The taxonomic status of Bufo species in East Asia,particularly those closely related to B.gargarizans,has been controversial for over half a century,and recent molecular studies further fueled the dispute.Whether B.an...The taxonomic status of Bufo species in East Asia,particularly those closely related to B.gargarizans,has been controversial for over half a century,and recent molecular studies further fueled the dispute.Whether B.andrewsi,B.bankorensis,B.minshanicus,B.sachalinensis,B.tibetanus,and B.tuberculatus constitute valid species or are geographic or ecological variants of B.gargarizans is at the centre of the discussion.Most of these forms possess morphological diagnostic traits,but those differences are minimal and often inconsistent.Molecular data produced many contradictory patterns.Lack of clear diagnostic characters,different usage or definition of various names,mis-identified specimens,limited amount of data or samples,and frequent inter-specific hybridization may all have contributed to the current controversies.Some contradiction may also result from differences between similarity-based clustering and gene tree-based branching patterns.Nevertheless,a dominant allopatric differentiation pattern within this species complex is apparent,and several regional population groups have been identified.Some of the groups resemble old names(e.g.B.minshanicus,B.tibetanus),while others are new.These groups may represent geographic or ecological variants,incipient species,or valid species,and the current data do not warrant a convincing taxonomic revision.To move forward,a single study with comprehensive sampling and genomic data is needed to establish a solid framework,which will define naturally existing population clusters/lineages,and detect their levels of divergence and approximate geographic boundaries.Morphological and ecological examination will follow to understand their biology,determine their taxonomic status,and identify consistent diagnostic characters if necessary.展开更多
Difference in body size between males and females(sexual size dimorphism:SSD)and its variation are a common phenomenon in animal kingdom.Rensch’s rule predicts that the degree of SSD variation increases with the enla...Difference in body size between males and females(sexual size dimorphism:SSD)and its variation are a common phenomenon in animal kingdom.Rensch’s rule predicts that the degree of SSD variation increases with the enlarged mean body size when males are larger than females and decreases when females are larger than males.Here,whether the patterns of variations in SSD in the Andrew’s toad(Bufo andrewsi)follow Rensch’s rule was tested using unpublished data from 14 populations and published data from 17 populations.Results show the reduced major axis regression of log10(male size)on log10(female size)across all populations displayed a significant hyperallometric relationship,which was consistent with inverse Rensch’s rule(the degree of SSD increased with enlarged mean body size).SSD could also be explained by sexual age difference(SAD)due to a positive SSD–SAD relationship among all populations.The findings suggest that the occurrence of inverse Rensch’s rule in B.andrewsi is likely to be a result of fecundity selection on increased reproductive investments in larger females.展开更多
Previous studies on the Bufo viridis complex, which is distributed broadly across Iran, are incomplete and restricted to a few regions or a few samples. In this paper a new detailed study on the B. viridis complex in ...Previous studies on the Bufo viridis complex, which is distributed broadly across Iran, are incomplete and restricted to a few regions or a few samples. In this paper a new detailed study on the B. viridis complex in southern of Iran （from West to East） is presented. The analysis of 18 morphometric characters with univariate and multivariate methods reveals significant differences between three members of the B. viridis complex namely B. variabilis, B. luristanicus, and B. surdus distributed in southern part of Iran. Our result help to resolve an old taxonomic problem about B. surdus subgroup （taxa closely related to B. surdus） confirming that B. luristanicus and B. surdus are distinct species. Moreover, for the first time we report and describe karyotype details of B. luristanicus and B. surdus which confirmed that they are diploid. Karyological studies demonstrate that all toads from three mentioned species have 2n = 22 chromosomes. These chromosomes are arranged into two groups. First group has six large chromosomes and the second group is composed of five small chromosomes. These chromosomes are metacentric or submetacentric. The number of submetacentric chromosomes is different in three mentioned species of B. viridis complex. Neither sexual heteromorphism, nor secondary constriction was observed in any pairs of chromosomes.展开更多
