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PubMed1988

Births following the transfer of cultured embryos obtained by in vitro and in vivo fertilization in the marmoset monkey (Callithrix jacchus).

The aim of this study was to determine whether marmoset monkeys are a suitable primate model for in vitro fertilization (IVF), embryo culture, and transplantation studies. A prostaglandin analogue given in early pregnancy and human chorionic gonadotropin given near the end of the ensuing follicular phase were used for controlling the reproductive cycle, timing oocyte collection, and synchronizing the cycles of oocyte donors and embryo recipients. Five embryos obtained by IVF were transferred at early stages to the uterus of three recipients, and two gave birth to live infants. Some of the embryos were cultured to advanced blastocyst stages. In vivo fertilized oocytes were also cultured and transferred to two recipients, and one gave birth. We concluded that the marmoset is one of the best primates for such investigations.

Animals↗
PubMed2001

Increased ACTH and cortisol secretion after interleukin-alpha injection in the common marmoset (Callithrix jacchus jacchus).

We have studied the effect of intravenous injection of interleukin-1 (dose range: from 0.25 to 4.5 microg/kg of body weight) on plasma ACTH and cortisol levels in the marmoset, a primate paradygm of peripheral glucocorticoid resistance. Blood sampling were collected and body temperature recorded 0, 15, 30, 60, 120, 180, 240 and 300 min after injection. Interleukin-1 stimulated secretion of ACTH in a dose-dependent fashion. Maximal secretion occurred 120 min after injection, and lasted up to 240 min. Plasma ACTH levels returned to baseline 300 min after interleukin-1 injection. Plasma cortisol levels were related to ACTH levels. Body temperature elevation, which occurred 10-15 min after injection was dose-dependent, and lasted 3 h. Results suggest that the pyrogenic effect of interleukin is associated, in the marmoset, with integrated activation of the hypothalamic-pituitary-adrenal axis. In light of the proneness of marmosets to hyperimmune disorders, our data are consistent with the hypothesized central biological role of IL-1, as well as the pathophysiological relevance of the neuro-endocrine-immune cross-talk during the acute phase response.

Adrenocorticotropic Hormone↗
PubMed1997

Psychosocial stress and urinary cortisol excretion in marmoset monkeys (Callithrix kuhli).

Activation of the hypothalamic-pituitary-adrenal (HPA) axis is one of the hallmarks of the physiological responses to psychosocial stressors. The most common method of assessing HPA function is via the measurement of plasma cortisol levels. However, venipuncture involves capture and restraint, which can modify HPA function. We validated a noninvasive procedure for monitoring HPA responses to stressors by measuring excretion of free urinary cortisol. Samples collected throughout the day displayed marked circadian variation, with low cortisol values in first-void samples, followed by a mid-morning peak in cortisol excretion. Concentrations of excreted cortisol declined throughout the day. Exposing marmosets to mild and moderate stressors (11 h isolation in a small cage and manual restraint) increased excreted cortisol concentrations in a dose-dependent fashion: isolation in a small cage led to elevated cortisol in afternoon samples, while manual restraint and isolation produced elevated cortisol in both morning and afternoon samples. The marmoset HPA is differentially sensitive to rather subtle variations in stressors, and these results show that urinary cortisol excretion is a valid and sensitive index of the HPA response to these stressors.

Animals↗
PubMed1997

Relationship between expression of integrins and granulosa cell apoptosis in ovarian follicles of the marmoset (Callithrix jacchus).

