Synaptic ribbon populations in the pineal gland of the rhesus monkey (Macaca mulatta)

Summary Pinealocytes of rhesus monkeys that had been ovariectomized and given intramuscular injections of 250 g estradiol-benzoate for 3 consecutive days tended to have more synaptic ribbons (SR) and exhibited a significantly greater size of ribbon fields (RF) compared to untreated animals. These data are consistent with hypotheses that pinealocyte function in primates is altered by hormonal imbalances and that the SR participates in this response. RF were positioned in various parts of the cytoplasm and along the plasma membrane. Participation of SR in direct cell-to-cell contacts was suggested by the formation of densities along the plasma membrane. It is postulated that large RF serve as storage organelles and that the formation of RF results from division of pre-existing SR in each field. Reconstructions made from serial thin sections revealed that profiles of RF comprised separate SR that were not folded and sectioned along various planes.     

Comparative study of the transfection efficiency of commonly used viral vectors in rhesus monkey (Macaca mulatta) brains

Viral vector transfection systems are among the simplest of biological agents with the ability to transfer genes into the central nervous system.In brain research,a series of powerful and novel gene editing technologies are based on these systems.Although many viral vectors are used in rodents,their full application has been limited in non-human primates.To identify viral vectors that can stably and effectively express exogenous genes within nonhuman primates,eleven commonly used recombinant adeno-associated viral and lentiviral vectors,each carrying a gene to express green or red fluorescence,were injected into the parietal cortex of four rhesus monkeys.The expression of fluorescent cells was used to quantify transfection efficiency.Histological results revealed that recombinant adeno-associated viral vectors,especially the serotype 2/9 coupled with the cytomegalovirus,human synapsin Ⅰ,or Ca2+/calmodulin-dependent protein kinase Ⅱ promoters,and lentiviral vector coupled with the human ubiquitin C promoter,induced higher expression of fluorescent cells,representing high transfection efficiency.This is the first comparison of transfection efficiencies of different viral vectors carrying different promoters and serotypes in non-human primates (NHPs).These results can be used as an aid to select optimal vectors to transfer exogenous genes into the central nervous system of non-human primates.     

Expression of SWAP-70 in the uterus and feto-maternal interface during embryonic implantation and pregnancy in the rhesus monkey (Macaca mulatta)

SWAP-70 is a unique signaling protein involved in multiple processes including lymphatic cell activation, migration, adhesion, and cytoskeleton organization. Its role in reproductive system remains to be unclear. In the present study, the spatial and temporal expression of SWAP-70 in the uterus during normal menstrual cycle as well as on the feto-maternal interface during pregnancy was investigated in the rhesus monkey by in situ hybridization and immunohistochemistry. It was shown that SWAP-70 was mainly expressed in glandular epithelial cells of uterine endometrium, and the level peaked at the mid-secretory stage. At the beginning of embryonic implantation, SWAP-70 was intensely expressed at the implantation site, mainly localized in glandular and luminal epithelial cells, as well as in primary trophoblasts and epithelial plaque. High level of SWAP-70 was observed in villous cytotrophoblast (VCT), syncytiotrophoblast (ST), column cytotrophoblast, trophoblast shell, interstitial trophoblast, and endovascular trophoblast during gestational days 15–25. From gestational day 50 to term, expression of SWAP-70 decreased evidently and was restricted in VCT cells. What’s more, SWAP-70 co-localized with F-actin on the feto-maternal interface, especially in highly motive extravillous trophoblasts. The data indicate that SWAP-70 may be involved in regulating motility of trophoblast cells during embryonic implantation and placentation.     

