To explore the effect of the extracellular ATP and its receptors on axon regeneration in the sciatic nerve defect in rats, 0.5 cm defect of the sciatic nerve was made and repaired with long arm silicon tube like a ...To explore the effect of the extracellular ATP and its receptors on axon regeneration in the sciatic nerve defect in rats, 0.5 cm defect of the sciatic nerve was made and repaired with long arm silicon tube like a 'Y' type. The single arm of the silicon tube was sutured to proximal of the sciatic nerve. 10 μl 1 mmol/L ATP in physiological saline was injected into the left chamber of the silicon tube (experimental group) and physiological saline injected into another silicon tube as a control group. In another model 1 mmol/L 10 μl ATP and 1 mmol/L 10 μl ATP+0.2 mg/ml suramin were injected respectively into two arms of the silicon tube. After 4 and 8 weeks the specimens were obtained from the silicon tube for examining the axon regeneration histologically and image analysis. All the regeneration axons grew into the silicon tube containing the ATP, but there was no axon regeneration in the silicon tube containing the ATP+Suramin or physiological saline. It was demonstrated that extracellular ATP had a powerful attraction to the regenerated axon of peripheral nerve. The suramin inhibited the axon induction of the extracellular ATP completely via the ATP receptors.展开更多
ATP was added to the cultured sensory neurons obtained from the dorsal root ganglia of the neonatal rats and PBS was added to serve as control. MTT assays were conducted to evaluate the survival and activity of the cu...ATP was added to the cultured sensory neurons obtained from the dorsal root ganglia of the neonatal rats and PBS was added to serve as control. MTT assays were conducted to evaluate the survival and activity of the cultured neurons. And the silicone regenerative chamber was used after the sciatic nerve incision of the mature SD rat. 1 mmol/L ATP was injected into the left chamber and 0.09 % natrium chloride was injected into the right chamber as controls. The changes of nitric oxide synthase (NOS) activity in the corresponding dorsal root ganglia were measured histochemically and image analysis was also performed 4 days after the sciatic nerve injury. The results showed that extracellular ATP could enhance the survival of the neurons and the number of NOS positive neurons were significantly different between the ATP and control groups ( P <0.05). It was suggested that extracellular ATP had neurotrophic effect on neurons survival and could inhibit the NOS activity of the sensory neurons after the peripheral nerve incision, hence exerting the protective effect on the neurons, which was valuable for nerve regeneration after nerve injury.展开更多
Cadmium(Cd)is one of the most widespread and toxic heavy metals to plants.Extracellular ATP(exATP)is thought to be an extracellular effector in regulating the physiological responses of plant cells to environmental st...Cadmium(Cd)is one of the most widespread and toxic heavy metals to plants.Extracellular ATP(exATP)is thought to be an extracellular effector in regulating the physiological responses of plant cells to environmental stresses.However,the function of exATP in Cd-stressed plant cells is much unknown.The present work showed that treating tobacco(Nicotiana tabacum L.cv.Bright Yellow-2)cell-suspension cultures with exogenous CdCl2 reduced the cell viability,exATP level,and Mg content.However,the production of reactive oxygen species(ROS),Cd content,and electrolyte leakage of the cells were enhanced by exogenous CdCl2.When the Cd-induced accumulation of ROS was decreased by the supplement with DMTU(dimethylthiourea,a scavenger of ROS),the Cd-induced increases of the electrolyte leakage and Cd content were alleviated,and the Cd-induced reductions of cell viability were partly rescued,suggesting that Cd-induced reduction of cell viability could be related to the ROS accumulation.Under the condition of Cd stress,when the reduction of exATP level was partly rescued by exogenous ATP(20μM),the increases of ROS production,electrolyte leakage,and Cd content were attenuated,and the reduction of cell viability was also alleviated.These observations indicate that exATP can regulate the cell viability in the Cd–stressed plant cells possibly by an ROS-associated mechanism.展开更多
In recent years, adenosine tri-phosphate （ATP） has been reported to exist in apoplasts of plant cells as a signal molecule. Extracellular ATP （eATP） plays important roles in plant growth, development, and stress t...In recent years, adenosine tri-phosphate （ATP） has been reported to exist in apoplasts of plant cells as a signal molecule. Extracellular ATP （eATP） plays important roles in plant growth, development, and stress tolerance. Here, extra- cellular ATP was found to promote stomatal opening of Arabidopsis thaliana in light and darkness. ADP, GTP, and weakly hydrolyzable ATP analogs （ATPγS, Bz-ATP, and 2meATP） showed similar effects, whereas AMP and adenosine did not affect stomatal movement. Apyrase inhibited stomatal opening. ATP-promoted stomatal opening was blocked by an NADPH oxidase inhibitor （diphenylene iodonium） or deoxidizer （dithiothreitol）, and was impaired in null mutant of NADPH ox- idase （atrbohD/F）. Added ATP triggered ROS generation in guard cells via NADPH oxidase. ATP also induced Ca^2＋ influx and H ＋ efflux in guard cells. In atrbohD/F, ATP-induced ion flux was strongly suppressed. In null mutants of the heterotrimeric G protein α subunit, ATP-promoted stomatal opening, cytoplasmic ROS generation, Ca^2＋ influx, and ^H＋ efflux were all sup- pressed. These results indicated that eATP-promoted stomatal opening possibly involves the heterotrimeric G protein, ROS, cytosolic Ca^2＋, and plasma membrane H＋-ATPase.展开更多
