PT  - JOURNAL ARTICLE
AU  - Heloisa Balan Assalin
AU  - José Antonio Rocha Gontijo
AU  - Patrícia Aline Boer
TI  - miRNAs, target genes expression and morphological analysis on the heart in gestational protein-restricted offspring
AID  - 10.1101/507038
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 507038
4099  - http://biorxiv.org/content/early/2018/12/27/507038.short
4100  - http://biorxiv.org/content/early/2018/12/27/507038.full
AB  - BACKGROUND Gestational protein restriction was associated with low birth weight, hypertension and higher prevalence of cardiac disorders in adults. Several mechanisms, including epigenetics, could be related with the cardiovascular phenotype on protein-restricted offspring. Thus, we investigated the morphological cardiac effects of gestational protein restriction and left ventricle miRNAs and target genes expression pattern in both 12-day and 16-week old gestational protein-restricted male offspring.METHODS Pregnant Wistar rats were allocated into two groups, according to protein supply during pregnancy: NP (normal protein diet-17%) or LP (low protein diet − 6%). The study evaluates the effects of maternal protein restriction on food consumption and body weight of both pregnant dams and offspring, systolic blood pressure in 16-wk old offspring and on cardiac morphometric and molecular parameters in both 12-d and 16-wk old offspring.RESULTS Dams on the gestational protein-restricted diet had lower body weight gain and higher food intake. Gestational protein-restricted offspring had low birth weight, followed by rapidly body weight recovery, hypertension, and increased myocytes crosssectional area and collagen fraction at 16-week old age. At 12-days old, miR-184, miR-192, miR-376c, miR-380-3p, miR-380-5p, miR-451, and miR-582-3p had increased expression, and miR-547 and miR-743 a had decreased expression in the gestational protein-restricted left ventricle. At 16-week old, let-7b, miR-125a-3p, miR-142-3p, miR-182 and miR-188-5p had increased expression and let-7g, miR-107, miR-127, miR-181a, miR-181c, miR-184, miR-324-5p, miR-383, miR-423-5p and miR-484 had decreased expression in gestational protein-restricted left ventricle. Target predicted gene expression analysis shown higher expression of Dnmt3a, Oxct1, Rictor and Trps1 and lower expression of Bbs1 and Calml3 in 12-day old protein-restricted offspring. 16-week old protein-restricted offspring had higher expression of Adrbk1, Bbs1, Dnmt3a, Gpr22, Inppl1, and Oxct1 genes.CONCLUSION Gestational protein restriction leads to offspring low birth weight, increased systolic blood pressure and morphological heart alterations that could be related to early heart miRNA expression changes that perpetuate into adulthood and which are associated with the regulation of essential genes involved in cardiovascular development, heart morphology, function, and metabolism.Authors’ contributionsHBA: Data curation, Investigation, Formal analysis, Methodology, Visualization, Writing-original draft; JARG: Formal analysis, Methodology, Visualization, Writing-review & editing; PAB: Conceptualization, Formal analysis, Funding acquisition, Methodology, Resources, Supervision, Visualization, Writing-original draft, Writing-review & editingThe authors thank Dr. Tom Fleming - University of Southampton for the collaboration with the paper review.List of abbreviations11β-HSD11beta-hydroxysteroid dehydrogenase2-way ANOVAtwo-way analysis of varianceAdrbk1 geneBeta adrenergic receptor kinase 1 geneBbs1 geneBardet-Biedl Syndrome 1 geneCalml3 geneCalmodulin-like protein 3 genecDNAcomplementary DNACOBEABrazilian College of Animal ExperimentationCSAcross-sectional areaDNAdeoxyribonucleic acidDnmt3a geneDNA methyltransferase 3 alpha geneGAPDHglyceraldehyde 3-phosphate dehydrogenaseGlut4type 4 glucose transporterGpr22 geneG-protein coupled receptor 22 geneHEhematoxylin and eosinIDT®Integrated DNA TechnologiesInppl1 geneInositol Polyphosphate Phosphatase Like 1 geneLPlow protein dietLVleft ventriclemiRNAs or mir-or miR–MicroRNAs (miR-mature form of the miRNA; mir-pre-miRNA)mRNAmessenger RNANIHNational Institutes of HealthNPnormal protein dietOxct1 gene3-Oxoacid CoA-Transferase 1 genePI3K-Aktphosphatidylinositol 3’-kinase (PI3K)-Akt signaling pathwayRictor geneRapamycin-insensitive companion of mammalian target of rapamycin geneRNAribonucleic acidShip2SH2 domain containing inositol 5-phosphatase 2Trps1 geneTranscriptional Repressor GATA Binding 1 geneUNICAMP–State University of Campinas