Species discrimination and concept formation by rhesus monkeys (Macaca mulatta)

There are 19 species in genusMacaca and some of them are living in sympatry (Fooden, 1980). Although inter-specific hybrids are relatively easy to produce under artificial conditions, hybridization does not occur naturally. What is preventing that among the species of genusMacaca? Three rhesus monkeys acquired a discrimination between pictures with rhesus monkeys and without rhesus monkeys. All subjects showed positive transfer of this discrimination to new pictures with rhesus monkeys and without rhesus monkeys. A further test showed that these monkeys could discriminate between pictures of rhesus monkeys and pictures of Japanese monkeys. The results suggest that rhesus monkeys recognize rhesus monkeys as a class, independent of the actual stimuli such as a picture or an individual monkey. The ability to recognize members of their own species and the opportunities for such learning may be an important factor preventing hybridization among the species of genusMacaca.     

Transplantation of fetal neocortex in rhesus monkey

A feasibility study of neural transplantation in adult rhesus monkey was undertaken. Fresh and preserved neocortex containing multiplying and maturing neurons obtained from 55–70 gestation days were transplanted into the striatum, cerebellum and cerebral cortex of adult monkeys. Tissues were preserved for 4 days either at subzero temperature in the freezer compartment of the ordinary refrigerator in Ringer lactate or incubated in culture medium. While 2 monkeys out of 5 injected with preserved tissue had successful transplants after 4 months, all the 10 monkeys injected with fresh tissue had no transplants. The size of the two surviving transplants was small. The neurons in the transplants were mainly in clusters. Many of the cells were immature and some showed early degenerative changes. Neuronal processes were restricted to the transplants and thus showed lack of morphological integration with the host tissue. Further studies are in progress to define the nature of the embryonic tissue of primate which can grow and survive and also the role of neural grafts in functional recovery following experimental lesions of the brain regions.     

Ultrastructure of synapses and cellular relationships in the oculomotor nucleus of the rhesus monkey

Summary The fine structure of synapses and of cellular relations was examined in the somatic efferent portion of the oculomotor nucleus of the adult rhesus monkey. Axosomatic and axodendritic synapses are characterized by distinct synaptic clefts which usually measure 20–30 nm between pre- and postsynaptic membranes. Cytoplasmic thickenings of pre- and postsynaptic membranes are often observed. Subjunctional bodies are present at both axosomatic and axodendritic synapses. Somatic and dendritic spine synapses are present. Serial synapses are also found, suggesting the operation of presynaptic inhibition in this nucleus. At some synapses the extracellular gap between pre- and postsynaptic membranes is reduced to 5–9 nm. However, junctions similar to the latter are also present between neurons and glia, and at the junctions between adjacent glial elements. The present results provide no evidence for a clear morphological substrate for electrotonic transmission in the somatic efferent portion of the primate oculomotor nucleus.Supported in part by grants from the National Institutes of Health (NS-15320), the Kroc Foundation, and by the Medical Research Service of the Veterans Administration (to S.G.W.), and from the National Science Foundation (BNS 77-28493) and the Muscular Dystrophy Association of America (to G.D.P.)     

Sebaceous gland adenoma in a rhesus monkey (Macaca mulatta)

A tumor mass was identified below the shoulder region of a 5-year-old male rhesus monkey (Macaca mulatta). The mass was excised and diagnosed as sebaceous gland adenoma based on the microscopic findings. Morphologically it appeared as an elevated, dome-shaped, circumscribed mass of 3.6 x 2.8 x 3.2 cm in dimension with tan speckled color. Histologically, the tumor was composed of mature, sebaceous cells (sebocytes), basal cells arranged in a mass of irregular shapes and sizes, with a characteristic appearance of poly or multilobular structure. Sebocytes were well differentiated with foamy cytoplasm in the center of the lobules and poorly or undifferentiated densely staining basal cells in the periphery of the lobules. Cellular changes in the adjacent lymph node included hyperplasia of plasma cells, macrophages and lymphoid elements with typical mitosis.     

