X-ray induced translocations in premeiotic germ cells of monkeys

The induction of reciprocal translocations by various X-ray exposures was studied in spermatogonial stem cells of rhesus monkeys (Macaca mulatta) and stump-tailed macaques (Macaca arctoides) by means of spermatocyte analysis many cell generations after irradiation. The yields of translocations recovered from irradiated stump-tailed macaques were lower than those observed in rhesus monkeys and represent in fact the lowest induction rates per Gy ever recorded for experimental mammals. In the rhesus monkey a humped dose-effect relationship was found with (a) a homogeneous response with (pseudo-)linear kinetics below 1 Gy, (b) much more variability at higher doses, and (c) no induction at all at doses of 4 Gy and above. It is suggested that the post-irradiation proliferation differentiation pattern of surviving rhesus monkey spermatogonial stem cells i mainly responsible for these characteristics of the dose-response curve.     

基于线粒体控制区序列的猕猴属系统发育研究

通过线粒体部分控制区DNA序列数据探讨7种猕猴属物种的分子系统发育关系。结果表明熊猴的核苷酸多样度最高,而藏酋猴核苷酸多样度较低。基于控制区序列数据所构建的最大似然树,不考虑食蟹猴的位置,7种猕猴物种可粗略地分为3个种组,即狮尾猴组(包括北平顶猴)、头巾猴组(包括红面猴、熊猴和藏酋猴)和食蟹猴组(包括恒河猴和台湾猴)。与前人(Fooden＆Lanyon,1989;Tosi et al,2003a;Deinard＆Smith,2001;Evans et al,1999;Hayasaka et al,1996;Morales＆Melnick,1998)的结果不同,我们的结果支持食蟹猴比北平顶猴分化早的假设;东部恒河猴(相对于台湾猴)和东部熊猴(相对于藏酋猴)出现并系。与Y染色体、等位酶、核基因以及部分形态学数据推测的结果(Delson,1980;Fooden＆Lanyon。1989;Fooden,1990;Tosi et al,2000,2003a,b;Deinard＆Smith,2001)一致,红面猴应归于头巾猴组,但此结论与前人(Hayasaka et al,1996;Morales＆Melnick,1998;Tosi et al,2003a)依据线粒体得到的结果有较大分歧。     

Placental transfer of phenylalanine in Macaca mulatta and Macaca fascicularis

We investigated placental transport mechanisms of phenylalanine in Macaca mulatta and Macaca fascicularis. In the beginning of the third trimester we administered i.v. phenylalanine and p-chlorophenylalanine to pregnant animals. Initial higher phenylalanine concentrations were observed followed by a rapid decrease in both rhesus mothers and fetuses when compared with phenylalanine levels in fascicularis mothers and fetuses. In general, however, placental transfer mechanisms of phenylalanine did not differ significantly between the two species.     

小鼠抗食蟹猴IgG单克隆抗体的制备

目的制备抗食蟹猴、恒河猴等非人灵长类实验动物免疫球蛋白二级抗体,开展对其传染病血清学快速诊断方法的建立。方法采用饱和硫酸铵盐析、Agarose-Protein G亲和层析技术,从食蟹猴血清中提纯IgG。经SDS-PAGE电泳鉴定,采用常规法免疫C57BL/6小鼠,三次免疫后取脾细胞与Sp2/0-Ag14骨髓瘤细胞通过PEG4000融合制备杂交瘤细胞,利用间接ELISA、Western blot等方法进行筛选、鉴定。结果得到5株阳性杂交瘤,分别命名为2B6、2B7、2D9、3B2、5E4,并且5株杂交瘤分泌的抗体均与恒河猴的IgG或血清发生交叉反应,而与其他物种如东北虎、犬等动物的IgG或血清无交叉反应。结论 5株杂交瘤产生的单克隆抗体（McAb）具有较好免疫活性,且能长期、稳定地分泌抗体。此项研究工作为后续研究食蟹猴、恒河猴传染病血清学诊断方法奠定基础。     

Blood groups of crab-eating macaques (Macaca fascicularis) demonstrated by isoimmune rhesus monkey (Macaca mulatta) sera.

