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Termination of pregnancy in common marmosets (Callithrix jacchus) following administration of antiserum to chicken riboflavin carrier protein.

The presence of protein(s) which is immunologically and functionally similar to the well characterised avian riboflavin carrier protein has been demonstrated in female common marmosets during pregnancy. The protein(s) that interacts with [14C]riboflavin has an immunological homology to chicken riboflavin carrier protein. The termination of pregnancy in four out of six marmosets following administration of antiserum to chicken riboflavin carrier protein supports the hypothesis that vitamin carrier protein ensures an uninterrupted supply of the vitamin to the growing embryo and that immunoneutralization of maternal riboflavin carrier protein leads to fetal death and termination of pregnancy.

Animals↗

The distribution of glial fibrillary acidic protein and vimentin in postnatal marmoset (Callithrix jacchus) brain.

Antisera to glial fibrillary acidic protein (GFAP) and vimentin were used to elucidate the distribution of these intermediate filament proteins in postnatal marmoset brains of various ages. The ependyma of the lateral ventricles was unique in being equally immunoreactive for both GFAP and vimentin at all ages. Vimentin alone was consistently demonstrated in endothelial and leptomeningeal cells at all ages. In neonates, vimentin immunoreactivity greatly exceeded that of GFAP and was located primarily in radial glia in the subependymal plate of the anterior cerebrum. Their vimentin-positive processes formed thick fascicles in the corpus callosum but separated into fine fibres on entering the cortex. GFAP immunoreactivity in these cells and processes was very limited. With age, GFAP-positive cells increased in number and displayed the typical stellate appearance of astroglia. The vimentin-positive radial glial population decreased considerably during this period and by 6 months had virtually disappeared. The GFAP reaction in adult brain was even more widespread, largely due to the increased number of positive astrocytes in the white matter. Vimentin immunoreactivity in the adult was greatly diminished and positive radial glia were not detectable. A major change in intermediate filament protein expression, therefore, occurs in the early postnatal period and probably reflects phases in the differentiation of radial glial precursors into astrocytes.

Animals↗

Cell proliferation in the subependymal layer of the postnatal marmoset, Callithrix jacchus.

Cells of the subependymal layer (SEL) have been shown to be capable of continued postnatal cell division throughout life in rodents. To determine if the primate brain behaves similarly, proliferative activity in the SEL of the marmoset has been investigated by tritiated thymidine autoradiography and bromodeoxyuridine immunocytochemistry. Both methods revealed the presence of DNA-synthesizing cells at all postnatal ages studied. The labelling index (LI), low at birth, reached a peak of almost 4% at one month but decreased gradually thereafter. In animals older than two years the LI was extremely low and labelled cells were rarely seen anywhere in the brain. The cell density of the SEL, in contrast to the low LI, was highest in neonates and decreased linearly with increasing age. Bromodeoxyuridine immunoreactivity revealed the distribution of proliferating cells in the SEL and neighbouring regions. Such cells were most abundant around the anterior lateral ventricle where the SEL was most evident. Proliferating cells were numerous in neonates, though not adjacent to the ependyma where counts for the LI were made, and were mainly located dorsally and ventrally at the junctions of the corpus callosum and caudate nucleus.

Aging↗

Effects of variable linoleate intake on aortic PGI2-like activity and fatty acid composition: difference between rat and marmoset (Callithrix jacchus).

Two animal species, the laboratory rat and a non-human primate (the common marmoset) were compared with respect to the influence of dietary lipid supplementation on both fatty acid composition and prostanoid synthetic capacity of aorta. Standard chow (REF. diet) was supplemented (12% w/w) with either sheep fat (SF) or sunflower seed oil (SSO). Differences were apparent in the aortic phospholipid fatty acids from both animal models, as a result of the diet, although the type and the extent of changes displayed a considerable species variation. For example, after the SSO diet, the proportion of linoleic acid in rat aorta displayed only a modest increase whilst in marmosets it was doubled. No change was apparent in the proportion of arachidonic acid in rats, whereas the marmosets showed a reduction (P less than 0.05) with both lipid supplements. Rats fed the SF diet showed a considerable increase in aortic PGI2-like activity compared to REF. And SSO supplemented groups. In contrast, no dietary induced changes were observed in the aorta from marmosets despite compositional changes. The results indicate no direct association between the synthesis of PGI2-like activity and membrane lipids and demonstrate the need for careful selection of the proper animal model, particularly in nutritionally orientated experiments.

Animals↗

Dietary lipids and adipose tissue fatty acids in the marmoset monkey (Callithrix jacchus).

