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Transcriptomic analysis of Andrias davidianus meat and experimental validation for exploring its bioactive components as functional foods

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Food Science and Human Wellness 2024年第1期13卷 166-172页

作者： Changge Guan Zhenglin Tan Shucheng Li Yi Wang Naoyuki Yamamoto Chong Zhang Songjun Wang Junjie Chen Xinhui Xing Key Laboratory for Industrial Biocatalysis Ministry of Education of ChinaDepartment of Chemical EngineeringTsinghua UniversityBeijing 100084China School of Life Science and Technology Tokyo Institute of TechnologyYokohama 226-0026Japan Sensing System research center National Institute of Advanced Industrial Science and TechnologyTsukuba 305-8565Japan center for Synthetic and Systems Biology Tsinghua UniversityBeijing 100084China Henan Giant Salamander Protection and Development Association Luoyang 471000China Intelligent Computing research center Harbin Institute of TechnologyShenzhen 518071China Institute of Biopharmaceutical and Health Engineering Tsinghua Shenzhen International Graduate SchoolShenzhen 518055China Institute of Biomedical Health Technology and Engineering Shenzhen Bay LaboratoryShenzhen 518067China

Andrias davidianus(Chinese giant salamander,CGS)is the largest and oldest extant amphibian species in the world and is a source of prospective functional food in ***,the progress of functional peptides mining was slow due to lack of reference genome and protein sequence *** this study,we illustrated full-length transcriptome sequencing to interpret the proteome of CGS meat and obtain 10703 coding DNA *** functional annotation and amino acid composition analysis,we have discovered various genes related to signal transduction,and 16 genes related to *** have also found vast variety of functional peptides through protein coding sequence(CDS)analysis by comparing the data obtained with the functional peptide ***-Pro-Ile predicted by the CDS analysis was released from the CGS meat through enzymatic hydrolysis,suggesting that our approach is *** study suggested that transcriptomic analysis can be used as a reference to guide polypeptide mining in CGS meat,thereby providing a powerful mining strategy for the bioresources with unknown genomic and proteomic sequences.

关键词： Chinese giant salamander Transcriptomic analysis Bioactive components Functional peptides mining

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Would behavioral thermoregulation enable pregnant viviparous tropical lizards to cope with a warmer world?

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Integrative Zoology 2017年第5期12卷 379-395页

作者： Saúl LÓPEZ-ALCAIDE Miguel NAKAMURA Eric N.SMITH Enrique MARTÍNEZ-MEYER Departament of Zoology Institute of BiologyNational Autonomous University of MexicoMexico CityMexico center for research in Mathematics GuanajuatoMexico amphibian and Reptile Diversity research center and Department of Biology University of Texas at ArlingtonArlingtonTexasUSA

Sceloporus lizards depend on external heat to achieve their preferred temperature(Tsel)for performing physiological *** both in the field and laboratory indicates that pregnant females of this Genus select body temperatures(Tb)lower than 34℃ as higher temperatures may be lethal to ***,thermoregulation is crucial for successful embryo *** the increase in global air temperature,it is expected that the first compensatory response of species that inhabit tropical climates will be behavioral *** tested whether viviparous Sceloporus formosus group lizards in the wild exhibited differences in thermoregulatory behavior to achieve the known Tsel for developing embryos regardless of local thermal *** quantified field active body temperature,thermoregulatory behavior mechanisms(time of sighting,microhabitat used and basking time)and available microhabitat thermal conditions(*** temperature)for 10 lizard species during gestation,distributed along an altitudinal *** applied both conventional and phylogenic analyses to explore whether Tb or behavioral thermoregulation could be regulated in response to different thermal *** species showed no significant differences in field Tb during gestation regardless of local thermal *** contrast,they exhibited significant differences in their behavioral thermoregulation associated with local environmental *** on these observations,the differences in thermoregulatory behavior identified are interpreted as compensatory adjustments to local thermal *** conclude that these species may deal with higher temperatures predicted for the tropics by modulating their thermoregulatory behavior.