A light and transmission electron microscope study of hepatic portal tracts in the rhesus monkey (Macacus rhesus)

This study reports on morphological features of hepatic portal tracts in the liver of a rhesus monkey. The light microscope shows that the number of each type of principal component comprising a portal tract varies but that there are usually one to five lymphatics, one bile ductule, one bile duct, one arteriolar and one arterial branch of the hepatic artery, and one hepatic portal vein. Bile ductules, in cross section, have 6-10 cells (mostly low pyramidal, but with a few cuboidal) bordering the lumen, an outside diameter of from about 20 to 25 microm, and a luminal diameter of from 2 to 10 microm. Bile ducts, in cross section, have more than 10 cells (about equal numbers of low pyramidal and cuboidal) bordering the lumen, an outside diameter greater than 25 microm and a luminal diameter of greater than 10 microm. The term "pyramidal" has not previously been applied to the cells of the ductules and ducts. The monkey tracts show several cytological features previously undescribed, viz., abortive cilia and basal bodies in the duct cells, abortive cilia in the ductule cells, and an occasional aggregation of ribosomes in arterial endothelial cells. They also show a major histological feature previously mentioned but not illustrated, viz., bundles of nerve processes which exhibit a preferential location, i.e., proximity to the arterioles and arteries.     

Acepromazine-ketamine anesthesia in the rhesus monkey (Macaca mulatta).

Six adult male rhesus monkeys (Macaca mulatta) were anesthetized using a combination of acepromazine maleate and ketamine hydrochloride administered by intramuscular injection. This combination produced a smooth induction to anesthesia requiring less than 5 minutes. The average duration of anesthesia was slightly less than 1 hour and was safely prolonged with additional doses of ketamine. The depth of anesthesia was sufficient for minor surgical procedures and for precise radiological studies. No deleterious side effects were noted, and animals recovered completely in a short time.     

Growth of the masseter muscle in rhesus monkeys (Macaca mulatta)

The growth of the masseter muscle in eight infant, juvenile, and adolescent female rhesus monkeys (M. mulatta) was examined over a 2.5 year period using serial radiographic cephalometric techniques with the aid of radiopaque muscle markers. The radiopaque markers, which are composed of small pieces of root canal broach inserted into the muscle belly, make it possible to determine longitudinal masseter muscle growth as well as migration of the masseter muscle relative to the mandible. It was found that the masseter muscle increased in length by 64% during the total growth period, most of which occurred between 6 and 18 months of age. Relative to the cranium, the masseter muscle grew markedly inferiorly and only slightly posteriorly. Relative to the mandible, the masseter migrated in a posterior and slightly superior direction, keeping pace with the ramus and condyle as they grew posteriorly and posterosuperiorly throughout the study period. It was concluded that: 1) radiopaque muscle markers are a valuable tool for analysis of muscle growth and alteration of muscle location; 2) the masseter muscle in the rhesus monkey undergoes elongation, probably due to addition of sarcomeres at the fiber-tendon junctions; and 3) posterior migration of the masseter muscle relative to the corpus of the mandible, probably due to the nature of its periosteal attachment, results in a stability of the anteroposterior position of the masseter muscle despite the anterior displacement of the mandible.     

Effect of cyproterone acetate on prolactin secretion in the female rhesus monkey

Summary Adult female rhesus monkeys were given cyproterone acetate orally in doses of 0.04, 0.4, 4 and 40mg per kg per day for 12 weeks. Its effects were assessed on serum prolactin (PRL) concentration, the morphology of the PRL cells, and the development of the mammary glands. Serum PRL was relatively unchanged in the control animals from the fourth through the twelfth weeks of the study. In contrast, PRL was significantly elevated in each group of drug-treated animals during the same time periods. There was no development of the mammary glands nor was there any evidence of milk secretion in the control animals; however, in the monkeys given cyproterone acetate the mammary glands had extensive lobuloalveolar growth and milk-like secretion that could be expressed as early as the fourth or fifth week of the study. By immunocytochemistry and differential light microscopic staining techniques, the PRL cells in the pituitary glands of the experimental animals were found to be more numerous and much larger than those present in the controls. They displayed a well developed Golgi complex and had an abundance of cytoplasmic RNA. These data suggest that PRL secretion is markedly enhanced by cyproterone acetate.Supported in part by USPHS Grant AM12583     