Twenty-one isoimmune sera produced in rhesus monkey (Macaca mulatta) containing type-specific antibodies for simian-type red cell antigens were tested for their cross-reactivity with red cells from crab-eating macaques (M. fascicularis). The majority of the antisera gave cross-reactions determining polymorphisms in the red cells of crab-eating macaques, homologous to those of rhesus monkeys. These results attest to the close taxonomic realationship between the two species of macaques, and have the practical implication that isoimmune sera produced for blood typing can also be used for typing red cells from related species, as has been also observed in studies on apes.     

Dose-response relationship for translocation induction in spermatogonia of the crab-eating monkey (Macaca fascicularis) by chronic gamma-ray-irradiation

The induction of reciprocal translocations in spermatogonia of the crab-eating monkey (Macaca fascicularis) by chronic gamma-irradiation (1.8 x 10(-5) Gy/min, about 0.024 Gy/22 h/day) was examined. The frequencies of translocation per cell were 0.15% at 0.3 Gy, 0.27% at 1.0 Gy and 0.33% at 1.5 Gy. The dose-response relationship for translocation yield was a linear one with a regression coefficient (b) of 0.16 x 10(-2). When the slope (b) of the regression line was compared with that at a high dose rate (0.25 Gy/min, b = 1.79 x 10(-2), it was clear that the induction rate of translocations after chronic gamma-irradiation was only about one-tenth of that after high-dose-rate irradiation. Thus, there was evidence for a pronounced dose-rate effect in the crab-eating monkey.     

Dose-response relationship of gamma-ray-induced reciprocal translocations at low doses in spermatogonia of the crab-eating monkey (Macaca fascicularis)

The yield of translocations induced by acute gamma-irradiation at low doses (0.25 and 0.50 Gy) in the crab-eating monkey's (Macaca fascicularis) spermatogonia was examined. The frequencies of translocations per cell were 0.53% at 0.25 Gy and 1.07% at 0.50 Gy. Over the low dose range from 0 to 1 Gy, the dose-response relationship for translocation yield was a linear one with a regression coefficient of 1.79 X 10(-2). To estimate the sensitivity to the induction of translocations in the crab-eating monkey's spermatogonia, the slope of the regression line was compared with those in other mammalian species. Consequently, over the low dose range below 1 Gy, the sensitivity of the crab-eating monkey's spermatogonia to translocation induction was similar to several mammalian species, the mouse. Chinese hamster, and the rabbit, but significantly higher than that of the rhesus monkey and lower than that of the marmoset.     

Ultrasonic assessment of pregnancy and fetal development in three species of macaque monkeys

Serial ultrasonic assessments of gestational sac (GS) and fetal biparietal diameter (BPD) were performed in the Japanese monkey (Macaca fuscata fuscata), rhesus monkey (Macaca mulatta), and crab-eating monkey (Macaca fascicularis). In the Japanese monkey, GS increased linearly for 3-8 weeks, whereas BPD increased in a linear-quadratic manner over 8 weeks to term. Ultrasonic assessments of spontaneously aborted fetuses with BPD growth retardation and diagnosis of a pelvic chocolate cyst also were reported.     

Ameloblastic odontoma in a cynomolgus monkey (Macaca fascicularis)

An ameloblastic odontoma occurred in an adult cynomolgus monkey (Macaca fascicularis). The neoplasm involved the upper left maxilla as a disfiguring, fleshy growth. The tissue and cellular changes were consistent with an ameloblastic odontoma which is rare in man, nonhuman primates (NHP), and other vertebrate animals. The monkey was one of 50 adult, single-caged housed cynomolgus monkeys. No additional clinical signs of disease were present. Hematology and serum chemistries were within normal limits. There have been three reports in the literature of ameloblastic odontomas in NHP; however, this is the first reported case of an ameloblastic odontoma in a cynomolgus monkey.     

