Genetic variation in the carbonic anhydrase isozymes of macaque monkeys. II. Inheritance of red cell carbonic anhydrase levels in different carbonic anhydrase I genotypes of the pig-tailed macaque,Macaca nemestrina

The inheritance of red blood cell levels of carbonic anhydrase isozymes (CA I and CA II) has been studied in different carbonic anhydrase I genotypes of the pig-tailed macaque, Macaca nemestrina. Quantitation of CA I isozymes in a series of animals indicates that the total CA I concentration is the sum of the average effects of each CA I structural allele and that the average effects are independent of the various allelic combinations. The relative average effects were 0.32:0.95:1.0 for the CA I ^a, CA I^b, and CA I ^c structural genes, respectively. It is also demonstrated that the level of CA II is related to the CA I genotypes. Multiple regression analysis demonstrated that each dose of CA I-deficiency gene present decreased the CA II concentration by approximately 30%, with this decrease in CA II level being solely related to the dose of CA I-deficiency gene and not to the level of CA I. The CA I-deficient animals produce CA I products that are similar to the common CA Ia, CA Ib, CA Ic electrophoretic types. Limited mating data indicate that the CA I components in CA I-deficient animals are inherited codominantly.Supported by U.S. Public Health Service Research Grant GM-15419.This report is a portion of a dissertation submitted to the University of Michigan in partial fulfillment of the requirements for the Doctor of Philosophy degree.U.S. Public Health Service Predoctoral Trainee (GM-71-14).     

Genetic variation in the carbonic anhydrase isozymes of macaque monkeys. IV. Degradation by heat and proteolysis of normal and variant carbonic anhydrase isozymes of Macaca nemestrina

Studies were undertaken on the heat denaturation and proteolytic degradation by alpha-chymotrypsin of the normal red cell carbonic anhydrase isozyme, CA II, and two electrophoretic variants of carbonic anhydrase I, CA Ia and CA Ib, of the pigtail macaque. The heat degradation results showed a difference of about 40-fold in the rate constants between CA Ia and CA Ib, which is due to the marked thermostability of CA Ib compared to CA Ia. The enthalpies and entropies of activation were calculated from the heat denaturation constants. These values were compared, on enthalpy-entropy compensation plots, with those values previously determined for the human CA I and CA II isozymes. They were highly correlated and clearly fell into two distinct clusters, separated by about 200 kJ mol-1; one group comprising the macaque and human CA I isozymes and the other the CA II isozymes. The proteolytic degradation results showed that CA Ia is degraded about 2.5 times more rapidly than CA Ib by alpha-chymotrypsin. Thus, the characteristic 3/1 ratio of CA Ib/CA Ia in mature red cells could be accounted for by the greater susceptibility of CA Ia to degradation at some stage in red cell development.     

Pneumatocele in a pig-tailed macaque (Macaca nemestrina)

Radiographic examination of a pig-tailed macaque (Macaca nemestrina) with pneumonia revealed a large pneumatocele. The pneumatocele, a thin-walled, partially fluid filled radiolucent area, occupied approximately one-third of the left thorax. Rapid resolution of the pneumatocele accompanied antimicrobial treatment of the pneumonia and coincided with clinical improvement. Severe pulmonary acariasis was found at postmortem 15 months later.     

Genetic aspects of quantitative variation in the carbonic anhydrases of the pig-tailed macaque,Macaca nemestrina. I. Response to thyroxine

Effects of thyroxine on incorporation of l-serine-C¹⁴ into four carbonic anhydrase isozymes (CA II, CA Ia, CA Ib, CA Ic) and hemoglobin were quantified in reticulocytes of Macaca nemestrina in vitro. Response to thyroxine differed significantly between CA Ia and two allelic variants (CA Ib and CA Ic) and the nonallelic isozyme (CA II). The effects of thyroxine on serine incorporation into hemoglobin and three of the carbonic anhydrase isozymes were shown to be nonlinear with thyroxine concentration.     