The fatty acid composition of marmoset monkey perirenal fat was examined after long-term feeding of different lipid supplemented diets. Saturated fatty acid supplementation significantly reduced linoleic acid; n-6 polyunsaturated fatty acid (PUFA) supplementation decreased oleic acid but did not reduce the saturated fatty acid content of perirenal fat. Dietary supplementation with fish oil also reduced the level of linoleic acid but increased the level of palmitic acid as well as that of the long-chain n-3 PUFAs. Perirenal fat levels of linoleic acid and docosahexaenoic acid are reliable measures of n-6 and n-3 PUFA intake respectively, but the levels of saturated fatty acids palmitic and stearic do not accurately reflect the marmosets dietary intake.

Adipose Tissue↗

A novel population of tyrosine hydroxylase immunoreactive neurones in the basal forebrain of the common marmoset (Callithrix jacchus).

We have observed in the basal forebrain of the common marmoset a group of neurones which display tyrosine hydroxylase immunoreactivity (THir) with three different polyclonal antibodies and one monoclonal antibody, and which express TH mRNA as shown by in situ hybridization histochemistry. The population of cells is composed of large multipolar neurones and is located predominantly in the substantia innominata and at the ventral, medial and lateral margins of the external segment of the globus pallidus. The cell morphology and the distribution of THir cells corresponds closely to the caudal portion of the nucleus basalis of Meynert. Adjacent sections demonstrate both THir and choline acetyltransferase immunoreactivity in the cells in this group, as well as strong acetylcholinesterase activity but not dopamine immunoreactivity. These observations indicate that many cholinergic neurones in the posterior nucleus basalis of Meynert of the marmoset contain tyrosine hydroxylase, and suggest that both acetylcholine and catecholamine may be synthesised as co-localised neurotransmitters within the same magnocellular neurones. We observe no THir cells in similar areas of the basal forebrain of either rhesus or talapoin monkeys.

Animals↗

Purification and properties of cytochrome P-450 from liver microsomes of phenobarbital-treated marmoset monkeys (Callithrix jacchus).

One form of cytochrome P-450 from phenobarbital-induced marmoset liver was purified to apparent electrophoretic homogeneity and compared with the major inducible form isolated from rat liver. Whereas spectral properties and molecular weights, as well as catalytic activities towards aminopyrine and ethylmorphine N-demethylation are quite similar, rates of O-dealkylation with enzymes from the two species are considerably different. While ethoxycoumarin deethylation for the marmoset cytochrome is about one-fortieth of that for the rat, ethoxyresorufin and even pentoxyresorufin dealkylations for the marmoset form are not detectable. By contrast, aldrin epoxidation as catalyzed by this cytochrome is about three times as high as that obtained from the rat.

Aldrin↗

Calcium-binding proteins in the spiral ganglion of the monkey, Callithrix jacchus.

Calcium-binding proteins can act as intermediaries between changing levels of free intracellular calcium ions and the physiological response of neurons. Some of these proteins, among them calbindin (CB), calretinin (CR) and parvalbumin (PV), can act as calcium buffers. A survey of previous studies in rodents and human fetuses leads to the impression that many spiral ganglion cells co-express CB, CR, and PV. The findings of the present study suggest that, in the adult marmoset, the expression of CB is restricted to a small number of cells, most likely type II ganglion cells, and that at least some of the numerous type I ganglion cells co-express CR and PV. In the neonate marmoset, large numbers of putative type I ganglion cells from the apical cochlear turn transiently expressed a light and granular labeling for CB-like immunoreactivity, in addition to the cells we believe to be type II ganglion cells exhibiting a strong and solid CB-like staining. The spiral ganglion cells in all developmental stages co-expressed the mitochondrial enzyme cytochrome oxidase. Furthermore, a small population of CB-LI axons of unknown origin was found to terminate near the CB-immunoreactive ganglion cells.

Animals↗

Quantification of three-dimensional exploration in the cylinder test by the common marmoset (Callithrix jacchus).