DNA fragmentation (apoptosis) was studied during the follicular, periovulatory and luteal phase in the marmoset monkey ovary by means of terminal transferase mediated in situ nick end labeling, and correlated with immunohistochemical localization of integrins (beta 1, alpha 2 and alpha 6 subunits). For this purpose a double-labeling technique was developed. During all phases, apoptosis of granulosa cells was exclusively restricted to tertiary follicles displaying advanced stages of atresia (as morphologically determined). During early stages of atresia in tertiary follicles, indicated by widened intercellular spaces, no apoptosis was seen. Staining intensities for integrins beta 1 and alpha 6 were strong in intact primordial/primary, secondary and tertiary follicles. Integrin expression of granulosa cells was weak in atretic tertiary follicles but not in atretic primary or secondary follicles. Double labeling revealed that DNA fragmentation was solely found in granulosa cells of tertiary follicles displaying faint or absent staining for both integrin subunits. During the periovulatory and the luteal phase, granulosa cells of atretic tertiary follicles bordering on the basal membrane, which were referred to as luteinizing cells, expressed the beta 1 subunit as well as the alpha 2-integrin subunit whereas granulosa cells neighboring to the antrum were apoptotic and negative for integrin immunoreactivity. In summary, early atresia of tertiary follicles is first characterized by morphological alterations as wide intercellular gaps, without any signs of granulosa cell apoptosis. Advanced stages of atresia, in tertiary follicles however, are accompanied by apoptosis of granulosa cells and a faint or absent staining for integrin subunits beta 1 and alpha 6. According to recent in vitro findings, our results point to a possible relation between granulosa cell apoptosis of tertiary follicles and integrin expression in ovarian marmoset follicles.

Animals↗
PubMed1999

The effects of spectacle wear in infancy on eye growth and refractive error in the marmoset (Callithrix jacchus).

We made a comprehensive study, involving observations on 45 marmosets, of the effects on ocular growth and refraction of wearing spectacles from the ages of 4-8 weeks. This period was within the period early in life when the eye grows rapidly and refraction changes from hyperopia to its adult value of modest myopia. In one series of experiments we studied the effect of lenses of powers -8, -4, +4 and +8D fitted monocularly. In another series of experiments we studied the effect of lenses of equal and opposite powers fitted binocularly, with the two eyes alternately occluded, so as to give an incentive to use both eyes, and in particular to accommodate, for at least part of each day, through the negative lens. The vitreous chamber of eyes that wore negative lenses of -4D or -8D, combined with alternate occlusion, elongated more rapidly than that of the fellow eye (negative lens eye-positive lens eye, 0.21 +/- 0.03 mm (S.E.M.), P < 0.01 and 0.25 +/- 0.06 mm, P < 0.05, respectively) and became relatively more myopic (2.8 +/- 0.26D, P < 0.01 and 2.4 +/- 0.61D, P < 0.05 respectively). Eyes that wore -4D lenses monocularly elongated more rapidly and became myopic than fellow eyes. Eyes that wore +4D or +8D lenses were less strongly affected: animals that wore +8D lenses monocularly (without alternate occlusion) developed a slight relative hyperopia (0.99 +/- 0.21D, P < 0.01), with the more hyperopic eyes also slightly shorter (0.09 +/- 0.05 mm) than their fellow eyes, but eyes wearing +4D lenses were not significantly different from their fellow eyes. Animals that wore -8D lenses monocularly (without alternate occlusion) developed a slight relative hyperopia after three weeks of lens-wear (0.85 +/- 0.26D, P < 0.05). These were the only eyes that responded in a non-compensatory direction to the optical challenge of spectacle wear, and we interpret this effect as one due to visual deprivation. After the removal of lenses, the degree of anisometropia slowly diminished in those groups of animals in which it had been induced, but in the three groups in which the largest effects had been produced by lens-wear the overall mean anisometropia (0.68 +/- 0.24D, P < 0.01) and vitreous chamber depth (VCD) discrepancy (0.09 +/- 0.03 mm, P < 0.01) were still significant at the end of the experiments, when the animals were 273 days old. The reduction of anisometropia in these groups was associated with an increase in the rate of elongation of the vitreous chamber in the eyes that had previously grown normally i.e. the less myopic eyes grew more rapidly than their fellow eyes: in the seven weeks following lens-wear these eyes became more myopic and longer than normal eyes (refraction P < 0.001; VCD P < 0.001). Control experiments showed that occlusion of one eye for 50% of the day had no effect on eye growth and refraction, and therefore that alternate occlusion itself had no effect.

Age Factors↗
PubMed1997

The common marmoset (Callithrix jacchus) as a model for neuroleptic-induced acute dystonia.

