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Control of lipolysis in intra-abdominal fat cells of nonhuman primates: comparison with humans.

The mechanisms that control lipolysis in intra-abdominal fat cells from various primate species, the marmoset (Callithrix jacchus), the baboon (Papio papio), and the macaque (Macaca fascicularis), were compared to those of human intraabdominal fat cells. Selective beta 1- or beta 2-adrenoceptor agonists induced lipolysis in all species. Selective beta 3-agonists (BRL 37344, CL 316243, and SR 58611) acted as partial agonists in marmoset but were inefficient in other primates, including humans. alpha 2-Adrenoceptor number ([3H]RX 8210002 binding) equalized (baboon) or exceeded (other primates) beta 1/beta 2-adrenoceptors ([3H]CGP 12177 binding). Baboon fat cell membranes expressed similar amounts of coupled beta- and alpha 2-adrenoceptors. In all species, norepinephrine- or epinephrine-induced lipolysis did not reach the lipolytic effect of isoproterenol but their effects were enhanced after alpha 2-adrenoceptor blockade. N6-phenylisopropyladenosine (PIA) induced a full antilipolytic effect in baboon, macaque, and human adipocytes through adenosine receptors ([3H]DPCPX binding). Peptide YY (PYY) weakly inhibited lipolysis in baboon. Adrenocorticotropic hormone (ACTH) was inactive whereas parathyroid hormone (PTH) partially stimulated lipolysis in primates. Histamine was partially lipolytic in marmoset only. This study emphasizes the similarities of the mechanisms controlling the lipolysis in nonhuman primate and in human adipocytes and suggests that the baboon and the macaque should provide unique models for the study of the regulation of lipolysis.

Abdomen↗

Marmosets in New World leishmaniasis research.

In leishmaniasis, the search for a suitable experimental animal model is highly recommended, especially in Leishmania (Viannia) braziliensis (L. (V) b) research. Diverse species of neotropical primates have increasingly been used as experimental hosts of Leishmania. This article describes aspects of parasite L(V)b and marmoset, Callithrix penicillata (Primates, Callithricidae), interaction and summarizes the biology of parasitism in this primate model. Results of recent studies on primary infections with several strains of L(V)b as well as parasitism evolution, clinical outcome patterns and immunoprotection experiments, are summarily discussed. The relevance of homologous reinfection experiments for vaccine development in leishmaniasis is discussed.

Animals↗

Evidence for the experimental transmission of cerebral beta-amyloidosis to primates.

The brains of three marmosets (Callithrix jacchus) injected intracerebrally 6-7 years earlier with brain tissue from a patient with early onset Alzheimer's disease were found to contain moderate numbers of amyloid plaques with associated argyrophilic dystrophic neurites and cerebral amyloid angiopathy but no neurofibrillary tangles. The plaques and vascular amyloid stained positively with antibodies to beta (A4)-protein. The brains of three age-matched control marmosets from the same colony did not show these neuropathological features. The brain of one of two marmosets injected with brain tissue from a patient with prion disease with concomitant beta-amyloid plaques and cerebral amyloid angiopathy also showed beta-amyloid plaques and angiopathy but no spongiform encephalopathy. An occasional plaque was found in the brains of two of four marmosets injected with brain tissue from three elderly patients with age-related pathology, two of whom had an additional diagnosis of possible prion disease. Neither plaques nor cerebral amyloid angiopathy were found in six other marmosets who were older than the injected animals, in 12 further marmosets who were slightly younger but who had been injected several years previously with brain tissue which did not contain beta-amyloid, or in 10 younger marmosets who had been subjected to various neurosurgical procedures. These results suggest that cerebral beta-amyloidosis may be induced by the introduction of exogenous amyloid beta-protein.

Aged↗

Hemorrhagic typhlocolitis associated with attaching and effacing Escherichia coli in common marmosets.