关键词： altitudinal gradient behavior global warming Sceloporus formosus thermal stress

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On the identification of paedomorphic and overwintering larval newts based on cloacal shape： review and guidelines

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Current Zoology 2017年第2期63卷 165-173页

作者： Mathieu DENOEL Laboratory of Fish and amphibian Ethology Behavioural Biology Unit FOCUS University of Liege 22 Quai van Benden 4020 Liege Belgium

Paedomorphosis is an alternative process to metamorphosis in which adults retain larval traits at the adult stage. It is frequent in newts and salamanders, where larvae reach sexual maturity with- out losing their gills. However, in some populations, larvae overwinter in water, while remaining immature. These alternative ontogenetic processes are of particular interest in various research fields, but have different causes and consequences, as only paedomorphosis allows metamor- phosis to be bypassed before maturity. It is thus relevant to efficiently identify paedomorphs ver- sus overwintering larvae. In this context, the aim of this article was 3-fold： firstly, to perform a meta-analysis of the identification procedures carried out in the literature; secondly, to determine the effectiveness of body size to make inferences about adulthood by surveying natural newt popu- lations of Lissotriton helveticus and Ichthyosaura alpestris, and thirdly, to propose easy guidelines for an accurate distinction between large larvae and paedomorphs based on an external sexual trait, which is essential for reproduction--the cloaca. More than half of the studies in the literature do not mention the diagnostic criteria used for determining adulthood. The criteria mentioned were the presence of mature gonads （10%）, eggs laid （4%）, courtship behavior （10%）, and external morphological sexual traits （39%） including the cloaca （24%）. Body-size thresholds should not be used as a proxy for paedomorphosis, because overwintering larvae can reach a larger size than paedomorphs within the same populations. In contrast, diagnosis based on cloacal external morphology is recommended, as it can be processed by the rapid visual assessment of all caught specimens, thus providing straightforward data at the individual level for both sexes.

关键词： amphibians cloacal morphology facultative paedomorphosis neoteny review secondary sexual traits sexual di-morphism sexual maturity.

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Reproductive Isolating Mechanisms in the Bangladesh Coastal Bullfrog Hoplobatrachus litoralis and Its Congeneric Species Revealed by Crossing Experiments and Examination on Spermatogenesis of the Hybrids

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Asian Herpetological research 2017年第1期18卷 27-38页

作者： Mahmudul HASAN Mohammed Mafizul ISLAM Md. Mukhlesur Rahman KHAN Ratanasate WANICHANON Atsushi KURABAYASHI Masayuki NUMIDA amphibian research center Hiroshima University 1-3-1 Kagamiyama Higashihiroshima 739-8526 Japan Department of Fisheries Biology and Genetics Bangladesh Agricultural University Mymensingh-2202 Bangladesh Department of Anatomy Phramonkutklao Medical College Rajawithi Rd. Bangkok 10400 Thailand 1-6-15 Ushitaasahi Higashi ku Hiroshima 732-0067 Japan

To elucidate reproductive isolating mechanisms in the Bangladesh coastal bullfrog Hoplobatrachus litoralis and its congeneric species, we performed crossing experiments using three species： H. litoralis, H. tigerinus and H. rugulosus. In addition, we conducted histological observations on spermatogenesis of the hybrids. The reciprocal hybrids between H. litoralis and H. tigerinus developed normally with somewhat lower viability at the metamorphosis stage compared with the controls. Most of the metamorphosed frogs became mature. On the other hand, almost all hybrids between female H. rugulosus and male H. litoralis or H. tigerinus died of underdevelopment at the tadpole stage, and only a few hybrids metamorphosed normally and survived to maturity. The inner structures of the testes of the control H. litoralis and H. tigerinus were completely normal, with seminiferous tubules filled with compact bundles of normal spermatozoa. Those of the reciprocal hybrids between H. litoralis and H. tigerinus were almost normal or slightly abnormal, with seminiferous tubules that contained pycnotic nuclei in addition to normal bundles of normal spermatozoa, which demonstrates slight abnormalities in spermatogenesis. In contrast, the hybrids between female H. rugulosus and male H. litoralis or H. tigerinus had no bundles of spermatozoa nor spermatids in the seminiferous tubules, which indicates entirely abnormal spermatogenesis. Meiotic chromosome figures in the reciprocal hybrids between H. litoralis and H. tigerinus showed slight abnormalities, with the occurrence of univalents and increase of rod-shaped bivalents. These results indicate that H. litoralis and H. tigerinus are not isolated from each other by hybrid inviability nor by hybrid sterility, although the hybrids showed somewhat abnormal spermatogenesis in hybrids and that H. rugulosus is isolated from both H. litoralis and H. tigerinus by incomplete hybrid inviability and complete hybrid sterility.