恒河猴与食蟹猴微卫星DNA多态性的比较分析

目的分析恒河猴和食蟹猴群体间的遗传多样性,确立一种对恒河猴和食蟹猴种群个体的遗传鉴别方法。方法利用聚合酶链反应（PCR）扩增技术采用15个多态性微卫星DNA位点对50只恒河猴和50只食蟹猴个体进行了DNA多态性的分析,对比两群体间等位基因数目差异。结果筛选的15个具有显著多态性的微卫星DNA位点对恒河猴和食蟹猴种群可以进行DNA多态性分析,其等位基因数目均在7个以上,且两群体间有11个位点的等位基因数存在一定的差异。结论利用这些多态性微卫星DNA位点建立一种有效鉴别恒河猴和食蟹猴种群遗传背景的方法具有一定的可行性。     

Longitudinal study of craniofacial growth in Macaca fascicularis

This paper is an analysis of normal craniofacial growth in adolescent crab-eating macaques (Macaca fascicularis). Eight female adolescent monkeys were used in this study. Their individual craniofacial growth was studied for a 24-month period utilizing tantalum implants and roentgenographic cephalograms. Throughout the observation period, each monkey consistently showed a class I molar relationship with a good overjet and overbite. The amount of anterior displacement of the maxilla and the mandible was significantly dominant compared to the vertical displacements at every observation period. The midface exhibited a maxillary differential growth pattern in which the premaxilla displaced superiorly and the posterior maxilla moved inferiorly, resulting in a counterclockwise rotation of the entire maxilla. Growth of the lower anterior teeth and alveolar bone compensated for the incremental vertical spaces which were induced by superior displacement of the premaxilla and inferior repositioning of the chin. In addition, the amount of anterior displacement of the upper and lower anterior teeth were significantly larger than that of the premaxilla and the chin. The dentocraniofacial growth pattern in Macaca fascicularis was quite similar to that seen in Macaca mulatta.     

Normal hematological indices, blood chemistry and histology and ultrastructure of pancreatic islets in the wild Indian bonnet monkeys (Macaca radiata radiata)

The present study is aimed at determining some haematological and biochemical parameters in the wild Indian bonnet monkeys as also the microscopic and ultrastructural characteristics of their pancreatic islets. Adult wild Indian bonnet monkeys (Macaca radiata radiata) of both sexes weighing between 2.5 and 4 kg were used in these experiments. Their platelet, reticulocyte and total leukocyte counts and the blood concentrations of hemoglobin and plasma proteins and the serum concentrations of aspartate amino transferase, alanine amino transferase and calcium are similar to the values reported for M. mulatta. Plasma glucose is lower when compared with reported values of M. mulatta and M. fascicularis. Insulin levels are comparable with those of M. mulatta and M. nigra. Histology of islets is similar to that of humans. Ovoid cell collections of islet cells are scattered throughout the pancreas. Ultrastructure of A, B and D cells is similar to humans. These findings suggest that this relatively underutilized macaques may be a suitable model for biomedical research.     

Gonadotropin-independent proliferation of the pale type A spermatogonia in the adult rhesus monkey (Macaca mulatta)