We describe the necropsy results from three common marmosets presented during an outbreak of hemorrhagic diarrhea in a colony of 230 common marmosets (Callithrix jacchus). Necropsy revealed consistent hemorrhagic typhlocolitis and variable ileitis associated with gram-negative bacilli closely adherent to enterocytes. Electron microscopy revealed bacilli attached intimately to shallow cup-like projections of enterocyte apical membranes with loss of microvilli (attaching and effacing). Escherichia coli isolates from affected marmosets were serogroup O26 and were positive for the E. coli attaching and effacing locus and negative for shiga-like toxin I and II, heat-stable enterotoxins a and b, and heat-labile enterotoxin by DNA probe hybridization. Results of a Vero cell cytotoxicity assay confirmed that the E. coli isolates did not produce shiga-like toxin. The increased availability of diagnostic probes for specific virulence factors such as attaching and effacing E. coli should lead to a greater understanding of the frequency of E. coli as a cause of diarrhea in nonhuman primates.

Animals↗

Biochemical and morphological characterization of vascular and lymphocytic interleukin-4 receptors.

The distribution of the interleukin (IL)-4 receptor in normal human and common marmoset (Callithrix jacchus) tissues was examined by immunofluorescence and flow cytometry using monoclonal antibodies specific for the human IL-4 receptor to gain further insight into IL-4-mediated inflammatory and immunological events. IL-4 receptor positivity was unequivocally demonstrated on lymphocytes, predominantly T cells, and on blood vessels in many tissues. Vascular IL-4 receptor immunofluorescence consisted of a strong smooth muscle cell positivity and weaker positive staining of capillary and venular endothelial cells. Subnanomolar concentrations of IL-4 induced a genistein-sensitive up-regulation of VCAM-1 in vascular cell cultures. Tumor necrosis factor-alpha induced a genistein-resistant up-regulation of VCAM-1. IL-4 strongly induced expression of the IL-4 receptor on splenocytes (T lymphocytes) but not on vascular smooth muscle or endothelial cell cultures. Receptor cross-linking to [125I]IL-4 revealed a 65- to 75-kDa accessory receptor subunit consistent with a recently cloned IL-13 receptor associated with the IL-4 receptor on both vascular endothelial and smooth muscle cells. The demonstration of a vascular distribution pattern for the IL-4 receptor in addition to expression on lymphocytes suggests that vascular functional alterations, transduced through a unique IL-4 receptor complex (the type II IL-4 receptor), may be of importance during immunological and allergic inflammatory events.

Animals↗

Evaluation of zona pellucida antigens as potential candidates for immunocontraception.

Antibodies directed against the zona pellucida can interrupt sperm-egg recognition in vitro. However, the mechanisms by which anti-zona antibodies exert this contraceptive effect in vivo remain uncertain. There is an accumulating body of evidence to suggest that active immunity against zona antigens not only induces infertility via an antibody-mediated interruption of sperm--egg interaction but also disrupts normal ovarian function. We have evaluated the consequence of inducing active immunity against purified recombinant human ZP3 (rec.hZP3) and human ZP3 peptides, using the marmoset monkey, Callithrix jacchus, as an animal model. Although infertility was established in animals that received rec.hZP3, it was associated with ovarian dysfunction characterized by suppression of folliculogenesis and depletion of the primordial follicle pool. Immunization with continuous human ZP3 peptides, identified by epitope mapping studies, did not induce ovarian pathology but the antibody titres were insufficient to suppress fertility significantly and consistently. Clearly, further research is required to identify and segregate epitopes on the zona glycoproteins that are capable of inducing infertility without any side effects.

Animals↗

Immunocytochemical localization of ZP3 in primordial follicles of rabbit, marmoset, rhesus monkey and human ovaries using antibodies against human ZP3.

