2021 Vol. 42, No. 3
2021, 42(3): 253-261. doi: 10.24272/j.issn.2095-8137.2020.368
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Parabiosis is a surgical method of animal modeling with a long history. It has been widely used in medical research, particularly in the fields of aging, stem cells, neuroscience, and immunity in the past two decades. The protocols for parabiosis have been improved many times and are now widely accepted. However, researchers need to consider many details, from surgical operation to perioperative management, to reduce mortality and maintain the parabiosis union. Although parabiosis has certain inevitable limitations, it still has broad application prospects as an irreplaceable animal model in the medical research field.
Construction of a chromosome-level genome assembly for genome-wide identification of growth-related quantitative trait loci in Sinocyclocheilus grahami (Cypriniformes, Cyprinidae)
2021, 42(3): 262-266. doi: 10.24272/j.issn.2095-8137.2020.321
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The Dianchi golden-line barbel, Sinocyclocheilus grahami (Regan, 1904), is one of the “Four Famous Fishes” of Yunnan Province, China. Given its economic value, this species has been artificially bred successfully since 2007, with a nationally selected breed (“S. grahami, Bayou No. 1”) certified in 2018. For the future utilization of this species, its growth rate, disease resistance, and wild adaptability need to be improved, which could be achieved with the help of molecular marker-assisted selection (MAS). In the current study, we constructed the first chromosome-level genome of S. grahami, assembled 48 pseudo-chromosomes, and obtained a genome assembly of 1.49 Gb. We also performed QTL-seq analysis of S. grahami using the highest and lowest bulks (i.e., largest and smallest size) in both a sibling and random population. We screened two quantitative trait loci (QTLs) (Chr3, 14.9–39.1 Mb and Chr17, 4.1–27.4 Mb) as the major growth-related locations. Several candidate genes (e.g., map2k5, stat1, phf21a, sox6, and smad6) were also identified, with functions related to growth, such as cell differentiation, neuronal development, skeletal muscle development, chondrogenesis, and immunity. These results built a solid foundation for in-depth MAS studies on the growth traits of S. grahami.
Despite 250 years of taxonomic classification and over 1.2 million species already catalogued, known species diversity is only a small part of true species diversity on Earth, and thus, the known species are only the tip of iceberg. Here, we investigated the genus Pholcus
Walckenaer, 1805 of the family Pholcidae C. L. Koch, 1850 in the Changbai Mountains, Northeast China, which provides an excellent case of high species diversity. Previously, only 14 endemic Pholcus spiders, all belonging to the P. phungiformes species group, and two introduced species P. manueli Gertsch, 1937 and P. zichyi Kulczyński, 1901 from the P. crypticolens species group, have been recorded from this area. Our study confirmed 11 new species of the P. phungiformes species group based on morphology and three methods of molecular species delimitation: P. gaizhou Yao & Li, sp. nov. , P. guanshui Yao & Li, sp. nov. , P. jiguanshan Yao & Li, sp. nov. , P. longxigu Yao & Li, sp. nov. , P. luoquanbei Yao & Li, sp. nov. , P. shenshi Yao & Li, sp. nov. , P. tianmenshan Yao & Li, sp. nov. , P. wangjiang Yao & Li, sp. nov. , P. xingqi Yao & Li, sp. nov. , P. yaoshan Yao & Li, sp. nov. , and P. yuhuangshan Yao & Li, sp. nov. This study brings the fauna of the P. phungiformes species group from the Changbai Mountains to 25 species, approximately two times more than previously known, which could indicate that species diversity in the area is underestimated for all arthropod fauna.
2021, 42(3): 272-279. doi: 10.24272/j.issn.2095-8137.2020.319
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A new species of the genus Atelopus, Atelopus fronterizo sp. nov. , from eastern Panama is described herein based on molecular, morphological, and bioacoustic evidence. The new species can be distinguished from its congeners occurring in the region by a combination of the following characters: (1) phalangeal reduction in thumb; (2) SVL (females only) (35.1–50.1; n=13), HW/SVL (0.23–0.34; n=59), EYND/HW (0.27–0.39; n=60), TIBL/SVL (0.41–0.56; n=58), and HAL/SVL (0.22–0.28; n=49); (3) dorsal color pattern with green or yellow background and extensive dark olive blotches forming transversal bands or mottling; (4) advertisement call duration 176–235 ms with 19–34 pulses, average pulse rate 131.69 pulses/s, and dominant frequency 2 422.50–2 606.50 Hz. The new species is nested within the Central American clade of Atelopus. The minimum Kimura‐2‐parameter (K2P) genetic divergence between Atelopus fronterizo sp. nov. and its most phylogenetically similar congeners (A. certus and A. glyphus) is >2.6% for 16S and >4.9% for COI (
Table 1). The phylogenetic relationship is strongly supported by ultrafast bootstrap values for the maximum-likelihood trees of both genetic markers (16S, 96; COI, 100, Figure 1A). Bayesian analysis of the concatenated sequences resulted in a tree with similar topology and high posterior probability support (0.99; Supplementary Figure S1). In addition, haplotype networks inferred from COI and 16S (Supplementary Figure S2) showed a well-separated clade containing the new species (two for COI, four for 16S). The number of mutational steps between haplotypes for the new species samples is very low (1–4 in 16S; one in COI), and the minimum number of mutational steps from the nearest species is nine for 16S (distance to A. certus) and 28 for COI (distance to A. glyphus).