To examine whether acute dystonia is induced by neuroleptic treatment, common marmosets were treated with haloperidol orally twice a week over 25 weeks until dystonic behavior was elicited. Movement disorders such as acute dystonia were observed 6 weeks after the initial treatment, and had appeared in all treated animals by 25 weeks. Once these movement disorders were induced, they consistently reappeared after further treatment with haloperidol, and once haloperidol dosing was discontinued, the episodes vanished. Then, various neuroleptic drugs (bromperidol, chlorpromazine, risperidone thioridazine, sulpiride, tiapride, and clozapine) or a nonneuroleptic drug (diazepam) were administered orally instead of haloperidol in the above animals. All the neuroleptic drugs except for clozapine elicited similar abnormal behavior, while diazepam failed to induce any dystonia. An anticholinergic drug, trihexyphenidyl, which is known to reduce acute dystonia in patients, was also given orally to the above haloperidol-sensitized animals, followed by further treatment with haloperidol 30 min later. This clearly suppressed the induction of dystonia by haloperidol. The similarity between these findings for haloperidol-pretreated common marmosets and clinical findings suggests that the present model is useful for predicting the potential of antipsychotics to induce acute dystonia in humans.

Acute Disease↗
PubMed1996

Inducibility of cytochromes P-450 by dioxin in liver and extrahepatic tissues of the marmoset monkey (Callithrix jacchus).

Cytochrome P-450 induction was investigated in the marmoset monkey, a non-human primate, using dioxins as inducing agents. Animals received a single subcutaneous dose of 1.6 nmol tetrachlorodibenzo-p-dioxin or tetrabromodibenzo-p-dioxin/kg body weight. Microsomal fractions were prepared from liver, lung and kidney, and homogenates were prepared from gut and adrenal glands. Anti-peptide antibodies which bind to CYP1A1, CYP1A2, CYP2B6 and CYP3A4 in human were used to identify related forms in the marmoset. The results indicate that CYP1A2 is constitutively expressed in liver, but not in lung, kidney, gut or adrenal gland and that CYP1A1 is not expressed in any of these tissues in untreated animals. Treatment with dioxin induced both CYP1A1 and CYP1A2 in liver, but only CYP1A1 in lung. No induction of CYP1A1 or CYP1A2 was found in kidney, small intestine or adrenal glands. Methoxy-, ethoxy-, pentoxy- and benzoyloxyresorufin O-dealkylases and high affinity phenacetin O-deethylase activities were induced in the liver, whereas ethoxycoumarin O-deethylase and aryl hydrocarbon hydroxylase activities were not affected by dioxin treatment. High-affinity phenacetin O-deethylase and CYP1A2 apoprotein were detected only in liver, consistent with this activity being specifically catalysed by CYP1A2. Furafylline was found to be a competitive inhibitor of methoxyresorufin O-demethylase activity with a Ki of 10 microM. In the lung the induction of CYP1A1 was accompanied by 15- and 23-fold increases in ethoxyresorufin O-deethylase and methoxyresorufin O-demethylase activities, respectively, suggesting that both activities are catalysed by CYP1A1. In contrast, there was no induction of aryl hydrocarbon hydroxylase activity in lung or liver showing that, unlike in many other species, marmoset CYP1A1 does not catalyse this reaction efficiently. The expression, distribution, induction and substrate specificities of marmoset monkey P-450 enzymes differ from the situation found in rodents and other species, demonstrating that caution has to be exercised when making cross-species extrapolations.

Animals↗
PubMed1999

Unipolar brush cells in the cochlear nuclei of a primate (Callithrix jacchus).

Unipolar brush cells (UBCs) have been recognized in the cerebella of many species including primates. They have also been identified in the cochlear nuclei (CN) of many mammals, however, not in any primate. Our immunohistochemical study in the marmoset demonstrates the presence of abundant calretinin immunoreactive (CR-ir) UBCs in the cerebellum, as in other mammals. In the marmoset CN, in contrast, CR-ir UBCs could hardly be identified. The reverse pattern was evident in sections immunostained for a metabotropic glutamate receptor (mGluR2/3-IR): beautifully stained UBCs were present in the CN, but the stained cerebellar structures were difficult to identify as UBCs. The present findings indicate that UBCs are present in the CN of primates, as in other mammals, but that cerebellar and CN UBCs might differ in their molecular equipment in primates.

Animals↗
PubMed1997

Calbindin immunoreactivity delineates the circadian visual centers of the brain of the common marmoset (Callithrix jacchus).