Advertisement calls play an important role in influencing female mate choice and are regarded as one of the key characters responsible for reproductive isolation and speciation events in frogs and toads. The black-spi...Advertisement calls play an important role in influencing female mate choice and are regarded as one of the key characters responsible for reproductive isolation and speciation events in frogs and toads. The black-spined toad, Bufo melanostictus, is one of the most common amphibians in Southeast Asia and is frequently observed in human habitations throughout its distributional range. The advertisement calls of the toad have not been well described. Here, we report the advertisement calls of this species from Lishui, Zhejiang, China during its breeding season. The advertisement calls of B. melanostictus are composed of single notes with a single harmonic frequency emitted sporadically. The call parameters recorded in this study were highly variable across recording times and with different body sizes. Dominant frequency was negatively correlated with body size. Comparing the calls of the B. melanostictus population from Lishui with the populations from five other localities(Bangkok, Thailand; Coorg, India; Burma, Myanmar; Bali, Indonesia; Halimun-Salak National Park, Indonesia), we found that call structure was similar among five different populations with no harmonics, while the Halimun-Salak National Park population contained two clear harmonics. Dominant frequency was similar between Bali(1.56 kHz) and Coorg(1.6 kHz) populations, while it was highly variable among other populations. Besides call duration, call interval and pulse/call rate may also be significantly different among the comparing populations. Pooling the available data, we observed some similarities and differences in call parameters among the six populations.展开更多
Microelectrode technique was used to record the spontaneous electrical activities of the neurons in olfactory bulb of the Bufo bufo gargarizans, both in hibernation and after hibemation. This study investigated the el...Microelectrode technique was used to record the spontaneous electrical activities of the neurons in olfactory bulb of the Bufo bufo gargarizans, both in hibernation and after hibemation. This study investigated the electrophysiological characteristics of amphibian olfactory bulb in the period of hibernation and after hibernation and its effects on the start of hibernation and spontaneous awakening. The research showed four forms of spontaneous firings： single spontaneous firing, burst spontaneous firing, irregular spontaneous firing and consecutive single spontaneous firing. The single spontaneous firing includes slow depolarized spontaneous firing and fast depolarized spontaneous firing, and the slow depolarized spontaneous firing occurs only during the hibernation period. In hibernation, the low amplitude and low frequency firing with a longer duration may be relevant to maintaining the tonicity of the central nervous system in toads that are in hibernation, and this kind of firing may also provide an excited basis for their arousal from hibernation. After hibernation, the amplitude and frequency of firing increase, but the firing duration gets shorter. This form of short-term firing, which may be a phenomenon of sensory neurons fast adapting, is one of the neuronal mechanisms for the arousal of hibernating animals.展开更多
In many anurans, the forelimb muscles of males are used to grasp females and are often heavier than those of females despite the larger female body size. Such sexual dimorphism in forelimb musculature is thought to re...In many anurans, the forelimb muscles of males are used to grasp females and are often heavier than those of females despite the larger female body size. Such sexual dimorphism in forelimb musculature is thought to result from sexual selection. In addition, the hindlimbs of frogs and toads play an important role in the reproductive process as amplectant males can expel rivals with robust hindlimbs through kicking. In this study, the sexual dimorphism in dry mass for six hindlimb muscles of the Asiatic toad （Bufo gargarizans） was investigated. The results showed that, when controlled for body size, the hindlimb muscle mass of males significantly exceeded that of females for every muscle. The hindlimb muscle mass of amplectant males was also significantly larger than that of non-amplectant males. These results suggested that if strong hindlimb muscles could improve mating success of males, sexual selection would promote the evolution of dimorphism in this character.展开更多
Lead has been recognized as a high risk toxic for most organisms including human. The effects of Pb in non-mammalian vertebrates are oprly known, particularly in anuran amphibians. The purpose of this study was to det...Lead has been recognized as a high risk toxic for most organisms including human. The effects of Pb in non-mammalian vertebrates are oprly known, particularly in anuran amphibians. The purpose of this study was to determine the effect of this metal on some hematological parameters of adult Bufo arenarum. It was found that all parameters remained unaltered within normal ranges, with the exception of reticulocyte counts which was significantly increased compared to the in controls (3. 7% vs. 0. 2 % ). It is suggested that the selective change found in reticulocyte count might be considered as an early response of a biomarker to sublethal exposition of Bufo arenarum to lead.展开更多
