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

2021 ◽  
Vol 12 ◽  
Author(s):  
Shubo Jin ◽  
Yuning Hu ◽  
Hongtuo Fu ◽  
Sufei Jiang ◽  
Yiwei Xiong ◽  
...  
Keyword(s):  
Sexual Development ◽  
Pyruvate Dehydrogenase ◽  
Quantitative Polymerase Chain Reaction ◽  
Cell Types ◽  
Macrobrachium Nipponense ◽  
Reading Frame ◽  
Testis Development ◽  
Positive Effects ◽  
Oriental River Prawn

Pyruvate dehydrogenase E1 (PDHE1) 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, Macrobrachium nipponense. In this study, we used rapid amplification of cDNA ends, quantitative polymerase chain reaction (qPCR), in situ hybridization, western blotting, RNA interference (RNAi), and histological analyses to assess the potential functions of Mn-PDHE1 in the sexual development of male M. nipponense. The full cDNA sequence of Mn-PDHE1 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 PDHE1 in male sexual development in M. nipponense and in the metamorphosis process. In situ hybridization and western blot results indicated that PDHE1 was involved in testis development, and RNAi analysis showed that PDHE1 positively regulated the expression of insulin-like androgenic gland factor in M. nipponense. 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 Mn-PDHE1 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 PDHE1 delayed testis development. Thus, PDHE1 has positive effects on male sexual development in M. nipponense. This study highlights the functions of PDHE1 in M. nipponense and its essential roles in the regulation of testis development.

scholarly journals Identification and Characterization of the Succinate Dehydrogenase Complex Iron Sulfur Subunit B Gene in the Oriental River Prawn, Macrobrachium nipponense

2021 ◽  
Vol 12 ◽  
Author(s):  
Shubo Jin ◽  
Yuning Hu ◽  
Hongtuo Fu ◽  
Sufei Jiang ◽  
Yiwei Xiong ◽  
...  
Keyword(s):  
Western Blot ◽  
Sexual Development ◽  
Developmental Stages ◽  
Quantitative Polymerase Chain Reaction ◽  
Full Genome Sequence ◽  
Macrobrachium Nipponense ◽  
Testis Development ◽  
Positive Effects ◽  
Dsrna Injection

Previous studies have revealed that SDHB has potential functions in the male sexual differentiation and development in M. nipponense through providing ATP. In this study, the functions of Mn-SDHB were further analyzed 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-SDHB was 54,608 bp at Chromosome 34, including 7 introns and 6 exons. The full-length cDNA sequence of Mn-SDHB was 1,268 base pairs (bp) long with an open reading frame of 807 bp, encoding for 268 amino acids. The highest expression level of Mn-SDHB in different tissues was observed in the testis, and male prawns at post-larval developmental stage 25 during different developmental stages, indicating that SDHB was potentially involved in the male sexual development in M. nipponense. In situ hybridization and western-blot analysis indicated that SDHB plays essential roles in the testis development. The in situ hybridization analysis also implies the potential roles of Mn-SDHB in ovarian development. The expressions of Mn-IAG were decreased after Mn-SDHB dsRNA injection, indicating SDHB has the positive regulatory effects on IAG in M. nipponese. Thus, SDHB was involved in the mechanism of the male sexual development. The testis development was inhibited, and sperms were rarely observed after 10 days of Mn-SDHB dsRNA injection, indicating SDHB has positive effects on the male sexual development in M. nipponense. This study highlights the functions of SDHB in M. nipponense, which provide new insights for the future studies of the male sexual development in other crustacean species.

scholarly journals Identification and Characterization of The Pyruvate Dehydrogenase E1 Gene in The Oriental River Prawn, Macrobrachium Nipponense

2021 ◽  
Author(s):  
Shubo Jin ◽  
Yuning Hu ◽  
Hongtuo Fu ◽  
Sufei Jiang ◽  
Yiwei Xiong ◽  
...  
Keyword(s):  
Western Blot ◽  
Sexual Development ◽  
Pyruvate Dehydrogenase ◽  
Quantitative Polymerase Chain Reaction ◽  
Full Genome Sequence ◽  
Control Group ◽  
Chromosome 11 ◽  
Additional Function ◽  
Positive Effects

Abstract 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.

scholarly journals Potential Functions of Gem-Associated Protein 2-Like Isoform X1 in the Oriental River Prawn Macrobrachium nipponense: Cloning, qPCR, In Situ Hybridization, and RNAi Analysis