关键词： Hoplobatrachus reproductive isolation interspecific hybrids spermatogenesis meiotic chromosomes

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A screening assay for thyroid hormone signaling disruption based on thyroid hormone-response gene expression analysis in the frog Pelophylax nigromaculatus

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Journal of Environmental Sciences 2015年第8期27卷 143-154页

作者： Yinfeng Zhang Yuanyuan Li Zhanfen Qin Huili Wang Jianzhong Li State Key Laboratory of Environmental Chemistry and Ecotoxicology Research Center for Eco-environmental Sciences Chinese Academy of Sciences Department of Environmental Bio-Technology Research Center for Eco-environmental Sciences Chinese Academy of Sciences

amphibian metamorphosis provides a wonderful model to study the thyroid hormone（TH）signaling disrupting activity of environmental chemicals, with Xenopus laevis as the most commonly used species. This study aimed to establish a rapid and sensitive screening assay based on TH-response gene expression analysis using Pelophylax nigromaculatus, a native frog species distributed widely in East Asia, especially in China. To achieve this, five candidate TH-response genes that were sensitive to T3 induction were chosen as molecular markers, and T3 induction was determined as 0.2 nmol/L T3 exposure for 48 hr. The developed assay can detect the agonistic activity of T3 with a lowest observed effective concentration of 0.001 nmol/L and EC50 at around 0.118–1.229 nmol/L, exhibiting comparable or higher sensitivity than previously reported assays. We further validated the efficiency of the developed assay by detecting the TH signaling disrupting activity of tetrabromobisphenol A（TBBPA）, a known TH signaling disruptor. In accordance with previous reports, we found a weak TH agonistic activity for TBBPA in the absence of T3,whereas a TH antagonistic activity was found for TBBPA at higher concentrations in the presence of T3, showing that the P. nigromaculatus assay is effective for detecting TH signaling disrupting activity. Importantly, we observed non-monotonic dose-dependent disrupting activity of TBBPA in the presence of T3, which is difficult to detect with in vitro reporter gene assays. Overall, the developed P. nigromaculatus assay can be used to screen TH signaling disrupting activity of environmental chemicals with high sensitivity.

关键词： Thyroid hormone signaling disruptor Pelophylax nigromaculatus TH-response gene expression Screening assay Tetrabromobisphenol A

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Species Diversity and Elevational Distribution of amphibians in the Xianxialing and Wuyishan Mountain Ranges,Southeastern China

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Asian Herpetological research 2020年第1期11卷 44-55页

作者： Zhiqiang CHEN Yun TANG Yingying WANG Yindie LI Longhui LIN Guohua DING Laboratory of amphibian Diversiry Investigation College of EcologyLishui UniversityLishui 323000 ZhejiangChina Hangzhou Key Lab for Animal Adaptation and Evolution College of Life and Environmental SciencesHangzhou Normal UniversityHangzhou 310036ZhejiangChina

The species diversity and altitudinal distribution of amphibians along an eleva tional gradient of 200-1600 m in the Xianxialing and Wuyishan Mountain Ranges in Southeastern China were investiga ted through time-constra ined visual surveys along 32 transect lines in 9 survey areas,in which the ha bitat types were also recorded.A total of 27 amphibian species belonging to 19 genera,7 families,and 2 orders were *** species diversity of the amphibians plateaued at low elevation,and the altitudinal boundary of their distribution was at 800 *** species compositions were dissimilar in the two mountain ranges probably beca use the annual average temperature and annual rainfall were different in both *** eleva tional Rapoport's rule demonstrated that the species range size of the amphibians expanded as the elevation increased in both mountain *** results of the cross species method supported the rule only when the influence of the low-frequency occurrence proba bility of an investigated species was excluded,whereas those of the Steven's method strongly corroborated the rule rega rdless of the incidental occurrence or absence of the species.