The goal of the present study was to examine the relative roles of testosterone (T) and FSH in the proliferation and differentiation of pale type A (Ap) spermatogonia in the rhesus monkey (Macaca mulatta). Twenty adult male monkeys were treated with daily injections of a GnRH-receptor antagonist, acyline, to suppress endogenous gonadotropin secretion during an experiment comprising three phases. Phase 1 established a chronic hypogonadotropic state marked by a profound decrease in testicular size. During phase 2, half the monkeys were implanted with T-filled capsules, and the other half received control implants. Treatment with T produced circulating T levels of approximately 15 ng/ml and normal testicular T content. At the end of phase 2, monkeys were fitted with indwelling i.v. catheters and housed in remote sampling cages for the final phase. During phase 3, five monkeys from the T- and non-T-treated groups were stimulated with recombinant human FSH. The remaining five monkeys from each group received an infusion of vehicle. On the last day of FSH or vehicle infusion, monkeys were bilaterally castrated after receiving an i.v. bolus of bromodeoxyuridine (BrdU). The BrdU labeling of Ap spermatogonia was robust in the hypogonadotropic group and was uninfluenced by treatment with T and FSH, either alone or in combination. In contrast, both T and FSH stimulated spermatogonial differentiation, and this effect was amplified by combined treatment. We conclude that marked Ap spermatogonial proliferation occurs constitutively and in a gonadotropin-independent manner and that differentiation of Ap into B spermatogonia is absolutely gonadotropin dependent and may be driven by either T or FSH.     

Observations on acute gastric dilatation in nonhuman primates.

In the years 1967-1977 we diagnosed 23 cases of acute gastric dilatation in monkeys. Fourteen of these animals were Macaca mulatta, five Macaca fascicularis, and one each of Macaca nemestrina, Aotus trivirgatus, Saimiri sciureus, and Colobus guereza. Fourteen of the animals were males, nine were females, and all were adults or subadults. Mortality was 78% (18 of 23 animals). Thirteen of the animals had received on anesthetic, immobilizing, or tranquilizing drug 1-2 days before developing acute gastric dilatation; seven monkeys were overfed, and two had been transferred from one area to another the day prior to developing the disease. Two animals were found dead in their cages with no apparent cause for the gastric dilatation. Five Macaca mulatta and three Macaca fascicularis recovered following treatment, but two Macaca mulatta subsequently succumbed to another episode of acute gastric dilatation. Treatment consisted of evacuation of the stomach, correction of blood volume deficits and acid-base disturbances by administration of appropriate fluids, and supportive therapy for shock.     

Bone Marrow Depletion as a Coincidental Finding to Hypothermia in Macaca mulatta

Three of 16 juvenile rhesus monkeys (Macaca mulatta) and 1 rhesus of 79 adult rhesus and cynomolgus monkeys (Macaca fascicularis) were found comatose in a state of profound hypothermia after a heating failure occurred in the room in which they were housed. One juvenile monkey died shortly thereafter. The three other monkeys were revived with gradual warming and supportive therapy but later experienced separate acute clinical crises manifesting shock and died at 19, 31, and 51 days after the initial episode. Histopathologic findings of severe bone marrow depletion were observed in each of the three monkeys that died after the initial episode.     

A taxonomic re-appraisal of Sarcocystis nesbitti (Protozoa: Sarcocystidae) from the monkey Macaca fascicularis in Yunnan, PR China

The first detection of Sarcocystis nesbitti Mandour, 1969 in the Chinese mainland is reported and the morphology of the sarcocyst is described in detail. The parasite was detected in the monkey, Macaca fascicularis, maintained on a monkey farm in Yunnan Province; the infection may have occurred via faecal contamination from local rats, mice and/or birds. S. nesbitti was characterized as follows: a macroscopic sarcocyst, length of the cyst up to 2 mm; cyst wall smooth, thin and no perpendicular protrusion is seen under the light microscope; border of cyst wall wavy, primary cyst wall thin (38-65 nm) and invaginated; ground substance about 0.5-0.76 microm thick with electron-dense granules and concentric spherical bodies. The cyst wall is described as type 1 by electron microscopy. It is suspected that S. nesbitti may utilize Macaca mulatta, M. fascicularis, Cercocebus atys, and Papio papionis, as well as human as intermediate hosts. The taxonomy of S. nesbitti is re-appraised in the light of a consideration of possible experimental artefacts and examination of the past literature. Evidence is presented that S. nesbitti may be one of the species infecting humans in South Asia and that the monkey may be a potential reservoir host.     