Three anti-keratin MAbs were used to identifykeratins expressed in early embryos of Bufo bufogargarizans.MAb AF5 recognized three polypeptides ofkeratin in oocytes,fertilized eggs,up to neurula withMr of 68,65 and 60K...Three anti-keratin MAbs were used to identifykeratins expressed in early embryos of Bufo bufogargarizans.MAb AF5 recognized three polypeptides ofkeratin in oocytes,fertilized eggs,up to neurula withMr of 68,65 and 60Kd respectively.At tailbud stage,three other keratins(62,58 and 54Kd)began to expressand could be detected by AF5.MAbs D10 and K12 gavedifferent results,both of them could identify four keratin-like molecules with unusual molecular weights(Mr 98,95,30 and 27 Kd).Moreover,D10 could also detect a 54 Kdkeratin in neurula and tailbud stage embryos,while K12could reveal,beside 54Kd keratin,other four more kera-tins(68,65,62 and 60 Kd).The possible interpretation ofthese results and their implications are discussed.展开更多
Relatively little is known about the relationship between Bufo gargarizans populations from Zhoushan Archipelago and nearby continental regions on the Pacific coast of eastern China.In this paper,155 new specimens of ...Relatively little is known about the relationship between Bufo gargarizans populations from Zhoushan Archipelago and nearby continental regions on the Pacific coast of eastern China.In this paper,155 new specimens of B.gargarizans from Zhoushan Archipelago and adjacent continents and 71 published specimens of B.gargarizans from China's Mainland were studied.Phylogeographical and dating analyses of B.gargarizans were performed using mitochondrial DNA sequencing with a length of 1436 bp.A mt DNA tree that indicated seven major clades was obtained.The earliest split in the mt DNA tree corresponding to the divergence of populations from the western highland region occurred approximately 4.0 million years ago（mya）.A subsequent clade occurred about 3.4 mya,with cladogenesis continuing toward the end of the Pleistocene.The continental clades were distributed in the western,central and northeastern regions of China.Zhoushan Archipelago clades consisted of two largely geographically overlapping subclades with the mt DNA divergence time of 0.73 mya.These results indicated there was extensive dispersal after vicariance.The B.gargarizans populations on Zhoushan Archipelago most probably originated from populations in nearby eastern continental regions of China.It was concluded that geological uplifting during the Pliocene and several sea-level changes in Pleistocene might have influenced the divergence and population demographical history of this species.展开更多
The present study was carried out to investigate the histological and histochemical changes in the liver and skin on different developmental stages of Egyptian toad Bufo regularis to be used as a histological key for ...The present study was carried out to investigate the histological and histochemical changes in the liver and skin on different developmental stages of Egyptian toad Bufo regularis to be used as a histological key for such species. Our experiment started when tadpoles began to feed. The adapted embryos are divided into 3 large tanks of 200 embryos each, collections of samples started from feeding age every three days. Both histological and histochemical results showed that the general architecture of the different organs was correlated with the state of development, i.e. larval, met-amorphic and post-metamorphic. They, therefore, displayed different characteristic features de-pending on the investigated developmental stage starting from the larval stage (stage 44) and ending with the post-metamorphic stage 66.展开更多
The present study aimed to investigate changes in the protein contents using SDS-PAGE, also using the random amplified polymorphic DNA (RAPD-PCR) assay to identify changes in DNA concentration in the embryos of Egypti...The present study aimed to investigate changes in the protein contents using SDS-PAGE, also using the random amplified polymorphic DNA (RAPD-PCR) assay to identify changes in DNA concentration in the embryos of Egyptian toad Bufo regularis. Our experiment started when tadpoles began to feed. The adapted embryos were divided into 3 large tanks of 200 embryos each, collections of samples started from feeding age every three days. Cluster method was used to indicate the distinct distance between the band patterns of different stages. An increase occurred in the concentration of protein fractions in these tadpoles upon metamorphosis and that when metamorphosis was completed. DNA concentration exhibited an evident low values at the premetamorphic stage 44 (105.85 ± 0.519 ng/μl) compared with the other more advanced developmental stages 55 and 56 (204.056 ± 0.651 ng/μl and 234.55 ± 2.325 ng/μl respectively). In conclusion, acquire information about the changes in the molecular content of Bufo regularis during metamorphosis has been estimated.展开更多
