Differentially expressed genes associated with adaptation to different thermal environments in three sympatric Cuban <i>Anolis</i> lizards

Akashi, Hiroshi; Díaz, Antonio Cádiz; Shigenobu, Shuji; Makino, Takashi; Kawata, Masakado

doi:10.1111/mec.13625

Cited by 38 publications

(40 citation statements)

References 32 publications

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“…We performed RT-PCR using primers that were mainly designed on the basis of RNA sequence data from A . sagrei [30], and we isolated HSF1 , HSF2 , and HSF3 cDNA clones from the hindlimb, and HSF4 cDNA clones from the eye [13]. Frog cDNA clones for HSF1 and HSF2 were isolated previously from the African clawed frog, Xenopus laevis , which has a tetraploid genome [35, 36].…”

Section: Resultsmentioning

confidence: 99%

“…The PCR primers shown in S1 Table were designed based on RNA sequence data from A . sagrei [30] and Ensembl genome sequence data from Anolis carolinensis (http://www.ensembl.org/index.html). The sequences of multiple cDNA clones from each HSF gene were verified using a BigDye Terminator v3.1 Cycle Sequencing Kit and 3500 Genetic Analyzer (Applied Biosystems), and one representative full-length cDNA clone per gene (pTOPO2.1-AsHSF1 to pTOPO2.1-AsHSF4) was selected for analysis.…”

Section: Methodsmentioning

confidence: 99%

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HSF1 and HSF3 cooperatively regulate the heat shock response in lizards

et al. 2017

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Cells cope with temperature elevations, which cause protein misfolding, by expressing heat shock proteins (HSPs). This adaptive response is called the heat shock response (HSR), and it is regulated mainly by heat shock transcription factor (HSF). Among the four HSF family members in vertebrates, HSF1 is a master regulator of HSP expression during proteotoxic stress including heat shock in mammals, whereas HSF3 is required for the HSR in birds. To examine whether only one of the HSF family members possesses the potential to induce the HSR in vertebrate animals, we isolated cDNA clones encoding lizard and frog HSF genes. The reconstructed phylogenetic tree of vertebrate HSFs demonstrated that HSF3 in one species is unrelated with that in other species. We found that the DNA-binding activity of both HSF1 and HSF3 in lizard and frog cells was induced in response to heat shock. Unexpectedly, overexpression of lizard and frog HSF3 as well as HSF1 induced HSP70 expression in mouse cells during heat shock, indicating that the two factors have the potential to induce the HSR. Furthermore, knockdown of either HSF3 or HSF1 markedly reduced HSP70 induction in lizard cells and resistance to heat shock. These results demonstrated that HSF1 and HSF3 cooperatively regulate the HSR at least in lizards, and suggest complex mechanisms of the HSR in lizards as well as frogs.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

HSF1 and HSF3 cooperatively regulate the heat shock response in lizards

et al. 2017

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“…The cDNA was synthesized using high‐capacity cDNA reverse transcription kit (ABI). Anolis trpa1 full‐length cDNAs were amplified by two‐step PCR, and the primers were designed on the 5′ and 3′ untranslated regions (UTR) according to the TRPA1 sequences obtained from the transcriptomic study performed on the three species (Akashi et al., ). For the second‐step PCR, we designed forward and reverse primers with additional sequences containing Bam HI and Not I restriction enzyme digestion sites, respectively.…”

Section: Methodsmentioning

confidence: 99%

Comparisons of behavioural and TRPA1 heat sensitivities in three sympatric Cuban Anolis lizards