2019 ◽  
Vol 20 (16) ◽  
pp. 3995 ◽  
Author(s):  
Shubo Jin ◽  
Yuning Hu ◽  
Hongtuo Fu ◽  
Sufei Jiang ◽  
Yiwei Xiong ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Sexual Development ◽  
Developmental Stages ◽  
Quantitative Polymerase Chain Reaction ◽  
Negative Effects ◽  
Macrobrachium Nipponense ◽  
Reading Frame ◽  
Gonad Differentiation ◽  
Qpcr Analysis

Gem-associated protein 2-like isoform X1 (GEM) was previously predicted to be involved in the sexual development of male Macrobrachium nipponense. In this study, we analyze the GEM functions in depth using quantitative polymerase chain reaction (qPCR), in situ hybridization, and RNA interference (RNAi). The full-length Mn-GEM cDNA sequence was 1018 base pairs (bp) long with an open reading frame of 777 bp encoding 258 amino acids. qPCR analysis of Mn-GEM in different tissues and developmental stages showed that Mn-GEM was highly expressed in the gonad and from post-larval developmental stage day 5 (PL5) to PL15, which indicated that GEM has potential roles in gonad differentiation and development in M. nipponense. In situ hybridization and qPCR analysis of various stages of the reproductive cycle of the testis and ovary indicated that GEM may promote spermatid development and gametogenesis in M. nipponense. After injecting with double-stranded RNA (dsRNA) of Mn-GEM, mRNA expression of Mn-insulin-like androgenic gland hormone (Mn-IAG) and the content of testosterone increased with the decrease of Mn-GEM expression, indicating that GEM has negative effects on the male sexual differentiation and development in M. nipponense. Results of this study highlight the functions of GEM in M. nipponense, which can be applied to future studies of male sexual development in M. nipponense and other crustacean species.

scholarly journals Investigation of novel functions of three genes in oriental river prawn,Macrobrachium nipponense: Molecular Cloning, Expression, andIn situHybridization Analysis

2018 ◽  
Author(s):  
Shubo Jin ◽  
Hongtuo Fu ◽  
Yuning Hu ◽  
Shengming Sun ◽  
Sufei Jiang ◽  
...  
Keyword(s):  
Amino Acids ◽  
Sexual Development ◽  
Cdna Sequence ◽  
The Novel ◽  
Androgenic Gland ◽  
Macrobrachium Nipponense ◽  
Testis Development ◽  
Gland Cells ◽  
Oriental River Prawn

AbstractThree genes were predicted to be potentially involved in the male sexual development inM.nipponense, including the Gem-associated protein 2-like isoform X1 (GEM), Ferritin peptide, and DNA polymerase zeta catalytic subunit (Rev3). In this study, we aimed to investigate their novel functions in depth. The full-length cDNA sequence of Mn-GEM was 1,018 bp, encoding 258 amino acids. The partial Mn-Rev3 cDNA sequence was 6,832 bp, encoding 1,203 amino acids. Tissue distribution indicated that all of these three genes have higher expression level in testis and androgenic gland, implying their novel functions in male sexual development. In situ hybridization analysis further confirmed the novel roles of these three genes. Rev3 promote the testis development during the whole reproductive cycle, while GEM and ferritin only promote the activation of testis development. Besides, these three genes play essential roles in funicular structure development surrounding the androgenic gland cells, which promote and support the formation of androgenic gland cells. The expression in hepatopancreas cells also suggested their role in immune system inM. nipponense.This study advances our understanding of male sexual development inM. nipponense, as well as providing the basis for further studies of male sexual differentiation and development in crustaceans.

scholarly journals Molecular Characterization of a Novel Cathepsin L in Macrobrachium nipponense and Its Function in Ovary Maturation

2022 ◽  
Vol 12 ◽  
Author(s):  
Sufei Jiang ◽  
Yiwei Xiong ◽  
Wenyi Zhang ◽  
Junpeng Zhu ◽  
Dan Cheng ◽  
...  
Keyword(s):  
Cathepsin L ◽  
Cysteine Proteases ◽  
Macrobrachium Nipponense ◽  
Reading Frame ◽  
Physiological Processes ◽  
Gonad Somatic Index ◽  
Oriental River Prawn ◽  
Ovary Maturation

