Objective-To use transient and stable transfection of Chinese hamster ovary cells to clone the gene encoding feline erythropoietin (feEPO) protein, characterize the expressed protein, and assess its biological activity. Sample Population-Cultures of Chinese hamster ovary or TF-1 cells. Procedure-The gene encoding feEPO was cloned into a eukaryotic expression plasmid. Chinese hamster ovary cells were transiently or stably transfected with the plasmid. Expressed recombinant feEPO (rfeEPO) protein was purified from transiently transfected cells. The protein was characterized by use of SDS gel electrophoresis and western blot analysis. Biological activity was assessed by measuring thymidine incorporation by TF-1 erythroleukemic cells. Results-Purified rfeEPO from supernatants of transiently transfected cells was determined to be 34 to 40 kilodaltons (kd) by use of SDS gel electrophoresis, whereas the molecular weight predicted from the amino acid sequence was 21.5 kd. The banding pattern and high molecular weight suggested the protein was glycosylated. The rfeEPO proteins derived from transient or stable transfections subsequently were determined to be biologically active in vitro. Conclusions and Clinical Relevance-The gene encoding feEPO can be transfected into eukaryotic cells, and the expressed rfeEPO protein is biologically active in vitro. Cats with chronic renal failure often are anemic as a result of reduced expression of erythropoietin (EPO). Treatment with human-derived EPO stimulates RBCs in anemic cats; however, treatment is often limited by the development of antibodies directed against the recombinant human protein, which can then cross-react with endogenous feEPO. Recombinant feEPO may prove beneficial for use in cats with chronic renal failure.

Terminal renal dysfunction (TRD) was induced in 2 groups of dogs by partial surgical ablation of the kidney. Dogs of a control group and of 1 of the TRD groups were maintained on a diet containing normal phosphorus concentration, whereas dogs of the other TRD group were maintained on a low-phosphorus diet. Mild anemia developed in dogs of both TRD groups and could not be attributed to iron deficiency, increased erthrocyte concentration of 2,3-diphosphoglycerate, or absolute deficiency of erythropoietin (EP). Subsequently, all dogs were acutely depleted of approximately 25% of their blood volume. Erythropoietin concentration in dogs of the TRD groups was lower than that of controls, however, erythroid regenerative capacity was comparable with that of control dogs when plasma parathyroid hormone (PTH) concentration was lowered by reduced dietary intake of phosphorus. The PCV in dogs of the chronic TRD goups had a slight positive correlation with serum EP concentration, and a significant (P lesss than 0.05) negative correlation with plasma PTH and serum phosphorus and creatinine concentrations, using a correlation matrix. There was no longer a significant correlation between plasma PTH concentration and PCV after controlling for serum creatinine concentration by use of a multiple linear regression analysis. A significant (P less than 0.05) negative correlation also was observed between plasma PTH and serum EP concentrations, but not between serum EP and phosphorus or or creatinine concentrations. Significance of the EP and PTH association was reduced when analyzed, using a multiple linear regression analysis that included serum creatinine values. According to these data, PTH does have an independent effect on EP concentration, but the relationship between PTH and EP is not sufficiently strong to account for the genesis of the anemia of chronic TRD.

Serum from dogs with surgically induced renal impairment was incorporated into the medium for erythroid bone marrow cultures. A significant correlation was found between serum activities of erythropoietin and numbers of erythroid colony-forming units grown in culture. Serum creatinine concentrations had no correlation, and serum parathyroid hormone activities had a negative correlation with numbers of erythroid colony-forming units that was below the level of significance. Purified 1-84 parathyroid hormone added to bone marrow cultures was found to be stimulatory to erythroid colony-forming unit growth in higher concentrations, but decreased the number of burst-forming units. Unmeasured substances in the canine serum appeared to have a greater effect on the canine erythroid bone marrow cultures than did creatinine or parathyroid hormone values.

Determination of influence of polymorphism of gene of erythropoetin receptor (EPOR) on productive qualities of sows and reproductive characters of boars of Belarusian meaty breed was realized in the conditions of the Republic of Belarus. Research results proved the possibility of EPOR gene application as a genetic marker which determined productive abilities of swine of Belarusian selection breed. Frequency of undesirable allel EPOR**c of EPOR gene in the analyzed swine stock balanced within the frames of wide range: from 44% for Belarusian meaty breed sows up to 78% for Belarusian Kripnaya Belaya breed sows. Analysis of influence of genotype with EPOR gene on multiple fetation of sows of Belarusian meaty breed showed that sows with the preferred genotype (EPOR**tt) statistically reliable (P less than 0,01) exceeded sows with undesirable genotype (EPOR**cc) in accordance with number of alive birth piglets on 1,7 heads. Among sows of Belarusian Kripnaya Belaya breed with genotype EPOR**ct there were stated multiple fetation indexes exceeding the sows with genotype EPOR**tt on 0,5 heads. Desirable EPOR**tt genotype irrespective of sow age rendered the positive influence on their multiple fetation. It was recommended to realize genetic testing on determination og polymorphism of gene of erythropoetin receptor among the animals of Belarusian meaty and Belarusian Kripnaya Belaya breeds. Selection of parental pairs of the selection nucleus was recommended to realize taking into consideration genotypes, giving preference to animals with EPOR**t allel