AUTHOR=Jin Shubo , Hu Yuning , Fu Hongtuo , Jiang Sufei , Xiong Yiwei , Qiao Hui , Zhang Wenyi , Gong Yongsheng , Wu Yan 

TITLE=Identification and Characterization of the Pyruvate Dehydrogenase E1 Gene in the Oriental River Prawn, Macrobrachium nipponense

JOURNAL=Frontiers in Endocrinology

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/endocrinology/articles/10.3389/fendo.2021.752501

DOI=10.3389/fendo.2021.752501

ISSN=1664-2392

ABSTRACT=<p>Pyruvate dehydrogenase E1 (<italic>PDHE1</italic>) is thought to play essential roles in energy metabolism, and a previous study suggested that it also has potential regulatory roles in male sexual development in the oriental river prawn, <italic>Macrobrachium nipponense</italic>. In this study, we used rapid amplification of cDNA ends, quantitative polymerase chain reaction (qPCR), <italic>in situ</italic> hybridization, western blotting, RNA interference (RNAi), and histological analyses to assess the potential functions of <italic>Mn-PDHE1</italic> in the sexual development of male <italic>M. nipponense</italic>. The full cDNA sequence of <italic>Mn-PDHE1</italic> was 1,614 base pairs long, including a 1,077 base pair open reading frame that encodes 358 amino acids. qPCR analysis revealed the regulatory functions of <italic>PDHE1</italic> in male sexual development in <italic>M</italic>. <italic>nipponense</italic> and in the metamorphosis process. <italic>In situ</italic> hybridization and western blot results indicated that <italic>PDHE1</italic> was involved in testis development, and RNAi analysis showed that <italic>PDHE1</italic> positively regulated the expression of insulin-like androgenic gland factor in <italic>M. nipponense</italic>. Compared with the cell types in the testes of control prawns, histological analysis showed that the number of sperm was dramatically lower after test subjects were injected with <italic>Mn-PDHE1</italic> dsRNA, whereas the numbers of spermatogonia and spermatocytes were higher. Sperm constituted only 1% of cells at 14 days after injection in the RNAi group. This indicated that knockdown of the expression of <italic>PDHE1</italic> delayed testis development. Thus, <italic>PDHE1</italic> has positive effects on male sexual development in <italic>M</italic>. <italic>nipponense</italic>. This study highlights the functions of P<italic>DHE1</italic> in <italic>M</italic>. <italic>nipponense</italic> and its essential roles in the regulation of testis development.</p>