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PubMed2003

1H-NMR spectroscopy combined with pattern recognition analysis reveals characteristic chemical patterns in urines of MS patients and non-human primates with MS-like disease.

Proton nuclear magnetic resonance (1H-NMR) spectroscopy in combination with pattern recognition techniques were used to investigate the composition of organic compounds in urines from patients with multiple sclerosis (MS), patients with other neurological diseases (OND) and healthy controls (H). Using a valid animal model of MS, namely the common marmoset (Callithrix jacchus) model of experimental autoimmune encephalomyelitis (EAE), the relation of disease progression and alteration of the urine composition was investigated. Urine samples were collected during different stages of EAE, either induced with whole human myelin or with the myelin protein MOG in complete adjuvant. The urine samples were analysed with 1H-NMR spectroscopy allowing simultaneous detection of an array of compounds. Spectral differences between urines from EAE-affected and healthy monkeys were assessed with multivariate analysis. Evidence is provided that development of EAE is associated with changes in the chemical composition of the urine, in particular of compounds with NMR peaks in the region of the spectrum between 0.5 and 3.50 ppm. In addition, we found preliminary evidence for differences between urines from MS, OND and H groups.

Animals↗
PubMed1997

Comparative study on age-dependent development of surface receptors on peripheral blood lymphocytes in children and young nonhuman primates (marmosets).

General conformity was found concerning age-dependent changes in the major subsets (CD4, CD8, CD2, CD20) and the adhesion molecules CD11a and CD29 on peripheral blood lymphocytes in children and young marmosets (Callithrix jacchus). Differences exist in the expression of the surface receptors CD45RA and CD56. Taking the total number of white blood cells into account, an age-related increase in the absolute numbers of peripheral blood lymphocytes was found in marmosets whereas in children, an age-dependent decrease was observed. The definition of reference ranges for different stages of maturation in children and young marmosets may serve as a basis on which comparative risk assessment of possible effects on lymphocyte subsets after pre- or perinatal drug exposure can be estimated.

Adolescent↗
PubMed1998

Lateralized use of the mouth in production of vocalizations by marmosets.

We have found that the common marmoset, Callithrix jacchus, displays a larger left hemimouth during production of fear expressions, with or without vocalization, and a larger right hemimouth when producing a social contact call. Thus, marmosets have right hemisphere specialization for the production of negative emotional expressions and vocalizations and left hemisphere specialization for the production of social contact communication. These hemispheric specializations for social communication in marmosets are the same as those found in humans for speech production and for the control of emotional expressions. We suggest that hemispheric specializations for communication in humans may well have precursors in primate evolution.

Animals↗
PubMed2001

Prolactin levels are elevated after infant carrying in parentally inexperienced common marmosets.

Alloparental behavior (parental behavior directed to nondescendant young) is pervasive among human cultures but rare among other mammals. New World primates of the family Callitrichidae, including common marmosets (Callithrix jacchus jacchus), dwell in large families and many family members carry and/or provision infants. This study experimentally characterized alloparenting in common marmosets by measuring infant retrieval and carrying outside of the context of the family group. Prolactin was measured in samples collected before and after infant exposure to determine whether elevated prolactin levels were predictive of alloparental responsiveness. Seventeen nonbreeding, parentally inexperienced, singly or peer-housed common marmosets (10 females, 7 males, 6-66 months of age) were tested repeatedly in an infant retrieval paradigm. Infant retrieval was shown by 5 out of 17 monkeys (29%) in their first test. The rate of retrieval increased to 10 monkeys (59%) by the fourth test. No significant differences in age, sex, or housing condition existed between monkeys that retrieved infants and those not retrieving. Prolactin concentrations were significantly elevated in serum obtained after testing only in monkeys that retrieved infants. Prolactin levels after infant exposure were positively related to carrying duration. A separate experiment verified that neither prolactin nor cortisol was significantly elevated in response to prolonged handling. Cortisol levels were inversely proportional to handling time. Prolactin levels were not significantly related to handling time. Our results indicate that alloparental behavior is expressed at high rates after minimal infant experience, and prolactin levels rise concomitantly with the expression of alloparental behavior in this species.

