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Sex determination in the common marmoset (Callithrix jacchus).

The general morphology of the external genitalia was examined in 43 common marmosets (Callithrix jacchus) ranging in age from newborn to adult. At birth, the scrotum was a small irregular fold of skin on either side of the caudal aspect of the penis. The testes were not present in the scrotum until 8--11 months of age. The scrotum covered the penis in the adult male, was devoid of fur, was covered by pearly while nodules, and presented a median raphe. In the young female, the pudendal pad closely resembled the scrotum of the male. The vulva was pendulous, and the small vestibular opening was located near its most ventral aspect and closely resembled the preputial opening of the male. In the adult female, the pudendal pad was pendulous, was studded with white nodules, and closely resembled the scrotum of the male. Sex determination by casual observation resulted in numerous errors. Accurate sex determination was based on differences in the preputial and vestibular openings, demonstration of the glans penis in the prepuce, and palpation of the testes in the scrotum or inguinal region.

Animals↗

Use of the common marmoset, Callithrix jacchus, in reproductive research.

A self-sustaining colony of common marmosets, Callithrix jacchus, was established for reproductive research. Methods of management were developed to ensure optimal breeding conditions for marmosets in captivity. Studies of the reproductive biology and endocrinology of this species suggest that the common marmoset may be a suitable model for the human in certain aspects of the ovarian cycle, pregnancy, puberty and the hypothalamic and pituitary control of luteinizing hormone secretion. The rapid rate of reproduction of C. jacchus in captivity allows a relatively large colony to be maintained at moderate cost and also makes this species a most suitable laboratory primate for longer term studies of the safety and efficacy of new contraceptive methods.

Aging↗

Reproduction of wild-caught and laboratory-born marmoset species used in biomedical research (Saguinus sp, Callithrix jacchus).

Reproductive performances of wild-caught and laboratory-born marmosets maintained in a paired-cage system were summarized. Wild-caught, white-lipped (WL) marmosets (Saguinus fuscicollis, S nigricollis) averaged 2.6 live births per pair per yr compared with 1.6 for cotton-topped (CT) marmosets (S oedipus); S fuscicollis illigeri pairs were the best producers. Of first and second generation laboratory-born animals, crossbred or hybrid WL pairs were the best producers; first generation pairs averaged 2.7 live births per pair per yr, comparable to the production of wild-caught WL pairs. In contrast, laboratory-born Sf illigeri pairs produced only 0.8 live births per pair per yr. No significant differences in reproductive performance were observed between first generation outbred vs inbred and between parent-reared vs hand-reared pairs. Based on limited data, the reproductive capabilities of wild-caught and laboratory-born Callithrix jacchus appeared superior to both WL and CT marmosets. Ten examples of marmoset models in biomedical research were discussed briefly.

Animals↗

The embryology of the common marmoset (Callithrix jacchus).

Earlier studies have noted the unusual placental anatomy of marmosets and the high frequency of biovular, synchorial twinning. The maintenance of a captive colony of Callithrix jacchus has enabled further studies to be made of embryonic development in this species. Thirty-six embryos from seventeen pregnancies were examined in detail and assigned to horizons on the basis of Streeter's classification of external form and internal structure. The specimens represented tha major part of the embryonic period and, with few exceptions, the structural development of the marmoset closely resembled that of the other primates that have been described. Marmoset embryos of a given horizon were consistently shorter than those of man. The timing of morphogenesis in C. jacchus was different from that observed in man and other primates. There was a considerable delay in early embryogenesis in the marmoset, although subsequent to stage XI, development proceeded at approximately the same rate as other primates. The available evidence suggested that this anomaly was not due to delayed implantation or embryonic diapause but to a generally slower rate of early development. The evolution of synchorial twinning is considered as a possible reason for this mode of development.

Abortion, Induced↗

Retinohypothalamic projections in the common marmoset (Callithrix jacchus): A study using cholera toxin subunit B.

Retinal projections in vertebrates reach the primary visual, accessory optic, and circadian timing structures. The central feature of the circadian timing system is the principal circadian pacemaker, the suprachiasmatic nucleus (SCN) of the hypothalamus. The direct projections from the retina to the SCN are considered the entrainment pathway of the circadian timing system. In this study, unilateral intravitreal injections of cholera toxin subunit B were used to trace the retinal projections to the marmoset hypothalamus. The retinohypothalamic tract reaches the ventral suprachiasmatic nucleus bilaterally, as anticipated from previous studies. However, labeled fibers were found in several other hypothalamic regions, such as the medial and lateral preoptic areas, supraoptic nucleus, anterior and lateral hypothalamic areas, retrochiasmatic area, and subparaventricular zone. These results reveal new aspects of retinohypothalamic projection in primates and are discussed in terms of their implications for circadian as well as noncircadian control systems.