Osmotic tolerance and membrane permeability characteristics of rhesus monkey (Macaca mulatta) spermatozoa

Biophysical characteristics of the plasma membrane, such as osmotic sensitivity and water and cryoprotectant permeability are important determinants of the function of spermatozoa after cryopreservation. A series of experiments was conducted with rhesus macaque spermatozoa at 23 degrees C to determine their: (1) cell volume and osmotically inactive fraction of the cell volume; (2) permeability coefficients for water and the cryoprotectants dimethyl sulfoxide, glycerol, propylene glycol, and ethylene glycol; (3) tolerance to anisosmotic conditions; and (4) motility after a one step addition and removal of the four cryoprotectants. An electronic particle counter and computer aided semen analysis were used to determine the cell volume and permeability coefficients, and motility, respectively. Rhesus spermatozoa isosmotic cell volume was 27.7+/-3.0 microm3 (mean+/-SEM) with an osmotically inactive cell fraction of 51%. Hydraulic conductivity in the presence of dimethyl sulfoxide, glycerol, propylene glycol, and ethylene glycol was 1.09+/-0.30, 0.912+/-0.27, 1.53+/-0.53, and 1.94+/-0.47 microm/min/atm, respectively. Cryoprotectant permeability was 1.39+/-0.31, 2.21+/-0.32, 3.38+/-0.63, and 6.07+/-1.1 (x10(-3)cm/min), respectively. Rhesus sperm tolerated all hyposmotic exposures. However, greater than 70% motility loss was observed after exposure to solutions of 600 mOsm and higher. A one step addition and removal of all four cryoprotectants did not cause significant motility loss. These data suggest that rhesus sperm are tolerant to hyposmotic conditions, and ethylene glycol may be the most appropriate cryoprotectant for rhesus sperm cryopreservation, as it has the highest permeability coefficient of the tested cryoprotectants.     

恒河猴糖尿病动物模型血清及组织中细胞因子的变化

目的探讨恒河猴糖尿模型病细胞因子的水平和变化规律。方法健康恒河猴5只,小剂量(30mg/kg)多次静脉注射链脲佐菌素(STZ),血糖稳定后3、9、12和19个月检测血清中白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、肿瘤坏死因子-α(TNF-α)的含量。安乐濒死状态的动物,取胰腺、肝脏、肾脏制成石蜡切片,用免疫组化染色法显示组织中IL-6、IL-10、TNF-α的表达,并对结果进行图像分析和统计学处理。结果造模后9个月动物血清中IL-10浓度显著降低(P<0.01)。IL-6、TNF-α水平在采样的各个时间点均显著高于造模前(P<0.01)。胰腺组织中IL-6中等强度着色,对照组弱阳性表达(P<0.05)。对照组IL-10强阳性表达,模型组弱阳性(P<0.01)。模型组TNF-α中等强度表达,对照组弱阳性(P<0.05)。模型组肝脏组织TNF-α中等阳性表达,对照组弱阳性(P<0.05)。结论小剂量多次注射STZ后恒河猴细胞因子的含量及变化与临床类似,可作为相关研究的动物模型。     

Localization of 3H-dihydrotestosterone in the pituitary gland of the rhesus monkey

Summary The uptake and retention of radiolabelled dihydrotestosterone by the pituitary gland was examined in the rhesus monkey. Two animals were given an intravenous injection of 1.0g/kg ³H-dihydrotestosterone (DHT) alone while one monkey received both the labelled androgen and 100g/kg of unlabelled steroid. One and a half hours later, they were sacrificed. The pituitary glands were removed and processed for autoradiography and immunocytochemistry. Autoradiographic localization of DHT was discernible in the partes nervosa, intermedia and distalis, albeit the highest concentration of radiolabelled cells was noted in the pars distalis. Immunocytochemical staining with antibodies to rat PRL, human TSH and ovine LH revealed a population of steroid-concentrating cells that contained TSH and a second group that contained LH. None of the cells that reacted with the anti-PRL serum were radiolabelled.     