In this study,to clarify the bioactive polypeptides included in the skins and secretions of Bufo,we screened the Japanese toad(Bufo japonicus formosus) skin cDNA library by colony polymerase chain reaction(PCR),and ob...In this study,to clarify the bioactive polypeptides included in the skins and secretions of Bufo,we screened the Japanese toad(Bufo japonicus formosus) skin cDNA library by colony polymerase chain reaction(PCR),and obtained a transcript of 1 075 bp consisting of 1 37 bp 5′ untranslated region(UTR),515 bp 3′ UTR and a 423 bp open reading frame(ORF) encoding a polypeptide of 140 amino acid residues(GenBank accession number: KF359945).Homolog analysis showed a 70%–96% homology with sterol carrier protein-2(SCP-2) present in other animals,which is implicated in lipid metabolism of other organisms.The gene SCP-2 of Chinese toad(B.gargarizans) was cloned from a first strand cDNA of Bufo skin(GenBank accession number: KF381341) via PCR,whose encoding polypeptide has only one amino acid difference from that of Japanese toad.Tissue distribution analysis showed that SCP-2 expressed in all organs tested,though in the liver and spleen it manifested lower expression than in other organs.These findings might indicate SCP-2 being one of the active ingredients in toad skin.These findings may in turn have implications for further drug development from traditional Chinese medicine sources.展开更多
There was a slow-relaxing tail of skeletal muscles in vitro upon the inhibition of Ca2+-pump by cyclopiazonic acid (CPA). Herein, a new linearly-combined bi-exponential model to resolve this slow-relaxing tail from th...There was a slow-relaxing tail of skeletal muscles in vitro upon the inhibition of Ca2+-pump by cyclopiazonic acid (CPA). Herein, a new linearly-combined bi-exponential model to resolve this slow-relaxing tail from the fast-relaxing phase was investigated for kinetic analysis of the isometric relaxation process of Bufo gastrocnemius in vitro, in comparison to the single exponential model and the classical bi-exponential model. During repetitive stimulations at a 2-s interval by square pulses of a 2-ms duration at 12 V direct currency (DC), the isometric tension of Bufo gastrocnemius was recorded at 100 Hz. The relaxation curve with tensions falling from 90% of the peak to the 15th datum before next stimulation was analyzed by three exponential models using a program in MATLAB 6.5. Both the goodness of fit and the distribution of the residuals for the best fitting sup- ported the comparable validity of this new bi-exponential model for kinetic analysis of the relaxation process of the control muscles. After CPA treatment, however, this new bi-exponential model showed an obvious statistical superiority for kinetic analysis of the muscle relaxation process, and it gave the estimated rest tension consistent to that by experimentation, whereas both the classical bi-exponential model and the single exponential model gave biased rest tensions. Moreover, after the treatment of muscles by CPA, both the single exponential model and the classical bi-exponential model yielded lowered relaxation rates, nevertheless, this new bi-exponential model had relaxation rates of negligible changes except much higher rest tensions. These results suggest that this novel linearly-combined bi-exponential model is desirable for kinetic analysis of the relaxation process of muscles with altered Ca2+-pumping activity.展开更多
文摘The taxonomic status of Bufo species in East Asia,particularly those closely related to B.gargarizans,has been controversial for over half a century,and recent molecular studies further fueled the dispute.Whether B.andrewsi,B.bankorensis,B.minshanicus,B.sachalinensis,B.tibetanus,and B.tuberculatus constitute valid species or are geographic or ecological variants of B.gargarizans is at the centre of the discussion.Most of these forms possess morphological diagnostic traits,but those differences are minimal and often inconsistent.Molecular data produced many contradictory patterns.Lack of clear diagnostic characters,different usage or definition of various names,mis-identified specimens,limited amount of data or samples,and frequent inter-specific hybridization may all have contributed to the current controversies.Some contradiction may also result from differences between similarity-based clustering and gene tree-based branching patterns.Nevertheless,a dominant allopatric differentiation pattern within this species complex is apparent,and several regional population groups have been identified.Some of the groups resemble old names(e.g.B.minshanicus,B.tibetanus),while others are new.These groups may represent geographic or ecological variants,incipient species,or valid species,and the current data do not warrant a convincing taxonomic revision.To move forward,a single study with comprehensive sampling and genomic data is needed to establish a solid framework,which will define naturally existing population clusters/lineages,and detect their levels of divergence and approximate geographic boundaries.Morphological and ecological examination will follow to understand their biology,determine their taxonomic status,and identify consistent diagnostic characters if necessary.
