The plateau zokor ( Eospalax baileyi) is a species of subterranean rodent endemic to the Tibetan Plateau. It is well adapted to the cold and hypoxic and hypercapnic burrow. To study the oxygenation properties of plateau zokor hemoglobins (Hbs), we measured intrinsic Hb-O2 affinities and their sensitivities to pH (Bohr effect); CO2; Cl−, 2,3-diphosphoglycerate (DPG); and temperature using purified Hbs from zokor and mouse. The optimal deoxyHb model of plateau zokor was constructed and used to study its structural characteristics by molecular dynamics simulations. O2 binding results revealed that plateau zokor Hbs exhibit remarkably high intrinsic Hb-O2 affinity, low CO2 effects compared with human and the relatively low anion allosteric effector sensitivities (DPG and Cl−) at normal temperature, which would safeguard the pulmonary Hb-O2 loading under hypoxic and hypercapnic conditions. Furthermore, the high anion allosteric effector sensitivities at low temperature and low temperature sensitivities of plateau zokor Hbs would facilitate the releasing of O2 in cold extremities and metabolic tissues. However, the high Hb-O2 affinity of plateau zokor is not compensated by high pH sensitivity as the Bohr factors of plateau zokor Hbs were as low as those of mouse. The results of molecular dynamics simulations revealed the reduced hydrogen bonding between the α1β1- and α2β2-dimer interface of deoxyHb in zokor compared with mouse. It may be the primary mechanism of the high intrinsic Hb-O2 affinities in zokor. Specifically, substitution of the 131Ser→Asn in the α2-chain weakened the connection between α1- and β2-subunit.
Ecological immunology involves the study of the immune function of wildlife, which is seldom compared with that of model animals. Here, we evaluated and compared the level of the innate immune response in the plateau zokor (Eospalax baileyi), an indigenous underground rodent from the Tibetan Plateau, with that in the bamboo rat (Rhizomys pruinosus) and Sprague‐Dawley (SD) rat (Rattus norvegicus). The spleen was observed by ordinary light and transmission electron microscopy, and the spleen index was calculated. After liposaccharide (LPS) challenge, the expression of Toll‐like receptor 2 (TLR2), TLR4, and hypoxia‐inducible factor 1α (HIF‐1α) in the spleen was detected by Western blot analysis and immunofluorescence. The expression of nuclear factor‐κB1 (NF‐κB1) and mitogen‐activated protein kinase 14 (MAPK14) in the spleen was detected by real‐time quantitative polymerase chain reaction, and the levels of interleukin 6 (IL‐6), tumor necrosis factor‐α (TNF‐α), and interferon‐β (IFN‐β) in the spleen were detected by enzyme‐linked immunoassay. The spleen index of the plateau zokor was lower than that of the bamboo rat and SD rat. The expression of TLR4, NF‐κB1, and MAPK14 and the levels of IL‐6 and TNF‐α in the spleen of the plateau zokor were lower than those of the bamboo rat and SD rat, while the expression of TLR2 and HIF‐1α and the level of IFN‐β were higher than those of the bamboo rat and SD rat. We speculate that suppression of the TLR4 signaling pathway in the plateau zokor is an adaptation to hypoxic tunnels that decreases antigenic risk and maintains immune homeostasis. Moreover, the spleen of the plateau zokor is reduced in size, reducing the innate immunity investment in the spleen. We also noted that high levels of HIF‐1α in the spleen of the plateau zokor suppressed crosstalk between HIF‐1α and TLR4, promoting the innate immune response.
Both the warming climate and ultraviolet-B radiation (UVBR) are considered to be notable environmental factors affecting amphibian population decline, with particular effects on tadpole development. However, the phenotypes of tadpoles living at high altitudes may be improved by moderately warming temperatures, reducing or eliminating the negative effects of oxidative damage caused by cool temperatures or strong UVBR at high altitudes. To verify this hypothesis, Rana kukunoris tadpoles, which live at high altitudes, were used to test the effect of the interaction of temperature and UVBR on their development and antioxidant systems in a fully factorial design. The tadpoles were exposed to UVBR and UVBR-free environments at 14 (cool temperature) and 22 °C (warm temperature), respectively. UVBR and a warm temperature had opposite influences on several traits of the tadpoles, including their survival, developmental rate, individual size, preferred body temperature, thermal tolerance temperature, oxidative damage, and enzymatic and nonenzymatic antioxidant systems. The moderate temperature could compensate for or override the negative effects of UVBR by increasing the tadpoles’ preferred body temperature and critical tolerance temperature, thus enhancing the locomotion ability and thermal sensitivity of their antioxidant systems. Furthermore, the dark skin coloration and aggregation behavior of R. kukunoris tadpoles may also be effective strategies for allowing them to resist UVBR and helping them to better adapt to a warming environment with stronger UVBR. Thus, it is possible that a moderate degree of warming may increase the capacity of living organisms to adapt to environmental changes and thus have positive effects on the development of tadpoles living at high altitudes.
Investigating how highland amphibians respond to changes in ambient temperature may be of great significance for their fate prediction and effective conservation in the background of global warming. Here, using field individuals as the control group, we investigated the influence of high temperatures (20.5 and 25.5°C) and heat wave (15–26.6°C) on the thermal preference, critical thermal limits, locomotor performance, oxidative stress, and antioxidant enzyme activities in high-altitude frog Nanorana pleskei (3,490 m) endemic to the Qinghai-Tibet Plateau (QTP). After 2 weeks of acclimation to high temperatures and heat wave, the thermal preference (Tpref), critical thermal maximum (CTmax), and range of tolerable temperature significantly increased, while the critical thermal minimum (CTmin) was significantly decreased. The total time of jump to exhaustion significantly decreased, and burst swimming speed significantly increased in frogs acclimated in the high temperature and heat wave groups compared with the field group. In the high temperature group, the level of H2O2 and lipid peroxide (malondialdehyde, MDA), as well as the activities of glutathione peroxidase (GPX), glutathione reductase (GR), catalase (CAT), superoxide dismutase (SOD), and total antioxidant capacity (T-AOC) significantly increased in the liver or muscle. However, in the heat wave group, the MDA content significantly decreased in the liver, and antioxidants activities decreased in the liver and muscle except for CAT activities that were significantly increased in the liver. These results indicated that N. pleskei could respond to the oxidative stress caused by high temperatures by enhancing the activity of antioxidant enzymes. The heat wave did not appear to cause oxidative damage in N. pleskei, which may be attributed to the fact that they have successfully adapted to the dramatic temperature fluctuations on the QTP.
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