The hypothalamic suprachiasmatic nucleus and the thalamic pregeniculate nucleus (which includes the intergeniculate leaflet) comprise the circadian visual system in the primate brain. In this study, we used intraocular injections of cholera toxin subunit B to identify those nuclei in the common marmoset brain, and demonstrated that calbindin D-28k immunoreactivity apparently labels most neurons in both the suprachiasmatic and pregeniculate nuclei. These data suggest that calbindin D-28k could represent a reliable neuronal marker for structures of the circadian visual system in marmosets and provide anatomical information on the primate equivalent of the rodent intergeniculate leaflet.

Animals↗
PubMed1997

The distribution of p75 neurotrophin receptor-immunoreactive cells in the forebrain of the common marmoset (Callithrix jacchus).

The distribution of neurones that could be stained immunohistochemically with antibody to the p75 neurotrophin protein was studied in the forebrain of the common marmoset. The p75-immunoreactive forebrain cells appear to correspond to choline acetyltransferase-immunoreactive (i.e., cholinergic) neurones. Two populations of cells could be distinguished on the basis of the intensity of p75 immunostaining. Moderately stained cells correspond to cholinergic interneurones of the caudate and putamen, while intensely stained cells correspond to the cholinergic neurones projecting to the cortex, amygdala, and hippocampus, located in the septum, diagonal band, and basal nucleus of Meynert. The distribution of cells of the diagonal band/basal nucleus complex is more extensive in the marmoset than in other primate species, extending into parts of the postcommissural fornix via the posterior septum, and by small projections dorsal to the anterior commissure and via the thalamic fasciculus from the basal nucleus; the posterior extent of the basal nucleus continues extensively into the lamina between the globus pallidus and the putamen.

Animals↗
PubMed1995

The interstitial space of the thyroid gland of marmosets (Callithrix jacchus).

The interstitial space of the thyroid gland of adult marmosets contains, like the stroma of other organs, cells and intercellular substance (matrix), blood vessels (predominantly capillaries), lymph vessels and unmyelinated nerves. It is demarcated from the follicular epithelium, the capillaries and Schwann cells by a basal lamina (BL). The perifollicular BL shows thickenings of up to 3 microns over long distances or a multilayered arrangement. These thickened segments exhibit numerous epithelial processes and ridges; in other words, the contour of the basal cell membrane is very irregular in these areas. Indentations of capillaries into the epithelium are rarely observed. The endothelium is only slightly porous. Lymph capillaries occur in large numbers. They originate freely in the interstitial space, show gaps or unspecific contacts between the thin endothelial cells; a basal lamina is missing. Bundles of 10-nm thick filaments (anchor filaments) extend to the endothelial cells of the lymph capillaries. Thin and very long (up to 8 microns) plate-like processes surround the capillaries or run parallel to the outer contour of the follicles. They originate at the poles of oval, fibroblast-like cells. Since these cells are FXIII- and C3bi-positive, they can be considered as dendritic cells. They obviously play a role in the frequently-observed autoimmune diseases of this species. In addition, monocytes and all transitional forms including macrophages, fibrocytes and lymphocytes as well as numerous mast cells occur. In the region of the BL, integrins of the beta 1-group (alpha 6) can be demonstrated immunohistologically in addition to the usual components (collagen type IV, laminin and heparan sulfate-proteoglycan). Of the fibrillar collagens type I does not occur, type III occurs only in small amounts, whereas types V and VI are observed in large amounts. The presented findings may serve as basis for more extensive experiments on these primates.

Animals↗
PubMed1997

Proliferation and apoptosis in follicles of the marmoset monkey (Callithrix jacchus) ovary.

