Differential expression analysis of components of the insulin signaling cascade in the canine corpus luteum after RNA sequencing
Bibliographic record
Abstract
The present study aims to characterize the expression of genes regulated by insulin in the canine corpus luteum (CL) along the diestrus. Hypothesis is that insulin plays a role in the regulation of steroidogenesis and the functions of lutein cells, consequently. Non-pregnant bitches were submitted to ovariosalpingohisterectomy every 10 days between 10 and 60 (n = 4 / group) after ovulation (po). CL collected were used for RNA sequencing (RNA-seq) and validated by real-time PCR, and protein analysis by Western blotting and immunofluorescence. Bioinformatics analysis detected the presence of 34.408 genes in the samples, of which 29.011 genes showed some level of gene expression in diestrus. The temporal gene expression revealed the presence of 5.116 genes differentially expressed at least one comparison between groups and 1.106 genes were not noted in the canine genome. Seven genes directly linked to insulin signaling were selected: the IRS1 (insulin substrate 1 receptor), involved with the onset of insulin signaling, the PI3KR3 (phosphoinositide-3-kinase, regulatory subunit 3 (gamma) and PI3KCG (phosphoinositide-3- kinase , catalytic, gamma polypeptide) that are PI3K isoforms involved in phosphorylation of AKT, the MAPK9 (mitogen-activated protein kinase 9), MAPK13 (mitogen-activated protein kinase 13), and MAPK14 (mitogenactivated protein kinase 14) involved in the cell proliferation and SOCS1 (suppressor of cytokine signaling 1), known as a positive regulator of insulin signaling initiation. The expression of IRS1, MAPK13 and MAP14 was higher in the second half of diestrus, concomitant with decreased expression of steroidogenic enzymes HSD3B and CYP7B1, suggesting a negative regulatory performed by these genes. Rather, PI3KCG, PI3KR3, MAPK9 and SOCS1 expression have maximum values at the beginning of estrus, similar to the CYP19A1 expression, suggesting a positive control. The results corroborate with studies in which the different components involved in the insulin signaling establish a direct interaction with the expression of steroidogenic enzymes, from low molecular weight mediators such as inositolphosphoglycan. After the association of insulin with its receptor, the mediators of this class are generated, internalized and alter the activities of some steroidogenic enzymes and cellular metabolism, in addition to acting in the cleavage of cholesterol side chain resulting in increased production of hormones. These data allow a better understanding of the regulation of luteal physiology in general and more specifically on the regulation of steroidogenesis through insulin signaling.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".