Aging↗
PubMed1998

Concurrent monitoring of EEG and performance in the common marmoset: a methodological approach.

A model has been developed in a nonhuman primate, the common marmoset (Callithrix jacchus), which should enable the study of long term effects of compounds with potentially psychoactive properties. The technique facilitates concurrent monitoring of both behavioral and electrophysiological parameters while animals remain in their home cages. Subjects were trained to perform tests from a neuropsychological test battery (The Cambridge Neuropsychological Test Automated Battery, CANTAB) in which they learned to discriminate between pairs of stimuli presented on a touch sensitive computer screen. Single channel cortical electroencephalography (EEG) by radiotelemetry was simultaneously recorded while behavioral testing took place.

Animals↗
PubMed1995

Demonstration of nitric oxide synthase (NOS) in marmosets by NADPH diaphorase (NADPH-d) histochemistry and NOS immunoreactivity.

Since species interdiversity often prevents the extrapolation of laboratory rodent data to man and similar problems may exist for nitric oxide synthase (NOS), NADPH-d activity and immunohistochemistry of NOS were investigated in the New World monkey Callithrix jacchus (marmoset), which has been shown to be close to the human situation in many respects. Using the NADPHd reaction with beta-NADPH and nitroblue tetrazolium (NBT) on acetone-chloroform pretreated cryosections, NBT formazan was found in many neural and non-neural (e.g. diverse epithelia, striated muscle fibers, vascular endothelium) cells in numerous tissues and organs. Prefixation with formaldehyde lowered the number of NADPH-d active sites and the amount of formazan with the exception of neuronal NADPH-d as did incubation of fresh or acetone-chloroform-pretreated sections for NADPH-d in the presence of 0.5% formaldehyde. When 1% formaldehyde or 0.5 mM permanganate were used significant amounts of formazan appeared only in central and peripheral neurons, vasal endothelial cells, small intestinal enterocytes, plasma membrane region of striated muscle fibers as well as arteriolar cells in the kidney; except for enterocytes, these observations were confirmed by NOS-immunohistochemistry which revealed in addition reactive cells in the thymus and intestinal lamina propria.

Acetone↗
PubMed2001

Behavioral effects of buspirone in the marmoset employing a predator confrontation test of fear and anxiety.

In order to further validate the recently developed marmoset (Callithrix penicillata) predator confrontation model of fear and anxiety, we investigated the behavioral effects of buspirone with this method. The apparatus consisted of three parallel arms connected at each end to a perpendicular arm, forming a figure-eight continuous maze. A taxidermized wild oncilla cat (Felis tigrina) was positioned facing a corner of the parallel arms, alternating between the left or right side of the maze among animals tested. All subjects were first submitted to seven 30-min maze habituation trials (HTs) in the absence of the predator, and then to five randomly assigned treatment trials (TTs) in the presence of the predator: three buspirone sessions (0.1, 0.5 and 1.0 mg/kg), saline and sham injection controls. Twenty minutes after treatment administration, the animal was released into the maze and had free access to the apparatus for 30 min. All trials were taped for later behavioral analysis. Buspirone significantly decreased the frequency of scent marking, while increasing the time spent in proximity to the 'predator' stimulus, indicating an anxiolytic effect. Neither locomotor activity, exposure to a novel environment, stimulus location and habituation, nor gender influenced the effects of the drug treatments. These results further validate this method and demonstrate the potential usefulness of this ethologically based paradigm to test anxiety and fear-induced avoidance in nonhuman primates and its susceptibility to anxiolytic pharmacological manipulations.

Animals↗
PubMed1997

Marmoset conspecific confrontation: an ethologically-based model of anxiety.