Detection of systemic amyloidosis in the pig-tailed macaque (Macaca nemestrina)

Secondary amyloidosis is a progressive systemic disease for which there is no reliable diagnostic assay, preventive measure, or treatment. In an attempt to elucidate an antemortem diagnosis, 30 female pig-tailed macaques (Macaca nemestrina) at the Washington National Primate Research Center were surveyed for amyloidosis. Amyloid was demonstrated histologically in 47% (14 of 30) of the animals. The distribution and severity of amyloid deposition was variable. Affected animals had a mean age (+/-1 standard deviation) of 13.2 +/- 4.9 y, which was significantly greater than the mean age of unaffected animals (9.3 +/- 4.1) y. Twelve tests were evaluated for detection of amyloidosis; the diagnostic value of each was determined through comparison of histologically positive and histologically negative animals. Diagnostic tests evaluated were endoscopic examination and biopsy of the stomach and colon, abdominal ultrasonography, hepatic radiology, serum amyloid A (SAA), endothelin 1, alpha-fetal protein, aspartate aminotransferase (AST), alanine aminotransferase, gamma-glutamyltransferase (GGT), alkaline phosphatase, cholesterol, blood urea nitrogen, total bilirubin, C-reactive proteins, and erythrocyte sedimentation rate. Amyloidotic animals demonstrated a distinctive serologic profile: elevated SAA, GGT, and AST in combination with decreased total protein and albumin. Radiology demonstrated hepatomegaly in animals with hepatic amyloid deposition. In the absence of known infection or trauma, an amyloidotic serologic profile and radiologic hepatomegaly are consistent with systemic amyloidosis in M. nemestrina.     

Genetic variation and evolution in the red cell carbonic anhydrase isozymes of macaque monkeys

The electrophoretic phenotypes of the two isozymes of red cell carbonic anhydrase, CA I and CA II, are described in nine species of macaque monkeys from southeast Asia and Japan. Twelve phenotypes of CA I, apparently under the control of seven alleles, and five phenotypes of CA II, under the control of three alleles, were found in the different macaque populations studied. Extensive electrophoretic polymorphisms of CA I were found in three species (Macaca nemestrina, Macaca speciosa, and Macaca fuscata), and polymorphisms at the CA II locus were found in Macaca irus, Macaca mulatta, and M. nemestrina. In addition to the electrophoretic polymorphisms at the CA I locus in M. nemestrina, an inherited deficiency of CA I was also discovered in which approximately 30% of the individuals in all populations of M. nemestrina tested showed the deficient phenotype. Although the recessive gene controlling this deficiency appears to be an allele of the CA I locus, it is postulated that the CA I deficiency could also be under the control of a closely linked gene. The comparative data on the extent of genetic variation observed in the two isozymes of red cell carbonic anhydrase in macaques appear to support the concept that CA I has evolved more rapidly than CA II in mammals.Supported by USPHS grant GM-15419 and NSF grants GF-253, GB-7426, and GB-15060 of the U.S.-Japan Cooperative Science and Systemic Biology Programs.     

Genetic variation in the carbonic anhydrase isozymes of macaque monkeys. I. The radioimmunosorbent assay

A radioimmunosorbent technique is described which is capable of independently detecting both isozymes of carbonic anhydrase, CA I and CA II, in concentrations as low as 1 ng/ml. The technique is used to quantitate the different electrophoretic variants of red cell CA I as well as levels of CA II in the pig-tailed macaque, Macaca nemestrina.Supported by U.S. Public Health Service research grant GM-15419.U.S. Public Health Service Predoctoral Trainee (GM-71-14).     

A periodic dosing model of fetal alcohol syndrome in the pig-tailed macaque (Macaca nemestrina)

