Dataset Information

Exploring molecular dynamics indicators associated with reproductive performance of Bos indicus cattle in plasma samples through data-independent acquisition mass spectrometry

ABSTRACT: Aim of the study was to profile the blood plasma proteome of healthy Bos indicus cattle and use it as a baseline against which to compare different conditions in Bos indicus cattle, such as disease, reproduction associated proteins and pregnancy specific proteins etc. Data-independent acquisition (DIA) bases quantitative proteomics approach coupled to LC-MS/MS methods were used to achieve this aim. An Orbitrap Fusion Tribrid mass spectrometer (Thermo Fischer Scientific, Bremen, Germany) connected to the Easy- nLC-1200 nanoflow liquid chromatography system (Thermo Scientific) was used for data acquisition.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Bos Taurus (bovine)

TISSUE(S): Blood Plasma

SUBMITTER: Keshava Prasad T. S.

LAB HEAD: T. S. Keshava Prasad

PROVIDER: PXD034103 | Pride | 2024-07-15

REPOSITORIES: Pride

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Publications

Exploring molecular dynamic indicators associated with reproductive performance of Bos indicus cattle in blood plasma samples through data-independent acquisition mass spectrometry.

Devadasan M Joel MJ Ramesha Kerekoppa P KP Ramesh Poornima P Kootimole Chinmaya Narayana CN Jeyakumar Sakthivel S Ashwitha A A Ammankallu Shruthi S Rai Akhila Balakrishna AB Kumaresan Arumugam A Vedamurthy Veerappa G VG Raju Rajesh R Das D N DN Kataktalware Mukund A MA Prasad Thottethodi Subrahmanya Keshava TSK

Journal of proteomics 20230613

Improving reproductive performance of cattle is of paramount importance for sustainable dairy farming. Poor reproduction performance (RP) hinders the genetic improvement of important Bos indicus cattle breeds. It is well known that incorporation of molecular information along with conventional breeding method is far better than use of conventional method alone for the genetic improvement of reproductive performance traits in cattle. Therefore, the present study sought to investigate the plasma p ...[more]

PMID: 37321300

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Project description:The current study was designed to investigate the circulating miRNA expression signature of dairy cows during several key time points around calving, and under different fatty acids (FA) treatments to get insights into different aspects of metabolic adaptations and their interaction with administrated FA. In this trial, 32 transition Holstein dairy cows were randomly assigned to one of the following treatments: 1-CTRL (n = 8; coconut oil, Bio-Kokosöl #665, Kräuterhaus Sanct Bernhard, KG, Bad Ditzenbach, Germany; 38 and 76 g/d in antepartum (AP) and postpartum (PP)), 2- Essential fatty acids (EFA, n=8, a combination of linseed oil (DERBY® Leinöl #4026921003087, DERBY Spezialfutter GmbH, Münster, Germany; 39 and 78 g/d in AP and PP) and safflower oil (GEFRO Distelöl, GEFRO Reformversand Frommlet KG, Memmingen, Germany; 2 and 4 g/d in AP and PP), 3- conjugated linoleic acid (CLA, n = 8, Lutalin® ; cis-9, trans-11, 10 g/d trans- 10, cis-12 CLA, BASF SE, Lampertheim, Germany; 19 and 38 g/d in AP and PP), and 4- EFA+CLA, a combination of linseed oil, safflower oil and Lutalin® from day 63 AP to day 63 PP. From each cow, four blood samples were collected individually on days -21, +1, +28, and +63 relative to calving, and the plasma fraction was used for RNA extraction. Total RNA was extracted from 128 plasma samples using the QIAGEN miRNeasy serum/plasma kit (Qiagen GmbH, Hilden, Germany), and checked for yield and purity using the small RNA kit on a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). We used the QIAseq miRNA library kit (QIAGEN, Hilden, Germany) for library preparation and Caliper (PerkinElmer, Waltham, MA) for quality check. NGS was performed on the libraries using the NovaSeq 6000 (Illumina, San Diego, CA) setting on single-end 100 bp mode on the respective platform of IGA Technology Services, Udine, Italy (https://igatechnology.com/). Raw-sequencing data was checked for quality (format conversion to FASTQ, demultiplexing, Adapter Trimming, and UMI Remova) using the Illumina Bcl2fastq2 Conversion Software (version 2.20). Detected sequences were analyzed with the miRDeep2 (miRDeep2.pl) software package (version 2.0.0.5) to detect known miRNA and predict putative novel miRNA (using human, ovine, and goat miRNA datasets). MiRDeep2 was fed with the Bos taurus miRNA collection available at miRBase (www.mirbase.org). Read counts for each known and novel miRNA were compiled using the HTSeq-count script. The quantified counts were normalized using the TMM method via the edgeR (version 3.12.0) package in R software (Version 4.0). Only miRNA with at least one count per million over at least 2 samples were taken into consideration for the analysis. Differentially expressed miRNA (DEM) during the time points were identified by performing generalized linear model (GLM) likelihood ratio tests (glmLRT) using the GLM approach in edgeR. The DEMs have been interpreted separately I-during different time points, and also II-between treatments, to gain insight into metabolic and immune adaptation, as well as the role of fatty acids at each time point. A time-affected DEM identified key signaling pathways that are involved in initiating and regulating metabolic shifts during the transition from gestation (non-lactation) to lactation. Although, fatty acid treatments had only minor effects on miRNA expression (insufficient DEM to perform pathway analysis).

