RESUMO
An unusual lipoprotein was detected and purified from the blood of some members of a large colony of baboons, Papio sp. This lipoprotein was found to be similar to human lipoprotein a in all respects and is therefore termed lipoprotein a. Baboon lipoprotein a had a density of 1.052 g/ml and was located between low- and high-density lipoproteins in a density gradient ultracentrifugation. However, despite its greater density, baboon lipoprotein a was larger than low-density lipoprotein, based on gradient gel electrophoresis and gel filtration. The lipoprotein contained a very large apolipoprotein (apolipoprotein-lipoprotein a) which was found to consist of an apolipoprotein B linked to another protein called apolipoprotein a by a disulfide bridge(s). In all these characteristics, baboon lipoprotein a was similar to human lipoprotein a.
Assuntos
Lipoproteínas/sangue , Papio/sangue , Animais , Apolipoproteínas B/análise , Centrifugação com Gradiente de Concentração , Cromatografia em Gel , Dissulfetos/análise , Eletroforese em Gel de Poliacrilamida , Lipoproteína(a) , Lipoproteínas LDL/sangueRESUMO
We have raised specific antibodies against the protein component of baboon lipoprotein (a) (Lp(a]. Apolipoprotein (apo) Lp(a) is a very large protein which separates into two distinct proteins, apo B and apo (a), when 2-mercaptoethanol is included during sample treatment for sodium dodecyl sulfate-electrophoresis. The antibodies were specific for baboon apo (a) and apo B. The presence of the two distinct antigens in the lipoprotein permitted the development of an enzyme-linked immunosorbent assay that was specific for Lp(a) particles in serum. The assay could detect less than 1 ng of Lp(a) protein per well and was useful in the range of 1-9 ng. The assay was specific for Lp(a) and did not respond to other lipoproteins, such as low density lipoprotein. Lp(a) could be accurately quantitated in serum frozen at -80 degrees C in plastic tubing segments. Using the Lp(a) assay, the mean serum level of 80 unrelated baboons was 4.7 mg/dl, with the distribution skewed toward the lower levels. These data further support the value of the baboon as a model of the atherogenic lipoprotein Lp(a).
Assuntos
Lipoproteínas/sangue , Papio/sangue , Animais , Especificidade de Anticorpos , Apolipoproteínas A/imunologia , Apolipoproteínas B/imunologia , Eletroforese das Proteínas Sanguíneas , Centrifugação com Gradiente de Concentração , Ensaio de Imunoadsorção Enzimática , Immunoblotting , Lipoproteína(a) , Lipoproteínas/imunologiaRESUMO
The serum protein transferrin (Tf) is a valuable marker for genetic studies of primates, because it is usually polymorphic, exhibiting as many as 13 allelic forms with high heterozygosity. The standard procedure to detect the different phenotypes requires vertical electrophoresis on polyacrylamide gels for 18 h at 4 degrees C. We have simplified the procedure by using the automated programmable PhastSystem for electrophoresis, which uses precast miniature gels and takes less than 2 h. Also, we have developed a pool of sera that displays the 6 most common alleles in rhesus monkeys. This pool allows for a more accurate assignment of phenotypes. Using this simplified procedure, we have typed over 3000 monkeys of various species and have confirmed the reliability of this technique for parentage determination.
Assuntos
Eletroforese em Gel de Poliacrilamida/métodos , Genótipo , Transferrina/genética , Alelos , Animais , Macaca mulatta/sangue , FenótipoRESUMO
One of the prime concerns at zoos and at primate breeding facilities is to maintain genetic variability. This can be accomplished by avoiding inbreeding. It is relatively easy to assess genetic variability and the level of inbreeding by using pedigree information and genetic markers. In this study we used genetic markers controlled by 6 independent polymorphic loci (GPI, PGD, CA2, MPI, DIA1, Tf) to ascertain genetic variation in two captive and one wild population of rhesus monkeys. Two other loci ADA and NP were also examined and found to be monomorphic in the three populations. F-statistics and contingency chi-square analyses indicated that there was significant genetic differentiation among the populations. We also found that the mean heterozygosities were very similar in the three populations, in spite of the diverse breeding strategies. These data are important because rhesus monkeys are frequently used for biomedical research; and the genetic markers provide useful information for genetic management of captive colonies of nonhuman primates. © 1992 Wiley-Liss, Inc.