A method of measuring confrontation-induced behavioural changes in common marmosets (Callithrix jacchus) together with automated monitoring of locomotor activity has been developed as a possible model of anxiety. Recording both affiliative and agonistic behaviours between male/female pairs of marmosets and using diazepam as a reference drug, it has been possible to define a profile of behavioural changes which could be regarded as representing an anxiolytic response. Unfamiliar male/female pairs of marmosets were brought into close (non-contact) proximity in a controlled environment, in which their locomotor activity was recorded automatically. Simultaneously, their interactive behaviour was assessed by an independent observer via closed-circuit television. The following behaviours were analysed: aggressive postures, allogrooming, scratching, anxiety-related behaviours, social contact and self-grooming. Administration of diazepam at 1 and 3.5 mg/kg PO induced a significant (compared to control) reduction in scratching, aggressive behaviours, anxiety-related behaviours and an increase in allogrooming without affecting locomotor activity during confrontation. Differing responses dependent on gender were not found, nor did gender influence the effect of treatment on behaviour. Habituation to repeated confrontation did not occur. The results from this study demonstrate that this method can be used to measure anxiolytic activity in an objective manner.

Aggression↗
PubMed2002

The contraceptive potential of ZP3 and ZP3 peptides in a primate model.

It has been known for some time that antibodies raised against ZP3, the major component of the glycoprotein shell that surrounds all mammalian oocytes, can successfully inhibit sperm-egg interaction in vitro. In our own studies using the non-human primate Callithrix jacchus, active immunisation was successfully achieved when homologous or heterologous ZP3 was used as an immunogen. However this long-term suppression of fertility was at the expense of ovarian function. An ovarian pathology was observed which was characterised by a disruption of folliculogenesis and depletion of the primordial follicle pool. Adverse auto-immune reactions have also been observed in mice following induction of immunity to mouse ZP3. Following careful selection of B-cell epitopes on mouse ZP3, peptide vaccines were formulated which could circumvent these adverse side effects and induce reversible infertility in actively immunised mice. To identify similar epitopes on primate ZP3, epitope mapping studies were performed and several candidate regions of the molecule were identified. These were incorporated into chimeric peptide vaccines and administered as single or triple peptide vaccines. Active immunisation successfully induced antibodies that bound exclusively to the zona pellucida of marmoset and human ovarian sections. These antibodies were able to suppress human sperm-egg binding by up to 60% in vitro. Encouragingly, no adverse side effects on ovarian function were observed following long-term immunisation however, no loss of fertility was consistently observed in vivo. Thus considerable research is still required to identify a combination of ZP epitopes that will induce reversible infertility in the absence of any ovarian dysfunction.

Animals↗
PubMed1997

Inhibition of allergic encephalomyelitis in marmosets by vaccination with recombinant vaccinia virus encoding for myelin basic protein.

A primary demyelinating form of experimental allergic encephalomyelitis (EAE) resembling human multiple sclerosis (MS) occurs in Callithrix jacchus marmosets following immunization with human white matter. Participation of a T-cell immune response against myelin basic protein (MBP) in this disease model is supported by observations of increased reactivity against MBP in PBMC and of adoptive transfer of an inflammatory form of EAE by MBP-reactive T-cells. To evaluate the effects of ectopic presentation of MBP on marmoset EAE, animals were vaccinated prior to induction of EAE by subcutaneous injection of attenuated strains of vaccinia virus genetically engineered to contain either the entire coding sequence for human MBP (vT15) or the equine herpes virus glycoprotein gH gene (vAbT249). Vaccination with vT15 was followed by transient cytoplasmic and surface membrane expression of MBP in circulating PBMC (15-45 days). The onset of clinical EAE after immunization (pi) was markedly delayed in vT15-vaccinated animals (37-97 days pi, n = 4) compared to vAbT249-vaccinated controls (14-18 days pi, n = 3). Proliferative responses against MBP but not against vaccinia antigens or phytohemagglutinin were suppressed in protected animals. Thus, development of attenuated live viruses carrying genes for myelin antigens could be useful for induction of immunologic tolerance and for modulation of autoimmune demyelination.