Project description:Puberty is a complex physiological event measured by various indicator traits in genetic improvement programs of beef cattle; thus, developing a more complete understanding of the genes and regulatory pathways and networks involved in puberty will provide knowledge to help improve genetic selection strategies. Herein, we characterized the transcriptome of five reproductive tissues (i.e. hypothalamus, pituitary gland, ovary, uterus, and endometrium) as well as tissues known to be relevant to growth and metabolism needed for cattle to achieve puberty (i.e., longissimus dorsi muscle, fat, and liver). These tissues were collected from pre (PRE)- and post (POST)-pubertal Brangus (3/8 Brahman; Bos indicus x 5/8 Angus; Bos taurus) heifers derived from a population of cattle used to identify QTL associated with fertility traits. In order to exploit the power of complementary omics analyses, PRE and POST puberty co-expression gene networks were constructed by combining the results from RNA-Seq, GWAS, and bovine transcription factors. RNA-Seq of 8 tissues among PRE and POST Brangus heifers revealed 1515 differentiallyexpressed and 943 tissue-specific genes within the 17,832 genes confirmed by metrics of RNA-Seq analysis. Combining the results from RNA-Seq and GWAS indentified a total of 25 QTL associated to heifer fertility. The hypothalamus experienced the most notable up-regulation of genes via puberty. Complementary omics procedures revealed 2,450 co-expressed genes across the 8 tissues relative to puberty. The PRE network had 372,861 connections whereas the POST network had 328,357 connections. A sub-network from this process revealed key transcriptional regulators (i.e., PITX2, FOXA1, TSG1D1, DACH2, LHX4, PROP1 and SIX6). Results from multiples sources of omics data will facilitate the design of breeding strategies to improve fertility in Bos indicus-influenced composite cattle. Sixty-one samples from PRE and POST pubertal composite beef heifers were analyzed with RNA-Seq. The transcriptome of five reproductive tissues (i.e. hypothalamus, pituitary gland, ovary, uterus, and endometrium) as well as tissues known to be relevant to metabolism andbody morphometrics needed for cattle to achieve puberty (i.e.,) was characterized. These tissues were collected from pre (PRE)- and post (POST)-pubertal Brangus (3/8 Brahman x 5/8 Angus) heifers derived from a population of cattle used to identify QTL associated with fertility. Total RNA was purified using a Trizol protocol (Invitrogen, Carlsbad, CA). Sequencing libraries were made using TruSeq RNA Sample Preparation kit of Illumina (San Diego, CA).

Project description:Low fertility remains a leading cause of poor productivity in dairy cattle. In this context, there is significant interest in developing novel tools for accurate early diagnosis of pregnancy. MicroRNAs (miRNAs) are short RNA molecules which are critically involved in regulating gene expression during both health and disease. MiRNAs have been shown to regulate ovarian function, uterine receptivity, embryonic development and placental function. Circulating miRNAs can provide useful biomarkers of tissue function and disease; importantly, differential miRNA profiles have been linked to pregnancy and preeclampsia in humans. This study sought to establish the potential of circulating miRNAs as biomarkers of early pregnancy in cattle. We applied Illumina small-RNA sequencing to profile miRNAs in plasma samples collected from eight non-pregnant heifers on Days 0, 8 and 16 of the oestrous cycle and 11 heifers on Days 16 and 24 of pregnancy. We sequenced a total of 46 samples and generated 9.2 million miRNA reads per sample. There were no differences in miRNA read abundance between any of the pregnant and non-pregnant time-points (FDRâ>â0.1). As a complementary approach, we analysed sample pools (3-4 samples/pool) corresponding to Days 0, 8 and 16 of the oestrous cycle and Day 24 of pregnancy (nâ=â3 pools/group) using Qiagen PCR arrays. A total of 16 miRNAs were differentially expressed (FDRâ<â0.1) in plasma between pregnant and non-pregnant animals. RT-qPCR validation using the same plasma samples confirmed that miR-26a was differentially upregulated on Day 16 pregnant relative to non-pregnant heifers (1.7-fold; Pâ=â0.043), whereas miR-1249 tended to be upregulated in Day 16 pregnant heifers (1.6-fold; Pâ=â0.081). Further validation in an independent group of heifers confirmed an increase in plasma miR-26a levels during early pregnancy, which was significant only on Day 24 (2.0-fold; Pâ=â0.027). Through genome-wide analyses we have successfully profiled plasma miRNA populations associated with early pregnancy in cattle. We have identified miR-26a as a potential circulating biomarker of early pregnancy. Sequencing of three sequencial samples from each of eight cycling animals (Days 0, 8 and 16, total 24 samples) and two samples from each of 11 pregnant animals (Days 16 and 24, total 22 samples). Main comparisons were between non-pregnant and pregnant groups

Dataset Information

Exploring molecular dynamics indicators associated with reproductive performance of Bos indicus cattle in plasma samples through data-independent acquisition mass spectrometry

Publications

Exploring molecular dynamic indicators associated with reproductive performance of Bos indicus cattle in blood plasma samples through data-independent acquisition mass spectrometry.

Similar Datasets

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