Cathepsin L genes, which belonged to cysteine proteases, were a series of multifunctional protease and played important roles in a lot of pathological and physiological processes. In this study, we analyzed the characteristics a cathepsin L (named Mn-CL2) in the female oriental river prawn, Macrobrachium nipponense which was involved in ovary maturation. The Mn-CL2 was1,582 bp in length, including a 978 bp open reading frame that encoded 326 amino acids. The Mn-CL2 was classified into the cathepsin L group by phylogenetic analysis. Real-time PCR (qPCR) analysis indicated that Mn-CL2 was highly expressed in the hepatopancreas and ovaries of female prawns. During the different ovarian stages, Mn-CL2 expression in the hepatopancreas and ovaries peaked before ovarian maturation. In situ hybridization studies revealed that Mn-CL2 was localized in the oocyte of the ovary. Injection of Mn-CL2 dsRNA significantly reduced the expression of vitellogenin. Changes in the gonad somatic index also confirmed the inhibitory effects of Mn-CL2 dsRNA on ovary maturation. These results suggest that Mn-CL2 has a key role in promoting ovary maturation.

scholarly journals Identification and Characterization of Four Autophagy-Related Genes That Are Expressed in Response to Hypoxia in the Brain of the Oriental River Prawn (Macrobrachium nipponense)

2019 ◽  
Vol 20 (8) ◽  
pp. 1856 ◽  
Author(s):  
Shengming Sun ◽  
Ying Wu ◽  
Hongtuo Fu ◽  
Xianping Ge ◽  
Hongzheng You ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Open Reading Frames ◽  
Dependent Manner ◽  
Macrobrachium Nipponense ◽  
Oriental River Prawn ◽  
Adverse Environmental Conditions ◽  
The Brain ◽  
Identification And Characterization

Autophagy is a cytoprotective mechanism triggered in response to adverse environmental conditions. Herein, we investigated the autophagy process in the oriental river prawn (Macrobrachium nipponense) following hypoxia. Full-length cDNAs encoding autophagy-related genes (ATGs) ATG3, ATG4B, ATG5, and ATG9A were cloned, and transcription following hypoxia was explored in different tissues and developmental stages. The ATG3, ATG4B, ATG5, and ATG9A cDNAs include open reading frames encoding proteins of 319, 264, 268, and 828 amino acids, respectively. The four M. nipponense proteins clustered separately from vertebrate homologs in phylogenetic analysis. All four mRNAs were expressed in various tissues, with highest levels in brain and hepatopancreas. Hypoxia up-regulated all four mRNAs in a time-dependent manner. Thus, these genes may contribute to autophagy-based responses against hypoxia in M. nipponense. Biochemical analysis revealed that hypoxia stimulated anaerobic metabolism in the brain tissue. Furthermore, in situ hybridization experiments revealed that ATG4B was mainly expressed in the secretory and astrocyte cells of the brain. Silencing of ATG4B down-regulated ATG8 and decreased cell viability in juvenile prawn brains following hypoxia. Thus, autophagy is an adaptive response protecting against hypoxia in M. nipponense and possibly other crustaceans. Recombinant MnATG4B could interact with recombinant MnATG8, but the GST protein could not bind to MnATG8. These findings provide us with a better understanding of the fundamental mechanisms of autophagy in prawns.

scholarly journals MnFtz-f1 Is Required for Molting and Ovulation of the Oriental River Prawn Macrobrachium nipponense

2021 ◽  
Vol 12 ◽  
Author(s):  
Huwei Yuan ◽  
Wenyi Zhang ◽  
Yin Fu ◽  
Sufei Jiang ◽  
Yiwei Xiong ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Significant Inhibition ◽  
Pcr Analysis ◽  
Base Pairs ◽  
Regulatory Relationship ◽  
Macrobrachium Nipponense ◽  
Reading Frame ◽  
Starting Point ◽  
Oriental River Prawn

Molting and ovulation are the basic processes responsible for the growth and reproduction of Macrobrachium nipponense; however, the molecular mechanisms of molting and ovulation in M. nipponense are poorly understood. The present study aimed to use MnFtz-f1 as the starting point to study the molting and ovulation phenomena in M. nipponense at the molecular level. The full-length MnFtz-f1 cDNA sequence was 2,198 base pairs (bp) in length with an open reading frame of 1,899 bp encoding 632 amino acids. Quantitative real-time PCR analysis showed that MnFtz-f1 was highly expressed in the ovary at the cleavage stage and on the fifth day after hatching. In vivo administration of 20-hydroxyecdysone (20E) showed that 20E effectively inhibited the expression of the MnFtz-f1 gene, and the silencing of the MnFtz-f1 gene reduced the content of 20E in the ovary. In situ hybridization (ISH) analysis revealed the localization of MnFtz-f1 in the ovary. Silencing of MnFtz-f1 by RNA interference (RNAi) resulted in significant inhibition of the expression of the vitellogenin (Vg), Spook, and Phantom genes, thus confirming that MnFtz-f1 had a mutual regulatory relationship with Vg, Spook, and Phantom. After RNAi, the molting frequency and ovulation number of M. nipponense decreased significantly, which demonstrated that MnFtz-f1 played a pivotal role in the process of molting and ovulation.