RESUMO
Baboons possess Lp[a] that is similar to human Lp[a], including the presence of the unique protein, apo[a]. Baboon apo[a] occurred in at least nine isoforms distinguishable by size. Isoforms were resolved by 3-12% polyacrylamide gradient gel electrophoretic separation of serum proteins, and were detected with baboon apo[a]-specific antibodies. Thirty one different apo[a] isoform phenotypes were detected in a population of 165 unrelated baboons. Identical isoform phenotypes were observed in different samples from individual baboons, and isoform phenotypes were unaffected by changes in diet. In one experiment, 16 baboons were fed a series of five diets differing in amounts of cholesterol and saturated or unsaturated fats. There was no significant effect of diet on serum Lp[a] levels. In another group of baboons (n = 70) controlled for age and dietary history, enrichment of the diet with cholesterol and saturated fat caused a small, but significant (P less than 0.005), increase (means = 0.6 mg/dl) in serum Lp[a] concentration. Analysis of two large sire families suggested that apo[a] isoform patterns and serum Lp[a] concentrations were inherited. Putative parental alleles responsible for specific isoform bands appeared to segregate randomly. Heritability (h2) of serum Lp[a] concentration was estimated to be 0.95 +/- 0.04. We conclude that apo[a] isoform phenotypes and serum Lp[a] concentrations are inherited, and that Lp[a] concentrations are only slightly influenced by diet.
Assuntos
Apolipoproteínas A/sangue , Gorduras na Dieta/administração & dosagem , Lipoproteínas/sangue , Papio/genética , Animais , Apolipoproteínas B/sangue , Centrifugação com Gradiente de Concentração , Colesterol/sangue , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Feminino , Immunoblotting , Lipoproteína(a) , Masculino , Papio/sangue , Linhagem , FenótipoRESUMO
Like humans, baboons possess serum lipoprotein(a) that contains apolipoprotein(a) (apo(a], and baboon apo(a) also occurs in distinguishable glycoprotein isoforms. To investigate the molecular basis for isoform size variation, we isolated hepatic RNA from baboons possessing different apo(a) isoforms for Northern blot analyses with rhesus apo(a) cDNA probes. In a survey of 22 baboons, we detected a variety of sizes of apo(a) transcripts (5.2-11.2 kilobases) that corresponded with relative mobilities and numbers of serum apo(a) isoforms. In unrelated baboons possessing apo(a) isoforms of similar mobilities, we detected apo(a) transcripts with similar mobilities. In related baboons possessing apo(a) isoforms that were identical by descent, apo(a) transcripts were also identical in size. Using data from 22 baboons included in this study, we compared the sizes of 20 apo(a) isoforms and corresponding transcripts. Linear regression analysis established a highly significant correlation (p less than 0.001) between estimated apo(a) transcript size and glycoprotein mass (r2 = 0.996). From these results, we conclude that apo(a) glycoprotein isoforms are due to structural differences in apo(a) transcripts. However, we also noted exceptions to correspondence between apo(a) transcripts and isoforms that suggest the action of additional post-transcriptional control of serum apo(a) levels.
Assuntos
Apolipoproteínas/genética , Papio/genética , Alelos , Animais , Apoproteína(a) , Northern Blotting , Glicoproteínas/genética , Lipoproteína(a) , Lipoproteínas/sangue , Fígado/fisiologia , Peso Molecular , RNA Mensageiro/genéticaRESUMO
We used 18 genetic loci including blood groups, isozymes, and a serum protein to evaluate our efforts to preserve genetic variability in a specific pathogen-free (SPF) colony of rhesus monkeys. We compared genetic variability in the SPF population to the virally contaminated, non-SPF population from which it was derived. There was no change in the average gene diversity between the SPF and non-SPF populations. However, gene diversity at blood group Q locus increased significantly in the SPF population, while blood group M locus showed an insignificant trend toward decreased gene diversity. Allele frequencies changed significantly at blood group Q locus, although no alleles were lost from the population. We hypothesized that this change was due to extensive overreproduction by a small number of founder males that possessed the initially rare allele, Q1. There was no evidence that this change was associated with genes involved in viral infection.
Assuntos
Variação Genética , Macaca mulatta/genética , Organismos Livres de Patógenos Específicos , Alelos , Animais , Antígenos de Grupos Sanguíneos/genética , Cruzamento , Simulação por Computador , Feminino , Frequência do Gene , Endogamia , MasculinoRESUMO
Monodelphis domestica, the gray short-tailed opossum, is used increasingly as an animal model in studies that require repeated blood sampling. Consequently, it is important to establish safe bleeding regimens. We investigated the effects of repeated blood loss on various hematologic values and on different organs in this species. Approximately 2 ml of blood were taken weekly from 20 animals for 13 weeks. The animals were then necropsied; members of an age- and sex-matched control group were bled (2 ml) once and necropsied immediately to obtain baseline data. Ultimately, each animal in the experimental group lost approximately three times its total blood volume. After the first bleeding in the chronically bled group, the red blood cell counts, hemoglobin, and hematocrit values decreased significantly but remained constant thereafter. In another experimental group bled only once, the hematologic values rose to higher than baseline levels after a rest of 2 weeks. Thereafter the values slowly returned to baseline levels. A notable increase in Howell-Jolly bodies occurred in the chronically bled group. Histologically, there was marked erythroid hyperplasia in the bone marrow and extramedullary hematopoiesis in the spleen, but none in the liver. Because there were no obvious detrimental physiologic effects, we conclude that M. domestica is markedly tolerant of chronic blood loss.