基金funded by the National Natural Sciences Foundation of China(31772451,31970393)the Key Project of Science and Technology of Sichuan Province(22NSFSC0011)。
文摘Difference in body size between males and females(sexual size dimorphism:SSD)and its variation are a common phenomenon in animal kingdom.Rensch’s rule predicts that the degree of SSD variation increases with the enlarged mean body size when males are larger than females and decreases when females are larger than males.Here,whether the patterns of variations in SSD in the Andrew’s toad(Bufo andrewsi)follow Rensch’s rule was tested using unpublished data from 14 populations and published data from 17 populations.Results show the reduced major axis regression of log10(male size)on log10(female size)across all populations displayed a significant hyperallometric relationship,which was consistent with inverse Rensch’s rule(the degree of SSD increased with enlarged mean body size).SSD could also be explained by sexual age difference(SAD)due to a positive SSD–SAD relationship among all populations.The findings suggest that the occurrence of inverse Rensch’s rule in B.andrewsi is likely to be a result of fecundity selection on increased reproductive investments in larger females.
文摘Previous studies on the Bufo viridis complex, which is distributed broadly across Iran, are incomplete and restricted to a few regions or a few samples. In this paper a new detailed study on the B. viridis complex in southern of Iran （from West to East） is presented. The analysis of 18 morphometric characters with univariate and multivariate methods reveals significant differences between three members of the B. viridis complex namely B. variabilis, B. luristanicus, and B. surdus distributed in southern part of Iran. Our result help to resolve an old taxonomic problem about B. surdus subgroup （taxa closely related to B. surdus） confirming that B. luristanicus and B. surdus are distinct species. Moreover, for the first time we report and describe karyotype details of B. luristanicus and B. surdus which confirmed that they are diploid. Karyological studies demonstrate that all toads from three mentioned species have 2n = 22 chromosomes. These chromosomes are arranged into two groups. First group has six large chromosomes and the second group is composed of five small chromosomes. These chromosomes are metacentric or submetacentric. The number of submetacentric chromosomes is different in three mentioned species of B. viridis complex. Neither sexual heteromorphism, nor secondary constriction was observed in any pairs of chromosomes.
基金supported by the grants from the National Natural Science Foundation of China(30770310,30970435)the Natural Science Foundation of Zhejiang,China(Y307453)
文摘Advertisement calls play an important role in influencing female mate choice and are regarded as one of the key characters responsible for reproductive isolation and speciation events in frogs and toads. The black-spined toad, Bufo melanostictus, is one of the most common amphibians in Southeast Asia and is frequently observed in human habitations throughout its distributional range. The advertisement calls of the toad have not been well described. Here, we report the advertisement calls of this species from Lishui, Zhejiang, China during its breeding season. The advertisement calls of B. melanostictus are composed of single notes with a single harmonic frequency emitted sporadically. The call parameters recorded in this study were highly variable across recording times and with different body sizes. Dominant frequency was negatively correlated with body size. Comparing the calls of the B. melanostictus population from Lishui with the populations from five other localities(Bangkok, Thailand; Coorg, India; Burma, Myanmar; Bali, Indonesia; Halimun-Salak National Park, Indonesia), we found that call structure was similar among five different populations with no harmonics, while the Halimun-Salak National Park population contained two clear harmonics. Dominant frequency was similar between Bali(1.56 kHz) and Coorg(1.6 kHz) populations, while it was highly variable among other populations. Besides call duration, call interval and pulse/call rate may also be significantly different among the comparing populations. Pooling the available data, we observed some similarities and differences in call parameters among the six populations.
基金Liaoning Provincial Department of Education-Funded Project, No. 20068040the Experimental Center of Shenyang Normal University Foundation, No. 2008007
文摘Microelectrode technique was used to record the spontaneous electrical activities of the neurons in olfactory bulb of the Bufo bufo gargarizans, both in hibernation and after hibemation. This study investigated the electrophysiological characteristics of amphibian olfactory bulb in the period of hibernation and after hibernation and its effects on the start of hibernation and spontaneous awakening. The research showed four forms of spontaneous firings： single spontaneous firing, burst spontaneous firing, irregular spontaneous firing and consecutive single spontaneous firing. The single spontaneous firing includes slow depolarized spontaneous firing and fast depolarized spontaneous firing, and the slow depolarized spontaneous firing occurs only during the hibernation period. In hibernation, the low amplitude and low frequency firing with a longer duration may be relevant to maintaining the tonicity of the central nervous system in toads that are in hibernation, and this kind of firing may also provide an excited basis for their arousal from hibernation. After hibernation, the amplitude and frequency of firing increase, but the firing duration gets shorter. This form of short-term firing, which may be a phenomenon of sensory neurons fast adapting, is one of the neuronal mechanisms for the arousal of hibernating animals.
