Dataset Information

Triggering acute RyR1 Ca2+ leak leads to improved mitochondrial remodelling and function

ABSTRACT: C2C12 myotubes at day 7 of differentiation were stimulated with a single session of SIT (6 x 30s with 4 min at rest) and were immediately after the stimulation treated or not with 10 uM S107 drug for 72h. The myotubes were then harvested at 72h post stimulation (for SIT condition) and 72h post stimulation and S107 treatment (for SIT S107 condition) for mass spectrometry analysis using the proteomics approach. N = 5 independent myotube wells per condition.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Skeletal Muscle Cell

SUBMITTER: Manfredo Quadroni

LAB HEAD: Nicolas Place

PROVIDER: PXD018409 | Pride | 2021-10-20

REPOSITORIES: pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	191211_Zanou_11877_01.raw	Raw
	191211_Zanou_11877_02.raw	Raw
	191211_Zanou_11877_03.raw	Raw
	191211_Zanou_11877_04.raw	Raw
	191211_Zanou_11877_05.raw	Raw

Items per page:

1 - 5 of 26

Similar Datasets

Project description:Adventitious root formation at the base of plant cuttings is an innate de novo organogenesis process that allows massive vegetative propagation of many economically and ecologically important species. The early molecular events following shoot excision are not well understood. Using whole-genome microarrays, we detected significant transcriptome remodeling during 48 hours following shoot removal in Populus softwood cuttings in the absence of exogenous auxin, with 27% and 36% of the gene models showing differential abundance between 0 and 6 hours, and 6 and 24 hours, respectively. During these two time intervals, gene networks involved in protein turnover, protein phosphorylation, molecular transport and translation were among the most significantly regulated. Transgenic lines expressing a constitutively active form of the Populus type-B response regulator PtRR13 (ÎDDKPtRR13) have a delayed rooting phenotype and cause misregulation of COV1, a negative regulator of vascularization; PDR9, an auxin efflux transporter; two AP2/ERF genes with sequence similarity to TINY1. Cytokinin action appeared to disrupt root development 24 hours after shoot excision, when root founder cells are hypothesized to be sensitive to the negative effects of cytokinin. Our results suggest that PtRR13 acts downstream of cytokinin to repress adventitious root formation in intact plants, and that reduced cytokinin signaling after shoot excision enables coordinated expression of ethylene, auxin and vascularization pathways leading to adventitious root development. Populus tremula x Populus alba INRA-clone No. 717-1-B4 plants expressing a constitutively active form of the Populus type-B response regulator PtRR13 (ÎDDKPtRR13) were generated via an Agrobacterium-mediated protocol developed by Han et al. (2000). Non-transgenic and ÎDDKPtRR13 line were grown to a height of 60 cm. A 14 cm tall apical cuttings were collected from the mother plants. Cuttings were placed in 25 cm2 pots Fafard mix #4 and placed on a mist bench with intermittent mist to prevent shoot desiccation. Samples were collected at the indicated time points and consisted of a 5 mm section measured up from the base of the cutting (one sample per cutting). Total RNA was extracted with the RNeasy mini kit (Qiagen USA) and DNase treated in-column with the RNase-Free DNase set (Qiagen USA). Double-stranded cDNA was synthesized using SuperScript Double Strand cDNA Synthesis Kit (Invitrogen USA, Carlsbad, CA) with oligo-dT primers following the manufacturerâs protocol except that the synthesis step was extended to 16 hours. Cy-3 labeling and hybridization steps were performed by NimbleGen using their standard procedures. A custom-designed microarray platform was used comprising single 60-mer probes designed against 55,793 annotated gene models from the sequenced genome of P. trichocarpa. Each 60-mer probe was chosen form a group of 6-7 non-overlapping probes designed against different parts of the gene model. The probe whose value was the most similar to the average of 6-7 experimental probes was assumed to be the most reliable for transcript level estimation. A total of 39 microarray chips were used in these experiments: 39 chips = 2 genotypes (NT and ÎDDK) x 4 time points (0, 6, 24 and 48 hours) x 5 biological replications, except for the 0 hour ÎDDK where 4 biological replications were used. Signal intensities were log2 transformed and quantile normalized (Sugiharto et al., 1992). Normalized signals were analyzed in SAS 9.1 (SAS Institute, Cary, NC) using a mixed model analysis of variance (ANOVA) with genotype and genotype by time interactions as fixed effects, and biological replication as a random effect.

Dataset Information

Triggering acute RyR1 Ca2+ leak leads to improved mitochondrial remodelling and function

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets