Data for "Sacchride Contents Change and the Role of PI3K-Akt Signal Transduction Pathway in Silkworm Eggs Treated with HCl to Prevent or Break Diapause"
Description
Acid soaking with HCl is a common way to prevent or break diapause of silkworm eggs and has been widely used in production. However, its mechanism remains exploring. Few literature reported in metabolism of mono-, or disaccharides, and the differential expression patterns of genes and proteins related to the PI3K/Akt signaling pathway across the stages of silkworm diapause eggs, non-diapause eggs, instant acid soaking eggs, and red bean color acid-soaked eggs (Rbcase). Silkworm 932 strains eggs at different stages were used to explore the contents of several saccharides and analyze the expression profiles of 4 diapause-related genes (BmAkt, BmGsk-3, BmGs, and CaMKII) in the PI3K-Akt signaling pathway via qPCR. The results showed that in addition to monosaccharides such as glucose and fructose, the contents of polysaccharides, maltose, trehalose, and sorbitol varied significantly at different stages and states of the eggs. The expression profiles of the 4 genes were differently low but increased in instant acid-soaked eggs and Rbcase before hatching. Western blotting results showed two bands at 51 kDa and 61 kDa for BmAkt. The ratio of p-Akt/Akt in diapause eggs and Rbcase was equivalent to each other but significantly higher in Rbcase after activation of PI3K-Akt signaling pathway. These findings revealed that saccharides are basic energic sources, and PI3K-Akt signaling pathway play important roles in acid soaking to prevent or break diapause eggs.
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Steps to reproduce
1. Using GS-MC (Agilent 7890A/5975C (GC/MS) from Thermofinnigan, USA) , the content of monosaccharides and disaccharides and repeated by iMark™ Microplate reader (No.168-1130, from Bio-Rad Laboratories, USA) were determinations were completed at the Argo-biological Gene Research Center, GDAAS. The measured data was normalized using 13Cribitol as an internal standard. 2. For qPCR, BmAkt was searched from UNIPROT (http://www.uniprot.nlm.nih.gov/), while BmGsk-3, BmGs, BmCaMKII related proteins were searched from NCBI (http://www.ncbi.nlm.nih.gov/), and miRBase (http://www. mirbase.org/). The CDS sequences encoding the relevant protein gene were downloaded: BmActin A3 (gi|187281836), BmAkt (protein_id="ABY50539.1"), BmGsk-3 (protein_id="ADM 32521.1"), BmGs (gi|101736417), and BmCaMKII (protein_id="001177296.1"). 3. Protein extracts were analyzed by Western blot (Wes™, from Bio-tech, S/N:WS-2494, USA) using Akt and p-Akt antibodies, with β-actin used as the internal reference protein. 4. Collected data of monosaccharide samples and relative expression level of qPCRwas analyzed by SPSS v28.0.1.1 software. Student’s t-test was used to evaluate the statistical differences between samples. All the data are presented as the mean ± standard error of the mean (SEM).
Institutions
- South China Agricultural UniversityGuangdong, Guangzhou
Categories
Funders
- National Sericulture Technology System Guangdong West Experimental StationGrant ID: CARs-022
- Public Welfare Industry (Agriculture) Research Special ProjectGrant ID: No.201403064