Animals↗

Social context affects phee call production by nonreproductive common marmosets (Callithrix jacchus).

Common marmosets produce two variants of their long call (phee call) in different situations. Intergroup calls are produced in territorial situations, and intragroup separation calls are produced by marmosets isolated from group members. Marmoset groups frequently include postpubertal, nonreproductive members; their roles in the spontaneous production of territorial vocalizations is unclear. This study analyzed the production of home cage phee calls by nonreproductive, postpubertal marmosets while they were housed in their natal groups and after pairing with an opposite-sex conspecific. Additionally, the production of the separation phee call variant was assessed in both social conditions. The results indicated that the marmosets rarely produced home cage, or territorial, phee calls while they were natally housed. In contrast, both males and females produced the territorial phee call at a much higher rate as early as 4 days after pairing. Agematched females remaining in their natal groups throughout the study produced home cage phee calls infrequently. Most marmosets produced separation phee calls at a high rate after separation from either their natal group or a partner, suggesting that the makeup of a social group has little effect on an animal's motivation to reunite with conspecifics. These results suggest that the social environment has an important influence on the production of territorial phee calls.

Animals↗

Behavioral discrimination between circumgenital odor from peri-ovulatory dominant and anovulatory female common marmosets (Callithrix jacchus).

Social peer groups of callitrichid monkeys [marmosets and tamarins] exhibit intrasexual dominance hierarchies in captivity. This laboratory study employed two-choice behavioral discrimination bioassys to test the hypothesis that scent from female common marmosets contains chemical cues that permit discrimination between dominant females in the periovulatory versus luteal phase of the ovarian cycle and females holding dominant versus subordinate status. When scent from only dominant females was presented, marmosets directed significantly greater amounts of investigatory behavior toward peri-ovulatory scent versus scent collected during the luteal phase of the ovarian cycle. Animals of both sexes demonstrated significant discriminatory behavior between scent deposited by dominant versus subordinate females, but only when the dominant female was in the peri-ovulatory phase of the ovarian cycle. Test animals directed equal amounts of investigative behavior toward scent from luteal-phase dominant females and subordinate females. Female test subjects deposited significantly more scent marks over presented scents than did male subjects, particularly when the scent had been donated by a peri-ovulatory female. Chemical odors specific to the periovulatory and luteal phases of the ovarian cycle may play a role in mediating behavioral interactions among marmosets.

Animals↗

Effects of separation and novelty on distress vocalizations and cortisol in the common marmoset (Callithrix jacchus).

In socially-bonding species, separation from familiar attachment figures is widely known to stimulate a physiological and behavioral stress response. This study investigated the hormonal and vocal responses of adult common marmosets to separation from familiar group members and to 24 hr of cohabitation with an unfamiliar opposite-sex conspecific. All subjects were removed from their home cages and placed into a novel environment for 20 min. In one group, marmosets were exposed to an unfamiliar, opposite-sex partner in the novel environment and remained paired with this partner for the 24 hr test period. In three other groups, marmosets experienced the novel environment alone and subsequently were returned to their original social- or single-housing condition, or kept separate from their social groups for a 24 h period. Blood samples were collected the day before, and at 30 min, 90 min, and at 24 h after separation. Cortisol responses were differentially affected by the length of separation and the presence of unfamiliar conspecifics. Brief separation followed by the return to the social group had minimal effect on plasma cortisol levels. All marmosets produced high levels of separation calls in the novel environment, but there was no apparent relationship between calling and cortisol levels. The lack of a temporal relationship between the production of distress vocalizations and serum cortisol has previously been noted in squirrel monkey and rhesus monkey infant separation studies; the behavioral and physiological responses to separation appear to be similarly dissociated in the marmoset. Further, the characteristics of a separation environment can differentially affect the hormonal response by adult marmosets without differentially affecting their behavioral response.

Animals↗

Context and sex differences exist in the acoustic structure of phee calls by newly-paired common marmosets (Callithrix jacchus).

