syndrom

Diacylglycerol is required for the formation of COPI vesicles in the Golgi-to-ER transport pathway

Diacylglycerol is necessary for trans-Golgi network (TGN) to cell surface transport, but its functional relevance in the early secretory pathway is unclear. Although depletion of diacylglycerol did not affect ER-to-Golgi transport, it led to a redistribution of the KDEL receptor to the Golgi, indicating that Golgi-to-ER transport was perturbed. Electron microscopy revealed an accumulation of COPI-coated membrane profiles close to the Golgi cisternae. Electron tomography showed that the majority of these membrane profiles originate from coated buds, indicating a block in membrane fission. Under these conditions the Golgi-associated pool of ARFGAP1 was reduced, but there was no effect on the binding of coatomer or the membrane fission protein CtBP3/BARS to the Golgi. The addition of 1,2-dioctanoyl-sn-glycerol or the diacylglycerol analogue phorbol 12,13-dibutyrate reversed the effects of endogenous diacylglycerol depletion. Our findings implicate diacylglycerol in the retrograde transport of proteins from Golgi to the ER and suggest that it plays a critical role at a late stage of COPI vesicle formation

Activació de les fosfolipases C i D per neurotransmissors

Els fosfolípids de les membranes cel·lulars compleixen un paper fonamental en el manteniment de 1'estructura i la compartició cel·lular, gràcies a la seva naturalesa molecular amfipàtica, i cada cop més són observats des de la perspectiva de la transducció de senyals com a precursors de missatgers intracel·lulars. Diverses fosfolipases, sensibles a l'activació per neurotransmissors i altres estímuls extracel·lulars, catalitzen la seva hidròlisi trencant enllaços ester o fosfoester, i l'acció, com a segons missatgers, dels productes que en poden derivar permet dibuixar una xarxa d'esdeveniments que se succeeixen i s'entrellacen configurant una part important del complex comandament del metabolisme i de les funcions cel·lulars. En aquest capítol tractarem alguns aspectes relacionats amb la intervenció de les fosfolipases C de fosfoinosítids (PLC) i de les fosfolipases D (PLD) en sistemes de transducció

Lipid phosphate phosphatase 3 participates in transport carrier formation and protein trafficking in the early secretory pathway

The inhibition of phosphatidic acid phosphatase (PAP) activity by propanolol indicates that diacylglycerol (DAG) is required for the formation of transport carriers at the Golgi and for retrograde trafficking to the ER. Here we report that the PAP2 family member lipid phosphate phosphatase 3 (LPP3, also known as PAP2b) localizes in compartments of the secretory pathway from ER export sites to the Golgi complex. The depletion of human LPP3: (i) reduces the number of tubules generated from the ER-Golgi intermediate compartment and the Golgi, with those formed from the Golgi being longer in LPP3-silenced cells than in control cells; (ii) impairs the Rab6-dependent retrograde transport of Shiga toxin subunit B from the Golgi to the ER, but not the anterograde transport of VSV-G or ssDsRed; and (iii) induces a high accumulation of Golgi-associated membrane buds. LPP3 depletion also reduces levels of de novo synthesized DAG and the Golgi-associated DAG contents. Remarkably, overexpression of a catalytically inactive form of LPP3 mimics the effects of LPP3 knockdown on Rab6-dependent retrograde transport. We conclude that LPP3 participates in the formation of retrograde transport carriers at the ER-Golgi interface, where it transitorily cycles, and during its route to the plasma membrane