Opsariichthys Bidens Research Articles

Three genome assemblies of Opsariichthys bidens from Yangzte River, Pearl River and Qiantang River basins

Opsariichthys bidens is an endemic minnow, mainly distributed in China as an emerging aquaculture species. In this study, O. bidens collected from Yangzte River (YR), Pearl River (PR) and Qiantang River (QR) basins were used for karyotypic study and genome sequencing. Using PacBio long reads and Hi-C data, we assembled high-quality O. bidens genomes of 852.41 Mb (YR), 843.11 Mb (PR) and 840.94 Mb (QR) with scaffold N50 lengths of 21.01 Mb, 23.62 Mb and 24.75 Mb, respectively, of which 90.39%, 95.67% and 99.01% were anchored to 37, 38 and 39 chromosomes, respectively. 26,556 (YR), 25,036 (PR) and 26,283 (QR) protein-coding genes were predicted, respectively. The karyotype of O. bidens from YR, PR and QR were 2 N = 74 (6 m + 6sm + 4st + 58t), 2 N = 76 (4 m + 6sm + 4st + 62t) and 2 N = 78 (4 m + 4sm + 4st + 66t), respectively. Collinearity analysis and telomere predictions indicated that the observed chromosomal evolution was driven by Robertsonian translocation. These genome assemblies facilitate cryptic species determination, evolutionary study and genetic breeding of genus Opsariichthys.

Mixture toxic mechanism of phoxim and prochloraz in the hook snout carp Opsariichthysbidens

Pesticides are usually found as mixtures in surface water bodies, even though their regulation in aquatic ecosystems is usually approached individually. In this context, this work aimed to investigate the enzymatic- and transcriptional-level responses after the mixture exposure of phoxim (PHX) and prochloraz (PRC) in the livers of hook snout carp Opsariichthys bidens. These data exhibited that co-exposure to PHX and PRC induced an acute synergistic impact on O. bidens. The activities of catalase (CAT), superoxide dismutase (SOD), carboxylesterase (CarE), and caspase3 varied significantly in most of the individual and combined challenges relative to basal values, indicating the activation of oxidative stress, detoxification dysfunction, as well as cell apoptosis. Besides, the transcriptional levels of five genes (gst, erα, mn-sod, cxcl-c1c, and il-8) exhibited more pronounced changes when subjected to combined pesticide exposure in contrast to the corresponding individual compounds. The findings revealed the manifestation of endocrine dysfunction and immune disruption. These results underscored the potential biochemical and molecular toxicity posed by the combination of PHX and PRC to O. bidens, thereby contributing to a deeper comprehension of the ecological toxicity of pesticide mixtures on aquatic organisms. Importantly, the concurrent presence of PHX and PRC might exacerbate hepatocellular damage in hook snout carps, potentially attributable to their synergistic toxic interactions. This study underscored the toxicological potency inherent in the co-occurrence of PHX and PRC in influencing fish development, thereby offering valuable insights for the risk assessment of pesticide mixtures and the safeguarding of aquatic organisms.

Effects of Environmental Enrichment on the Growth, Gonadal Development, and Welfare of the Chinese Hooksnout Carp (Opsariichthys bidens)

To investigate the effects of environmental enrichment on the growth and welfare level of the Chinese hooksnout carp (Opsariichthys bidens), an experiment was carried out for 56 d with four types of enrichment environments: low-density physical enrichment, high-density physical enrichment, low-density social enrichment, and high-density social enrichment. The experimental results revealed that both physical and social enrichment treatments positively affected the growth and gonadal development of O. bidens. The physical enrichment treatment was found to increase the antioxidants’ content, such as CAT, SOD, and GSH, in the liver of O. bidens. Moreover, this treatment was able to lead to a reduction in MDA content and enhancement of the fish’s antioxidant capabilities. Behavioral observations, cortisol level analysis, and pcna gene expression analysis indicated that the physical enrichment treatment redused the clustering, aggression, and stress behaviors of O. bidens, maintaining them in a low-stress state. In most respects, the high-density physical enrichment has more positive effects than the low-density one. In contrast, the social enrichment treatments resulted in higher stress levels for O. bidens.