关键词： amphibian diversity altitudinal distribution Rapoport’s rule cross-species method Steven’s method

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Resurvey After 20 Years:Updating the Distribution,Population,and Potential Threats of the Chinting Alpine Toad

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Asian Herpetological research 2024年第2期15卷 73-81页

作者： Yuezheng FAN Qingfeng CHEN Yuwen CHENG Kaicheng TANG Ling SUN Yonghao HUANG Lan ZHAO Dong LIANG Xiaoyi WANG Junhua HU Chengdu Institute of Biology Chinese Academy of SciencesChengdu 610041SichuanChina University of Chinese Academy of Sciences Beijing 100049China The State-owned Forest Farm of Hongya County Meishan 620360SichuanChina Sichuan Mt.Emei Forest Ecosystem National Observation and research Station Leshan 614201SichuanChina

Considering the increased anthropogenic impacts,species with a limited range and low detectability often lack fundamental information and conservation actions,placing them at a high risk of *** Chinting alpine toad Scutiger chintingensis is a rare mountain amphibian endemic to the eastern margin of the Qinghai-Xizang Plateau in *** its whole distribution range,only three known populations(Wolong,Emei,and Wawu)exist and no recent population status report has been documented for this species over the past two *** 2020 to 2023,we investigated the species distribution,and assessed the risk factors for the main *** recorded this species in all distribution areas,and updated a new distribution site with a lower elevation *** relative population density was 0.024±0.012 ind./m^(2)on Mount Emei,whereas only 0.008±0.017 ind./m^(2)on Mount *** significant difference was observed in the number of individuals between the two populations;however,the relative population density was significantly *** and waste discharge resulting from the construction of scenic areas,as well as disturbances from tourism,were the primary anthropogenic factors that influenced the survival of this *** results provide the updated information on the distribution and population status of the Chinting alpine toad,and suggest that unrecorded populations,as well as a wider elevation range,may exist for this *** findings emphasise the importance of timely updates of species distribution and population information and offer a basis for the future conservation of endangered amphibians.

关键词： distribution endemic amphibian population size Scutiger chintingensis threat factor

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High-elevation hypoxia impacts perinatal physiology and performance in a potential montane colonizer

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Integrative Zoology 2020年第6期15卷 544-557页

作者： Jérémie SOUCHET Eric J.GANGLOFF Gaëlle MICHELI Coralie BOSSU Audrey TROCHET Romain BERTRAND Jean CLOBERT Olivier CALVEZ Albert MARTINEZ-SILVESTRE Elodie DARNET Hugo LE CHEVALIER Olivier GUILLAUME Marc MOSSOLL-TORRES Laurent BARTHE Gilles POTTIER HervéPHILIPPE Fabien AUBRET Station d’Ecologie Théorique et Expérimentale du Centre National de la Recherche Scientifique MoulisFrance Department of Zoology Ohio Wesleyan UniversityDelawareOhioUSA Catalonia Reptile and amphibian Rescue center(CRARC) MasquefaBarcelonaSpain Bomosa Pl.Parc de la MolaLes EscaldesAndorra Pirenalia EncampAndorra Nature En Occitanie ToulouseFrance Département de Biochimie Centre Robert-CedergrenUniversitéde MontréalMontréalCanada

Climate change is generating range shifts in many organisms,notably along the elevational gradient in mountainous ***,moving up in elevation exposes organisms to lower oxygen availability,which may reduce the successful reproduction and development of oviparous *** test this possibility in an upwardcolonizing species,we artificially incubated developing embryos of the viperine snake(Natrix maura)using a split-clutch design,in conditions of extreme high elevation(hypoxia at 2877 m above sea level;72%sea-level equivalent O2 availability)or low elevation(control group;*** at 436 m above sea level).Hatching success did not differ between the two *** developing at extreme high elevation had higher heart rates and hatched earlier,resulting in hatchlings that were smaller in body size and slower swimmers compared to their siblings incubated at lower ***,post-hatching reciprocal transplant of juveniles showed that snakes which developed at extreme high elevation,when transferred back to low elevation,did not recover full performance compared to their siblings from the low elevation incubation *** results suggest that incubation at extreme high elevation,including the effects of hypoxia,will not prevent oviparous ectotherms from producing viable young,but may pose significant physiological challenges on developing offspring in *** early-life performance limitations imposed by extreme high elevation could have negative consequences on adult phenotypes,including on fitness-related traits.