et al. 2018

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Thermal tolerances of organisms play a role in defining geographic ranges and occurrence of species. In Cuba, three sympatric species of Anolis lizards (Anolis allogus, Anolis homolechis and Anolis sagrei) inhabit different thermal microhabitats. A previous study found that these species showed distinct gene expression patterns in response to temperature stimuli, suggesting the genetically distinct thermal physiology among species. To investigate whether the Anolis species inhabiting locally distinct thermal habitats diverge their thermal tolerances, we first conducted behavioural experiments to analyse the temperatures at which the three Anolis species escape from heat source. Then, for each of the three species, we isolated cDNA encoding a putative molecular heat sensor, transient receptor potential ion channel ankyrin 1 (TRPA1), which has been suggested to play a role on eliciting behavioural responses to heat stimuli. We performed electrophysiological analysis to quantify activation temperature of Anolis TRPA1 to see whether the pattern of divergence in TRPA1 responses is congruent with that of divergence in behavioural responses. We found that temperatures triggering behavioural and TRPA1 responses were significantly lower for shade-dwelling species (A. allogus) than for sun-dwelling species (A. homolechis and A. sagrei). The ambient temperature of shade habitats where A. allogus occurs stays relatively cool compared to that of open habitats where A. homolechis and A. sagrei occur and bask. The high temperature thresholds of A. homolechis and A. sagrei may reflect their heat tolerances that would benefit these species to inhabit the open habitats.

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“…Moreover, biochemical, and molecular genetic studies in female P. sinensis have discovered a number of apoptosis-related genes in which some critical genes were also demonstrated to be involved in spermatogenesis (Zhang et al, 2013; Liu et al, 2016b). Recently, the RNA sequencing (RNA-Seq) technique serves as an accurate and appropriate approach when combined with digital gene expression (DGE) tag profiling, and it has been successfully used for the expression analysis of transcriptional changes in some reptile species, including brown treesnake ( Boiga irregularis ), anolis lizard ( Anolis allogus ), alligator ( Alligator mississippiensis ), and western painted turtle ( Chrysemys picta bellii ; Mcgivern et al, 2014; Akashi et al, 2016; Jiang et al, 2016; Yatsu et al, 2016). Nevertheless, there is no report concerning the global expression profiling of apoptosis-related genes during the process of dissociated spermatogenesis, and the mechanism underlying spermatogenesis was largely unexplored in P. sinensis .…”

Section: Introductionmentioning

confidence: 99%

Molecular and Cellular Mechanisms of Apoptosis during Dissociated Spermatogenesis

et al. 2017

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Apoptosis is a tightly controlled process by which tissues eliminate unwanted cells. Spontaneous germ cell apoptosis in testis has been broadly investigated in mammals that have an associated spermatogenesis pattern. However, the mechanism of germ cell apoptosis in seasonally breeding reptiles following a dissociated spermatogenesis has remained enigmatic. In the present study, morphological evidence has clearly confirmed the dissociated spermatogenesis pattern in Pelodiscus sinensis. TUNEL and TEM analyses presented dynamic changes and ultrastructural characteristics of apoptotic germ cells during seasonal spermatogenesis, implying that apoptosis might be one of the key mechanisms to clear degraded germ cells. Furthermore, using RNA-Seq and digital gene expression (DGE) profiling, a large number of apoptosis-related differentially expressed genes (DEGs) at different phases of spermatogenesis were identified and characterized in the testis. DGE and RT-qPCR analysis revealed that the critical anti-apoptosis genes, such as Bcl-2, BAG1, and BAG5, showed up-regulated patterns during intermediate and late spermatogenesis. Moreover, the increases in mitochondrial transmembrane potential in July and October were detected by JC-1 staining. Notably, the low protein levels of pro-apoptotic cleaved caspase-3 and CytC in cytoplasm were detected by immunohistochemistry and western blot analyses, indicating that the CytC-Caspase model might be responsible for the effects of germ cell apoptosis on seasonal spermatogenesis. These results facilitate understanding the regulatory mechanisms of apoptosis during spermatogenesis and uncovering the biological process of the dissociated spermatogenesis system in reptiles.

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Differentially expressed genes associated with adaptation to different thermal environments in three sympatric Cuban Anolis lizards

Cited by 38 publications

References 32 publications

HSF1 and HSF3 cooperatively regulate the heat shock response in lizards

HSF1 and HSF3 cooperatively regulate the heat shock response in lizards

Comparisons of behavioural and TRPA1 heat sensitivities in three sympatric Cuban Anolis lizards

Molecular and Cellular Mechanisms of Apoptosis during Dissociated Spermatogenesis

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