Identification of candidate genes from androgenic gland in Macrobrachium nipponense regulated by eyestalk ablation

Abstract

The eyestalk of crustaceans, such as Macrobrachium nipponense, contains many neurosecretory hormones affecting the process of reproduction, molting, metabolism of glucose, and other functions. In this study, important metabolic pathways and candidate genes involved in male sexual development were selected from M. nipponense. The methodology involved performing long-read and next generation transcriptome sequencing of genes from the androgenic gland after eyestalk ablation. qPCR analysis revealed that the mRNA expression of Mn-IAG was significantly increased after ablation of both the single-side (SS) and double-side (DS) eyestalk, compared with the control group (CG). The long-read transcriptome generated 49,840 non-redundant transcripts. A total of 1319, 2092 and 4351 differentially expressed genes (DEGs) were identified between CG versus SS, SS versus DS and CG versus DS, respectively. These data indicated that ablation of the double-sided eyestalk played stronger regulatory roles than the single-side ablation on male sexual development in M. nipponense. This was consistent with the qPCR analysis. Cell Cycle, Cellular Senescence, Oxidative Phosphorylation, Glycolysis/Gluconeogenesis and Steroid Hormone Biosynthesis were the primary enriched metabolic pathways in all three comparisons, and the important genes from these metabolic pathways were also selected. qPCR permitted secondary confirmation of ten DEGs identified through RNA-seq. RNAi-mediated silencing analyses of Hydroxysteroid dehydrogenase like 1 (HSDL1) revealed that HSDL1 has a positive regulatory effect on testes development. This study provides valuable insight into male sexual development in M. nipponense, including metabolic pathways and genes, paving the way for advanced studies on male sexual development in this species and in other crustaceans.

Identification of Important Genes Involved in the Sex-Differentiation Mechanism of Oriental River Prawn, Macrobrachium nipponense, During the Gonad Differentiation and Development Period

“…The genes participated in the mechanism of gonad differentiation and development and were predicted to be found in cell, cell part, cellular process, binding, and organelle in the GO assignment and the functional groups of general function prediction only, signal transduction mechanisms, posttranslational modification, protein turnover, and chaperones in the COG classification ( Figure 3 ). This finding is consistent with previous studies ( Jin et al, 2013 ; Jin B et al, 2021 ). A total of 482, 98, 240, and 208 DEGs were identified between PL5 vs. PL10, PL10 vs. PL15, PL15 vs. PL25F, and PL15 vs. PL25M, respectively.…”

“…The gonad was identified to be differentiated and developed from PL10 to PL25. Many previous studies reported that immune-related and energy-related metabolic pathways played essential roles in the gonad development in M. nipponense ( Jin et al, 2020 ; Jin B et al, 2021 ), which is consistent with this study. In addition, a total of 29 genes were considered as the strong candidate genes, involved in the mechanism of gonad differentiation and development of M. nipponense , based on gene annotation and differentially expressed pattern.…”

“…The transcriptome profiling analysis was performed on an Illumina High-seq 2500 sequencing platform. Previously published studies have described well the detailed procedures for RNA-Seq and analysis ( Jin et al, 2013 ; Jin B et al, 2021 ). The Trinity program (version: trinityrnaseq_r20131110) was used to assemble the clean data into nonredundant (Nr) transcripts ( Grabherr et al, 2011 ).…”

Effects of Luteinizing Hormone Releasing Hormone A2 on Gonad Development in Juvenile Amur Sturgeon, Acipenser schrenckii, Revealed by Transcriptome Profiling Analysis

“…The Illumina High-seq 2500 sequencing platform, which is widely used in transcriptome studies, was used to perform transcriptome profiling analysis. Previous studies have described the detailed procedures for RNA-seq and analysis ( Jin et al, 2013 ; Jin et al, 2021 ). The Trinity program (version: trinityrnaseq_r20131110) was used to assemble the clean data into non-redundant transcripts ( Grabherr et al, 2011 ).…”

Identification and Expression Analysis of Dsx and Its Positive Transcriptional Regulation of IAG in Black Tiger Shrimp (Penaeus monodon)

“…Since IAG was first identified in a decapod (the red-clawed crayfish Cherax quadricarinatus ) in 2007, it has also been found in many crustacean species such as Macrobrachium rosenbergii [ 3 ]; Macrobrachium nipponensis [ 4 , 5 ]; Penaeus monodon [ 6 ]; Fenneropenaeus chinensis [ 7 ], and Portunus pelagicus [ 8 ]. The mechanism by which IAG mediates male sexual differentiation and gonad development has been demonstrated in crustaceans [ 9 , 10 ]. For example, silencing of IAG expression with siRNA [ 11 ] and dsRNA [ 12 ] in M. rosenbergii causes functional sex reversal and stagnation of testicular spermatogenesis.…”