scholarly journals Parainfluenza Virus 5 Infection in Neurological Disease and Encephalitis of Cattle

2020 ◽  
Vol 21 (2) ◽  
pp. 498 ◽  
Author(s):  
Melanie M. Hierweger ◽  
Simea Werder ◽  
Torsten Seuberlich
Keyword(s):  
Neurological Disease ◽  
Cell Types ◽  
High Viral Load ◽  
Brain Cell ◽  
Parainfluenza Virus ◽  
Reading Frame ◽  
In Situ Techniques ◽  
Parainfluenza Virus 5

The etiology of viral encephalitis in cattle often remains unresolved, posing a potential risk for animal and human health. In metagenomics studies of cattle with bovine non-suppurative encephalitis, parainfluenza virus 5 (PIV5) was identified in three brain samples. Interestingly, in two of these animals, bovine herpesvirus 6 and bovine astrovirus CH13 were additionally found. We investigated the role of PIV5 in bovine non-suppurative encephalitis and further characterized the three cases. With traditional sequencing methods, we completed the three PIV5 genomes, which were compared to one another. However, in comparison to already described PIV5 strains, unique features were revealed, like an 81 nucleotide longer open reading frame encoding the small hydrophobic (SH) protein. With in situ techniques, we demonstrated PIV5 antigen and RNA in one animal and found a broad cell tropism of PIV5 in the brain. Comparative quantitative analyses revealed a high viral load of PIV5 in the in situ positive animal and therefore, we propose that PIV5 was probably the cause of the disease. With this study, we clearly show that PIV5 is capable of naturally infecting different brain cell types in cattle in vivo and therefore it is a probable cause of encephalitis and neurological disease in cattle.

scholarly journals Transcriptome Profiling Analysis of the Testis After Eyestalk Ablation for Selection of the Candidate Genes Involved in the Male Sexual Development in Macrobrachium nipponense

2021 ◽  
Vol 12 ◽  
Author(s):  
Shubo Jin ◽  
Yin Fu ◽  
Yuning Hu ◽  
Hongtuo Fu ◽  
Sufei Jiang ◽  
...  
Keyword(s):  
Candidate Genes ◽  
Sexual Development ◽  
Metabolic Pathways ◽  
Transcriptome Profiling ◽  
Negative Effects ◽  
Macrobrachium Nipponense ◽  
Eyestalk Ablation ◽  
Testis Development ◽  
Single Side ◽  
Profiling Analysis

The eyestalk of crustacean species secretes many hormones, affecting the process of reproduction, molting, metabolism of glucose, and other functions in crustaceans. In this study, important metabolic pathways and candidate genes involved in the male sexual development were identified through performing the transcriptome profiling analysis of the testis after the ablation of eyestalk from Macrobrachium nipponense. The histological observations revealed that the testis development became vigorous after eyestalk ablation, indicating that the hormones secreted by the eyestalk have negative effects on the testis development in M. nipponense. Transcriptome profiling analysis revealed that 1,039, 1,226, and 3,682 differentially expressed genes (DEGs) were identified between normal prawns (CG) vs single-side eyestalk ablation prawns (SS), SS vs double-side eyestalk ablation prawns (DS), and CG vs DS, respectively, indicating that the ablation of double-side eyestalk has more significant regulatory roles on male sexual development than that of single-side ablation, which was consistent with the histological observations. Lysosome, Apoptosis, Glycolysis/Gluconeogenesis, and Insulin signaling pathway were the main enriched metabolic pathways in all of these three comparisons, and the important genes from these metabolic pathways were also selected. The qPCR verifications of 10 DEGs from these metabolic pathways were the same as those of RNA-seq. The qPCR, in situ hybridization, and RNA interference analysis of Mn-NFkBα revealed that NFkBα has a positive regulatory effect on testis development. This study provided new insights on male sexual development in M. nipponense, promoting the studies on male sexual development in other crustaceans as well.

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