Animals↗
PubMed1998

Molecular cloning in the marmoset shows that semenogelin is not the precursor of the TRH-like peptide pGlu-Glu-Pro amide.

Two peptides with similar structures to thyrotropin-releasing hormone (TRH), pGlu-Glu-Pro amide and pGlu-Phe-Pro amide, have been identified in human seminal fluid and it has been shown that one of these peptides, pGlu-Glu-Pro amide, has the ability to increase the capacitation of sperm cells, consistent with a role in fertility. In order to select a species in which there is a high degree of expression of the genes that code for 'TRH-like' peptides, we have determined the levels of these peptides in the prostate, pancreas and thyroid of a range of species including rat, rabbit, ox, marmoset, macaque and man. The peptides were extracted from the tissues and purified before determination by RIA with TRH antibody. In addition, trypsin digestion and TRH RIA was used to investigate the presence of N-extended forms. The highest concentrations of TRH-immunoreactive peptides were found in the tissues of the marmoset, Callithrix jacchus. Ion-exchange chromatography demonstrated that marmoset thyroid contained principally authentic TRH, the pancreas contained both TRH and TRH-like peptides while the prostate contained TRH-like peptides alone. Further purification by HPLC showed that the main TRH-immunoreactive peptide in marmoset prostate was pGlu-Glu-Pro amide and a second component was identified as pGlu-Phe-Pro amide. The results indicate that the biosynthesis of these peptides could be studied to advantage in the marmoset. The biosynthetic precursors of the TRH-like peptides have not been identified. To examine whether pGlu-Glu-Pro amide might originate from semenogelin, we determined the sequence of semenogelin in the marmoset. It exhibited a high degree of homology with human semenogelin-I, but in place of the Lys-Gln-Glu-Pro sequence that might give rise to pGlu-Glu-Pro amide, marmoset semenogelin possessed the sequence Ser-Gln-Asp-Gln which cannot serve as a precursor for a TRH-like peptide. Further evidence was obtained by Northern blot analysis of a range of marmoset tissues. The results showed that semenogelin is not present in marmoset prostate. It is concluded that pGlu-Glu-Pro amide originates from a precursor distinct from semenogelin, both in marmoset and in man.

Amino Acid Sequence↗
PubMed2000

A new primate model for multiple sclerosis in the common marmoset.

Experimental autoimmune encephalomyelitis (EAE) in outbred marmoset monkeys (Callithrix jacchus) is a recently developed nonhuman primate model of multiple sclerosis. Here, Bert 't Hart and colleagues compare this model to EAE in rhesus monkeys, highlighting autoimmune mechanisms in CNS inflammation and demyelination, including the role of major histocompatibility complex restriction and preclinical evaluation of innovative immunotherapies.

Acute Disease↗
PubMed2001

Mechanisms of estradiol inactivation in primate endometrium.

In uterine endometrium, the level of estradiol is controlled by oxidative 17beta-hydroxysteroid dehydrogenase (17HSD) activity which converts the bioactive hormone to the less active compound estrone. At least three different types of 17HSD (types 2, 4 and 8) use estradiol as their preferred substrate and may contribute to the overall rate of estradiol-inactivation in the uterus. In this study the marmoset monkey (Callithrix jacchus) was used for the investigation of the particular contribution of each type of 17HSD. Northern Blots revealed essentially the same tissue distribution as in the human. Likewise, uterine 17HSD enzyme activity increases in the secretory phase of the reproductive cycle, in parallel to the rise in circulating progesterone levels. Northern analysis of uteri from defined time points of the reproductive cycle showed that only the level of 17HSD2 expression is strongly upregulated in the secretory phase, whereas 17HSD4 and 17HSD8 seem to be expressed constitutively.

17-Hydroxysteroid Dehydrogenases↗
PubMed2000

L-DOPA produces strong induction of c-fos messenger RNA in dopamine-denervated cortical and striatal areas of the common marmoset.

