Plasma glucose and immunoreactive insulin (IRI) concentrations and activities of enzymes related to glucose metabolism in livers were measured in dogs and cats. Nucleotide sequences of the conserved region of glucokinase (GK) cDNA that contained ATP- and glucose-binding domains were determined in canine liver and feline pancreas for design of the species-specific oligonucleotide primers for reverse transcription-polymerase chain reaction (RT-PCR) analysis. There were no significant differences in plasma glucose and IRI concentrations between dogs and cats. In feline liver, although GK activities were not detected, activities of hexokinase, fructokinase, pyruvate kinase, glucose-6-phosphate dehydrogenase, fructose-1,6-bisphosphatase and glucose-6-phosphatase were significantly higher than those in canine liver. The partial sequences of canine liver GK and feline pancreas GK cDNA were respectively 88% and 89% identical with the rat liver GK cDNA. Expression of GK gene was observed in canine liver and pancreas and feline pancreas with RT-PCR using species specific primers based on the cDNA sequences.
Activities of enzymes related to energy metabolism and isoenzyme patterns of lactate dehydrogenase (LDH) were determined in peripheral leukocytes and livers of Holstein dairy cows and Institute of Cancer Research (ICR) mice. In dairy cow liver, activities of enzymes in glycolysis, malate-aspartate shuttle and lipogenesis were lower, but activities of glucose-6-phosphatase in gluconeogenesis were higher than those in mouse liver. Glucokinase activities were below detection limit in leukocytes and liver of the cows. Dairy cow leukocytes and liver showed the isoenzyme patterns with dominance of LDH-1, -2 and-3, whereas mouse leukocytes and liver showed that LDH-5 was dominant. The LDH isoenzyme patterns were very similar between leukocytes and liver in each animal species. Some enzymes in leukocytes may reflect those enzymes activities in liver and be a useful indicator for energy metabolism in animals.
ABSTRACT. Concentrations of plasma glucose, immunoreactive insulin (IRI) and free fatty acid (FFA) and activities of enzymes related to energy metabolism and lactate dehydrogenase (LDH) isoenzyme pattern in plasma and leukocytes were investigated in lactating Hols tein cows (dairy cattle) and fattening Japanese Black Wagyu x Holstein steers (beef cattle). IRI concentrations and LDH and malate dehydrogenase (MDH) activities in the plasma of beef cattle were significantly higher than those in dairy cattle. The cytosolic rat io of MDH/ LDH activity in the leukocytes of beef cattle was significantly higher than that of dairy cattle. These findings might be associated with the different energy metabolism between dairy and beef cattle. KEY WORDS: beef cattle, insulin, malate dehydrogenase.J. Vet. Med. Sci. 65(11): 1241-1243, 2003 It has been reported that activities of enzymes related to energy metabolism including ATP production in peripheral leukocytes are very variable in animal species [23,24]. In particular, activities of enzymes in the malate-aspartate shuttle, which has a very important role in the transfer from cytosolic NADH to mitochondrial NADH [13], change remarkably according to variation in energy metabolism of animals [2]. Lactate dehydrogenase (LDH) is a tetrameric molecule made up of four subunits of the two parent molecules, the H and M subunits, and each tissue has a characteristic composition of isoenzymes [8]. The isoenzyme patterns of LDH are variable in plasma and leukocytes of each animal species [24]. Activities of enzymes related to energy metabolism in leukocytes are considered to reflect the metabolic states in animal tissues [2,3,24]. On the other hand, plasma glucose and insulin concentrations are different between dairy and beef cattle [12,15,20,22]. It has been reported that Japanese Black heifers have higher plasma insulin level at 18 mo of age than at or before 12 mo of age, whilst Holstein heifers have very similar values at all stages [19].In the present study, activities of enzymes in the malateaspartate shuttle and LDH and LDH isoenzyme pattern were determined in plasma and peripheral leukocytes of the lactating dairy cattle and the fattening beef cattle to clarify one aspect of the differences in energy metabolism between the dairy and beef cattle.Eight lactating dairy Holstein cows (5 to 8 years old) maintained with good quality hay and concentrates in the Fuji Animal Farm of our university and four fattening Japanese Black Wagyu × Holstein steers (2 years old) kept with high concentrate feed at a farm in Shimane Prefecture were used. Blood was withdrawn from the jugular vein into heparinized tubes, with the animals at rest and unfasted. The plasma was recovered by centrifugation at 4°C. Leukocytes were isolated by centrifugation with a lymphocyte-isolating solution (ICN Biochemical Inc., Aurora, OH, U.S.A.). Cytosolic and mitochondrial fractions of leukocytes were prepared by the method described previously [23]. The activities of the enzymes were measured by using reported met...
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