Active immunization with zona pellucida (ZP) proteins leads to depletion of primordial and recruited follicles in the ovary by a yet unknown mechanism. Small amounts of ZP present on primordial follicles or granulosa cells may be one of the reasons for this ovarian pathology. This study was undertaken in an attempt to identify the presence of ZP in or on primordial follicles and granulosa cells in ovaries of rabbits, common marmosets (Callithrix jacchus), rhesus monkeys (Macacas mulatta) and humans. Therefore, monoclonal antibodies (mAbs) and rabbit and mouse antisera against recombinant human ZP3 (hZP3) were produced. All these antibodies bound to the ZP of native intact human oocytes. Several fixatives have been tested on pieces of rabbit ovaries. With 4% (w/v) paraformaldehyde in PBS (4% PFA) followed by paraffin embedding and cryostat sections postfixed with 4% PFA, the most intense staining of ZP with one of the mAbs was obtained and the morphology was well preserved. In humans, besides the ZP, the oocyte cytoplasm and granulosa cells of most primordial and recruited follicles were also stained with these antibodies. In rhesus monkey ovaries, all primordial oocytes were also stained, in addition to some granulosa cells of primary follicles. In marmosets, small dots of immunoreactive ZP were found on 60% of the primordial follicles but granulosa cells were not stained. In rabbits, only minor staining of primordial follicles was observed. After passive immunization of rabbits with mAb 4, antibodies were found on both primordial and recruited follicles. These results clearly show the presence of ZP3 on primordial follicles and in granulosa cells, and this could explain the ZP-induced ovarian pathology after active immunization with ZP3.

Animals↗

Determinant spreading associated with demyelination in a nonhuman primate model of multiple sclerosis.

Definition of the immune process that causes demyelination in multiple sclerosis is essential to determine the feasibility of Ag-directed immunotherapy. Using the nonhuman primate, Callithrix jacchus jacchus (common marmoset), we show that immunization with myelin basic protein and proteolipid protein determinants results in clinical disease with significant demyelination. Demyelination was associated with spreading to myelin oligodendrocyte glycoprotein (MOG) determinants that generated anti-MOG serum Abs and Ig deposition in central nervous system white matter lesions. These data associate intermolecular "determinant spreading" with clinical autoimmune disease in primates and raise important issues for the pathogenesis and treatment of multiple sclerosis.

Adjuvants, Immunologic↗

Molecular phylogeny of the New World monkeys (Platyrrhini, primates) based on two unlinked nuclear genes: IRBP intron 1 and epsilon-globin sequences.

Nuclear sequences of the 1.8 kilobase (kb) long intron 1 of the interstitial retinol-binding protein gene (IRBP), previously determined for 11 of the 16 extant genera of New World monkeys (superfamily Ceboidea, infraorder Platyrrhini), have now been determined for the remaining 5 genera. The maximum parsimony trees found, first with IRBP sequences alone and then with tandemly combined IRBP and epsilon-globin gene sequences from the same species, supported a provisional cladistic classification with the following clusters. Subtribes Callitrichina (Callithrix, Cebuella), Callimiconina (Callimico), Leontopithecina (Leontopithecus) and Saguina (Saguinus) constitute subfamily Callitrichinae, and subfamilies Callitrichinae, Aotinae (Aotus), and Cebinae (Cebus, Saimiri) constitute family Cebidae. Subtribes Chiropotina (Chiropotes, Cacajao) and Pitheciina (Pithecia) constitute tribe Pitheciini; and tribes Pitheciini and Callicebini (Callicebus) constitute subfamily Pitheciinae. Subtribes Brachytelina (Brachyteles, Lagothrix) and Atelina (Ateles) constitute tribe Atelini, and tribes Atelini and Alouattini (Alouatta) constitute subfamily Atelinae. The parsimony results were equivocal as to whether Pitheciinae should be grouped with Atelinae in family Atelidae or have its own family Pitheciidae. The cladistic groupings of extant ceboids were also examined by different stochastic evolutionary models that employed the same stochastic process of nucleotide substitutions but alternative putative phylogenetic trees on which the nucleotide substitutions occurred. Each model, i.e., each different tree, predicted a different multinomial distribution of nucleotide character patterns for the contemporary sequences. The predicted distributions that were closest to the actual observed distributions identified the best fitting trees. The cladistic relationships depicted in these best fitting trees agreed in almost all cases with those depicted in the maximum parsimony trees.

Animals↗

Functional and adaptive significance of primate pads and claws: evidence from New World anthropoids.