A Comparative Study on the Muscle and Gut Microbiota of Opsariichthys bidens from Rice Field and Pond Culture Breeding Modes.

To investigate difference in the quality of the different parts (back, tail muscles, and fish skin) of Opsariichthys bidens from pond and rice field cultures, a comparative study was conducted in terms of nutritional composition, volatile flavor profiles and gut microbiota. In detail, the texture, free amino acids, fatty acids were further assessed. The results suggested that the moisture content, crude protein and crude fat content in the skin of O. bidens are higher than those in the back and tail muscles, regardless of breeding modes. The fish cultured in the rice field had a higher protein content than those from the pond culture, while the fat content of the rice field-cultured fish was significantly low compared to the fish from the pond culture, especially in the back and tail parts. A total of 43 volatile components were detected by Gas Chromatography-Mass Spectrometry (GC-MS), with a maximum of 18 types of aldehydes and the highest concentration being nonanal. Compared to pond cultures, the fish from the rice field cultures showed more abundant flavor composition and odor-active compounds. The total content of DHA (Docosahexaenoic Acid) and EPA (Eicosapentaenoic Acid) in the rice field-cultured fish was higher than that of the pond group, while no significant disparity in amino acid composition was observed (p > 0.05). Comparative and clustering analyses of gut microbiota revealed notable discrepancies in the gut microbiota of O. bidens from two aquaculture systems. However, an inherent correlation between the gut microbiome and meat quality would be further emphasized in further studies. This study can offer a theoretical reference for the development of high-quality aquatic products by selecting the appropriate aquaculture models.

Synergistic toxic effects and mechanistic insights of beta-cypermethrin and pyraclostrobin exposure on hook snout carp (Opsariichthys bidens): A biochemical, transcriptional, and molecular approach

The extensive utilization of pesticides results in their frequent detection in aquatic environments, often as complex mixtures, posing risks to aquatic organisms. The hook snout carp (Opsariichthys bidens) serves as a valuable bioindicator for evaluating the impacts of environmental pollutants in aquatic ecosystems. However, few studies examined the toxic effects of pesticides on O.bidens, let alone the characterization of the combined effects resulting from their mixtures. This study aims to elucidate the toxic effects of beta-cypermethrin and pyraclostrobin on O.bidens, individually and in combination, focusing on biochemical, transcriptional, and molecular responses. By organizing and analyzing the toxicogenomic databases, both pesticides were identified as a contributor to processes such as apoptosis, oxidative stress, and inflammatory responses. The acute toxicity test revealed comparable acute toxicity of beta-cypermethrin and pyraclostrobin on O.bidens, with LC50 being 0.019 and 0.027 mg/L, respectively, whereas the LC50 decreased to 0.0057 and 0.0079 mg/L under the combined exposure, indicating potential synergistic effects. The activities of enzymes involved in oxidative stress and detoxification were significantly altered after exposure, with superoxide dismutase (SOD) and catalase (CAT) increasing, while malondialdehyde (MDA) levels decreased. The activity of CYP450s was significantly changed. Likewise, the expression levels of genes (mn-sod, p53, esr, il-8) associated with oxidative stress, apoptosis, endocrine and immune systems were significantly increased. Combined exposure to the pesticides significantly exacerbated the aforementioned biological processes in O.bidens. Furthermore, both pesticides can modify protein activity by binding to the surface of SOD molecules and altering protein conformation, contributing to the elevated enzyme activity. Through the investigation of the synergistic toxic effects of pesticides and molecular mechanisms in O.bidens, our findings highlight the importance of assessing the combined effects of pesticide mixtures in aquatic environments.

