IEgg production in laying hens gradually declines after reaching the peak, a phenomenon likely influenced by changes in the expression of key genes associated with the hypothalamic-pituitary-gonadal (HPG) axis. In this study, specific primers were designed for reproductive genes, and their expression levels were evaluated at both peak and post-peak egg production intervals (35 and 51 weeks) in laying hens. Nucleotide sequences of the target genes were retrieved from genomic databases, and primers were designed using bioinformatics software. The characteristics of these primers—including melting temperature, product length, and amplification efficiency—were assessed. Total RNA was then extracted from the hypothalamus and pituitary tissues; after verifying RNA quality and quantity, cDNA synthesis was conducted. The expression of the target genes was analyzed using Real-Time PCR at two time intervals following peak egg production. Data were normalized and analyzed using the ΔΔCt method. Results indicated that the designed primers for GnRH, FSH, and LH genes displayed high specificity and efficiency. Furthermore, significant differences in gene expression were observed between the two time points, potentially reflecting physiological changes associated with declining egg production after the peak (P < 0.05). This study demonstrates that specific primer design and Real-Time PCR are effective tools for investigating reproductive gene expression in laying hens. The observed changes in gene expression post-peak egg production provide a foundation for future research aimed at enhancing reproductive performance in laying hens.
Type of Study:
Applicable |
Subject:
Subject 02 Received: 2025/03/8 | Accepted: 2025/07/14 | Published: 2025/10/7