Preliminary investigation demonstrating the GHITM gene probably involved in apoptosis and growth of the golden apple snail (Pomacea canaliculata)

Wenchao Yu,Hongce Song,Qiuyun Jiang,Nannan Zhao,Cheng He,Xiaotong Wang,Wen Guo,Lingling Li,Lei Wei,Rui Zhan,Baolu Zhang,Xiaona Wang

doi:10.1186/s12864-019-6434-2

Abstract

BackgroundGrowth hormone inducible transmembrane protein (GHITM) is a highly conserved transmembrane protein. This study was conducted to investigate the role of GHITM gene in the apoptosis and growth of the golden apple snail Pomacea canaliculate.ResultsThe complete cDNA of this gene was cloned using the rapid amplification of cDNA ends (RACE) method and subjected to bioinformatics analysis. The full-length cDNA was 2242 bp, including an open reading frame of 1021 bp that encoded a protein of 342 amino acid residues. The mRNA expression profiles of GHITM gene in different tissues (liver, kidney, gonad and foot) and different growth phases (6-months old and 2-years old) showed that it was expressed in various tissues and different growth phases. Silencing of the GHITM gene by RNAi (RNA interference) experiments revealed that the GHITM gene possibly plays a role in inhibiting apoptosis through detecting the Caspase (Cysteine-requiring Aspartate Protease)-3 activity. In addition, the aperture width and body whorl length of the snail was significantly affected by RNAi, suggesting that this gene plays a significant role in promoting the growth of the organism.ConclusionsThese results demonstrated that the GHITM gene was involved in apoptosis and growth in golden apple snail.

Highlights

Growth hormone inducible transmembrane protein (GHITM) is a highly conserved transmembrane protein
Bioinformatics analysis of GHITM The identified open reading frame of GHITM encoded a putative protein of 342 amino acids with a predicted molecular mass of 36,373.87 Da and theoretical isoelectric point of 9.98
The results showed that GHITM gene was expressed in different tissues and growth stages, and widely throughout individuals, which was basically consistent with previous studies on other species [23, 32]

Summary

Results

Characterization of the GHITM cDNA sequence The full-length cDNA sequence of GHITM included a 5′- UTR (Untranslated Region) of 324 bp, an open reading frame of 1029 bp, encoding a protein of 342 amino acids, and a 3′-UTR of 890 bp. Expression level of GHITM in different tissues and different growth phases of the Golden apple snail To explore the expression pattern of the GHITM gene in different tissues, qPCR was performed using total RNA from liver, kidney, gonad and foot. To detect the expression pattern of the GHITM gene during different growth phases, RT-qPCR was performed with total RNA from young (6-months old) and adult (2-years old) golden apple snails. The GHITM transcript levels were not significantly different between 6-month-old and 2-year-old golden apple snails (Fig. 8). The change in the growth of the control group was greater than those of the two experimental groups, and the aperture width and body whorl length of the control group were significantly greater than those of the C group (Fig. 11)

Background

Discussion

Conclusions

Methods