Extracellular adenosine 5’-triphosphate(ATP) is a key signaling molecule present in the central nervous system(CNS),and now is receiving greater attention due to its role as a messenger in the CNS during different ph...Extracellular adenosine 5’-triphosphate(ATP) is a key signaling molecule present in the central nervous system(CNS),and now is receiving greater attention due to its role as a messenger in the CNS during different physiological and pathological events. ATP is released into the extracellular space through vesicular exocytosis or from damaged and dying cells. Once in the extracellular environment,ATP binds to the specific receptors termed P2,which mediate ATP effects and are present broadly in both neurons an...展开更多
Hydrogen sulfide （H2S） is a newly discovered gaseous signaling molecule and involved in ethylene and ABA-induced stomatal closure. As an important factor, extracellular ATP （eATP） was believed to participate in re...Hydrogen sulfide （H2S） is a newly discovered gaseous signaling molecule and involved in ethylene and ABA-induced stomatal closure. As an important factor, extracellular ATP （eATP） was believed to participate in regulation of stomatal closing. However, the mechanism by which eATP mediates HES-regulated stomatal closure remains unclear. Here, we employed Arabidopsis wild-type and mutant lines of ATP-binding cassette transporters （Atmrp4, Atmrp5 and their double mutant Atmrp4/5） to study the function of eATP in H_2S-regulated stomatal movement. Our results indicated that H_2S affected stomatal closing through stimulating guard cell outward K^＋ current. Moreover, we found that HES induced eATP generation by regulating the activity of an ABC transporter. The inhibitor of ABC transporters, glibenclamide （Gli）, could impair H_2S-regulated stomatal closure and reduce H_2S-dependent eATP accumulation in Atmrp4 and Atmrp5 mutants. In addition, the promotion effect of H_2S on outward K^＋ currents was diminished in Atmrp4/5 double mutant. Our data suggested that hydrogen peroxide （H_2O_2） is required for H_2S-induced stomatal closure, and the production of H_2O_2 is regulated by eATP via NADPH oxidase. Based on this work, we conclude that H_2S-induced stomatal closure requires ABC transporter-dependent eATP pro- duction and subsequent NADPH oxidase-dependent H_2O_2 accumulation.展开更多
文摘To explore the effect of the extracellular ATP and its receptors on axon regeneration in the sciatic nerve defect in rats, 0.5 cm defect of the sciatic nerve was made and repaired with long arm silicon tube like a 'Y' type. The single arm of the silicon tube was sutured to proximal of the sciatic nerve. 10 μl 1 mmol/L ATP in physiological saline was injected into the left chamber of the silicon tube (experimental group) and physiological saline injected into another silicon tube as a control group. In another model 1 mmol/L 10 μl ATP and 1 mmol/L 10 μl ATP+0.2 mg/ml suramin were injected respectively into two arms of the silicon tube. After 4 and 8 weeks the specimens were obtained from the silicon tube for examining the axon regeneration histologically and image analysis. All the regeneration axons grew into the silicon tube containing the ATP, but there was no axon regeneration in the silicon tube containing the ATP+Suramin or physiological saline. It was demonstrated that extracellular ATP had a powerful attraction to the regenerated axon of peripheral nerve. The suramin inhibited the axon induction of the extracellular ATP completely via the ATP receptors.
文摘ATP was added to the cultured sensory neurons obtained from the dorsal root ganglia of the neonatal rats and PBS was added to serve as control. MTT assays were conducted to evaluate the survival and activity of the cultured neurons. And the silicone regenerative chamber was used after the sciatic nerve incision of the mature SD rat. 1 mmol/L ATP was injected into the left chamber and 0.09 % natrium chloride was injected into the right chamber as controls. The changes of nitric oxide synthase (NOS) activity in the corresponding dorsal root ganglia were measured histochemically and image analysis was also performed 4 days after the sciatic nerve injury. The results showed that extracellular ATP could enhance the survival of the neurons and the number of NOS positive neurons were significantly different between the ATP and control groups ( P <0.05). It was suggested that extracellular ATP had neurotrophic effect on neurons survival and could inhibit the NOS activity of the sensory neurons after the peripheral nerve incision, hence exerting the protective effect on the neurons, which was valuable for nerve regeneration after nerve injury.
基金the tobacco cell culture.This work was supported by the n ational n atural Science Foundation of China(n O.31870246,31560059,and 31260059)the Fundamental Research Funds for the Gansu Universities of Gansu Provincial Department of Finance,the University Scientific Research Project of Gansu Province(n o.2015A-007)+1 种基金the Key Research and Development Project of Gansu Province(n o.18YF1 n A051)the Youth Teacher Scientific Research Ability Promotion Plan Innovation Team Project of n orthwest n ormal University.