A nonhuman primate on a periodic ethanol dosing schedule should provide a model of fetal alcohol syndrome (FAS) most relevant to the majority of pregnant women who are “social drinkers” and can exercise reasonable control over their ethanol intake. In this pilot study, four pregnant pig-tailed macaques (Macaca nemestrina) received ethanol once a week from 40 days' gestation. Doses were 2.5 gm/kg for three moderate-dose animals (MDAs) and 4.1 gm/kg for one high-dose animal (HDA). Peak blood ethanol levels reached a mean of 240–256 mg/dl for the MDAs and averaged 379 mg/dl for the HDA. Peak acetaldehyde did not vary with dose. One MDA aborted after the first dose. The other three pregnancies were compared with eight to ten control pregnancies, and the infants' development over the first six months was compared with that of the control offspring. Nutritional status of the pregnant females was normal. The fetal heart rate response to maternal restraint was absent in the HDA. Gestational duration and simian Apgar scores were normal. All three infants were abnormally large, and two were also abnormally heavy, with body weight appropriate to skeletal size. Skeletal maturation, judged by ossification and tooth eruption, was not accelerated. The high-dose infant (HDI) was scaphocephalic, with an underdeveloped cranial base and midface, and its brain was small and dysplastic; its reflex, motor, and cognitive development were retarded. One moderate-dose infant (MDI) had some brain abnormalities; it was hyperkinetic and showed developmental retardation on several behavioral measures. The other MDI was normal. We conclude that the periodic model offers an effective means of investigating FAS in M. nemestrina. Furthermore, when nutrition is maintained, intermittent intake of ethanol by the pregnant primate does not necessarily reatard fetal growth.     

Complete amino acid sequence of the gamma chain from the major fetal hemoglobin of the pig-tailed macaque, Macaca nemestrina.

The complete primary structure of the gamma chain of the major fetal hemoglobin from the pig-tailed macaque, Macaca nemestrina, was obtained by the automated sequencing of fragments produced by three nonenzymatic cleavage reactions. About two-thirds of the sequence was established from the amino terminus of the intact chain and two of the three fragments produced by cleavage at methionyl residues by cyanogen bromide. Acid clevage at the single aspartyl-prolyl linkage and cleavage at tryptophanyl residues in intact chains yielded the two fragments necessary to complete the sequence. This gamma chain, the first from a nonhuman primate to be sequenced, differes from the human G gamma and A gamma chains at but 4 and 5 positions, respectively. All substitutions are conservative and unlikely to produce alterations in the oxygen-binding properties of tetrameric fetal hemoglobin. Consideration of the data presented herein, together with published observations made on portions of other primate gamma chains, provides some insight into the evolutionary history of the multiple gamma-globin chains observed in several anthropoid primates.     

Subacute necrotizing encephalopathy in a pig-tailed macaque (Macaca nemestrina) that resembles mitochondrial encephalopathy in humans

A male pig-tailed macaque (Macaca nemestrina), approximately 5 years old, was found to be vision-impaired and to have profound behavioral abnormalities, including hyperactivity and self-injurious behavior that was not amenable to amelioration by environmental enrichment. Facial and skeletal dysmorphisms also were noted. Magnetic resonance imaging (MRI) and positron emission tomography (PET) scanning revealed areas of possible infarction in the occipital lobes and megaventriculosis. At necropsy, following euthanasia for humane reasons, severe polio- and leukoencephalomalacia accompanied by megaventriculosis were seen in both occipital lobes and in several sulci of the parietal and frontal lobes. Light microscopic findings included loss of neocortical structure, with necrosis, neuronal loss, astrogliosis, vascular proliferation, mild spongiosis, and demyelination. The extent and severity of lesions were most pronounced in the occipital lobes and were greater in the left than in the right hemisphere. Other lesions included mild bilateral atrophy of the optic nerves, thymic involution, necrotizing dermatitis due to trauma, and a spectrum of spermatozoal abnormalities. The imaging and gross and light microscopic changes found in this animal resemble the mitochondrial encephalopathies of humans; this was corroborated by results of immunohistochemical analysis demonstrating decreased expression of enzymes of the mitochondrial oxidative complex ([OC]-I, -III, and -IV) in brain and muscle, and detection of fibrinogen immunoreactivity in neurons and glial cells. The spermatozoal defects may represent yet another aspect of a mitochondrial defect.     

