bioRxiv Science⌕ Search

Biology subjects

Dalmasso, C.

Publications and source records attributed to Dalmasso, C..

2 recordsLinked to original sources

Sex-specific sequels of early life stress on serine/threonine kinase activity in visceral adipose tissue from obese mice

Adverse childhood experiences (ACEs) are an established independent risk factor for chronic disease development including obesity and hypertension; however, only women exposed to ACEs show a positive relationship with BMI. Our lab has reported that maternal separation and early weaning (MSEW), a mouse model of early life stress, induces sex-specific mechanisms underlying greater blood pressure response to a chronic high fat diet (HF). While female MSEW mice fed a HF display exacerbated perigonadal white adipose tissue (pgWAT) expansion and a metabolic syndrome phenotype compared to controls, male MSEW mice display similar levels of adiposity compared to control counterparts but display neurogenic hypertension. Thus, this study aimed to determine a pgWAT whether there is a sex-specific serine/threonine kinase (STKA) activity associated with early life stress. Frozen pgWAT was collected from MSEW and Control, male and female mice fed a HF to assess STKA activity using the Pamstation12 instrument. Overall, MSEW induces significant reduction of 7 phosphokinases (|Z| >=1.5) in females (QIK, MLK, PKCH, MST, STE7, PEK, FRAY) and 5 in males (AKT, SGK, P38, MARK, CDK), while 15 were downregulated in both sexes (DMPK, PKA, PKG, RSK, PLK, DYRK, NMO, CAMK1, JNK, PAKA, RAD53, ERK, PAKB, PKD, PIM, AMPK). This data provides new insights into the sex-specific dysregulation of the molecular network that regulates cellular phosphorylation signals in visceral adipose tissue and identifies possible target phosphokinases important in adipocyte hypertrophy as a result of exposure to early life stress combined with an unhealthy. Identifying functional metabolic signatures associated with early life stress is critical to elucidate the underlying molecular mechanisms behind the sex-specific obesity risk.

molecular biology↗

Comprehensive analysis of RNA-seq kits for standard, low and ultra-low quantity samples

High-throughput RNA-sequencing has become the gold standard method for whole-transcriptome gene expression analysis, and is widely used in numerous applications to study cell and tissue transcriptomes. It is also being increasingly used in a number of clinical applications, including expression profiling for diagnostics and alternative transcript detection. However, despite its many advantages, RNA sequencing can be challenging in some situations, for instance in cases of low input amounts or degraded RNA samples. Several protocols have been proposed to overcome these challenges, and many are available as commercial kits. In this study, we comprehensively test three recent commercial technologies for RNA-seq library preparation (TruSeq, SMARTer and SMARTer Ultra-Low) on human reference tissue preparations, using standard (1g), low (100 and 10 ng) and ultra-low (< 1 ng) input amounts, and for mRNA and total RNA, stranded or unstranded. The results are analyzed using read quality and alignment metrics, gene detection and differential gene expression metrics. Overall, we show that the TruSeq kit performs well with an input amount of 100 ng, while the SMARTer kit shows degraded performance for inputs of 100 and 10 ng, and the SMARTer Ultra-Low kit performs relatively well for input amounts < 1 ng. All the results are discussed in detail, and we provide guidelines for biologists for the selection of a RNA-seq library preparation kit.

genomics↗