The present study was carried out to investigate the possible etiology and pathology of cystic ovarian follicles by determining the expression levels of mRNA for LH-r gene in follicular cells of dominant follicles in comparison with cystic ovarian follicles in Iraqi cows. This study were performed in two steps, firstly aspiration of follicular fluids from dominant follicles and cystic follicles and stored in (-20 ℃) until estradiol and progesterone assay, secondary section of the follicular wall in to two hemispheres, and stored at -70 ℃ to -80 ℃ to molecular study. The macroscopic examination of the ovaries revealed that the numbers of dominant follicles are [1] samples while cystic follicles were in [2] samples. There was a significant difference (P<0.05) in the diameter of cystic ovarian follicles (37.56 ± 0.64 mm) compared with dominant follicles (19.93 ± 0.32 mm).Results of hormonal assay showed higher estradiol-17β (865.96±10.64 ng/ml) and progesterone (84.8±1.35 ng/ml) concentrations in follicular fluids of cystic ovarian follicles, which were significantly higher (P<0.05) in comparison with those of dominant follicles which were (314.39±2.55 ng/ml) and (50.25±1.57 ng/ml) respectively.Molecular study, to evaluate the relative quantification of LH-r gene in dominant and cystic follicular cells, has been done by extraction of the total RNA and assay its concentration from these cells, synthesis data of the complementary DNA (cDNA), that done by reverse transcription PCR (q-RT-PCR) technique, of target gene and compared of the gene expression in dominant and cystic follicular cells, the our results referred to down regulation of LH-r gene expression in follicular cells of the cystic ovarian follicles, that may be assistance to understand the etiology and pathology of this case (disease), thus the up-regulation of the this gene in cells of dominant follicles may explain the important role of the LH in ovulation mechanism and increasing follicular ovulation chance.
KeywordsGene Expression; Cows; Ovarian Follicles; LH-r Gene
Identification of the Expression Level to LH-r Gene In dominant and Cystic Ovarian Follicles Cells of the CowsCitation: Al-Delemi DHJ, Al-Gewary AKA, Ali Jeddoa ZM (2014)
Materials and Methods
Quantitative Reverse Transcriptase Real-Time PCR KitsAll kits which used in quantification of gene expression levels by qRT-PCR, and hormonal kit which used to RIA method, with their companies and countries of origin ( Table 1).
PrimersTwo set of primers are used in this study, first primer used for GAPDH gene as Housekeeping gene and other primer used for LH-r gene as target gene. These primers were designed by using NCBI-Gene Bank data base and Primer 3 design online, the primers used in quantification of gene expression using quantitative (real time-PCR) techniques based SYBER Green DNA binding dye, and supported from (Bioneer, Korea) company (Table 2).
Collection of the FolliclesThe presence study depends on collection of ovaries from sexually mature cows (4-6 years) old with unknown repr...
Cockscomb (Celosia argentea L.) is one of the most important ornamental plants in Iraq. In August 2021, severe root rot of cockscomb plants was observed with an incidence of 30% in all of the nurseries surveyed in Karbala and Babylon Provinces, Iraq. The symptoms appeared as severe leaf chlorosis and wilting with brown discoloration of the stem vascular bundle associated with dark brown lesions on the root that extended to the root collar and became extensive rot. Morphological features of 50% of the fungal colonies were identical and similar to Fusarium spp. The internal transcribed spacer, translation elongation factor, RNA polymerase I, and II, beta-tubulin 1, and the mitochondrial small-subunit ribosomal RNA genes were amplified, and sequenced. BLASTn analysis of these genetic marker’s sequences exhibited 99.84 to 100 % similarity with the corresponding global sequences of the Fusarium solani (Mart.). This identification was confirmed by the phylogenetic analysis of the concatenated sequences of the genetic markers. Although F. solani has caused economic damage to a broad range of commercially important plants in Iraq, to our knowledge, this is the first report of F. solani causing root rot of Cockscomb (C. argentea) in Iraq.
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