This study tests predicted morphoclines in fingertip morphology among four small-bodied (<1 kg) New World monkeys (Saimiri sciureus, Leontopithecus rosalia, Callithrix jacchus, and Saguinus oedipus) in order to test previous functional and adaptive explanations for the evolution of flattened nails, expanded apical pads, and grasping extremities within the Order Primates. Small-bodied platyrrhines which frequently forage among small-diameter substrates are expected to possess 1) relatively expanded apical pads, 2) well-developed epidermal ridges, 3) distally broad terminal phalanges, and 4) reduced flexor and extensor tubercles compared to those species which use large-diameter arboreal supports more frequently for their locomotor and postural behaviors. Results show that as the frequency of small-branch foraging increases among taxa within this sample, relative distal phalanx breadth also increases but distal phalanx length, height, and flexor tubercle size decrease. Moreover, epidermal ridge development becomes more pronounced as the frequency of small-branch foraging increases. Terminal phalanx breadth and epidermal ridge complexity are both positively correlated with apical pad size. The large, flexible apical pad increases stability of the hand and foot on small-diameter arboreal supports because the pad can contact the substrate in several planes which, in turn, enables the pad to resist disruptive forces from different directions by friction and interlocking (Hildebrand, 1995). The observed morphoclines demonstrate that a gradient in form from claw- to nail-like tegulae exists among these taxa. Thus, the distinction between claw- and nail-bearing platyrrhines is essentially arbitrary. These observations corroborate Cartmill's (1972) functional and adaptive model for the loss of claws in primates: namely, expanded apical pads are required for habitual locomotor and postural behaviors on small-diameter supports whereas claws are more useful for positional behaviors on large-diameter substrates. Finally, results from this study support previous suggestions that the keeled tegulae of callitrichines represent a derived postural adaptation rather than a primitive retention from an ancestral eutherian condition.

Adaptation, Physiological↗

Development of an enzyme-immunoassay (EIA) for the measurement of follicle-stimulating-hormone (FSH) in Callitrichid primates using a monoclonal antibody against the human-FSH-beta-subunit.

Despite the importance of Callitrichid primates in both biomedical and conservation research, practical and reliable immunoassays for the measurement of follicle-stimulating hormone (FSH) have not yet been described. A panel of monoclonal antibodies against specific peptide fragments within either the alpha or beta subunit of human FSH was evaluated for their ability to recognize FSH from Callitrichid and other New World primates. One of these, monoclonal antibody 46.3h6.b7 raised against human FSH, was selected due to its ability to recognize marmoset monkey FSH and its low crossreactivity with other gonadotrophins. The antibody formed the basis of an enzymeimmunoassay using a highly purified human urinary FSH (Metrodin, Serono) preparation coupled to biotin as label and unmodified as standard. After 24 h incubation, antibody bound label was visualized by addition of streptavidin-peroxidase followed by the appropriate substrate. Parallelism was obtained between the standard and dilutions of pituitary extracts, urine and plasma from the common marmoset (Callithrix jacchus) as well as from two tamarin species (Saguinus fuscicollis and S. oedipus) and one squirrel monkey (Saimiri sciureus). Profiles of plasma and urinary FSH during the follicular phase are shown for two individual marmosets. The ability to measure FSH in Callitrichidae provides new opportunities for studies of the reproductive biology of these New World primate species.

Animals↗

Rate of metabolism in the smallest simian primate, the pygmy marmoset (Cebuella pygmaea).

Rate of metabolism was measured with six adult pygmy marmosets (Cebuella pygmaea) at regulated ambient temperatures ranging between 20 degrees C and 35 degrees C. A novel combined nest box and metabolic chamber was designed to allow nighttime measurements on immobile animals in their home cage without disturbance. The basal rate of metabolism (BMR) was 98 ml O2 h-1, representing 74% of the value expected from the equation of McNab [Quarterly Review of Biology 63:25-54, 1988] relative to body mass. The thermoneutral zone was approximately 27-34 degrees C. Below the lower critical temperature (27-28 degrees C), thermal conductance (12.9 ml O2 h-1 degree C-1) was close to the predicted value. Body temperature ranged between 34.9 degrees C and 35.5 degrees C at night. When two animals rested together overnight in the nest box, the lower critical temperature was slightly lowered, and individual energy expenditure at 20-21 degrees C was reduced by about 34%. The basal rate of metabolism of C. pygmaea is much lower than reported in an earlier study based on daytime measurements but agrees with values reported from a more recent study conducted at night with a classical metabolic chamber. In order to compare the BMR of C. pygmaea with that of other primates, 23 species were included in a comparative study taking into account both phylogeny and body mass (independent contrasts approach). The scaling exponent of BMR to body mass obtained was indistinguishable from that published for eutherian mammals in general. Cebuella and Callithrix exhibit the lowest basal rates known for simians. This trait may possibly be linked to the natural diet, which includes a large proportion of gums that are difficult to digest, but additional metabolic studies on primates are needed for further examination of its adaptive significance.