The increasing trend in use of marmoset monkeys in behavioral studies has necessitated a greater understanding of their natural behavior, especially within confined spatial environments. One way to approach this issue is to examine locomotor and exploratory strategies using the cylinder test, as done recently for the rat by Gharbawie et al. [Gharbawie OA, Whishaw PA, Whishaw IQ. The topography of three-dimensional exploration: a new quantification of vertical and horizontal exploration, postural support, and exploratory bouts in the cylinder test. Behav Brain Res 2004;151:125-35]. We have used this paradigm in conjunction with Eshkol-Wachmann movement analysis in marmoset monkeys. We provide evidence that marmosets display systematic changes in both horizontal and vertical progressions during exploration. Quantitative analysis and comparison with the extant data in rats showed both similarities and differences. For example, both species display horizontal and vertical scanning movements, although the variability is considerably greater for the marmoset. The horizontal progressions consist of head scans, turning, and stepping. The amplitude of these progressions oscillates and decreases with trial duration. Vertical progressions, which consist of rears, head scans, and descent, also strongly oscillate but with constant mean amplitude during a trial. The difference between the two species is most evident in locomotor behavior. Rats use their forepaws primarily for postural support and follow forequarter movements while body weight shifts are generated by the hind limbs. Marmosets, however, can hold a vertical position without hand support and display more complex movement characteristics during exploration. The data from this study should be of use in designing experiments in which marmosets are engaged in free-roaming behavior within a confined space.

Animals↗

Retinal projections to the midline and intralaminar thalamic nuclei in the common marmoset (Callithrix jacchus).

In this study, we report the identification of a hitherto not reported direct retinal projection to midline and intralaminar thalamic nuclei in the marmoset brain. After unilateral intravitreal injections of cholera toxin subunit B (CTb), anterogradely transported CTb-immunoreactive fibers and presumptive terminals were seen in the following thalamic midline nuclei: paraventricular, rhomboid, interanteromedial, and reuniens, and thalamic intralaminar nuclei: central medial, central lateral, central dorsal, and parafascicular. Studies employing sensitive tracers in other primate species are needed in order to verify the possible universality of these projections. Some of the possible functional correlates of the present data are briefly discussed. The present results may contribute to the elucidation of the anatomical substrate of the functionally demonstrated involvement of this midline/intralaminar thalamic nuclear complex in several domains that include arousal and awareness, besides specific cognitive, sensory, and motor functions.

Animals↗

Multiple vaccine and pyridostigmine bromide interactions in the common marmoset Callithrix jacchus: immunological and endocrinological effects.

Following active service during the 1990/1991 Gulf Conflict, a number of UK and US veterans presented with a diverse range of symptoms, collectively known as Gulf Veterans Illnesses (GVI). The administration of vaccines and/or the pretreatment against possible nerve agent poisoning, pyridostigmine bromide (PB), given to Armed Forces personnel during the Gulf Conflict has been implicated as a possible factor in the aetiology of these illnesses. The possibility that adverse health effects may result from the administration of these vaccines (anthrax, pertussis, plague, yellow fever, polio, typhoid, tetanus, hepatitis B, meningococcal meningitis and cholera) and/or PB, have been investigated over an eighteen month period, in a non-human primate model, the common marmoset. This study reports immunological indices, including leukocyte phenotypes, intracellular cytokines IFN-gamma and IL-4 and antibody responses against vaccine antigens. Using human isotyping reagents previously shown to cross react with marmoset immunoglobulins (ibid) it was shown that marmosets responded strongly against anthrax PA and pertussis and weakly against killed whole cell plague, cholera and typhoid. At the end of the study the immune response to a previously unseen T-cell dependent antigen, keyhole limpet haemocyanin (KLH), was examined in order to determine whether immune function had been compromised by the compounds administered. Statistically equivalent, robust antibody responses were measured against KLH in all treatment groups indicating that the immune system had not been compromised by any of the treatments. In addition, urinary cortisol was measured at key points throughout the study as an index of physiological stress which may have been induced by the treatments. There were no effects of treatment on urinary cortisol secretion. With respect to the other immunological indices measured, there were no statistical differences between the treatment groups during the period of the study.

Animals↗

Development of methods to measure humoral immune responses against selected antigens in the common marmoset (Callithrix jacchus) and the effect of pyridostigmine bromide administration.

This methodological study was carried out in preparation for a major long term study, also reported in this volume, which was designed to investigate whether the combination of vaccines and pyridostigmine bromide (PB) could have been responsible for adverse signs and symptoms reported by a number of veterans of the 1990/1991 Gulf conflict. In this context, the marmoset has been used to model aspects of the human immune system. The purposes of this methodological study were to select appropriate immunochemical reagents to measure humoral responses induced in marmosets in response to selected health and hygiene and biological warfare vaccines and to initially assess the effects of PB on the responses recorded. Vaccines were administered at 1/5th of a human dose, and also investigated in combination with the nerve agent pretreatment compound PB. PB dosing was selected to induce an inhibition of erythrocyte acetylcholinesterase by 30%. In order to assess the functionality of the immune system, antibody responses to a neo-antigen (keyhole limpet haemocyanin--KLH), administered some 2 months following the completion of the vaccination schedule, were measured. The present study identified appropriate isotyping reporter reagents which cross-reacted with equivalent marmoset immunoglobulins. Robust antibody responses were identified against anthrax protective antigen (PA), whole cell pertussis vaccine and KLH, while weaker responses were measured against cholera and typhoid vaccines. The killed whole cell plague vaccine induced a response which was at the limit of detection of the assay. Coadministered PB had no discernable effect on immunological responses in this study.

