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.
Highlights
The eyestalk of crustaceans, such as Macrobrachium nipponense, contains many neurosecretory hormones affecting the process of reproduction, molting, metabolism of glucose, and other functions
A series of transcriptomes from the androgenic gland have been constructed in M. nipponense[16,17,18], and a series of important genes from the androgenic gland have been implicated as having an essential role in male sexual development[19,20,21,22]
The mRNA expression of Mn-insulin-like androgenic gland hormone (IAG) was measured in three groups: control group (CG), single-side ablation of eyestalk (SS), and double-side ablation of eyestalk (DS) (Fig. 1)
Summary
The eyestalk of crustaceans, such as Macrobrachium nipponense, contains many neurosecretory hormones affecting the process of reproduction, molting, metabolism of glucose, and other functions. 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. IAG was shown to promote male sexual differentiation and development in many crustacean s pecies[8,9,10] It is the most important male sex-related gene acknowledged in many crustacean species. The X-organ–SG complex (XO–SG) was identified as a principal neuroendocrine gland located in the eyestalk[23] It stores and releases the crustacean hyperglycemic hormone (CHH) superfamily of neurohormones, including CHH, ion transport peptides (ITP), gonad-inhibiting hormone (GIH), molt inhibiting hormone (MIH) and mandibular organ-inhibiting hormone (MOIH), playing essential roles in r eproduction24–26, molting[27,28,29], metabolism of g lucose[30,31] and other function[32,33,34]. CHH has been shown to promote testes development in M. nipponense[37]
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