Proliferation and apoptosis were studied in ovarian follicles of immature and pubertal marmosets and in mature marmosets during the follicular, periovulatory and luteal phases. Proliferation was evaluated using a Ki 67 antibody and apoptosis was assessed by in situ detection of DNA fragmentation. In the immature animals only small follicles were present, and the expression of Ki 67 was restricted to the granulosa cells of follicles localised near the medulla. There was no evidence of DNA fragmentation. In pubertal and adult animals Ki 67 expression was found in the granulosa cells of some but not all primordial and primary follicles. In the secondary and tertiary follicles immunoreactivity was localized in theca cells and granulosa cells. In atretic follicles (morphologically classified) the number of Ki 67 positive granulosa cells varied. In corpora lutea as well as in corpora lutea accessoria, staining was seen in the nuclei of some luteal cells. During all phases of the cycle, follicles from the secondary stage onwards were proliferating, whereas granulosa cells of primary follicles were only stained during the follicular phase. During all phases of the ovarian cycle apoptosis was restricted to the granulosa cells of tertiary follicles. With regard to proliferation and apoptosis, follicles exhibiting morphological signs of atresia can be classified as follows: (1) granulosa cells showing strong Ki 67 expression; (2) granulosa cells with reduced expression of Ki 67; (3) granulosa cells devoid of Ki 67 immunoreactivity and of apoptotic signs; (4) granulosa cells heavily stained for DNA fragmentation and not stained for Ki 67; (5) granulosa cells close to the antrum showing DNA fragmentation but luteinizing Ki 67 positive granulosa cells close to the basement membrane. In summary, it was shown that atresia of tertiary follicles is characterised by three consecutive stages: morphological alterations, cessation of proliferation and finally apoptosis in tertiary follicles. Thus, our results indicate that early atresia as evidenced by the morphological signs is not necessarily related to DNA fragmentation, since apoptosis is exclusively found in the granulosa cells of advanced atretic tertiary follicles.

Animals↗
PubMedDate not supplied

Distribution of the alpha1 subunit of the GABA(A) receptor on midget and parasol ganglion cells in the retina of the common marmoset Callithrix jacchus.

The inhibitory neurotransmitter gamma aminobutyric acid (GABA) has been shown to influence the responses of ganglion cells in the mammalian retina. Consistently, GABA(A) receptor subunits have been localized to different ganglion cell types. In this study, the distribution of the alpha1 subunit of the GABA(A) receptor on the dendrites of midget and parasol ganglion cells was investigated quantitatively in the retina of a New World monkey, the marmoset. Ganglion cells were injected with Neurobiotin in a live in vitro retinal whole-mount preparation. Retinal pieces were then processed with an antibody against the alpha1 subunit of the GABA(A) receptor. Strong punctate immunoreactivity indicative of synaptic localization is present in the ON and OFF sublamina of the inner plexiform layer. Many of the immunoreactive puncta coincide with the dendrites of both midget and parasol ganglion cells. Immunoreactive puncta are present on distal and proximal dendrites of ON and OFF cells of both ganglion cell types. On average, parasol cells show a slight increase in the spatial density of immunoreactive puncta with distance from the soma, whereas the density of immunoreactive puncta on midget cells stays even. Parasol ganglion cells show a slightly higher average density of immunoreactive puncta (0.083 puncta/microm dendrite) than midget cells (0.054 puncta/microm dendrite).

Animals↗
PubMedDate not supplied

Interaction between rod and cone signals in responses of lateral geniculate neurons in dichromatic marmosets (Callithrix jacchus).

Parvocellular (P-) and magnocellular (M-) cells in the marmoset LGN can receive prominent rod input up to relatively high illuminance levels (Kremers et al., 1997b). In the present paper, we quantify rod and cone input strengths under different retinal illuminance levels. The stimulus was based on the so-called "silent substitution" method. The activities of P- and M-cells of dichromatic animals were recorded extracellularly. We were able to adequately describe the response amplitudes and phases by a vector summation of rod and cone signals. At low retinal illuminance levels, the cells' responses were determined by rod and cone inputs. With increasing illuminances the strength of the cone input increased relative to the rod strength. But, we often found significant rod inputs up to illuminances equivalent to 700 td in the human eye or more. Rod input strength was more pronounced in cells with receptive fields at large retinal eccentricities. The phase differences between rod and cone inputs suggest that the rod signals lag about 45 ms behind the cone signals.

Animals↗
PubMedDate not supplied

Analysis of two types of cone bipolar cells in the retina of a New World monkey, the marmoset, Callithrix jacchus.