关键词： developmental plasticity embryonic metabolism high-elevation hypoxia locomotor performance Natrix maura

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Application of Xenopus laevis in ecotoxicology （Ⅰ）--Introduction and quality control of laboratory animal

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Chinese Science Bulletin 2006年第11期51卷 1273-1280页

作者： QIN Zhanfen XU Xiaobai State Key Laboratory of Environmental Chemistry and Ecotoxicology Research Center for Eco-environmental Sciences Chinese Academy ofSciences Beijing 100085 China

The aim of the series of papers is to discuss the application of Xenopus laevis, as model animal in biology, in ecotoxicology. X. laevis as model animal is wildly used in biological study and has pro- vided a lot of relating data because of many advan- tages, such as living in water and being easily main- tained, laying eggs in the whole year, and externally fertilizing and developing. Embryos and larvae of X. laevis like other amphibians are directly exposed in the aquatic environment and sensitive to pollutants. In addition, sex differentiation and sex organ devel- opment of X. laevis are sensitive to sex hormones and endocrine disruptors with sex hormone activities, which enable X. laevis to be used in studies on sex hormone disruption and reproductive toxicity of en- docrine disruptors. Metamorphic development of X. laevis is very sensitive to thyroid hormones and thy- roid disruptors, which enables X. laevis to be used for evaluating thyroid disruptors. Also, X. laevis ecotoxi- cology can be linked with amphibian population de- clines and malformed frog occurrence, being one of the hotspots in ecology. Thus, more and more labo- ratories have introduced X. laevis to ecotoxicological study. The quality of laboratory animals correlates with scientificity and reliability of results from animal experiments. It is especially important for toxicology. Quality control of X. laevis involving several factors such as water and food is discussed in this paper.

关键词：有爪蟾蜍生态毒理学实验动物质量控制

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Effects of triclosan on gonadal differentiation and development in the frog Pelophylax nigromaculatus

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Journal of Environmental Sciences 2018年第2期30卷 157-165页

作者： Juan Chen Tan Meng Yuanyuan Li Kun Gao Zhanfen Qin State Key Laboratory of Environmental Chemistry and Ecotoxicology Research Center for Eco-Environmental Sciences Chinese Academy of Sciences Beijing 100085 China University of Chinese Academy of Sciences Beijing 100049 China

Previous studies have reported that triclosan（TCS） could possess an androgenic *** aimed to investigate the effects of TCS on gonadal differentiation and development in the frog Pelophylax nigromaculatus, a sensitive amphibian species to androgenic chemicals.P. nigromaculatus tadpoles at stage 24 were exposed to TCS（3, 30, and 300 nmol/L） to stage46 in a semi-static exposure system. At the end of exposure, gonadal morphology and histology, sex ratio and gonadal expression of sex-biased genes were examined in P. nigromaculatus. In each TCS treatment group, we found several individuals whose gonads exhibited morphological and/or histological abnormalities. Gonadal histological abnormalities were characterized by few oocytes and many somatic cells. Although the percentage of the individuals with abnormal gonads was low（7.8%） among all animals treated with TCS,statistical test revealed the sex ratios in the 3 and 300 nmol/L TCS treatment groups were significantly different from the solvent control. In the 30 nmol/L TCS treatment group,abnormal gonads were also observed, although the sex ratio was not changed compared with the solvent control, which was possibly due to the smaller sample size in this group. In all the TCS treatment groups, the sex ratios were not obviously male-biased, but the expression levels of some sex-biased genes were significantly altered by TCS. Altogether,our results suggest that TCS, even at environmentally relevant concentrations, could disrupt gonadal differentiation and development in P. nigromaculatus, but we are not sure whether the disrupting effects were associated with masculinization or feminization.

关键词： Triclosan Pelophylax nigromaculatus Gonadal abnormality Androgenic activity

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