Comprehensive multi-omics investigation of sub-chronic toxicity induced by Cadmium and Triazophos Co-exposure in hook snout carps (Opsariichthys bidens)

The coexistence of heavy metals and pesticides poses a critical challenge in agricultural ecosystems. Traditional toxicity assessments often focus only on the individual impacts of either pesticides or heavy metals. Here, the untargeted metabolomics and 16 S rRNA sequencing were used to assess the individual and combined effects of cadmium (Cd) and triazophos (TRI) on hook snout carps (Opsariichthys bidens). Cd caused much more serious impacts on hepatic metabolism and gut microbiota than those in TRI. Combined Cd and TRI exposure synergistically affected hepatic metabolism, causing mitochondrial dysfunction and even oxidative damage. Simultaneously, 16 S rRNA sequencing highlighted significant variations in the composition and abundance of gut microbiota. A noteworthy connection emerged between these distinct microbiota profiles and disruptions in energy metabolism, ultimately leading to disorders in metabolites. These findings enhanced the understanding of risks posed by heavy metals and pesticides, providing insights for better environmental risk assessments of aquatic organisms.

Co-exposure to cadmium and triazophos induces variations at enzymatic and transcriptional levels in Opsariichthys bidens

Heavy metals and pesticides are significant pollutants in aquatic environments, often leading to combined pollution and exerting toxic effects on aquatic organisms. With the rapid growth of modern industry and agriculture, heavy metal cadmium (Cd) and pesticide triazophos (TRI) are frequently detected together in various water bodies, particularly in agricultural watersheds. However, the combined toxic mechanisms of these pollutants on fish remain poorly understood. This experiment involved a 21-day co-exposure of Cd and TRI to the hook snout carp Opsariichthys bidens to investigate the toxic effects on liver tissues at both enzymatic and transcriptional levels. Biochemical analysis revealed that both individual and combined exposures significantly increased the content or activity of caspase-3 (CASP-3) and malondialdehyde (MDA). Moreover, the impact on these parameters was greater in the combined exposure groups compared to the corresponding individual exposure groups. These findings suggested that both individual and combined exposures could induce mitochondrial dysfunction and lipid peroxidation damage, with combined exposure exacerbating the toxicological effects of each individual pollutant. Furthermore, at the molecular level, both individual and combined exposures upregulated the expression levels of cu-sod, cat, and erβ, while downregulating the expression of il-1. Similar to the patterns observed in the biochemical parameters, the combined exposure group exhibited a greater impact on the expression of these genes compared to the individual exposure groups. These results indicated that exposure to Cd, TRI, and their combination induced oxidative stress, endocrine disruption, and immunosuppression in fish livers, with more severe effects observed in the combined exposure group. Overall, the interaction between Cd and TRI appeared to be synergistic, shedding light on the toxic mechanisms by which fish livers responded to these pollutants. These findings contributed to the understanding of mixture risk assessment of pollutants and were valuable for the conservation of aquatic resources.

Dazl and dnd Identify Both Embryonic and Gonadal Germ Cells in Chinese Hook Snout Carp (Opsariichthys bidens)

In recent years, the Chinese hook snout carp (Opsariichthys bidens) is becoming increasingly popular due to its high nutritional value and delicious taste. However, anthropogenic influences have caused a rapid reduction in its population. Considering long-term development sustainability, it is essential to conduct research on its reproductive development. In this study, the germ-cell related genes dazl (Obdazl) and dnd (Obdnd) were cloned, and their expression patterns were revealed. RT-PCR analysis indicated that Obdazl and Obdnd were specifically expressed in gonads. In the testis, Obdazl and Obdnd RNAs have a similar expression pattern: their transcripts were abundant in spermatogonia, gradually decreased, and eventually disappeared during spermatogenesis. Early oocytes in the ovary exhibited strong expression of Obdazl and Obdnd RNAs and reduced remarkably after peak expression at III oocytes. Surprisingly, Obdazl RNA was dynamically found in perinuclear spots that finally aggregated into the Balbiani body (Bb) in the early oocytes. Moreover, Obdazl 3′ UTR enabled the reporter gene to be expressed stably in medaka PGCs, suggesting that dazl was identified as the germ cell marker in O. bidens, which may provide perspectives for further studies on the development of PGCs in the future.

