Effects of Autophagy Inhibition(CQ)and Autophagy Potentiation(RAPA)on Autophagy,Apoptosis,and Sex-Marker Genes in Testis and Ovary of Sepiella japonica
Chloroquine(CQ)and rapamycin(RAPA)are commonly used autophagy inhibitors and poten-t iators in the laboratory.Studies in model organisms such as mice and zebrafish have found that altering the level of autophagy can have an effect on spermatogenesis and oogenesis.In this study,we used Sepiella japonica to systematically compare the effects of autophagy(LC3,BECN1),apoptosis(p53,Cas3),invertebrate gap function protein gene(Inx4),male marker(Sox3,Sox30)and female marker(Nanos-1,FTZ-F1)gene expression after treatment with different concentrations of CQ and RAPA in testis and ovaries by establishing in vitro au-tophagy-inhibiting(CQ)and autophagy-potentiating(RAPA)tissue-culture models in testis and ovary.The re-sults showed that CQ treatment caused a significant inhibition of autophagy level in testis and ovary of S.japonica,a significant increase of autophagy marker gene LC3 and a significant decrease of BECN1;the trend of change of Inx4 was basically consistent with that of autophagy marker genes,suggesting that there was a close relationship between Inx4 and autophagy;apoptosis marker genes p53 and Cas3 were significantly elevat-ed,indicating that the The expression changes of male marker genes Sox4 and Sox30 lagged behind those of autophagy and apoptosis-related genes,indicating that autophagy inhibition also had a significant effect on ga-metogenesis.Autophagy potentiation(RAPA)likewise caused significant changes in autophagy marker genes(LC3,BECN1),invertebrate gap function protein gene(Inx4),apoptosis genes(p53,Cas3),male marker genes(Sox3,Sox30)and female marker genes(Nanos-1,FTZ-F1),suggesting that autophagy exerted a significant ef-fect on S.japonica spermatogenesis and oogenesis.The results of this study reveal for the first time in marine invertebrates the close relationship between Inx4 and autophagy,and the direct influence of both on gametoge-nesis,which provides important basic data for further research on the regulation of autophagy and Inx4 on ga-metogenesis in marine invertebrates including cephalopods.
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