基金supported by the National Natural Science Foundation of China(31101633)the Innovative Team Foundation of China West Normal University
文摘In many anurans, the forelimb muscles of males are used to grasp females and are often heavier than those of females despite the larger female body size. Such sexual dimorphism in forelimb musculature is thought to result from sexual selection. In addition, the hindlimbs of frogs and toads play an important role in the reproductive process as amplectant males can expel rivals with robust hindlimbs through kicking. In this study, the sexual dimorphism in dry mass for six hindlimb muscles of the Asiatic toad （Bufo gargarizans） was investigated. The results showed that, when controlled for body size, the hindlimb muscle mass of males significantly exceeded that of females for every muscle. The hindlimb muscle mass of amplectant males was also significantly larger than that of non-amplectant males. These results suggested that if strong hindlimb muscles could improve mating success of males, sexual selection would promote the evolution of dimorphism in this character.
文摘Lead has been recognized as a high risk toxic for most organisms including human. The effects of Pb in non-mammalian vertebrates are oprly known, particularly in anuran amphibians. The purpose of this study was to determine the effect of this metal on some hematological parameters of adult Bufo arenarum. It was found that all parameters remained unaltered within normal ranges, with the exception of reticulocyte counts which was significantly increased compared to the in controls (3. 7% vs. 0. 2 % ). It is suggested that the selective change found in reticulocyte count might be considered as an early response of a biomarker to sublethal exposition of Bufo arenarum to lead.
文摘Three anti-keratin MAbs were used to identifykeratins expressed in early embryos of Bufo bufogargarizans.MAb AF5 recognized three polypeptides ofkeratin in oocytes,fertilized eggs,up to neurula withMr of 68,65 and 60Kd respectively.At tailbud stage,three other keratins(62,58 and 54Kd)began to expressand could be detected by AF5.MAbs D10 and K12 gavedifferent results,both of them could identify four keratin-like molecules with unusual molecular weights(Mr 98,95,30 and 27 Kd).Moreover,D10 could also detect a 54 Kdkeratin in neurula and tailbud stage embryos,while K12could reveal,beside 54Kd keratin,other four more kera-tins(68,65,62 and 60 Kd).The possible interpretation ofthese results and their implications are discussed.
基金supported by the National Natural Science Foundation of China(Grant Nos.41541002 and 31372183)Natural Science Foundation of Zhejiang Province(Grant No.LY13C030002),China
文摘Relatively little is known about the relationship between Bufo gargarizans populations from Zhoushan Archipelago and nearby continental regions on the Pacific coast of eastern China.In this paper,155 new specimens of B.gargarizans from Zhoushan Archipelago and adjacent continents and 71 published specimens of B.gargarizans from China's Mainland were studied.Phylogeographical and dating analyses of B.gargarizans were performed using mitochondrial DNA sequencing with a length of 1436 bp.A mt DNA tree that indicated seven major clades was obtained.The earliest split in the mt DNA tree corresponding to the divergence of populations from the western highland region occurred approximately 4.0 million years ago（mya）.A subsequent clade occurred about 3.4 mya,with cladogenesis continuing toward the end of the Pleistocene.The continental clades were distributed in the western,central and northeastern regions of China.Zhoushan Archipelago clades consisted of two largely geographically overlapping subclades with the mt DNA divergence time of 0.73 mya.These results indicated there was extensive dispersal after vicariance.The B.gargarizans populations on Zhoushan Archipelago most probably originated from populations in nearby eastern continental regions of China.It was concluded that geological uplifting during the Pliocene and several sea-level changes in Pleistocene might have influenced the divergence and population demographical history of this species.