Animals↗

Hormonal monitoring of age at sexual maturation in female Goeldi's monkeys (Callimico goeldii) in their family groups.

The aim of this study was to determine whether or not sexual maturation is attained in the family group in captive-born Goeldi's monkey (Callimico goeldii) and if so, at what age and body weight. To monitor ovarian activity in 14 female Goeldi's monkeys, urinary content of pregnanediol-3alpha-glucuronide (PdG) was determined using radioimmunoassay. Urinary samples were collected between the ages of 6 and 70 weeks. Subjects became sexually mature while still housed in their family groups, at a median age of 57 weeks (48-< 70 weeks). Median body weight at the age of sexual maturity was 473 g (N=10; 420-543 g). This corresponded to 90% of the median non-pregnant body weight of breeding females in our colony (526 g, N=8). Therefore, Goeldi's monkey is similar to Leontopithecus but different from Cebuella, Callithrix, and Saguinus, in terms of daughters ovulating in the family group and at a relatively young age.

Aging↗

Transplacental and direct exposure of mouse and marmoset to ethylnitrosourea: analysis of chromosome aberrations.

The effect of ethylnitrosourea (ENU) on chromosomes of mouse bone marrow cells and transplacentally exposed embryonic liver cells was investigated. Chromosome aberrations were found to be dose- and time-dependent. The maximum damage was seen 6 h after the exposure. Chromosome aberrations were also induced in bone marrow cells and lymphocytes of marmosets (Callithrix jacchus) directly exposed to ENU. Aberrations did not, however, occur in offspring whose mothers were treated with ENU before conception. Furthermore, chronic transplacentally exposed offspring have been analyzed. The frequency of chromosome aberrations was not increased in their lymphocytes and fibroblasts. The elimination of chromosome aberrations during embryogenesis is discussed.

Animals↗

Abundance, diversity, and patterns of distribution of primates on the Tapiche River in Amazonian Peru.

This work presents data on the relative diversity, abundance, and distribution patterns of primates in a 20 km2 area of the Tapiche River in the Peruvian Amazon. Population data were collected while the study area was both inundated and dry (March to September 1997) using conventional line-transect census techniques. Survey results reflected the presence of 11 primate species, but population parameters on only eight of the species will be presented, including saddleback tamarins (Saguinus fuscicollis), Bolivian squirrel monkeys (Saimiri boliviensis), brown capuchins (Cebus apella), white-fronted capuchins (Cebus albifrons), monk sakis (Pithecia monachus), red titi monkeys (Callicebus cupreus), red uakaris (Cacajao calvus), and red howler monkeys (Alouatta seniculus). Woolly monkeys (Lagothrix lagotricha), night monkeys (Aotus nancymaae), and pygmy marmosets (Callithrix pygmaea) were also seen in the area. The data for the smaller-bodied primates is similar to that reported almost 18 years earlier, but the data for the larger-bodied primates reflect a loss in the number of animals present in the area. Pressure from hunters and the timber industry may account for declining numbers of large-bodied primates, while it appears that natural features peculiar to the conservation area contribute to the patchy pattern of distribution.

Animals↗

Does the milk of Callitrichid monkeys differ from that of larger anthropoids?