Animals↗

Orientation selectivity in the common marmoset (Callithrix jacchus): the periodicity of orientation columns in V1 and V2.

Orientation selectivity is a ubiquitous property of the primary visual cortex of mammals. Within the primate, orientation selectivity is arranged into vertical columns that are organized into a regular patchy pattern. Previous studies, in old world primates, have noted an anisotropy in this arrangement that appears to be due to the presence of ocular dominance columns within the same tissue. In addition, orientation selective responses appear to be arranged into bands of activity within the adjoining extrastriate region V2. Little is known about the precise arrangement of orientation columns within V2. In this study, we examined the layout of orientation columns within both V1 and V2 of a new world primate, the common marmoset, using optical imaging. New world primates have the advantage that, unlike the macaque, V2 exists on the cortical surface, a requirement for this form of optical mapping. We found the arrangement of orientation columns to be isotropic within marmoset V1 with an average repeat distance of around 575 mum, smaller than the repeat distance previously reported for the macaque. We found no evidence of ocular dominance within the animals tested supporting the claim that ocular dominance columns when present distort the mapping of orientation in V1. In V2 we found that orientation columns were larger and as in other primates were represented in discrete bands throughout V2. Orientation columns were spaced on average around 1 mm apart. This suggests that, at least in the marmoset, the visual system maps orientation at a different scale within V1 and V2.

Animals↗

Suppression of prolactin does not reduce infant care by parentally experienced male common marmosets (Callithrix jacchus).

High levels of prolactin have been found to correlate with the expression of paternal care in a variety of taxa. However, in mammals, there is little experimental evidence that prolactin is causally involved in the stimulation or maintenance of paternal care. Here, we suppressed prolactin production in paternally experienced common marmoset fathers in their family groups during the first 2 weeks after their infants were born. Circulating prolactin levels were suppressed using cabergoline (Dostinex: Pfizer), a long acting dopamine (D2) agonist with minimal behavioural side-effects. A within-subject design was used to compare behavioural and hormonal data on 5 paternally experienced fathers during two consecutive births. Cabergoline reduced prolactin to negligible levels in all fathers without effecting testosterone, DHT and cortisol and without adverse side-effects. However, lowering prolactin had no significant effect on the expression of majority of the behaviour patterns associated with paternal care. These included infant carrying, infant grooming and the frequency with which fathers retrieved and rejected infants. The only infant-related behaviour to be affected was the frequency with which fathers touched, licked and investigated infants. We noted a marginally significant increase in this behaviour during cabergoline treatment. Despite the lack of effect on paternal care, cabergoline did exert an effect on the affiliative/sexual behaviour of fathers as there was a significant increase in the grooming behaviour fathers directed at and received from their mates during drug treatment. This study showed that experienced male marmosets can express paternal behaviour in the absence of the high prolactin levels normally seen after infants are born.

Animals↗

Repeated parental deprivation in the infant common marmoset (Callithrix jacchus, primates) and analysis of its effects on early development.

BACKGROUND: We describe a successful demonstration that repeated early deprivation of parental care (ED), as used to study effects of early life stress in rats, can be performed in a primate and that it constitutes an early life stressor. METHODS: Seven breeding pairs of marmoset monkeys each provided control twins (CON) and twins that were subjected to ED for 30-120 min/day on postnatal days (PND) 2-28. Urine samples were obtained to monitor the acute effect of ED on cortisol and catecholamine levels. Behavior samples were obtained in the home cage to monitor the effects of ED on infant and infant-parent behavior. RESULTS: Early deprivation of parental care caused acute increases in cortisol, epinephrine, and norepinephrine. At PND 28, basal cortisol was reduced in ED compared with CON infants, and ED infants were smaller than CON infants. Early deprivation infants tended to spend more time in the suckling position than did CON. Early deprivation infants demonstrated more distress vocalization than CON infants, even though the parental care they received in the home cage was similar. Early deprivation infants tended to play less socially than did CON. CONCLUSIONS: To our knowledge, this is the first demonstration that a repeated early life stressor of the type developed in rats can also be applied to a primate species.

