Analyzing the structure and function of the brain's neural network is critical for identifying the working principles of the brain and the mechanisms of brain diseases.Recombinant rabies viral vectors allow for th...Analyzing the structure and function of the brain's neural network is critical for identifying the working principles of the brain and the mechanisms of brain diseases.Recombinant rabies viral vectors allow for the retrograde labeling of projection neurons and cell type-specific trans-monosynaptic tracing,making these vectors powerful candidates for the dissection of synaptic inputs.Although several attenuated rabies viral vectors have been developed,their application in studies of functional networks is hindered by the long preparation cycle and low yield of these vectors.To overcome these limitations,we developed an improved production system for the rapid rescue and preparation of a high-titer CVS-N2c-ΔG virus.Our results showed that the new CVS-N2c-ΔG-based toolkit performed remarkably:(1)N2cG-coated CVS-N2c-ΔG allowed for efficient retrograde access to projection neurons that were unaddressed by rAAV9-Retro,and the efficiency was six times higher than that of rAAV9-Retro;(2)the trans-monosynaptic efficiency of oG-mediated CVS-N2c-ΔG was 2–3 times higher than that of oG-mediated SAD-B19-ΔG;(3)CVS-N2c-ΔG could delivery modified genes for neural activity monitoring,and the time window during which this was maintained was 3 weeks;and(4)CVS-N2c-ΔG could express sufficient recombinases for efficient transgene recombination.These findings demonstrate that new CVS-N2c-ΔG-based toolkit may serve as a versatile tool for structural and functional studies of neural circuits.展开更多
Potted rice seedlings independently treated with N,P,and NP were continuously^(13)CO_(2) labeled to investigated the influence of N and P application on the contribution of photosynthesized C to the rhizosphere versus...Potted rice seedlings independently treated with N,P,and NP were continuously^(13)CO_(2) labeled to investigated the influence of N and P application on the contribution of photosynthesized C to the rhizosphere versus bulk soil and particulate organic matter(POM)versus mineral fraction(MIN).N and NP enhanced net assimilated^(13)C on day 14(D14),with maximum C assimilation occurring on day 22(D22)under NP.Aboveground biomass retained more^(13)C than belowground biomass for all treatments.^(13)C incorporation into the rhizosphere exceeded that in bulk soil,with the maximum(6-10%)found under N addition.Newly assimilated^(13)incorporated into POM increased in the rhizosphere under N and NP conditions,whereas MIN remained largely unaffected.^(13)C-MBC proportion in the total microbial biomass C(MBC)pool revealed that N and NP stimulated microbial activity to a greater degree than P.The main portion of^(13)C in the rhizosphere and bulk soil was found in POM on D14,which decreased over time due to microbial utilization.Contrastingly,root-derived ^(13)C in the MIN remained unchanged between sampling days,which indicates that the stabilization of rhizodeposits in this fraction might be the potential mechanism underlying SOM sequestration in paddy soils.展开更多
基金supported by the National Natural Science Foundation of China,Nos.32100899(to KZL),31830035(to FQX),31771156(to FQX),21921004(to FQX)the National Science and Technology Innovation 2030,No.2021ZD0201003(to FQX)+2 种基金the Key-Area Research and Development Program of Guangdong Province,No.2018B030331001(to FQX)the Strategic Priority Research Program of the Chinese Academy of Sciences,No.XDB32030200(to FQX)the Shenzhen Key Laboratory of Viral Vectors for Biomedicine,No.ZDSYS20200811142401005(to FQX)。
文摘Analyzing the structure and function of the brain's neural network is critical for identifying the working principles of the brain and the mechanisms of brain diseases.Recombinant rabies viral vectors allow for the retrograde labeling of projection neurons and cell type-specific trans-monosynaptic tracing,making these vectors powerful candidates for the dissection of synaptic inputs.Although several attenuated rabies viral vectors have been developed,their application in studies of functional networks is hindered by the long preparation cycle and low yield of these vectors.To overcome these limitations,we developed an improved production system for the rapid rescue and preparation of a high-titer CVS-N2c-ΔG virus.Our results showed that the new CVS-N2c-ΔG-based toolkit performed remarkably:(1)N2cG-coated CVS-N2c-ΔG allowed for efficient retrograde access to projection neurons that were unaddressed by rAAV9-Retro,and the efficiency was six times higher than that of rAAV9-Retro;(2)the trans-monosynaptic efficiency of oG-mediated CVS-N2c-ΔG was 2–3 times higher than that of oG-mediated SAD-B19-ΔG;(3)CVS-N2c-ΔG could delivery modified genes for neural activity monitoring,and the time window during which this was maintained was 3 weeks;and(4)CVS-N2c-ΔG could express sufficient recombinases for efficient transgene recombination.These findings demonstrate that new CVS-N2c-ΔG-based toolkit may serve as a versatile tool for structural and functional studies of neural circuits.
基金This work was funded by the National Natural Science Foundation of China[41877104,41950410565,41811540031]Hunan Province Base for Scientific and Technological Innovation Cooperation[2018WK4012]+6 种基金Natural Science Foundation of Hunan Province[2019JJ10003,2019JJ30028]the Youth Innovation Team Project of the Institute of Subtropical Agriculture,Chinese Academy of Sciences[2017QNCXTD_GTD]Talented Young Scientist Program(TYSP)supported by China Science and Technology Exchange Center(CSTEC)the Chinese Academy of Sciences President’s International Fellowship Initiative awarded to Anna Gunina[2019VCC0003]Tin Mar Lynn[2018PC0078]China National Key R&D Program[2019YFC0605004]Jiangxi Province Scienc and Technology Planned Project[20202BBG73007,20203BBG73068].
文摘Potted rice seedlings independently treated with N,P,and NP were continuously^(13)CO_(2) labeled to investigated the influence of N and P application on the contribution of photosynthesized C to the rhizosphere versus bulk soil and particulate organic matter(POM)versus mineral fraction(MIN).N and NP enhanced net assimilated^(13)C on day 14(D14),with maximum C assimilation occurring on day 22(D22)under NP.Aboveground biomass retained more^(13)C than belowground biomass for all treatments.^(13)C incorporation into the rhizosphere exceeded that in bulk soil,with the maximum(6-10%)found under N addition.Newly assimilated^(13)incorporated into POM increased in the rhizosphere under N and NP conditions,whereas MIN remained largely unaffected.^(13)C-MBC proportion in the total microbial biomass C(MBC)pool revealed that N and NP stimulated microbial activity to a greater degree than P.The main portion of^(13)C in the rhizosphere and bulk soil was found in POM on D14,which decreased over time due to microbial utilization.Contrastingly,root-derived ^(13)C in the MIN remained unchanged between sampling days,which indicates that the stabilization of rhizodeposits in this fraction might be the potential mechanism underlying SOM sequestration in paddy soils.
