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Modulatory effects of pioglitazone as a ligand for the peroxisome proliferator-activated receptor on semen quality and fertility potential of broiler breeder roosters.

ABSTRACT: Fertility potential in roosters is a crucial topic in broiler breeder reproduction which is thought to be associated with age. This study aims to investigate effects of 2 levels of pioglitazone (PIO) supplementation on peroxisome proliferator-activated receptor gamma (PPAR-γ) expression, semen quality, and fertility parameters of aged broiler breeder roosters. The efficacy of PIO was divided into 2 sections: receptor-dependent and receptor-independent. Expression of PPAR-γ mRNA and protein was assessed in sperm to monitor receptor-dependent actions. Sperm motility, velocity parameters, viability, mitochondrial activity, and apoptosis were assessed for the receptor-independent actions. Broiler breeder roosters were randomly assigned to 3 groups: 1) control received a basal diet (CTRL); 2) PIO-5 received a basal diet supplemented with 5 mg PIO/bird/day, and 3) PIO-10 received a basal diet supplemented with 10 mg PIO/bird/day. In addition, semen samples were collected from 24 Ross broiler breeder roosters at 30, 43, and 53 wk of age. Effects of PIO were significant in terms of total motility, straight-line velocity, mitochondrial activity, and apoptosis (P ≤ 0.05). Total motility, straight-line velocity and mitochondrial activity improved in both PIO groups (P ≤ 0.05) along with a significant reduction in early and late apoptosis in the PIO groups (P ≤ 0.05). Pioglitazone addition affected total motility, mitochondrial activity, early apoptosis and late apoptosis in a linearly and quadratically manner (P < 0.05). PPAR-γ mRNA and protein expression were not significantly upregulated by the different doses of PIO (P > 0.05). Similarly, fertility performance was not significantly changed in the PIO groups (P > 0.05). Moreover, PIO improved mitochondrial activity and decreased the apoptosis rate in the sperm of aged broiler breeder roosters. These improvements were associated with the receptor-independent actions of PIO and the mechanism of action of PIO did not appear to be affected by the PPAR-γ receptor in broiler breeder roosters.

SUBMITTER: Pourazadi L

PROVIDER: S-EPMC8965139 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature

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Modulatory effects of pioglitazone as a ligand for the peroxisome proliferator-activated receptor on semen quality and fertility potential of broiler breeder roosters.

Pourazadi Laya L Sharafi Mohsen M Torshizi Mohammad Amir Karimi MAK Shahverdi Abdolhossein A Alizadeh AliReza A

Poultry science 20220223 5

Fertility potential in roosters is a crucial topic in broiler breeder reproduction which is thought to be associated with age. This study aims to investigate effects of 2 levels of pioglitazone (PIO) supplementation on peroxisome proliferator-activated receptor gamma (PPAR-γ) expression, semen quality, and fertility parameters of aged broiler breeder roosters. The efficacy of PIO was divided into 2 sections: receptor-dependent and receptor-independent. Expression of PPAR-γ mRNA and protein was a ...[more]

PMID: 35349953

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Project description:Our aim was to investigate if the peroxisome proliferator-activated receptor (PPAR)-gamma agonist pioglitazone modulates inflammation through PPARalpha mechanisms.The thiazolidinediones (TZDs) pioglitazone and rosiglitazone are insulin-sensitizing PPARgamma agonists used to treat type 2 diabetes (T2DM). Despite evidence for TZDs limiting inflammation and atherosclerosis, questions exist regarding differential responses to TZDs. In a double-blinded, placebo-controlled 16-week trial among recently diagnosed T2DM subjects (n = 34), pioglitazone-treated subjects manifested lower triglycerides and lacked the increase in soluble vascular cell adhesion molecules (sVCAM)-1 evident in the placebo group. Previously we reported PPARalpha but not PPARgamma agonists could repress VCAM-1 expression. Since both triglyceride-lowering and VCAM-1 repression characterize PPARalpha activation, we studied pioglitazone's effects via PPARalpha.Pioglitazone effects on known PPARalpha responses--ligand binding domain activation and PPARalpha target gene expression--were tested in vitro and in vivo, including in wild-type and PPARalpha-deficient cells and mice, and compared with the effects of other PPARgamma (rosiglitazone) and PPARalpha (WY14643) agonists.Pioglitazone repressed endothelial TNFalpha-induced VCAM-1 messenger ribonucleic acid expression and promoter activity, and induced hepatic IkappaBalpha in a manner dependent on both pioglitazone exposure and PPARalpha expression. Pioglitazone also activated the PPARalpha ligand binding domain and induced PPARalpha target gene expression, with in vitro effects that were most pronounced in endothelial cells. In vivo, pioglitazone administration modulated sVCAM-1 levels and IkappaBalpha expression in wild-type but not PPARalpha-deficient mice.Pioglitazone regulates inflammatory target genes in hepatic (IkappaBalpha) and endothelial (VCAM-1) settings in a PPARalpha-dependent manner. These data offer novel mechanisms that may underlie distinct TZD responses.

Project description:BackgroundHumans are exposed to a complex mixture of environmental chemicals that impact bone and metabolic health, and traditional exposure assessments struggle to capture these exposure scenarios. Peroxisome proliferator activated receptor-gamma (PPARγ) is an essential regulator of metabolic and bone homeostasis, and its inappropriate activation by environmental chemicals can set the stage for adverse health effects. Here, we present the development of the Serum PPARγ Activity Assay (SPAA), a simple and cost-effective method to measure total ligand activity in small volumes of serum.MethodsFirst, we determined essential components of the bioassay. Cos-7 cells were transfected with combinations of expression vectors for human PPARγ and RXRα, the obligate DNA-binding partner of PPARγ, along with PPRE (DR1)-driven luciferase and control eGFP reporter constructs. Transfected cells were treated with rosiglitazone, a synthetic PPARγ ligand and/or LG100268, a synthetic RXR ligand, to characterize the dose response and determine the simplest and most efficacious format. Following optimization of the bioassay, we assessed the cumulative activation of PPARγ by ligands in serum from mice treated with a PPARγ ligand and commercial human serum samples.ResultsCos-7 cells endogenously express sufficient RXR to support efficacious activation of transfected PPARγ. Co-transfection of an RXR expression vector with the PPARγ expression vector did not increase PPRE transcriptional activity induced by rosiglitazone. Treatment with an RXR ligand marginally increased PPRE transcriptional activity in the presence of transfected PPARγ, and co-treatment with an RXR ligand reduced rosiglitazone-induced PPRE transcriptional activity. Therefore, the final bioassay protocol consists of transfecting Cos-7 cells with a PPARγ expression vector along with the reporter vectors, applying rosiglitazone standards and/or 10 μL of serum, and measuring luminescence and fluorescence after a 24 h incubation. Sera from mice dosed with rosiglitazone induced PPRE transcriptional activity in the SPAA in a dose-dependent and PPARγ-dependent manner. Additionally, human serum from commercial sources induced a range of PPRE transcriptional activities in a PPARγ-dependent manner, demonstrating the ability of the bioassay to detect potentially low levels of ligands.ConclusionsThe SPAA can reliably measure total PPRE transcriptional activity in small volumes of serum. This system provides a sensitive, straightforward assay that can be reproduced in any cell culture laboratory.

Dataset Information

Modulatory effects of pioglitazone as a ligand for the peroxisome proliferator-activated receptor on semen quality and fertility potential of broiler breeder roosters.

Publications

Modulatory effects of pioglitazone as a ligand for the peroxisome proliferator-activated receptor on semen quality and fertility potential of broiler breeder roosters.

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