Animals↗

Laminar expression of neurofilament protein in the superior colliculus of the marmoset monkey (Callithrix jacchus).

The expression profile of the monoclonal antibody SMI-32 was examined in the superior colliculus of adult marmosets. This antibody recognises subunits of the non- and dephosphorylated neurofilament protein, labelling predominantly neuronal perikarya and dendrites. The densest cellular label was observed in the intermediate layers (primarily, the stratum griseum intermediale), consisting of large multi- or bipolar neurones which were preferentially located within cytochrome oxidase-rich regions. The morphological characteristics of neurones showing heavy staining resemble those of extrinsic projection cells, suggesting a correlation between neurofilament content and axonal length.

Animals↗

Antifertility effect of an octapeptide, a fragment of FSH binding inhibitor, in the common marmoset (Callithrix jacchus).

The follicle-stimulating hormone (FSH) binding inhibitor (FSHBI) has been identified as one of the factors present in follicular fluid exerting autocrine/paracrine effects on FSH actions in the ovary. Our group has isolated FSHBI from human ovarian follicular fluid and deduced its partial amino acid sequence from the N-terminal region. A synthetic peptide corresponding to this sequence also demonstrated FSH binding inhibitory activity in vitro. The objective of the present study was to elucidate the effect of the octapeptide on ovarian cyclicity and pregnancy in the common marmoset. For the study, three groups of postpartum marmosets were treated with the octapeptide during the follicular phase. Administration of the octapeptide from days 6-10 postpartum (pp) predominantly induced luteal insufficiency in two of seven and two of five marmosets at a dose of 100 microg/day (group 1) and 300 microg/day (group 2), respectively. This was confirmed by the presence of small regressing corpora lutea on day 13 pp. Pregnancy was terminated prematurely in one animal from each of these groups. The treatment impaired fertility by 43% and 60% in the marmosets of groups 1 and 2, respectively. In view of the fact that FSH levels peak on day 2 and day 6 of the follicular phase in marmosets, a third group was administered 200 microg/day octapeptide from days 1-8 pp. The treatment induced luteal insufficiency in one out of four marmosets, while premature termination of pregnancy occurred in two other marmosets of this group, demonstrating a 75% effect on pregnancy. Thus, treatment of marmosets with the octapeptide, a fragment of the FSHBI, predominantly induced luteal insufficiency as well as resorption of the fetus leading to impairment of fertility.

Animals↗

New hyperprolactinemia and anovulation model in common marmoset (Callithrix jacchus) and effect of cabergoline.

We aimed to develop an anovulation model, using sulpiride-induced hyperprolactinemia in common marmosets. The serum prolactin level gradually increased during the twice-daily administration of sulpiride and reached a plateau after 4 days. Sulpiride produced as big a response at 10 mg kg(-1) as at 50 mg kg(-1). In this study, the length of the ovarian cycle was approximately 30 days in normal common marmosets. Serum progesterone and estradiol levels showed no consistent change during the first 2 months of treatment with sulpiride. When treatment with sulpiride had been continued for more than 2 months, serum progesterone and estradiol levels fell to within the range seen in the follicular phase of the normal cycle and absence of ovulation was recognized by laparoscopy. A single oral administration of cabergoline (at doses between 0.01 and 0.1 mg kg(-1)) dose dependently reduced the elevated serum prolactin level. Bromocriptine (at an oral dose of 10 mg kg(-1)) also reduced the serum prolactin level at 4 and 8 h after its administration. With bromocriptine, the prolactin level had recovered at 24 h, but with cabergoline at doses of 0.05 mg kg(-1) or more, it had still not recovered at 48 h. In anovulatory common marmosets, oral administration of cabergoline at a daily dose of 0.05 mg kg(-1) restored ovarian function and resulted in ovulation in 100% of the group (following a reduction in the serum prolactin level). Bromocriptine at a daily oral dose of 10 mg kg(-1) resulted in ovulation in 67% of the group, but this dose was about 200 times higher than the dose of cabergoline. We could produce an anovulatory model induced by sulpiride repeatedly administered over a long time period. It is suggested that, in this anovulatory model in common marmosets, cabergoline has a potent and long-lasting action as a dopamine D2 receptor agonist, and thus could be a useful drug for the treatment of galactorrhea and hyperprolactinemic amenorrhea and/or anovulation.

Animals↗