Differential gene expression in the liver of the African lungfish, Protopterus annectens, after 6 months of aestivation in air or 1 day of arousal from 6 months of aestivation

Abstract

The African lungfish, Protopterus annectens, can undergo aestivation during drought. Aestivation has three phases: induction, maintenance and arousal. The objective of this study was to examine the differential gene expression in the liver of P. annectens after 6 months (the maintenance phase) of aestivation as compared with the freshwater control, or after 1 day of arousal from 6 months aestivation as compared with 6 months of aestivation using suppression subtractive hybridization. During the maintenance phase of aestivation, the mRNA expression of argininosuccinate synthetase 1 and carbamoyl phosphate synthetase III were up-regulated, indicating an increase in the ornithine-urea cycle capacity to detoxify ammonia to urea. There was also an increase in the expression of betaine homocysteine-S-transferase 1 which could reduce and prevent the accumulation of hepatic homocysteine. On the other hand, the down-regulation of superoxide dismutase 1 expression could signify a decrease in ROS production during the maintenance phase of aestivation. In addition, the maintenance phase was marked by decreases in expressions of genes related to blood coagulation, complement fixation and iron and copper metabolism, which could be strategies used to prevent thrombosis and to conserve energy. Unlike the maintenance phase of aestivation, there were increases in expressions of genes related to nitrogen, carbohydrate and lipid metabolism and fatty acid transport after 1 day of arousal from 6 months aestivation. There were also up-regulation in expressions of genes that were involved in the electron transport system and ATP synthesis, indicating a greater demand for metabolic energy during arousal. Overall, our results signify the importance of sustaining a low rate of waste production and conservation of energy store during the maintenance phase, and the dependence on internal energy store for repair and structural modification during the arousal phase, of aestivation in the liver of P. annectens.

Fig 1. Quantitative RT-PCR results of selected genes that were differentially expressed in the SSH libraries.

Relative quantification of mRNA expression (fold change) of (A) betaine-homocysteine S-methyltransferase 1 (bhmt1, JZ575536), (B) fumarate hydratase (fh, JZ575565), (C) argininosuccinate synthetase 1 (ass1, JZ575533), (D) carbamoyl-phosphate synthetase III (cpsIII, JZ575539), (E) superoxide dismutase 1 (sod1, JZ575606), (F) ceruloplasmin (cp, JZ575541), (G) acyl-CoA desaturase (acd, JZ575387), (H) ferritin light chain (ftl, JZ575418) and (I) glyceraldehyde-3-phosphate dehydrogenase (gapdh, JZ575429), using β-actin as the reference gene, in the liver of Protopterus annectens after 6 months (mon) of aestivation as compared with the freshwater control (A-F), or 1 day (d) of arousal from 6 mon aestivation as compared with fish aestivated for 6 mon (G-I). Results represent mean + S.E.M. (N = 6). *Significantly different from the corresponding control (P<0.05).