Placental transfer of synthetic and endogenous estrogen conjugates in the rhesus monkey (Macaca mulatta)

The placental transfer of conjugated and nonconjugated estrogens was compared in the pregnant rhesus monkey. Placement of catheters in the maternal and fetal circulation allowed for the sampling of blood after the administration of radiolabeled naturally occurring and synthetic estrogens to mother or fetus. In all cases, nonconjugated-estrogen placental transfer was greater than conjugated-estrogen transfer. Comparison of the conjugated estrogens suggested that diethylstilbestrol-monoglucuronide (DESG) was transferred less efficiently than the estrone-sulfate (E, S). High-performance liquid chromatog-raphy (HPLC) of selected plasma samples revealed that 50–90% of the E, S observed in the maternal circulation was cleaved after fetal administration. In contrast, HPLC of maternal or fetal plasma samples after DESG administration revealed only intact DESG. These results emphasize differences in the placental transfer of the synthetic and naturally occurring estrogen hormones.     

Flexible fiberoptic bronchoscopy of the rhesus monkey (Macaca mulatta).

Using a flexible fiberoptic bronchoscope, 10 rhesus monkeys were bronchoscoped 30 times without complication. The bronchoscopic anatomy was found to be similar to that of the human except for the presence of a right cardiac lobe in the monkey.     

Laboratory assessment of temperament and environmental enrichment in rhesus monkey infants (Macaca mulatta)

This study investigated the combined effects of early temperamental characteristics and environmental enrichment on a variety of developmental measures in nursery-reared rhesus monkey infants. Twenty-three infants, reared in either standard laboratory cages or enriched environments, were tested during the 1st month of life for interactive, motor, and temperamental capabilities and characteristics. At 8 months of age, all subjects were assessed on a second series of tests designed to measure their problem-solving skills, motor capabilities, and temperamental responses under challenge. Results indicated that enrichment was associated with higher scores on subsequent problem-solving and motor tests. However, such effects were found to combine with early temperament ratings. Specifically, individuals performing best on the 8-month tests had not only been reared in enriched environments, but also had been rated low on fearfulness during the early assessment. In addition, individuals scoring poorest had been rated as fearful initially in addition to being reared without enrichment. Results indicated that while high ratings on early laboratory assessments of fearfulness may be predictive of poorer problem-solving performance under challenging conditions, these adverse effects may be partially attenuated by environmental enrichment.     

A description of the upper thoracic autonomic nervous system in the rhesus monkey (Macaca mulatta)

The purpose of this study was to describe the autonomic innervation of the carotid sinus and heart in the rhesus monkey. Nine male rhesus monkeys (Macaca mulatta) and one male crab-eating macaque (M. fascicularis) were carefully dissected from the origin of the vagus nerves and superior cervical ganglia to the level of the fourth thoracic ganglion. The specimens were either freshly killed or obtained no later than 24 hours post mortem. The macaque monkeys were found to possess an innervation pattern that displayed features common to dog (connections between the vagus nerves and middle cervical ganglia), baboon (distinct cervical sympathetic and cervical vagal nerve trunks), and man (nerves projecting from the middle cervical and stellate ganglia to the heart). Distinct inferior cervical and first thoracic ganglia were never seen, but rather, large and well defined stellate ganglia were found. The macaque innervation pattern, when considered as a whole, most closely resembled the baboon.     

Amniocentesis and antenatal sex determination in the rhesus monkey (Macaca mulatta).

Antenatal sex was correctly identified in 87.5% of the rhesus monkeys studied by evaluation of sex chromatin bodies in the exfoliated cells present in amniotic fluid. Fluid samples were obtained by either amniocentesis, amnioscopy, or by cesarean section at 80--120 days of gestation. Amniotic fluid concentrations of progesterone, total estrogen, and testosterone were correlated with fetal sex. Concentrations of these hormones decreased over the time period examined. Amniocentesis is a simple procedure which provides for routine fetal sex determination.