Genome-wide analysis reveals the contributors to fast molecular evolution of the Chinese hook snout carp (Opsariichthys bidens)

Variations in molecular evolutionary rate have been widely investigated among lineages and genes. However, it remains an open question whether fast rate of molecular evolution is driven by natural selection or random drift, and how the fast rate is linked to metabolic rate. Additionally, previous studies on fast molecular evolution have been largely restricted to concatenated matrix of genes or a few specifically selected genes, but less is known for individual genes at the genome-wide level. Here we addressed these questions using more than 5000 single-copy orthologous (SCO) genes through comparative genomic and phylogenetic analyses among fishes, with a special focus on a newly-sequenced clupeocephalan fish the Chinese hook snout carp Opsariichthys bidens. We showed O. bidens displays significantly higher mean substitution rate and more fast-evolving SCO genes (2172 genes) than most fishes studied here. The rapidly evolving genes are enriched in highly conserved and very basic functions such as translation and ribosome that are critical for biological fitness. We further revealed that ∼25 % of these fast-evolving genes exhibit a constant increase of substitution rate from the common ancestor down to the present, suggesting a neglected but important contribution from ancestral states. Model fitting showed that ∼85 % of fast-evolving genes exclusive to O. bidens and related species follow the adaptive evolutionary model rather than random-drift model, and 7.6 % of fast-evolving genes identified in O. bidens have experienced positive selection, both indicating the reflection of adaptive selection. Finally, metabolic rate was observed to be linked with substitution rate in a gene-specific manner. Overall, our findings reveal fast molecular evolution of SCO genes at genome-wide level in O. bidens, and uncover the evolutionary and ecological contributors to it.

Parentage Determination of Chinese Hooksnout Carp (Opsariichthys bidens) Based on Microsatellite DNA Markers

In this study, a panel of 14 microsatellite markers was screened to determine the parentage of 144 progenies in 12 Chinese hooksnout carp (Opsariichthys bidens) full-sib families. The combined exclusion probabilities for 14 loci were 97.6% and 99.9%, respectively, when there was no available parent information or only one parent information was provided. Simulation analysis demonstrated that the power of five loci to exclude false parents exceeded 99.0%, while that of eight loci reached 99.9% based on allele frequency data obtained from full-sib families (168 individuals). Moreover, the cumulative assignment success rate reached 100% with known parental and filial information when utilizing 13 or more loci, even in cases where no parent information was available. The results showed that this set of microsatellite markers proved to be a reliable and efficient tool for parentage determination of O. bidens.

Toxic mechanisms of imazalil, azoxystrobin and their mixture to hook snout carp (Opsariichthys bidens)

While combinations of pesticides better represent actual conditions within aquatic ecosystems, the specific toxic effects of these combinations have not been determined yet. The objective of this research was to assess the combined impact of imazalil and azoxystrobin on the hook snout carp (Opsariichthys bidens) and delve into the underlying causes. Our findings indicated that the 4-day LC50 value for imazalil (1.85 mg L−1) was greater than that for azoxystrobin (0.90 mg L−1). When imazalil and azoxystrobin were combined, they presented a heightened effect on the species. Enzyme activities like SOD, CAT, GST, and CarE, along with androgen and estrogen levels, displayed marked differences in most single and combined treatments in comparison to the baseline group. Moreover, four genes (mn-sod, cu-sod, il-1, and esr) related to oxidative stress, immunity, and the endocrine system exhibited more pronounced expression changes when exposed to combined pesticides rather than individual ones. Our tests revealed that the combined use of imazalil and azoxystrobin had more detrimental effect on aquatic vertebrates than when evaluated individually. This finding suggested that future ecological hazard analyses based only on individual tests might not sufficiently safeguard our aquatic ecosystems.

