Transfer RNAs(tRNA)are crucial adaptor molecules between messenger RNA(mRNA)and amino acids.Recent evidence in plants suggests that dicistronic tRNA-like structures also act as mobile signals for mRNA transcripts to m...Transfer RNAs(tRNA)are crucial adaptor molecules between messenger RNA(mRNA)and amino acids.Recent evidence in plants suggests that dicistronic tRNA-like structures also act as mobile signals for mRNA transcripts to move between distant tissues.Co-transcription is not a common feature in the plant nuclear genome and,in the few cases where polycistronic transcripts have been found,they include non-coding RNA species,such as small nucleolar RNAs and microRNAs.It is not known,however,the extent to which dicistronic transcripts of tRNA and mRNAs are expressed in field-grown plants,or the factors contributing to their expression.We analysed tRNA–mRNA dicistronic transcripts in the major horticultural crop grapevine(Vitis vinifera)using a novel pipeline developed to identify dicistronic transcripts from high-throughput RNA-sequencing data.We identified dicistronic tRNA–mRNA in leaf and berry samples from 22 commercial vineyards.Of the 124 tRNA genes that were expressed in both tissues,18 tRNA were expressed forming part of 19 dicistronic tRNA–mRNAs.The presence and abundance of dicistronic molecules was tissue and geographic sub-region specific.In leaves,the expression patterns of dicistronic tRNA–mRNAs significantly correlated with tRNA expression,suggesting that their transcriptional regulation might be linked.We also found evidence of syntenic genomic arrangements of tRNAs and protein-coding genes between grapevine and Arabidopsis thaliana,and widespread prevalence of dicistronic tRNA–mRNA transcripts among vascular land plants but no evidence of these transcripts in non-vascular lineages.This suggests that the appearance of plant vasculature and tRNA–mRNA occurred concurrently during the evolution of land plants.展开更多
Overnutrition causes hyperactivation of mTORC1-dependent negative feedback loops leading to the downregulation of insulin signaling and development of insulin resistance.In osteoblasts(OBs),insulin signaling plays a c...Overnutrition causes hyperactivation of mTORC1-dependent negative feedback loops leading to the downregulation of insulin signaling and development of insulin resistance.In osteoblasts(OBs),insulin signaling plays a crucial role in the control of systemic glucose homeostasis.We utilized mice with conditional deletion of Rptor to investigate how the loss of mTORC1 function in OB affects glucose metabolism under normal and overnutrition dietary states.Compared to the controls,chow-fed Rptorob−/−mice had substantially less fat mass and exhibited adipocyte hyperplasia.Remarkably,upon feeding with high-fat diet,mice with pre-and post-natal deletion of Rptor in OBs were protected from diet-induced obesity and exhibited improved glucose metabolism with lower fasting glucose and insulin levels,increased glucose tolerance and insulin sensitivity.This leanness and resistance to weight gain was not attributable to changes in food intake,physical activity or lipid absorption but instead was due to increased energy expenditure and greater whole-body substrate flexibility.RNA-seq revealed an increase in glycolysis and skeletal insulin signaling pathways,which correlated with the potentiation of insulin signaling and increased insulin-dependent glucose uptake in Rptorknockout osteoblasts.Collectively,these findings point to a critical role for the mTORC1 complex in the skeletal regulation of wholebody glucose metabolism and the skeletal development of insulin resistance.展开更多
Laboratory automation technologies have revolutionized biomedical research.However,the availability of automation solutions at the single-cell level remains scarce,primarily owing to the inherent challenges of handlin...Laboratory automation technologies have revolutionized biomedical research.However,the availability of automation solutions at the single-cell level remains scarce,primarily owing to the inherent challenges of handling cells with such small dimensions in a precise,biocompatible manner.Here,we present a single-cell-level laboratory automation solution that configures various experiments onto standardized,microscale test-tube matrices via our precise ultrasonic liquid sample ejection technology,known as PULSE.PULSE enables the transformation of titer plates into microdroplet arrays by printing nanodrops and single cells acoustically in a programmable,scalable,and biocompatible manner.Unlike pipetting robots,PULSE enables researchers to conduct biological experiments using single cells as anchoring points(e.g.,1 cell vs.1000 cells per“tube”),achieving higher resolution and potentially more relevant data for modeling and downstream analyses.We demonstrate the ability of PULSE to perform biofabrication,precision gating,and deterministic array barcoding via preallocated droplet-addressable primers.Single cells can be gently printed at a speed range of 5–20 cell⋅s−1 with an accuracy of 90.5–97.7%,which can then adhere to the substrate and grow for up to 72 h while preserving cell integrity.In the deterministic barcoding experiment,95.6%barcoding accuracy and 2.7%barcode hopping were observed by comparing the phenotypic data with known genotypic data from two types of single cells.Our PULSE platform allows for precise and dynamic analyses by automating experiments at the single-cell level,offering researchers a powerful tool in biomedical research.展开更多
基金This study was funded through a Pilot Program in Genomic Applications in Agriculture and Environment Sectors jointly supported by the University of Adelaide and the Australian Genome Research Facility Ltd.P.J.F.was supported by Graduate Research Scholarships from Wine Australia(PH1503)the University of Adelaide.N.S.was supported by a summer scholarship from the ARC Centre of Excellence in Plant Energy Biology(CE1400008).