Pregnancy outcomes after weekly oral administration of ethanol during gestation in the pig-tailed macaque (Macaca nemestrina)

Ethanol was orally administered once per week to gravid pig-tailed macaques (Macaca nemestrina) in doses of 0.3, 0.6, 1.2, 1.8, 2.5, 3.3, or 4.1 g/kg. A control group received a sucrose solution, isocaloric and isovolemic to the highest ethanol dose. Pregnancy was followed after 116 possible conceptions in 54 females. Peak plasma ethanol concentrations (PPECs) ranged from 24 +/- 6 mg/dl at the 0.3 g/kg dose to 549 +/- 71 mg/dl at the 4.1 g/kg dose. An increased rate of spontaneous abortion was related to ethanol exposure at and above 1.8 g/kg (mean PPEC = 205 mg/dl). Pregnancy failure in the first 30 days of gestation increased at doses above 2.5 g/kg. The effect on pregnancy outcome of weekly exposure to ethanol in this nonhuman primate is comparable to available data on humans. The methodology of this study represents an effective model for studying ethanol teratogenesis in a nonhuman primate.     

A longitudinal study of the growth pattern of the maxillary sinus in the pig-tailed macaque (Macaca nemestrina)

The ontogeny of sexual dimorphism in maxillary sinus size in a nonhuman primate was studied longitudinally for a period of 8 years in 25 female and 25 male Macaca nemestrina via lateral cephalograms. The maxillary sinus was traced and its area digitized. The growth of female maxillary sinuses was described with a Gompertz model; the best fit to the male data was obtained by the logistic model. Growth curves and confidence intervals revealed that the sinuses grew in a similar fashion for 3-4 years in both sexes. After this, female sinuses achieved a plateau in their development while male sinuses continued to grow. Confidence intervals suggested that size dimorphism appeared at the age of 6.3 years. Lowess regression indicated growth spurts in both sexes. Females experienced an earlier and smaller spurt than males. Sexual dimorphism in maxillary sinus size seems to represent a combination of differences in velocity and length of growth. This study indicates that growth of the maxillary sinus follows closely the growth in body size. Nevertheless, due to the variation in sinus size in Macaca, it is questionable if body size is the main determinant of maxillary sinus size. It is suggested that Macaca, with its wide geographic range and different environments, is an especially appropriate genus to use to test hypotheses about the evolution of skull pneumatization in primates.     

Blood groups of pig-tailed macaques (Macaca nemestrina).

The human-type A-B-O blood groups of 57 pig-tailed macaques (Macaca nemestrina) were determined and the calculated gene frequencies, O = 0.8908, A = 0.0825 and B = 0.0267, gave excellent fit with the hypothesis of inheritance by triple allelic genes. In tests for simian-type blood groups with rhesus, baboon and crab-eating macaque immune antisera, it was shown that the red cells of pig-tailed macaques are polymorphic for several simian-type specificities defined by those cross-reacting sera. Pig-tailed macaques share with other macaque species the complex Drh-graded blood group system, which seems to occupy a special role among red cell antigens of macaques. Normal sera of three female macaques contained spontaneous isoagglutinins which selectively agglutinated the red cells of some pig-tailed as well as stump-tailed macaques.     

Intraspecific red cell enzyme variation in the pigtailed macaque (Macaca nemestrina)

The erythrocytes of 350 pigtailed macaques (Macaca nemestrina) were examined for electrophoretic variation of hemoglobin and 26 enzymes. Seven enzymes showed variation in more than 1% of individuals: phosphoglucose isomerase, phosphoglucomutase-1, soluble NADP-dependent isocitric dehydrogenase, peptidase A, peptidase C, 2,3-diphosphoglycerate mutase, and acid phosphatase. Variation with lesser frequency was found in soluble glutamic-oxalacetic transaminase, phosphoglycerate kinase, lactic dehydrogenase, and hemoglobin. Only eight samples were tested for esterase D, and one of these had a variant phenotype. Enzymes with no clear variation were adenylate kinase, adenosine deaminase, phosphofructokinase, hexokinase, pyruvate kinase, glyceraldehyde 3-phosphate dehydrogenase, aldolase, phosphoglycerate mutase, phosphopyruvate hydratase (enolase), phosphoglucomutase-3, and superoxide dismutase. There was father-to-son transmission of PGI, PGM-1, peptidase C, 6PGD, 2,3-DPGAM, NADP-ICD, and acid phosphatase variants, suggesting that these loci are autosomal as in man.