Dataset Information

Molecular Characterization and Expression Profiling of Three Transformer-2 Splice Isoforms in the Redclaw Crayfish, Cherax quadricarinatus.

ABSTRACT: Sex determination/sex differentiation is determined by genetics, environmental factors, or the interactions of the two. The Transformer-2 (Tra-2) gene plays an important role in the sex determination cascade signal pathway in insects. In this study, the Tra-2 gene was isolated and characterized from the cDNA library of gonad tissues in the redclaw crayfish, Cherax quadricarinatus. Three splice variants were identified, designated as CqTra-2A, CqTra-2B, and CqTra-2C, and sequence analysis showed that they had a highly conserved RRM domain. Phylogenetic analysis was performed by the NJ method, and the results revealed that the Tra-2 protein of the redclaw crayfish was very closely related to those of Macrobrachium rosenbergii, Fenneropenaeus chinensis, and Macrobrachium nipponense. Real-time PCR analysis showed that the three isoforms were predominantly expressed in the ovary and gradually increased with embryonic development. Additionally, the expression pattern of CqTra-2 at different developmental stages was analyzed by qPCR and revealed that the phase of having a body length of 3 cm may be the key period for the sex differentiation of C. quadricarinatus. RNAi-targeting gene silencing further confirmed the function of CqTra-2 in sexual differentiation in redclaw crayfish. Our experimental data will contribute to understanding the mechanism of sex determination in crustaceans.

SUBMITTER: Cai L

PROVIDER: S-EPMC7363937 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Publications

Molecular Characterization and Expression Profiling of Three Transformer-2 Splice Isoforms in the Redclaw Crayfish, Cherax quadricarinatus.

Cai Lina L Zheng Jianbo J Jia Yongyi Y Gu Zhimin Z Liu Shili S Chi Meili M Cheng Shun S

Frontiers in physiology 20200709

Sex determination/sex differentiation is determined by genetics, environmental factors, or the interactions of the two. The Transformer-2 (Tra-2) gene plays an important role in the sex determination cascade signal pathway in insects. In this study, the Tra-2 gene was isolated and characterized from the cDNA library of gonad tissues in the redclaw crayfish, Cherax quadricarinatus. Three splice variants were identified, designated as CqTra-2A, CqTra-2B, and C ...[more]

PMID: 32733260

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Project description:Systemic acid-base balance and osmotic/ionic regulation in decapod crustaceans are in part maintained by a set of transport-related enzymes such as carbonic anhydrase (CA), Na+/K+-ATPase (NKA), H+-ATPase (HAT), Na+/K+/2Cl- cotransporter (NKCC), Na+/Cl-/HCO[Formula: see text] cotransporter (NBC), Na+/H+ exchanger (NHE), Arginine kinase (AK), Sarcoplasmic Ca+2-ATPase (SERCA) and Calreticulin (CRT). We carried out a comparative molecular analysis of these genes in three commercially important yet eco-physiologically distinct freshwater crayfish, Cherax quadricarinatus, C. destructor and C. cainii, with the aim to identify mutations in these genes and determine if observed patterns of mutations were consistent with the action of natural selection. We also conducted a tissue-specific expression analysis of these genes across seven different organs, including gills, hepatopancreas, heart, kidney, liver, nerve and testes using NGS transcriptome data. The molecular analysis of the candidate genes revealed a high level of sequence conservation across the three Cherax sp. Hyphy analysis revealed that all candidate genes showed patterns of molecular variation consistent with neutral evolution. The tissue-specific expression analysis showed that 46% of candidate genes were expressed in all tissue types examined, while approximately 10% of candidate genes were only expressed in a single tissue type. The largest number of genes was observed in nerve (84%) and gills (78%) and the lowest in testes (66%). The tissue-specific expression analysis also revealed that most of the master genes regulating pH and osmoregulation (CA, NKA, HAT, NKCC, NBC, NHE) were expressed in all tissue types indicating an important physiological role for these genes outside of osmoregulation in other tissue types. The high level of sequence conservation observed in the candidate genes may be explained by the important role of these genes as well as potentially having a number of other basic physiological functions in different tissue types.

Project description:BackgroundThe Australian redclaw crayfish (Cherax quadricarinatus, von Martens), is native to Australasia, but has been widely translocated around the world due to aquaculture and aquarium trade. Mostly as a result of escape from aquaculture facilities, this species has established extralimital populations in Australia and alien populations in Europe, Asia, Central America and Africa. In South Africa, C. quadricarinatus was first sampled from the wild in 2002 in the Komati River, following its escape from an aquaculture facility in Swaziland, but data on the current status of its populations are not available.MethodsTo establish a better understanding of its distribution, rate of spread and population status, we surveyed a total of 46 sites in various river systems in South Africa and Swaziland. Surveys were performed between September 2015 and August 2016 and involved visual observations and the use of collapsible crayfish traps.ResultsCherax quadricarinatus is now present in the Komati, Lomati, Mbuluzi, Mlawula and Usutu rivers, and it was also detected in several off-channel irrigation impoundments. Where present, it was generally abundant, with populations having multiple size cohorts and containing ovigerous females. In the Komati River, it has spread more than 112 km downstream of the initial introduction point and 33 km upstream of a tributary, resulting in a mean spread rate of 8 km year-1 downstream and 4.7 km year-1 upstream. In Swaziland, estimated downstream spread rate might reach 14.6 km year-1. Individuals were generally larger and heavier closer to the introduction site, which might be linked to juvenile dispersal.DiscussionThese findings demonstrate that C. quadricarinatus is established in South Africa and Swaziland and that the species has spread, not only within the river where it was first introduced, but also between rivers. Considering the strong impacts that alien crayfish usually have on invaded ecosystems, assessments of its potential impacts on native freshwater biota and an evaluation of possible control measures are, therefore, urgent requirements.

Dataset Information

Molecular Characterization and Expression Profiling of Three Transformer-2 Splice Isoforms in the Redclaw Crayfish, Cherax quadricarinatus.

Publications

Molecular Characterization and Expression Profiling of Three <i>Transformer-2</i> Splice Isoforms in the Redclaw Crayfish, <i>Cherax quadricarinatus</i>.

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