Receptor-mediated New Selective Autophagy Degrades The Golgi Apparatus

Aim: It has been recorded that selective autophagy was involved in quality control of some organelles such as mitochondria.Studies indicated that the selective autophagy of the Golgi apparatus may exist.This paper is going to prove a new selective autophagy of the Golgi apparatus and GOLPH3 was able to act as a novel cargo receptor to mediate Golgiphagy.Apelin-13/APJ induced cardiomyocytes hypertrophy through Golgiphagy.Methods: 1.Use fluorescence microscopy to observe the co-localization of GM130–RFP(the cis-Golgi marker)and LC3B-GFP.2.Use fluorescence microscopy to observe the co-localization of TGN46-RFP(the trans-Golgi marker)and LC3B-GFP.3.Use transmission electron microscopy to observe the occurrence of Golgiphagy.4.Use the co-immunoprecipitation assay to detect the interaction of GOLPH3 and LC3 B.5.Use siRNA to knock down GOLPH3 to observe the Golgiphagy.6.Overexpressing APJ receptor to observe the Golgiphagy.7.Use the immunohistochemical assay to detect the expression of GOLPH3 in myocardium of patients with cardiomyocytes hypertrophy.8.Use automated Cell Counter to detect the cell diameter and volume of H9c2 cells.Results: 1.Starvation promoted the co-localization of GA marker proteins and LC3 B in H9c2 cells,HUVECs,HA-VSMCs and HEK293 T cells.2.Hypoxia facilitated the co-localization of GA marker proteins and LC3 B in H9c2 cells,HUVECs,HA-VSMCs and HEK293 T cells.3.Rapamycin triggered the co-localization of GA marker proteins and LC3 B in H9c2 cells,HUVECs,HA-VSMCs and HEK293 T cells.4.Golgi stress inducers(BelfeldinA,Nocodazole,Monensin,Nigericin)induced the co-localization of GA marker proteins and LC3 B in H9c2 cells,HUVECs,HA-VSMCs and HEK293 T cells.5.Inducers of cardiac hypertrophy(Apelin-13,APJ-overexpression,Ang II and Static pressure)were able to promote the co-localization of GA marker proteins and LC3 B in H9c2 cells.6.Transmission electron microscopy images showed that starvation induced the occurrence of the Golgiphagy in H9c2 cells,HUVECs and HA-VSMCs.7.Transmission electron microscopy images also showed that inducers of cardiac hypertrophy(Apelin-13,Ang II and Static pressure)facilitated the Golgiphagy in H9c2 cells.8.Starvation,hypoxia and rapamycin promoted the interaction of GOLPH3 and LC3 B in H9c2 cells,HUVECs and HA-VSMCs.9.Golgi stress inducers triggered the interaction of GOLPH3 and LC3 B in H9c2 cells,HUVECs and HA-VSMCs.10.Inducers of cardiac hypertrophy(Apelin-13,APJ-overexpression,Ang II and Static pressure)facilitated the interaction of GOLPH3 and LC3 B in H9c2 cells.11.Fluorescence microscopyimages showed that si-GOLPH3 inhibited the co-localization of GA marker proteins with LC3 B in H9c2 cells,HUVECs,HA-VSMCs and HEK293 T cells.12.Transmission electron microscopy images also showed that si-GOLPH3 depressed the Golgiphagy in H9c2 cells.13.Apelin-13/APJ promoted the expression of GOLPH3 in H9c2 cells.14.Apelin-13/APJ induced cardiomyocytes hypertrophy through Golgiphagy.15.The expression of GOLPH3 increased in myocardium of patients with cardiomyocytes hypertrophy.Conclusion: This study unveiled a new selective autophagy of the Golgi apparatus and GOLPH3 was able to act as a novel cargo receptor to mediate Golgiphagy.Apelin-13/APJ induced cardiomyocytes hypertrophy through Golgiphagy.