文摘Transfer RNAs(tRNA)are crucial adaptor molecules between messenger RNA(mRNA)and amino acids.Recent evidence in plants suggests that dicistronic tRNA-like structures also act as mobile signals for mRNA transcripts to move between distant tissues.Co-transcription is not a common feature in the plant nuclear genome and,in the few cases where polycistronic transcripts have been found,they include non-coding RNA species,such as small nucleolar RNAs and microRNAs.It is not known,however,the extent to which dicistronic transcripts of tRNA and mRNAs are expressed in field-grown plants,or the factors contributing to their expression.We analysed tRNA–mRNA dicistronic transcripts in the major horticultural crop grapevine(Vitis vinifera)using a novel pipeline developed to identify dicistronic transcripts from high-throughput RNA-sequencing data.We identified dicistronic tRNA–mRNA in leaf and berry samples from 22 commercial vineyards.Of the 124 tRNA genes that were expressed in both tissues,18 tRNA were expressed forming part of 19 dicistronic tRNA–mRNAs.The presence and abundance of dicistronic molecules was tissue and geographic sub-region specific.In leaves,the expression patterns of dicistronic tRNA–mRNAs significantly correlated with tRNA expression,suggesting that their transcriptional regulation might be linked.We also found evidence of syntenic genomic arrangements of tRNAs and protein-coding genes between grapevine and Arabidopsis thaliana,and widespread prevalence of dicistronic tRNA–mRNA transcripts among vascular land plants but no evidence of these transcripts in non-vascular lineages.This suggests that the appearance of plant vasculature and tRNA–mRNA occurred concurrently during the evolution of land plants.
基金the National Health and Medical Research Council of Australia(APP1109207,awarded to ACWZ,PMB,and CGP)Australian Research Council(DP160100454,awarded to ACWZ and PMB)+1 种基金Diabetes Australia Research Program(awarded to ACWZ,SF and SM)an Australia Postgraduate Award(PT).
文摘Overnutrition causes hyperactivation of mTORC1-dependent negative feedback loops leading to the downregulation of insulin signaling and development of insulin resistance.In osteoblasts(OBs),insulin signaling plays a crucial role in the control of systemic glucose homeostasis.We utilized mice with conditional deletion of Rptor to investigate how the loss of mTORC1 function in OB affects glucose metabolism under normal and overnutrition dietary states.Compared to the controls,chow-fed Rptorob−/−mice had substantially less fat mass and exhibited adipocyte hyperplasia.Remarkably,upon feeding with high-fat diet,mice with pre-and post-natal deletion of Rptor in OBs were protected from diet-induced obesity and exhibited improved glucose metabolism with lower fasting glucose and insulin levels,increased glucose tolerance and insulin sensitivity.This leanness and resistance to weight gain was not attributable to changes in food intake,physical activity or lipid absorption but instead was due to increased energy expenditure and greater whole-body substrate flexibility.RNA-seq revealed an increase in glycolysis and skeletal insulin signaling pathways,which correlated with the potentiation of insulin signaling and increased insulin-dependent glucose uptake in Rptorknockout osteoblasts.Collectively,these findings point to a critical role for the mTORC1 complex in the skeletal regulation of wholebody glucose metabolism and the skeletal development of insulin resistance.
基金support from the National Institutes of Health(Grant numbers:R01HD103727,UH3TR002978,R01GM141055,R44OD024963,R44HL140800,and R44AG063643).
文摘Laboratory automation technologies have revolutionized biomedical research.However,the availability of automation solutions at the single-cell level remains scarce,primarily owing to the inherent challenges of handling cells with such small dimensions in a precise,biocompatible manner.Here,we present a single-cell-level laboratory automation solution that configures various experiments onto standardized,microscale test-tube matrices via our precise ultrasonic liquid sample ejection technology,known as PULSE.PULSE enables the transformation of titer plates into microdroplet arrays by printing nanodrops and single cells acoustically in a programmable,scalable,and biocompatible manner.Unlike pipetting robots,PULSE enables researchers to conduct biological experiments using single cells as anchoring points(e.g.,1 cell vs.1000 cells per“tube”),achieving higher resolution and potentially more relevant data for modeling and downstream analyses.We demonstrate the ability of PULSE to perform biofabrication,precision gating,and deterministic array barcoding via preallocated droplet-addressable primers.Single cells can be gently printed at a speed range of 5–20 cell⋅s−1 with an accuracy of 90.5–97.7%,which can then adhere to the substrate and grow for up to 72 h while preserving cell integrity.In the deterministic barcoding experiment,95.6%barcoding accuracy and 2.7%barcode hopping were observed by comparing the phenotypic data with known genotypic data from two types of single cells.Our PULSE platform allows for precise and dynamic analyses by automating experiments at the single-cell level,offering researchers a powerful tool in biomedical research.