2021, 42(3): 280-286. doi: 10.24272/j.issn.2095-8137.2020.377
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DNA studies of endangered or extinct species often rely on ancient or degraded remains. The majority of ancient DNA (aDNA) extraction protocols focus on skeletal elements, with skin and hair samples rarely explored. Similar to that found in bones and teeth, DNA extracted from historical or ancient skin and fur samples is also extremely fragmented with low endogenous content due to natural degradation processes. Thus, the development of effective DNA extraction methods is required for these materials. Here, we compared the performance of two DNA extraction protocols (commercial and custom laboratory aDNA methods) on hair and skin samples from decades-old museum specimens to Iron Age archaeological material. We found that apart from the impact sample-specific taphonomic and handling history has on the quantity and quality of DNA preservation, skin yielded more endogenous DNA than hair of the samples and protocols tested. While both methods recovered DNA from ancient soft tissue, the laboratory method performed better overall in terms of DNA yield and quality, which was primarily due to the poorer performance of the commercial binding buffer in recovering aDNA.
Does the Dzungarian racerunner (Eremias dzungarica Orlova, Poyarkov, Chirikova, Nazarov, Munkhbaatar, Munkhbayar & Terbish, 2017) occur in China? Species delimitation and identification with DNA barcoding and morphometric analyses
2021, 42(3): 287-293. doi: 10.24272/j.issn.2095-8137.2020.318
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The Eremias multiocellata-przewalskii species complex is a viviparous group in the genus Eremias, and a well-known representative of taxonomically complicated taxa. Within this complex, a new species – E. dzungarica (
Orlova et al., 2017) – has been described recently from western Mongolia and eastern Kazakhstan, with an apparent distribution gap in northwestern China. In this study, we used an integrative taxonomic framework to address whether E. dzungarica indeed occurs in China. Thirty specimens previously classified as E. multiocellata were collected in eastern Kazakhstan and the adjacent Altay region in China. The cytochrome c oxidase I (COI) barcodes were sequenced and compiled with those from Orlova et al. (2017) and analyzed with the standard and diverse barcoding techniques. We detected an absence of a barcoding gap in this complex, which indicates potential cryptic species in Eremias sp. 3 with high intraspecific diversity and multiple recently evolved species in Clade A. Both BIN and GMYC suggested an unrealistically large number of species (23 and 26, respectively), while ABGD, mPTP and BPP indicated a more conservative number of species (10, 12, and 15, respectively), largely concordant with the previously defined species-level lineages according to phylogenetic trees. Based on molecular phylogeny and morphological examination, all 30 individuals collected in this study were reliably identified as E. dzungarica – a distinct species – confirming the occurrence of this species in the Altay region, Xinjiang, China. Potentially owing to the larger sample size in this study, our morphological analyses revealed many inconsistencies with the original descriptions of E. dzungarica, which were primarily associated with sexual dimorphism and a broader range of values for various traits.
During a terrestrial vertebrate survey of the Dabie Mountains in Anhui Province, eastern China, we collected four Asian shrew mole specimens (hereafter, shrew moles). Based on published literature and comparison with previously collected materials, the four specimens were similar to shrew moles from the mountains of Southwest China; however, no species in this group has been previously recorded from the Dabie Mountains. The genetic and morphological characteristics of the specimens were analyzed, based upon which a new species of shrew mole is described, named Uropsilus dabieshanensis sp. nov.
