Pandiyan G.D.V.,TANUVAS Veterinary College and Research Institute |
Krupakaran R.P.,TANUVAS Veterinary College and Research Institute |
Balamurugan T.C.,TANUVAS Veterinary College and Research Institute |
Arunkumar S.,TANUVAS Veterinary College and Research Institute |
Perumal P.,TANUVAS Veterinary College and Research Institute
Indian Journal of Animal Sciences | Year: 2015
Matrix metalloproteinase -9 (MMP-9) is one of the most important metalloproteinases involved in degradation of cellular matrix during several physiological as well as pathological process including ovulation, fertilization, implantation and parturition. A study was conducted to confirm the presence of MMP-9 gene in blood samples of Murrah buffalo cows at different reproductive physiological stages. Healthy Murrah buffalo cows (18), aged about 3-6 years was divided into 3 groups, each group consisting of 6 animals, viz. pregnant (group 1), oestrous (group 2) and anoestrous (group 3) buffalo cows were selected. Experimental animals were managed and fed as per the farm schedule. Blood samples were collected in vacutainer containing sodium citrate and the blood samples were transferred immediately to the laboratory and were stored at -20°C in deep freezer till further use. Total RNA was isolated from the blood samples and reverse transcription- polymerase chain reaction (RT-PCR) was carried out. Primers targeting catalytic domain of the MMP-9 was designed. The total cellular RNA was obtained from 500 μL of blood was 0.108 μg and the concentration of the RNA was 0.216 μg/mL. The ratio of A260/A280 was 1.81 indicating that isolated RNA was reasonably pure. RT-PCR was carried out in 2 steps as and the first step was to synthesize cDNA and the second step to amplify the desired genes from cDNA by PCR. The RT-PCR products were subjected to 1% agarose gel electrophoresis and the expected sizes of 1080 bp for catalytic domain was observed in all the three groups of Murrah buffalo cows. © 2015, Indian Council of Agricultural Research. All rights reserved.
Prakash Krupakaran R.,TANUVAS Veterinary College and Research Institute |
Balamurugan T.C.,National Research Center on Mithun |
Durga Lakshmi R.,National Research Center on Mithun |
Sheeba A.,National Research Center on Mithun |
Perumal P.,National Research Center on Mithun
Indian Journal of Animal Sciences | Year: 2016
A study was conducted to evaluate the presence of matrix metalloproteinases (MMP) in the serum of domestic animal species. The serum samples were collected from four healthy male animals of each species, viz. goat, cattle, horse, rabbit, sheep, pig and 4 tumor affected dogs in a heparinzed vacutainer, during early morning before feeding the animals. All the serum samples were subjected to gelatin zymography. The major bands were observed at 220, 92 kDa of MMP-9 and 72 kDa of MMP-2 in all the species with minor variations in rabbit and goat. It was observed that these bands indicated the normal physiological state of the animals and in tumour samples, the intensity of both MMP-9 and MMP-2 was 2-3 times higher. The level of expression of latent form of MMP-9 band was comparable in goat, cattle, horse, sheep and pig and also they were as expressed in human, on contrast there was low level of expression in rabbit as it clearly indicated these MMP proteins were in low concentration in the serum of rabbit. The thickness of Pro-MMP-9 (92 kDa) band in horse serum was alike as in the human marker and it might be related to human protein. There was a faded band below 72 kDa band in all the species but it was absent in human serum as it could be the active form of MMP-2 (62 kDa). MMP-2 band in cattle and horse serum were correlated. The concentration of the MMP-2 band in sheep serum was higher than in the other species used in this study but it was lesser than the activity of protein isolated from canine tumor. It was concluded that MMP plays a significant role in normal physiological functions of every species and its activity was 4-5 times higher in tumor samples due to greater gelatinolytic activity. Thus, it was concluded that tumor samples exhibit greater gelatinolytic activity because of higher concentration of MMP proteins. © 2016, Indian Council of Agricultural Research. All rights reserved.