Common marmosets (Callithrix jacchus) with near-complete unilateral 6-hydroxydopamine denervation of the dopaminergic input received a single injection of saline or L-DOPA (15mg/kg plus 6.25mg/kg benserazide). Using in situ hybridization, the effects of these treatments on c-fos messenger RNA expression in the cerebral cortex, the striatal complex and the external layer of the pallidum were studied. Moreover, receptor autoradiography was used to determine the levels of dopamine D(1) and D(2) receptors in these areas. In the cerebral cortex, animals treated with L-DOPA displayed a high expression of c-fos messenger RNA restricted to the dopamine-denervated hemisphere. No changes in the levels of cortical D(1) and D(2) receptors were found in the dopamine-denervated hemisphere. L-DOPA treatment also induced a strong expression of c-fos messenger RNA in the striatal complex in the dopamine-denervated hemisphere. The levels of striatal D(2), but not D(1), receptors were increased in the dopamine-denervated hemisphere. In the external pallidum, the major terminal region for D(2) dopamine receptor-containing striatal projection neurons, L-DOPA treatment induced c-fos messenger RNA expression in both the intact and the dopamine-denervated hemispheres.Thus, using c-fos messenger RNA as a biochemical marker of postsynaptic neuronal activation, these results provide evidence that near-complete dopamine depletion causes a profound supersensitization to L-DOPA treatment in the cerebral cortex and in the striatal complex, but not in the external layer of the pallidum, of the primate brain. The cortical response may be unique to the primate brain, but c-fos messenger RNA activation within the striatum has also been reported in the rodent. The effects of L-DOPA probably depend both on a direct activation of supersensitized dopamine receptors by dopamine produced in the few remaining, but hyperactive, dopaminergic nerve terminals and in serotonergic nerve terminals, as well as on indirect actions of L-DOPA related to activation of circuitries connecting cerebral cortex and basal ganglia structures. These results provide novel information on the mechanisms underlying L-DOPA's action in the cerebral cortex, striatum and external pallidum in a primate model of Parkinson's disease.

Animals↗
PubMed2002

Characterization of calbindin-positive cones in primates.

The aim of this study is to characterize calbindin-positive photoreceptors and their opsin content in the retina of nocturnal prosimians (Microcebus murinus), New World monkeys (Callithrix jacchus), Old World monkeys (Macaca fascicularis), and humans. To identify the calbindin and opsin content of cones, combined multiple labeling with different fluorescent probes, antibodies directed against calbindin, short, and mid-long wavelength opsins, and lectin peanut agglutinin cytochemistry were used. With the exception of Microcebus, calbindin is present in the cones of all primates but is absent from rods. The distribution of calbindin is similar in human and macaque cones, with dense label in the inner segment, cell body, axon and cone pedicle. Cones in marmoset also show dense staining in the cell body, axon and pedicle but only light label in the inner segment. Primate cone outer segments do not contain calbindin. In the primates studied, three patterns of calbindin and opsin localization are observed. In macaque and marmoset all short and mid-long wavelength cones contain calbindin. In humans, all mid-long wavelength cones contain calbindin whereas all short wavelength cones are devoid of calbindin as confirmed by confocal microscopy. In the nocturnal prosimian Microcebus none of the mid-long or short wavelength cones contain calbindin. In addition to primates, calbindin is absent in cones of other nocturnal species but is present in cones of diurnal species suggesting a difference in the role of calbindin possibly related to the adaptational states or other photoreceptor properties.

Adult↗
PubMed1997

Learning impairments following injection of a selective cholinergic immunotoxin, ME20.4 IgG-saporin, into the basal nucleus of Meynert in monkeys.