Enzymatic and transcriptional level changes induced by the co-presence of lead and procymidone in hook snout carp (Opsariichthys bidens)

Understanding the interactions between different environmental pollutants is necessary in ecotoxicology since environmental contaminants never appear as single components but rather in combination with other substances. Heavy metals and pesticides are commonly detected in the environment, but the characterization of their mixture toxicity has been inadequately explored. This research aimed to elucidate the mixture impacts of the heavy metal lead (Pb) and the pesticide procymidone (PCM) on the hook snout carp (Opsariichthys bidens) using an array of biomarkers. The data showed that Pb and PCM possessed almost equivalent acute toxicity to the animals, with 4-days LC50 values of 120.9 and 85.15 mg L−1, respectively. Combinations of Pb and PCM generated acute synergistic effects on O. bidens. The contents of malondialdehyde (MDA), antioxidative (SOD), apoptotic (caspase-9), and detoxifying enzymes glutathione S-transferase (GST) and cytochrome P450 (CYP450) significantly changed after most of the mixture exposures compared with the baseline level and the corresponding individual exposures. This suggests the induction of oxidative stress, cell damage, and detoxification dysfunction. The expressions of eight genes (mn-sod, cu-sod, p53, cas3, erβ1, esr, ap, and klf2α) associated with oxidative stress, cell apoptosis, immune response, and hormonal functions exhibited pronounced changes when challenged with the mixture compared to the individual treatments. This indicates the occurrence of immune dysregulation and endocrine disorder. These findings provide an overall understanding of fish upon the challenge of sublethal toxicity between Pb and PCM and can be adopted to evaluate the complicated toxic mechanisms in aquatic vertebrates when exposed to heavy metal and pesticide mixtures. Additionally, these results might guide environmental regulation tactics to protect the population of aquatic vertebrates in natural ecosystems.

Effects of Acute Ammonia Exposure on the Antioxidant System and Anti‐Inflammatory Factors in Opsariichthys bidens

The present experiment was performed to reveal the mechanisms of toxic effects of ammonia nitrogen on Chinese hook snout carp (Opsariichthys bidens) based on the levels of antioxidant enzyme activities and the expression of 70‐kDa heat shock protein gene (HSP70) and tumor necrosis factor‐α (TNF‐α) after 96 h of acute toxic ammonia exposure to investigate its adaptability. The result showed that activities of antioxidant enzymes and lipid peroxidation in the intestine, liver, brain, and gills showed a general trend of increasing and then decreasing when exposed to ammonia nitrogen. The intestinal tissue structure was damaged to varying degrees, the expression of the HSP70 in the brain and gills increased at 0–12 h, and the expression of the TNF‐α in brain tissues decreased at 0–12 h. The expression of TNF‐α in the liver was continuously upregulated in the late stages of stress, and it did not decrease until the stress was released. The above results indicate that high concentrations of ammonia nitrogen caused varying degrees of damage to the neurological, respiratory, and metabolic‐related organs of the O. bidens.

Effects of exogenous steroid hormones on growth, body color, and gonadal development in the Opsariichthys bidens.