Cataract-causing allele in CRYAA (Y118D) proceeds through endoplasmic reticulum stress in mouse model
2021, 42(3): 300-309. doi: 10.24272/j.issn.2095-8137.2020.354
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As small heat shock proteins, α-crystallins function as molecular chaperones and inhibit the misfolding and aggregation of β/γ-crystallins. Genetic mutations of CRYAA are associated with protein aggregation and cataract occurrence. One possible process underlying cataract formation is that endoplasmic reticulum stress (ERS) induces the unfolded protein response (UPR), leading to apoptosis. However, the pathogenic mechanism related to this remains unexplored. Here, we successfully constructed a cataract-causing CRYAA (Y118D) mutant mouse model, in which the lenses of the CRYAA-Y118D mutant mice showed severe posterior rupture, abnormal morphological changes, and aberrant arrangement of crystallin fibers. Histological analysis was consistent with the clinical pathological characteristics. We also explored the pathogenic factors involved in cataract development through transcriptome analysis. In addition, based on key pathway analysis, up-regulated genes in CRYAA-Y118D mutant mice were implicated in the ERS-UPR pathway. This study showed that prolonged activation of the UPR pathway and severe stress response can cause proteotoxic and ERS-induced cell death in CRYAA-Y118D mutant mice.
A review of the Cypriniform tribe Yunnanilini Prokofiev, 2010 from China, with an emphasis on five genera based on morphologies and complete mitochondrial genomes of some species
2021, 42(3): 310-334. doi: 10.24272/j.issn.2095-8137.2020.229
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The loach tribe Yunnanilini from China is reviewed here using morphological characters and complete mitochondrial genomes of select species. Molecular data suggest that the tribe Yunnanilini is not monophyletic and can be divided into three clades. Species of the Yunnanilus nigromaculatus group form an independent genus and are placed in Eonemachilus. In the phylogenetic tree, Y. jinxiensis clusters with Paranemachilus genilepis, and Y. pulcherrimus clusters with Micronemacheilus cruciatus, indicating that Y. jinxiensis and Y. pulcherrimus belong to Paranemachilus and Micronemacheilus, respectively. Based on morphological data, Y. bailianensis and Y. longibarbatus are placed in Heminoemacheilus, while Y. jinxiensis and Y. pulcherrimus, are placed in Paranemachilus and Micronemacheilus, respectively. Yunnanilus niulanensis and Y. qujinensis are treated as junior synonyms of Eonemachilus caohaiensis. Eonemachilus, Micronemacheilus, and Yunnanilus are show short separation between anterior and posterior nostrils. The genera can be distinguished from each other by mouth structure, lateral line and cephalic lateral-line canals, and papillae on median part of both lips. The anterior and posterior nostrils of Heminoemacheilus and Paranemachilus are closely set. Paranemachilus and Micronemacheilus are distinguished by cheeks covered with scales and lips with papillae, respectively. Our phylogenetic tree and morphological characters support Traccatichthys as a valid genus, which can be distinguished from Micronemacheilus by anterior and posterior nostrils closely set (vs. clearly separated). Four species are placed in Traccatichthys.
Particulate matter exposure exacerbates susceptibility to SARS-CoV-2 infection in humanized ACE2 mice
2021, 42(3): 335-338. doi: 10.24272/j.issn.2095-8137.2021.088
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The global outbreak of coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), as of 8 May 2021, has surpassed 150 700 000 infections and 3 279 000 deaths worldwide. Evidence indicates that SARS-CoV-2 RNA can be detected on particulate matter (PM), and COVID-19 cases are correlated with levels of air pollutants. However, the mechanisms of PM involvement in the spread of SARS-CoV-2 remain poorly understood. Here, we found that PM exposure increased the expression level of angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) in several epithelial cells and increased the adsorption of the SARS-CoV-2 spike protein. Instillation of PM in a hACE2 mouse model significantly increased the expression of ACE2 and Tmprss2 and viral replication in the lungs. Furthermore, PM exacerbated the pulmonary lesions caused by SARS-CoV-2 infection in the hACE2 mice. In conclusion, our study demonstrated that PM is an epidemiological factor of COVID-19, emphasizing the necessity of wearing anti-PM masks to cope with this global pandemic.
Chronic pollution in aquatic ecosystems can lead to many adverse effects, including a greater susceptibility to pathogens among resident biota. Trifloxystrobin (TFS) is a strobilurin fungicide widely used in Asia to control soybean rust. However, it has the potential to enter aquatic ecosystems, where it may impair fish resistance to viral infections. To explore the potential environmental risks of TFS, we characterized the antiviral capacities of fish chronically exposed to TFS and subsequently infected with spring viraemia of carp virus (SVCV). Although TFS exhibited no significant cytotoxicity at the tested environmental concentrations during viral challenge, SVCV replication increased significantly in a time-dependent manner within epithelioma papulosum cyprini (EPC) cells and zebrafish exposed to 25 μg/L TFS. Results showed that the highest viral load was more than 100-fold that of the controls. Intracellular biochemical assays indicated that autophagy was induced by TFS, and associated changes included an increase in autophagosomes, conversion of LC3-II, accumulation of Beclin-1, and degradation of P62 in EPC cells and zebrafish. In addition, TFS markedly decreased the expression and phosphorylation of mTOR, indicating that activation of TFS may be associated with the mTOR-mediated autophagy pathway. This study provides new insights into the mechanism of the immunosuppressive effects of TFS on non-target aquatic hosts and suggests that the existence of TFS in aquatic environments may contribute to outbreaks of viral diseases.
Coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus (SARS-CoV-2), has become an unprecedented global health emergency. At present, SARS-CoV-2-infected nonhuman primates are considered the gold standard animal model for COVID-19 research. Here, we showed that northern pig-tailed macaques (Macaca leonina, NPMs) supported SARS-CoV-2 replication. Furthermore, compared with rhesus macaques, NPMs showed rapid viral clearance in lung tissues, nose swabs, throat swabs, and rectal swabs, which may be due to higher expression of interferon (IFN)-α in lung tissue. However, the rapid viral clearance was not associated with good outcome. In the second week post infection, NPMs developed persistent or even more severe inflammation and body injury compared with rhesus macaques. These results suggest that viral clearance may have no relationship with COVID-19 progression and SARS-CoV-2-infected NPMs could be considered as a critically ill animal model in COVID-19 research.
2021, 42(3): 354-361. doi: 10.24272/j.issn.2095-8137.2021.033
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Scientific research provides essential information for conservation of threatened species. Data deficiency due to insufficient research impedes the design of conservation plans, and research bias may mistakenly direct limited resources to low biodiversity regions or less threatened species. Here, we conducted a systematic review of published papers, grants, and graduate student training on carnivorans in China to identify species bias and research gaps. Furthermore, we collected intrinsic and extrinsic features of carnivorans, and identified features that impact research intensity using generalized linear models. We found that the amount of research on carnivorans increased markedly after 2000, but species bias existed. Bears and big cats received the greatest research attention, while most small- and medium-sized carnivorans received little attention, thus showing the 80-20 phenomenon. Species with a higher level of endemism and protection under Chinese law received more consideration. As an animal conservation icon in China, the giant panda (Ailuropoda melanoleuca) attracted more than 50% of overall carnivoran research resources. However, the giant panda also showed spillover effects, i.e., post-doctoral graduates who studied the giant panda shifted their research focus to other species after graduation, which may help improve research on other species. Thus, to improve and strengthen Carnivora research and conservation, we suggest investing greater effort in species of less concern, training of more graduate students, and reinforcing academic exchange. If such actions are not taken, many carnivoran species will continue being data deficient and threatened.
2021, 42(3): 362-376. doi: 10.24272/j.issn.2095-8137.2021.044
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Intermuscular bones (IBs) are slender linear bones embedded in muscle, which ossify from tendons through a process of intramembranous ossification, and only exist in basal teleosts. IBs are essential for fish swimming, but they present a choking risk during human consumption, especially in children, which can lead to commercial risks that have a negative impact on the aquaculture of these fish. In this review, we discuss the morphogenesis and functions of IBs, including their underlying molecular mechanisms, as well as the advantages and disadvantages of different methods for IB studies and techniques for breeding and generating IB-free fish lines. This review reveals that the many key genes involved in tendon development, osteoblast differentiation, and bone formation, e.g., scxa, msxC, sost, twist, bmps, and osterix, also play roles in IB development. Thus, this paper provides useful information for the breeding of new fish strains without IBs via genome editing and artificial selection.
LIN28A, an RNA-binding protein, plays an important role in porcine induced pluripotent stem cells (piPSCs). However, the molecular mechanism underlying the function of LIN28A in the maintenance of pluripotency in piPSCs remains unclear. Here, we explored the function of LIN28A in piPSCs based on its overexpression and knockdown. We performed total RNA sequencing (RNA-seq) of piPSCs and detected the expression levels of relevant genes by quantitative real-time polymerase chain reaction (qRT-PCR), western blot analysis, and immunofluorescence staining. Results indicated that piPSC proliferation ability decreased following LIN28A knockdown. Furthermore, when LIN28A expression in the shLIN28A2 group was lower (by 20%) than that in the negative control knockdown group (shNC), the pluripotency of piPSCs disappeared and they differentiated into neuroectoderm cells. Results also showed that LIN28A overexpression inhibited the expression of DUSP (dual-specificity phosphatases) family phosphatases and activated the mitogen-activated protein kinase (MAPK) signaling pathway. Thus, LIN28A appears to activate the MAPK signaling pathway to maintain the pluripotency and proliferation ability of piPSCs. Our study provides a new resource for exploring the functions of LIN28A in piPSCs.