Assuntos
Sangria/veterinária , Gambás/sangue , Animais , Peso Corporal , Eritropoese/fisiologia , Feminino , Hematopoese Extramedular/fisiologia , Masculino , Gambás/fisiologia , Tamanho do Órgão , Especificidade de Órgãos , Baço/anatomia & histologiaRESUMO
Previously we reported that the South American gray short-tailed opossum, Monodelphis domestica, had an MHC class-I locus similar to that of eutherian species. In addition to the detection of lymphocyte antigens by cytotoxic antisera, we concluded that this marsupial rejected allogeneic skin grafts, as would be expected of animals with MHC class-I polymorphism. However, this conclusion was based on a limited number of skin transplants that were assayed for only a short period. Here we report the results of 22 reciprocal skin grafts made between individuals of known genetic relationships. On the basis of gross inspection of the grafts and histologic examination, we found that the average time of the onset of graft rejection was about 19 days and that the average time for complete graft rejection was about 31 days. In general, it took longer for the onset of graft rejection among pairs of genetically related animals than among less related animals. These results indicate unequivocally that this marsupial species has a high degree of class-I polymorphism and rejects allogeneic skin transplants in a manner similar to but more slowly than eutherian mammals.
Assuntos
Gambás/imunologia , Transplante de Pele/imunologia , Transplante de Pele/veterinária , Animais , Feminino , Rejeição de Enxerto/veterinária , Sobrevivência de Enxerto , Masculino , Transplante de Pele/estatística & dados numéricos , Estatísticas não Paramétricas , Cauda , Fatores de Tempo , Transplante Homólogo/estatística & dados numéricos , Transplante Homólogo/veterináriaRESUMO
Genetic markers are indispensable for molecular and statistical genetic research involving nonhuman primates. Genetic markers must be used to ascertain parentage and to confirm the accuracy of pedigrees based solely on housing or demographic records; otherwise, the results of pedigree, linkage, or quantitative genetic analyses may be unreliable. Until recently, most genetic markers used in nonhuman primates were plasma proteins or isozyme polymorphisms, which were required in large numbers, because levels of genetic variation revealed by these markers were rather low. We compared the newer, PCR-amplified short tandem repeat markers (STRs) with a panel of classical biochemical polymorphic markers, for paternity determination among captive-bred rhesus monkeys. The STR markers exhibited an average genetic diversity of 64% and an expected paternity exclusion probability of 0.443. Both of these were greater than the average 54.5% genetic diversity and 0.298 exclusion probability exhibited by the biochemical markers. The STRs were much more efficient than the biochemical markers for parentage determination, since they required only half the amount of genetic typing data to resolve an average paternity case. Thus, the results of applying these two classes of genetic markers in paternity tests were somewhat different than expected on the basis of theoretical exclusion probabilities. These differences were probably due to inbreeding and other genetic differences among breeding colonies. Because they are more informative and provide rapid and efficient genetic data, STRs are now the method of choice for parentage determination and pedigree corroboration among nonhuman primates.
Assuntos
Macaca mulatta/genética , Paternidade , Polimorfismo Genético , Sequências de Repetição em Tandem , Animais , Biomarcadores , Proteínas Sanguíneas/genética , Anidrases Carbônicas/genética , Di-Hidrolipoamida Desidrogenase/genética , Marcadores Genéticos , Masculino , Mutação , Plasminogênio/genética , Transferrina/genéticaRESUMO
Previously we reported preliminary results suggesting that the marsupial Monodelphis domestica fails to exhibit a mixed lymphocyte reaction with allogeneic lymphocytes. To test whether this observation is simply a matter of a response too weak to detect, but capable of being augmented by immunization, we performed mixed lymphocyte culture tests on 23 of these animals that had been immunized with lymphocytes. Despite the fact that all recipients were sensitized to the lymphocytes of the donors, none of the animals had a substantial mixed lymphocyte response. Significant stimulation was noted with the mitogen concanavalin A; thus, the T cells were immunologically competent. It seems likely that the failure of this species to exhibit a significant mixed lymphocyte response is due to T cells whose ontogeny differs from that of the T cells of eutherian mammals.