To investigate the effects of exogenous steroid hormones on growth, body color, and gonadal development in the Opsariichthys bidens (O. bidens), synthetic methyltestosterone (MT) and 17β-estradiol (E2) were used for 28 days' treatment of 4-month-old O. bidens before the breeding season. Our results suggested that MT had a significant growth-promoting effect (P < 0.05), whereas E2 played an inhibitory role. On the body surface, the females in the MT group showed gray stripes, and the fish in other groups showed no obvious stripes. The males with MT treatment displayed brighter blue-green stripes compared to the CK and E2 groups. The histological analysis showed that the MT significantly promoted testes development in males, blocked oocyte development, and caused massive apoptosis in females, whereas the E2 group promoted ovarian development and inhibited testes development. Based on qRT-PCR analysis, in females, the expression of igf-1, dmrt1, and cyp19a1a genes revealed that E2 treatment resulted in down-regulation of igf-1 expression and up-regulation of cyp19a1a expression. In males, igf-1 and dmrt1 were significantly up-regulated after MT treatment, and E2 treatment led to down-regulation of igf-1. Therefore, this study demonstrates that MT and E2 play an important role in reversing the morphological sex characteristics of females and males.

Length-Weight Relationships for Ten Fish Species from Rivers in the Qinling Mountains, China

Length-weight relationships (LWRs) were reported for ten fish species (Abbottina rivularis, Hemibarbus labeo, Hemibarbus maculatus, Onychostoma macrolepis, Opsariichthys bidens, Rhynchocypris lagowskii, Schizothorax prenanti, Sinibrama macrops, Zacco platypus, and Siniperca scherzeri). Fishes were collected from 62 sampling stations covering an area of ∼7000 km2 in the Qinling Mountains, characterized by unique fauna and distribution pattern, using gillnets and ground cages during June to December 2022. The intercept a ranged from 0.013 for S. prenanti to 0.060 for H. labeo, and the slope b ranged from 2.537 for H. labeo to 3.079 for Z. platypus.

Transcriptomic analysis of colour dimorphism of Opsariichthys bidens provides insights into the mechanism of male colour

Opsariichthys bidens (O. bidens) is a fish species native to China. It has become a popular ornamental fish because of its interesting colour dimorphism. Its colour pattern is unique; however, with the exception of model species, such as zebrafish, the molecular mechanisms of most fish colour patterns remain unclear. In the present study, we compared and analysed the transcriptomic data of the skin and anal fins of male and female O. bidens. A total of 9028 differentially expressed genes (DEGs) were identified, of which 5259 were upregulated and 3769 were downregulated in males. Compared with known pigmentation genes, Mitf, Tyr, Tyrp1, Dct, Pnp4a, Alx4a, Gch1, and Csf1r were identified as potential genes related to colour dimorphism. Second, we tracked the process of colour development in male O. bidens and found that there were three stages in the formation of body colour. We explored the distribution, migration, and expression patterns of genes related to pigment cells at each stage using microscopic imaging, transmission electron microscopy, enzyme-linked immunosorbent assay, and high-performance liquid chromatography. Melanophores, xanthophores, and iridophores were found to be distributed on the skin or anal fins of O. bidens. It was inferred that this unique body colour pattern is caused by the differentiation and migration of pigment cells at specific stages. This discovery provides a basis for studying the molecular mechanisms of pigmentation in ornamental fish and will contribute to the genetic selection and breeding of O. bidens in ornamental fish cultures in the future.

Length-Weight Relationships of Eight Fish Species in the World’s Largest Water Diversion Project

The study provides the length-weight relationships (LWRs) for eight fish species, viz. Hemiculter bleekeri, H. leucisculus, Opsariichthys bidens (Xenocyprididae), Carassius auratus, Cyprinus carpio (Cyprinidae), Pseudorasbora parva (Gobionidae), Acanthorhodeus chankaensis, and Rhodeus ocellatus (Acheilognathidae), in the world’s largest water diversion project, namely, the middle route of South-to-North Water Diversion Project (SNWDP). Fish specimens were sampled by benthic fyke-nets, gillnets, multimesh gillnets, and electrofishing equipment seasonally from 2015 to 2020. The coefficient of determination (r2) was ranging from 0.964 (R. ocellatus) to 0.986 (H. leucisculus), and the b values for the eight species were within range of 2.5–3.5. Also, our study updates the information on the maximum total length for four species (H. bleekeri, H. leucisculus, A. chankaensis, and O. bidens).