Testicular steroidogenesis in the rhesus monkey (Macaca mulatta.

Studies were undertaken to investigate testicular steroidogenesis in the Rhesus monkey Macaca mulatta. Testicular fragments (50 mg) were incubated for 3 hr with pregnenolone-7-3H or with progesterone-7-3H. The major metabolite of pregnenolone was progesterone (70.1%), with a lesser conversion to 17-hydroxyprogesterone (1.6%), androstenedione (3.3%), and testosterone (7.2%). The delta-5 intermediates 17-hydroxypregnenolone (4.6%) and dehydroepiandrosterone (8.6%) were also identified in the pregnenolone incubates. A majority of the progesterone substrate was not metabolized by the testicular fragments (80.1%), while some conversion to 17-hydroxyprogesterone (3.4%), androstenedione (4.8%), and testosterone (11.7%) occurred in the incubates. These results suggest that testicular fragments from the Rhesus monkey may convert pregnenolone to testosterone through both the delta-4 and the delta-5 pathways.     

Phylogenetic relationships among Japanese, rhesus, Formosan, and crab- eating monkeys, inferred from restriction-enzyme analysis of mitochondrial DNAs [published erratum appears in Mol Biol Evol 1988 Sep;5(5):601]

Mitochondrial DNA (mtDNA) polymorphisms in four species of macaques, i.e., Japanese monkey (Macaca fuscata), rhesus monkey (M. mulatta), Formosan monkey (M. cyclopis), and crab-eating monkey (M. fascicularis), were analyzed to study phylogenetic relationships. When 17 restriction enzymes of 6-bp recognition were used, 42-49 sites were observed in the samples. The estimated number of nucleotide substitutions per site among Japanese, rhesus, and Formosan monkeys ranges from 0.0318 to 0.0396, and that between the crab-eating monkey and the other monkeys from 0.0577 to 0.0653. These findings suggest that the crab-eating monkey diverged from the other three approximately 1.5-3.0 Myr before the present (Mybp) and that the Japanese, rhesus, and Formosan monkeys diverged approximately 0.9-1.8 Mybp, although the branching order cannot be determined conclusively.      

Technique for culturing Macaca mulatta peripheral blood lymphocytes for fluorescence in situ hybridization of whole chromosome paints

The rhesus monkey (Macaca mulatta) has long been an important model in biomedical and behavioral research. The biomedical importance of M. mulatta is due to its 93% genetic similarity with humans and its complex social behavior. The recent sequencing of the M. mulatta genome has enhanced its role in biological research. However, the use of the macaque as an experimental model in cytogenetic assays has been problematic due to difficulties in obtaining large numbers of well-spread cells in metaphase without the use of extremely toxic mitogens such as staphylococcal enterotoxin A (SEA). Here we describe a technique for culturing and producing sufficient numbers of cells in metaphase using the common mitogens phytohemagglutinin (PHA), concanavalin A (ConA), and T-cell growth factor (TCGF) which act synergistically to induce M. mulatta T-lymphocyte division. Using this method we have obtained a mitotic index in 48 h cultures of 12.0+/-2.2 metaphase cells/100 cells (n=5 animals). Fluorescence in situ hybridization with whole chromosome painting of M. mulatta cells was performed with human whole-chromosome probes that labeled the following chromosomes for human (M. mulatta): 1(1), 2q(12), 2p(13), 4(5) pairs in red, and 3(2), 5(6) and 6(4) pairs in green. In humans this probe combination simultaneously paints 3 chromosome pairs in red and 3 in green, whereas in M. mulatta 4 chromosome pairs are labeled in red and 3 pairs are labeled in green. Using this method we show a baseline frequency of 0.026 translocations per 100 whole-genome cell equivalents in peripheral blood lymphocytes obtained from unexposed adolescent non-human primates. This method will add to the usefulness of M. mulatta as an animal model in biomedical research.