ABSTRACT
Automated blood cell counters can distinguish cells based on their size and the presence or absence of a nucleus. However, most vertebrates have nucleated blood cells that cannot be counted automatically. We established an alternative automatic method for counting peripheral blood cells by staining cells with the fluorescent dye acridine orange (AO) and analysing cell populations using flow cytometry (FCM). As promising new animal models, we chose Xenopus laevis and three inbred strains of X. tropicalis. We compared the haematological phenotypes, including blood cell types, cell sizes, cellular structure, and erythrocyte lifespans/turnover rate among X. laevis and the three inbred strains of X. tropicalis. Each cell type from X. laevis was sorted according to six parameters: forward- and side-scattered light emission, AO red and green fluorescence intensity, and cellular red and green fluorescence. Remarkably, the erythrocyte count was the highest in the Golden line, suggesting that genetic factors were associated with the blood cells. Furthermore, immature erythrocytes in anaemic X. laevis could be separated from normal blood cells based on red fluorescence intensity. These results show that FCM with AO staining allows for an accurate analysis of peripheral blood cells from various species.
Subject(s)
Blood Cells , Cell Separation/methods , Flow Cytometry/methods , Staining and Labeling/methods , Xenopus laevis/blood , Acridine Orange/chemistry , Animals , Animals, Inbred Strains/blood , Animals, Wild/blood , Blood Cell Count/methods , Fluorescent Dyes/chemistry , Models, Animal , Species SpecificityABSTRACT
The objective of this study was to assess differences in serum protein expression profiles of Przewalski's (Mongolian wild horse) and thoroughbred horses using proteome analysis. The serum proteins were separated by two-dimensional electrophoresis (2-DE) and five different gene products were identified. Proteins represented by the five spots were identified by matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry (MS)/MS technology. The identities of all proteins were deduced based on their similarity to proteins in the human plasma protein database. Three proteins (a haptoglobin-2 alpha glycoprotein and two haptoglobin-2beta glycoproteins with different accession numbers) were downregulated in Przewalski's horse sera compared to thoroughbred horse sera. Moreover, two proteins (tetraspanin-18 and pM5) were upregulated in Przewalski's horses compared to thoroughbred horses. Haptoglobin-2 alpha and haptoglobin-2beta may serve as candidate molecules in future studies of inflammation, coagulation, immune modulation and pro-oxidant and antioxidant activity with consequential effects on the entire metabolism of the horse.
Subject(s)
Animals, Inbred Strains/blood , Animals, Wild/blood , Blood Protein Electrophoresis , Blood Proteins/isolation & purification , Electrophoresis, Gel, Two-Dimensional , Horses/blood , Animals , Humans , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
Serum samples collected from 119 (72 male and 47 female) mastomys (Praomys coucha) of 2 specific-pathogen-free inbred strains (RI4 and RI7) were analyzed for 12 serum biochemical parameters. Sex-related differences (p < 0.01) were noted in alkaline phosphatase and glucose; the both higher in females than in males. Age-related changes (p < 0.01) were observed in total protein, albumin, total cholesterol, and alkaline phosphatase, with higher values for the first three parameters in the older group (200-250 days of age) than in the younger group (90-140 days of age). Four out of 12 parameters showed strain-related differences (p < 0.01), consistent with the large amount of genetic heterogeneity reported in this species. These serum biochemical reference values should provide information for the use of mastomys in laboratory research.
Subject(s)
Animals, Inbred Strains/blood , Muridae/blood , Animals , Blood Chemical Analysis , Female , Male , Reference Values , Species Specificity , Specific Pathogen-Free OrganismsABSTRACT
We determined the effects of sex, age, and environment (inbred, captive-wild, and wild animals) on selected blood parameters of the cotton rat (Sigmodon hispidus) in central Oklahoma (USA) from 1990 to 1994. Male and female cotton rats had similar blood profiles. Age-related differences were confined to differential white blood cell counts where adults possessed greater numbers of neutrophils and lower numbers of lymphocytes compared to juveniles. Environment had a strong influence on many hematological parameters. Hematocrit, hemoglobin concentration, RBC count, and eosinophil number were generally greater for wild stocks compared to inbred animals, and differences were more pronounced for adults than juveniles.