The generalization that anthropoid primates produce dilute milks that are low in protein and energy is based primarily on data from large monkeys of the families Cebidae and Cercopithecidae, as well as humans. The marmosets and tamarins (Callitrichidae) are not only much smaller in body size, but also typically raise multiple offspring during a relatively brief lactation. We hypothesized that selection for small body size and high reproductive rate might favor secretion of milk of higher energy and protein concentrations. To test this hypothesis, 46 milk samples collected from 10 common marmosets (Callithrix jacchus, ca. 350 g) were assayed for dry matter (DM), crude protein (CP), fat, and sugar; and gross energy (GE) was calculated from these constituents. We also assayed five samples collected from three golden lion tamarins (Leontopithecus rosalia, ca. 700 g) and six samples collected from a single pygmy marmoset (Cebuella pygmaea, ca. 150 g) over two lactation periods. All samples were collected between days 10 and 57 post partum, representing mid lactation for these species. The milks of these three species were similar, containing 14.0%, 16.1%, and 13.7% DM; 2.7%, 2.6%, and 2.9% CP; 3.6%, 5.2%, and 3.7% fat; 7.4%, 7.2%, and 7.8% sugar; and 0.76, 0.90, and 0.82 kcal/g for common marmosets, golden lion tamarins, and the pygmy marmoset, respectively. These species produced milks with energy values that were within the range reported for large anthropoids, albeit with slightly higher protein concentration. However, milk composition did vary substantially among individual common marmoset females, especially in the proportion of milk energy derived from fat. In contrast, CP as expressed as a percent of GE was remarkably constant among common marmoset females. Callitrichid milk appeared to be similar to that of larger anthropoid primates in GE, but was higher in CP and in the proportion of GE from CP. However, the small sample sizes for the golden lion tamarin and the pygmy marmoset, and the wide variation in milk composition found among common marmoset females cautions against definitively characterizing the milks of callitrichids from these data.

Animals↗

Quantitative data on training new world primates to urinate.

This study assessed the effectiveness of operant conditioning in training three species of captive callitrichid primates (Leontopithecus rosalia, Callithrix geoffroyi, and Saguinus imperator) to urinate on demand. There were three goals to the study: 1) to develop a system for quantitatively assessing positive reinforcement training; 2) to ascertain whether or not positive reinforcement techniques can be used to train callitrichid monkeys to urinate on demand, and if so, how many training sessions are required; and 3) to determine the effect on urination behavior of the trainer entering the cage to collect a urine sample. Positive reinforcement with a continuous reinforcement schedule was used to capture a natural behavior: urination. Training sessions (30 min each) were conducted at dawn thrice weekly during five consecutive phases: habituation, control, training (animals were rewarded for urinating), maintenance (animals had reached a defined training criteria and continued to be rewarded for urinating), and collection (animals were rewarded for urinating, and the trainer entered the cage to collect the sample). The numbers of 30-min training sessions required to train the three monkey species (L. rosalia, C. geoffroyi, and S. imperator) were five, six, and eight, respectively. For the three species, the mean number of urinations per animal was significantly greater during the training, maintenance, and collection phases compared to the control phase. However, the three species differed significantly in the manner in which the rates of urination changed across the five phases. A higher proportion of subjects urinated during the training, maintenance, and collection phases compared to the control phase. Latency to first urination varied significantly across the five phases, with significantly reduced latencies to urinate during the training, maintenance, and collection phases compared to the control phase. The entry of the trainer into the cage to collect the urine sample did not appear to alter urination behavior. We demonstrate that operant conditioning techniques, which typically incur minimal cost, time investment, and disturbance, can be used to increase the quantity of urine samples collected for physiological analysis, the proportion of animals that urinate, and the speed of sample collection.

Animals↗

Arterial vascularization of the mandible and maxilla of neotropical primates.

The objective of the present investigation was to conduct a comparative macroscopic study of the arterial vascularization of the mandible and maxilla of neotropical primates of the genera Cebus, Alouatta, Callithrix, and Leontopithecus. After vinyl was injected into the arterial system of the head of each specimen, the pieces were macerated and corroded. The level of the bifurcation of the common carotid artery into the internal and external carotids varied between the first and third cervical vertebrae. The external carotid artery accounts for most of the vascularization of the facial structures. The actual vessels responsible for the supply of this region are the sublingual, facial, angular, lingual, submandibular, submental, inferior and superior labial, maxillary, inferior alveolar, infraorbital, superior posterior alveolar, palatine major, and sphenopalatine arteries. We conclude that although the arterial vascular pattern was similar in all the genera studied, and resembles the human pattern, there are notable variations in the vasculature of the mandible and maxilla among these four neotropical genera.

Animals↗