Four groups of monkeys (Callithrix jacchus) were injected with saline or increasing amounts of the immunotoxin, ME20.4 IgG-saporin, directly into the basal nucleus of Meynert via a frontal trajectory which avoided damage to the overlying basal ganglia. ME20.4 IgG binds to the primate p75 low-affinity neurotrophin receptor, when the saporin derivitized antibody is injected into the basal forebrain, it selectively destroys the magnocellular neurons of the basal nucleus of Meynert which are the cells of origin of the cholinergic projection to the neocortex. The highest dose of ME20.4 IgG-saporin produced a significant impairment on acquisition of a perceptually difficult visual discrimination. There was no significant effect on retention of tasks learnt before or after surgery, nor on concurrent acquisition of several perceptually easy discriminations or serial reversal of an easy discrimination. These results suggest that the impairment is not due to visual, motor or motivational difficulties and does not consist of difficulties with the formation of reward associations. Rather the impairment is largely confined to acquisition of perceptual discriminations. There was a significant correlation between the density of ME20.4 immunostaining in the basal nucleus of Meynert and the density of acetylcholinesterase histochemical staining in the frontal and temporal cortex and an inverse correlation between both of these and the degree of learning impairment in the animals. Lesioned animals also showed significant impairment on acquisition and reversal of perceptually easy discriminations when treated with a dose of scopolamine which did not impair performance in control animals. These results provide further evidence that cortical cholinergic neurotransmission contributes to certain forms of learning. The availability of a selective cholinergic immunotoxin effective in primates provides an important new tool for the study of cholinergic function and its involvement in ageing, Alzheimer's disease and other pathological states.

Animals↗
PubMed2004

Social suppression of cortisol in female marmoset monkeys: role of circulating ACTH levels and glucocorticoid negative feedback.

Behaviorally subordinate female common marmoset monkeys (Callithrix jacchus) exhibit pronounced, chronic reductions of circulating cortisol levels. Cortisol suppression in these animals is mediated in part by adrenocortical hyporesponsiveness to adrenocorticotropic hormone (ACTH). In addition, we hypothesized that social subordination may activate a central, neurally mediated mechanism to further inhibit hypothalamo-pituitary-adrenal function. In this study, therefore, we evaluated basal plasma cortisol and ACTH concentrations, as well as cortisol and ACTH responses to dexamethasone (DEX), in dominant and subordinate females to initially characterize such a mechanism. Morning plasma cortisol and ACTH levels were determined before, and 1, 2, and 3 days following administration of DEX (0.5, 1.0, or 5.0 mg/kg, IM) or saline. Baseline cortisol concentrations prior to DEX treatment were significantly lower in subordinate females than in dominants, as previously reported. However, ACTH concentrations in the same blood samples did not differ between the two groups. Furthermore, dominant and subordinate females showed similar cortisol and ACTH responses to DEX. These results indicate that reduced circulating cortisol levels in subordinate females are not associated with either altered circulating ACTH concentrations or enhanced responsiveness to glucocorticoid negative feedback. However, the finding that basal ACTH levels are not elevated in subordinate females as compared to dominants, in spite of low circulating cortisol concentrations, suggests that ACTH secretion in subordinate females is restrained by a steroid-independent inhibitory mechanism operating at the level of the brain or pituitary.

Adaptation, Physiological↗
PubMed1999

Age-related changes in the brainstem auditory evoked potentials of the marmoset.

Changes in brain-stem auditory evoked potentials (BAEPs) with age were recorded in common marmosets (Callithrix jacchus) at the age of 1-2, 6-8 and 10-12 years. The auditory function was assessed by thresholds, latencies and amplitudes of BAEPs evoked by use of tone burst stimulations with audible frequencies ranging from 1 to 99 kHz. Prolongation of the latencies of later waves was observed in the animals at the age of 6-8 and 10-12 years at high frequencies, suggesting that aging in marmosets, as reported previously in humans and other animals, may cause earlier hearing loss at high frequency than at low frequency within the hearing range. At 10-12 years of age, the elevations of BAEP thresholds and the declines of BAEP amplitudes in older animals were also observed. As the differences in the parameters are small, it was suggested that only a moderate hearing loss occurred with onset late in life in common marmoset similar to that in CBA/Ca mice. Based on the results obtained in this study, BAEP latencies appear to be more sensitive indicators than BAEP thresholds and amplitudes for the early detection of hearing impairment.

Aging↗