Two types of cone bipolar cells, the blue cone bipolar cell and the diffuse bipolar cell (DB3), were labelled immunohistochemically and investigated in the retina of a New World monkey, the marmoset. Blue cone bipolar cells were labelled with an antiserum against cholecystokinin. Short-wavelength-sensitive (SWS) cones were labelled with an antiserum against the SWS cone opsin. The DB3 cells were labelled with antibodies to calbindin. Blue cone bipolar cells in marmoset do not form a regular mosaic but instead follow the random distribution of the SWS cones. Nevertheless, the SWS cone to blue cone bipolar cell connectivity in marmoset is very similar to that previously described for macaque. In contrast to the blue cone bipolar cells, the DB3 cells form a regular mosaic. The synaptic connectivity of DB3 cells in the inner plexiform layer was analyzed. They make output synapses onto ganglion cells and amacrine cells, and gap junctions with each other. Our results provide further evidence for the existence of parallel bipolar cell pathways in the primate retina and support the view that the retinae of Old World and New World primates have common neuronal connectivity. The random distribution of SWS cones and blue cone bipolar cells is an exception to the general rule of a regular mosaic distribution of cell populations in the retina.

Animals↗
PubMed2000

Intraretinal axon diameters of a New World primate, the marmoset (Callithrix jacchus).

PURPOSE: Previously, measurements of retinal ganglion cell axon diameter have been used to make inferences about the physiology and clinical pathology of the visual pathway. However, few of these studies were able to unequivocally relate axon diameter to retinal ganglion cell type and other associated measurements. In this and our previous study we have examined intraretinal axon diameters to determine if differences in axon diameter may help to explain conduction velocity measurements found previously. METHODS: Individual retinal ganglion cells of a New World primate, the common marmoset (Collithrix jacchus) were injected iontophoretically with 2% Lucifer yellow and 4% neurobiotin. Labelled cells were visualized by horseradish peroxidase immunohistochemistry and diaminobenzidine and then retinae were mounted vitreal side up on a glass slide. Cell measurements were made with the aid of a camera lucida attachment and computer-aided morphometry Axons were photographed under x 100 oil immersion and measured at a final magnification of x 4600. RESULTS: A sample of 62 parasol cells, 22 midget cells, 16 hedge cells and 11 small bistratified cells were analysed. Dendritic field diameter of the different cell classes showed only moderate (non-significant) increases with eccentricity. Only the parasol cells demonstrated a significant increase in mean axon diameter with eccentricity. When the parasol class was examined more closely, it was found that only parasol cells of the superior, inferior and temporal retina (SIT group) showed significant positive correlations between different cell parameters (mean axon diameter, soma diameter, dendritic field diameter, eccentricity). Soma and dendritic field diameters of the SIT group were significantly larger than those of the nasal parasol cells. However, mean axon diameters of the SIT cells were not significantly different from nasal parasol cells. Axon diameters of nasal parasol cells were very variable and overlapped those of the midget and hedge cell classes to a large extent. CONCLUSIONS: The present data show that for marmoset parasol cells there may not be a clearly defined distinction between nasal and superior, inferior and temporal parasol cells on the basis of axon size. Of particular interest in the present analysis is the clear separation of superior, inferior and temporal parasol cells and nasal parasol cells when comparing soma and dendritic field diameters which is not reflected in the distribution of axon diameters. We suggest that changes in diameter along the length of an axon, differences between retinal quadrants and the variability between cells may be related to minimization of spatiotemporal dispersion necessary for accurate perception of motion within the visual world.

Animals↗
PubMed2001

Female marmoset monkeys (Callithrix jacchus) can be identified from the chemical composition of their scent marks.

The present study analyzed 42 organic solvent extracts of scent mark pools from five dominant female common marmosets by gas chromatography (GC) and combined GC and mass spectrometry. We determined whether there were qualitative or quantitative differences between the chemical composition of scent marks from individual females. Gas chromatography and mass spectral analysis detected the same 162 chemicals in 86% (36/42) of scent mark pools from five dominant females. This near identical chemical composition of scent marks suggested there were few, if any, qualitative differences between the chemical composition of scent marks from individual females. Instead, quantitative differences in scent may provide the key factor distinguishing individual females. Using the relative concentration of highly volatile chemicals detected by GC in scent marks, linear discriminant analysis classified scent mark pools to their correct donor approximately 91% of the time. Such highly reliable statistical matching of scent to donor suggested that each individual female common marmoset has a unique ratio of highly volatile chemicals in their scent marks which may permit individual identification of females from odors in their scent alone.

Animals↗