Archive | 2021
Identification and Characterization of The Pyruvate Dehydrogenase E1 Gene in The Oriental River Prawn, Macrobrachium Nipponense
Abstract
\n Pyruvate dehydrogenase E1 (PDHE1) was predicted to be involved in the male sexual development in M. nipponense in the previous study. In this study, the functions of Mn-PDHE1 were further analysed in depth using quantitative polymerase chain reaction (qPCR), in situ hybridization, western-blot and RNA interference (RNAi), combined with the histological observations. The full-genome sequence of Mn- PDHE1 was 65,630 base pairs (bp) on Chromosome 11, including 15 introns and 14 exons. The full cDNA sequence of Mn-PDHE1 was 1614 bp, including a 1077 bp open reading frame, encoding for 358 amino acids. qPCR analysis revealed that Mn- PDHE1 showed the highest expression level in the testis, and male prawns at post-larval developmental stage 25 (PL25♂) was higher than PL25♀, indicating that PDHE1 was potentially involved in the male sexual development in M. nipponense. An additional function of Mn-PDHE1 was predicted to be involved in the metamorphosis process. In situ hybridization and western-blot indicated that PDHE1 was involved in the whole testis development. The Mn-IAG mRNA was decreased with the decrease of Mn- PDHE1. This indicated PDHE1 has the positive regulatory effects on IAG in M. nipponese. The testis and androgenic gland development were delayed after the injection of Mn- PDHE1 dsRNA, compared with those of control group, indicating PDHE1 has positive effects on the male sexual development in M. nipponense. This study highlights the functions of PDHE1 in M. nipponense, which can be applied to future studies of the male sexual development in other crustacean species.