文摘Cadmium(Cd)is one of the most widespread and toxic heavy metals to plants.Extracellular ATP(exATP)is thought to be an extracellular effector in regulating the physiological responses of plant cells to environmental stresses.However,the function of exATP in Cd-stressed plant cells is much unknown.The present work showed that treating tobacco(Nicotiana tabacum L.cv.Bright Yellow-2)cell-suspension cultures with exogenous CdCl2 reduced the cell viability,exATP level,and Mg content.However,the production of reactive oxygen species(ROS),Cd content,and electrolyte leakage of the cells were enhanced by exogenous CdCl2.When the Cd-induced accumulation of ROS was decreased by the supplement with DMTU(dimethylthiourea,a scavenger of ROS),the Cd-induced increases of the electrolyte leakage and Cd content were alleviated,and the Cd-induced reductions of cell viability were partly rescued,suggesting that Cd-induced reduction of cell viability could be related to the ROS accumulation.Under the condition of Cd stress,when the reduction of exATP level was partly rescued by exogenous ATP(20μM),the increases of ROS production,electrolyte leakage,and Cd content were attenuated,and the reduction of cell viability was also alleviated.These observations indicate that exATP can regulate the cell viability in the Cd–stressed plant cells possibly by an ROS-associated mechanism.
基金This work was supported by the National Science Foundation of China，the Program for New Century Excellent Talents in University，the State Key Laboratory of Plant Cell and Chromosome Engineering，No conflict of interest declared
文摘In recent years, adenosine tri-phosphate （ATP） has been reported to exist in apoplasts of plant cells as a signal molecule. Extracellular ATP （eATP） plays important roles in plant growth, development, and stress tolerance. Here, extra- cellular ATP was found to promote stomatal opening of Arabidopsis thaliana in light and darkness. ADP, GTP, and weakly hydrolyzable ATP analogs （ATPγS, Bz-ATP, and 2meATP） showed similar effects, whereas AMP and adenosine did not affect stomatal movement. Apyrase inhibited stomatal opening. ATP-promoted stomatal opening was blocked by an NADPH oxidase inhibitor （diphenylene iodonium） or deoxidizer （dithiothreitol）, and was impaired in null mutant of NADPH ox- idase （atrbohD/F）. Added ATP triggered ROS generation in guard cells via NADPH oxidase. ATP also induced Ca^2＋ influx and H ＋ efflux in guard cells. In atrbohD/F, ATP-induced ion flux was strongly suppressed. In null mutants of the heterotrimeric G protein α subunit, ATP-promoted stomatal opening, cytoplasmic ROS generation, Ca^2＋ influx, and ^H＋ efflux were all sup- pressed. These results indicated that eATP-promoted stomatal opening possibly involves the heterotrimeric G protein, ROS, cytosolic Ca^2＋, and plasma membrane H＋-ATPase.
文摘Extracellular adenosine 5’-triphosphate(ATP) is a key signaling molecule present in the central nervous system(CNS),and now is receiving greater attention due to its role as a messenger in the CNS during different physiological and pathological events. ATP is released into the extracellular space through vesicular exocytosis or from damaged and dying cells. Once in the extracellular environment,ATP binds to the specific receptors termed P2,which mediate ATP effects and are present broadly in both neurons an...
基金supported by the National Natural Science Foundation of China(31170237)the National Key Laboratory Program of China Agricultural University(SKLPPBKF11001)Shandong Taishan Scholar Program
文摘Hydrogen sulfide （H2S） is a newly discovered gaseous signaling molecule and involved in ethylene and ABA-induced stomatal closure. As an important factor, extracellular ATP （eATP） was believed to participate in regulation of stomatal closing. However, the mechanism by which eATP mediates HES-regulated stomatal closure remains unclear. Here, we employed Arabidopsis wild-type and mutant lines of ATP-binding cassette transporters （Atmrp4, Atmrp5 and their double mutant Atmrp4/5） to study the function of eATP in H_2S-regulated stomatal movement. Our results indicated that H_2S affected stomatal closing through stimulating guard cell outward K^＋ current. Moreover, we found that HES induced eATP generation by regulating the activity of an ABC transporter. The inhibitor of ABC transporters, glibenclamide （Gli）, could impair H_2S-regulated stomatal closure and reduce H_2S-dependent eATP accumulation in Atmrp4 and Atmrp5 mutants. In addition, the promotion effect of H_2S on outward K^＋ currents was diminished in Atmrp4/5 double mutant. Our data suggested that hydrogen peroxide （H_2O_2） is required for H_2S-induced stomatal closure, and the production of H_2O_2 is regulated by eATP via NADPH oxidase. Based on this work, we conclude that H_2S-induced stomatal closure requires ABC transporter-dependent eATP pro- duction and subsequent NADPH oxidase-dependent H_2O_2 accumulation.