Species composition and distribution of fishes at the shallow waters of the low and middle Amur river in summer, 2018 and 2019

Based on the data of seine surveys (June 2018 and 2019, 57 and 38 catches), species composition and structure of fish communities of the middle and low Amur River were studied. The species richness of fish in the low Amur River is higher compared to the middle Amur River, 37 and 22 species, respectively. Upstream of the Amur River, fish biomass falls. At the beginning of summer, its average value in the low Amur was equal 1,354±0,209 g/m2, in the middle Amur – 0,324±0,058 g/m2. In the middle Amur River, the fish biomass was dominated by Hemiculter spp. (29,1%), Opsariichthys bidens (28,0%), Leuciscus waleckii (22,1%), Squalidus spp. (7,5%) and Tachysurus sinensis (3,9%). In the catches on the low Amur River, the fish biomass was dominated by Hemiculter spp. (26,2%), Leuciscus waleckii (21,0%), Hypomesus olidus (18,1%), Rhynchocypris lagowskii (7,6%), Carassius gibelio (7,2%), Rhodeus sericeus (5,0%) and Xenocypris macrolepis (4,6%). Local maxima of fish abundance were determined by the ecological preferences of most common fish species (in particular, their reopreferendum).

An evaluation method for the priority protection of fishes based on multiple ecological dimensions

It’s essential to identify the priority conservation species in the habitat restoration and biodiversity conservation. Generally, people select the valuable fishes or the commercial fishes as target species in basins, according to experts’ experience rather than the scientific evaluation. With the influence of urban development, construction of water conservancy facilities and human activities, the river habitat of the Shaying River (Xuchang section) is poor. Fish conservation studies play an important part in the habitat restoration of this river section. Thus, we studied the evaluation method for the priority protection of fishes based on multiple ecological dimensions. The method including six multidimensional indicators, namely, habitat suitability, ecological stability, species endangerment criteria, intensity of human interference, genetic value, and species value. Then, we determined the indicator weights by the analytic hierarchy process (AHP), and assessed the protection priority of fishes based on various assessment criteria and comprehensive qualitative or quantitative analyses. The result shows that Ctenopharyngodon idellus, Opsariichthys bidens, and Culter alburnus Basilewsky have priority for protection, coinciding with the status in the basin. Overall, this study complements the existing indicator species selection methods, and is more scientific and comprehensive, providing an effective method for priority protection species and river habitat restoration projects.

Transcriptomes of Testes at Different Developmental Stages in the Opsariichthys bidens Predict Key Genes for Testis Development and Spermatogenesis

Testis development is a complex process involving multiple genes, and the molecular mechanisms underlying testis development in Opsariichthys bidens remain unclear. We performed transcriptome sequencing analysis on a total of 12 samples of testes from stages II, III, IV, and V of O. bidens and obtained a total of 79.52Gb clean data, as well as 288,573 transcripts and 116,215 unigenes. Differential expression analysis showed that 22,857 differentially expressed genes (DEGs) were screened in six comparison groups (III vs. II, IV vs. II, V vs. II, IV vs. III, V vs. III, and V vs. IV). Kyoto Encyclopedia of Genes and Genomes enrichment analysis of DEGs showed that six comparison groups were significantly enriched for a total of 20 significantly up- or down-regulated pathways, including six pathways related to signal transduction, three pathways related to energy metabolism, five pathways related to disease, and two pathways related to ribosomes. Furthermore, our investigation revealed that DEGs were enriched in several important functional pathways, such as Huntington's disease signaling pathway, TGF-β signaling pathway, and ribosome signaling pathway. Protein-protein interaction network analysis of DEGs identified 63 up-regulated hub genes, including 9 kinesin genes and 2 cytoplasmic dynein genes, and 39 down-regulated hub genes, including 13 ribosomal protein genes. This result contributes to the knowledge of spermatogenesis and testis development in O. bidens.