17 1Mahmood Ul Hassan 2M. S. Mughal 3M. I. Sohail 1M. M. Javed Journal of Animal and Plant Sciences JAPS 20162016/12/01 26 6 1872-1878 1018-7081 NA <p>Tumor necrosis factor alpha (TNF-&alpha;) an imperative cytokine is secreted from various immune cells, involved in regulation of inflammation process, leukocytes functioning and secretion of other cytokines. TNF&ndash;&alpha; has been identified and studied in many bony fishes. In Pakistan,&nbsp;<em>Cirrhinusmrigala</em>is an important farm fish. Being a novel organism, it was selected for the present studies. After acclimation, fish were divided into five groups i.e. control receiving no stimulation, experimental B,C,D and E receiving LPS and Poly I.C, 2 mg and 20 mg/kg body weight respectively through intra-peritoneal injection. These were dissected after 4, 8 and 12 h of injection. Spleen and head kidney were removed and processed for RNA isolation, quantified and used for cDNA synthesis. PCR was performed with TNF-&alpha; primers taking cDNA as template. Amplified products were run on 1.2% agarose gel and intensity was calculated by Quality One v.4.5.9 software (BIO-RAD). TNF-&alpha; expression was observed significantly up-regulated with 20 mg dose after 4 h of stimulation with LPS but with Poly I.C it was observed after 12 h. Spleen showed elevated expression than head kidney. So the expression level of TNF-&alpha; is not only dependent on time and dose of the stimulants but different organs also.</p> TNF-α, Cirrhinusmrigala, head kidney, spleen, LPS, Intra-peritoneal injection Pakistan Agricultural Scientists Forum https://thejaps.org.pk/AbstractView.aspx?mid=2016-JAPS-244