文摘The present study was carried out to investigate the histological and histochemical changes in the liver and skin on different developmental stages of Egyptian toad Bufo regularis to be used as a histological key for such species. Our experiment started when tadpoles began to feed. The adapted embryos are divided into 3 large tanks of 200 embryos each, collections of samples started from feeding age every three days. Both histological and histochemical results showed that the general architecture of the different organs was correlated with the state of development, i.e. larval, met-amorphic and post-metamorphic. They, therefore, displayed different characteristic features de-pending on the investigated developmental stage starting from the larval stage (stage 44) and ending with the post-metamorphic stage 66.
文摘The present study aimed to investigate changes in the protein contents using SDS-PAGE, also using the random amplified polymorphic DNA (RAPD-PCR) assay to identify changes in DNA concentration in the embryos of Egyptian toad Bufo regularis. Our experiment started when tadpoles began to feed. The adapted embryos were divided into 3 large tanks of 200 embryos each, collections of samples started from feeding age every three days. Cluster method was used to indicate the distinct distance between the band patterns of different stages. An increase occurred in the concentration of protein fractions in these tadpoles upon metamorphosis and that when metamorphosis was completed. DNA concentration exhibited an evident low values at the premetamorphic stage 44 (105.85 ± 0.519 ng/μl) compared with the other more advanced developmental stages 55 and 56 (204.056 ± 0.651 ng/μl and 234.55 ± 2.325 ng/μl respectively). In conclusion, acquire information about the changes in the molecular content of Bufo regularis during metamorphosis has been estimated.
基金Foundation items： This study was supported by the National Natural Science Foundation of China （31071181, 31372149） and the Students＇ Innovative Training Program of ZAFU （20120207, 20120213）
文摘In this study,to clarify the bioactive polypeptides included in the skins and secretions of Bufo,we screened the Japanese toad(Bufo japonicus formosus) skin cDNA library by colony polymerase chain reaction(PCR),and obtained a transcript of 1 075 bp consisting of 1 37 bp 5′ untranslated region(UTR),515 bp 3′ UTR and a 423 bp open reading frame(ORF) encoding a polypeptide of 140 amino acid residues(GenBank accession number: KF359945).Homolog analysis showed a 70%–96% homology with sterol carrier protein-2(SCP-2) present in other animals,which is implicated in lipid metabolism of other organisms.The gene SCP-2 of Chinese toad(B.gargarizans) was cloned from a first strand cDNA of Bufo skin(GenBank accession number: KF381341) via PCR,whose encoding polypeptide has only one amino acid difference from that of Japanese toad.Tissue distribution analysis showed that SCP-2 expressed in all organs tested,though in the liver and spleen it manifested lower expression than in other organs.These findings might indicate SCP-2 being one of the active ingredients in toad skin.These findings may in turn have implications for further drug development from traditional Chinese medicine sources.
基金Project supported by the National Natural Science Foundation of China (No. 30472139)the Education Commission for the First Batch of Excellent Young Teachers in Universities of Chongqing City, China
文摘There was a slow-relaxing tail of skeletal muscles in vitro upon the inhibition of Ca2+-pump by cyclopiazonic acid (CPA). Herein, a new linearly-combined bi-exponential model to resolve this slow-relaxing tail from the fast-relaxing phase was investigated for kinetic analysis of the isometric relaxation process of Bufo gastrocnemius in vitro, in comparison to the single exponential model and the classical bi-exponential model. During repetitive stimulations at a 2-s interval by square pulses of a 2-ms duration at 12 V direct currency (DC), the isometric tension of Bufo gastrocnemius was recorded at 100 Hz. The relaxation curve with tensions falling from 90% of the peak to the 15th datum before next stimulation was analyzed by three exponential models using a program in MATLAB 6.5. Both the goodness of fit and the distribution of the residuals for the best fitting sup- ported the comparable validity of this new bi-exponential model for kinetic analysis of the relaxation process of the control muscles. After CPA treatment, however, this new bi-exponential model showed an obvious statistical superiority for kinetic analysis of the muscle relaxation process, and it gave the estimated rest tension consistent to that by experimentation, whereas both the classical bi-exponential model and the single exponential model gave biased rest tensions. Moreover, after the treatment of muscles by CPA, both the single exponential model and the classical bi-exponential model yielded lowered relaxation rates, nevertheless, this new bi-exponential model had relaxation rates of negligible changes except much higher rest tensions. These results suggest that this novel linearly-combined bi-exponential model is desirable for kinetic analysis of the relaxation process of muscles with altered Ca2+-pumping activity.