Captive common marmosets of all ages robustly produce a "separation" phee call during brief separations from their group. In contrast, a second structural variant, which may function as an intergroup call, is produced in the home cage primarily by the reproductive adults. A previous study found that postpubertal but nonreproductive offspring rarely produce phee calls when in the home cage with the natal group, yet these marmosets call frequently after pairing with an opposite-sex partner. The sudden increase in home cage phee calls may indicate the rapid onset of intergroup calling. Alternatively, marmosets may be producing the separation phee variant as a result of separation from the natal group. The present study investigated whether phee calls produced by recently paired individuals in the home cage were structurally distinguishable from their calls recorded in a separation paradigm. We also tested whether sex differences, known to exist in the calls of mature adults, could be found in calls recorded from younger, nonreproductive animals separated from their natal groups. We analyzed 18 acoustic parameters of phee calls produced in the home cage after pairing and of calls produced during separation both from the natal group and from a new mate. Discriminant function analyses found that home cage calls were clearly discriminable from separation calls (average 91.7% correctly classified), indicating that the rapid increase in home cage phee call production shortly after pairing is not a consequence of separation from the family group. Postpubertal marmosets appear to show a rapid behavioral adjustment to separation from their natal groups. Additionally, sex was clearly discriminable in calls recorded both before and after pairing (average 86.8% correctly classified). Like calls recorded from well-established paired marmosets, phee calls produced by recently paired, postpubertal marmosets are discriminable by context and sex.

Animals↗

Fertilization-promoting peptide in reproductive tissues and semen of the male marmoset (Callithrix jacchus).

Fertilization-promoting peptide (FPP) is present in the prostate gland and semen of some mammals, and has been shown to enhance the fertilizing ability of both epididymal mouse and ejaculated human spermatozoa. The novel peptide may prove of importance for the treatment of some cases of male infertility, and a suitable animal model would be useful to test this hypothesis. To this end, we examined reproductive tissues and semen of the male marmoset for the presence of FPP. Peptides were extracted from seminal plasma, testes, prostate, and bulbourethral glands of intact and castrated male marmosets. The peptides were identified by ion-exchange chromatography followed by radioimmunoassay. The mean concentration of FPP immunoreactivity in semen from intact males was 58.7 nM (SE +/- 9.9 nM, n = 10), and anion-exchange chromatography revealed FPP as the only immunoreactive peptide present. Analysis of tissues revealed that FPP in semen was likely to be derived from the prostate gland, which contained this peptide as the major source of immunoreactivity (10.86 pmol/gland; SE +/- 4.39 pmol/gland, n = 4). Only low concentrations of FPP were detectable in the bulbourethral glands, and the peptide was undetectable in the testis. Surprisingly, FPP was readily detectable in the seminal plasma from one castrated marmoset and was present in the prostate gland from 3 castrates at levels which did not differ significantly from those in intact animals (5.47 pmol/gland, SE +/- 1.64 pmol/gland, n = 3). Plasma testosterone measurements indicated that residual circulatory androgens remained after castration, which may be consistent both with the maintenance of mating behavior and the presence of prostatic FPP. We conclude that FPP is present within the prostate gland and seminal plasma of the marmoset at concentrations consistent with a role in male fertility in this species.

Animals↗

Non-invasive collection of ejaculates from the common marmoset (Callithrix jacchus) using penile vibrostimulation.

Penile vibrostimulation (PVS), a noninvasive repeatable method, has been shown in the squirrel monkey to yield semen of higher quality than rectal probe electro-ejaculation (RPE). The present study aimed at establishing the conditions for PVS to collect ejaculates from marmoset monkeys. Ten adult males were trained on the appropriate handling before each was subject to six to 12 PVS tests. Ejaculation was stimulated using a FertiCare personal vibrator fitted with a 2 cm x 0.5 cm i.d. glass tube. The stimulus was repeatedly applied over a frequency of 75-95 Hz and amplitude of 1-2 mm for up to 20 min. Ejaculates were analyzed for volume, total sperm number, sperm concentration, and proportion of living and motile sperm. Ejaculates were obtained in 31 of 88 PVS tests; 87.1% of the ejaculations occurred at 80-85 Hz frequency and 1-1.5 mm amplitude. In 18 tests ejaculates were produced within 49.7 seconds. Ejaculates were characterized by (mean values): volume 31.9 microl, total sperm number 34.2 x 10(6)/ejaculate, concentration 1,154.2 x 10(6) sperm/ml, live sperm 74.6%, motile sperm 59.6%. Total number and concentration of spermatozoa were significantly enhanced in singly living males. PVS yielded three to four times more spermatozoa than comparable previously published values for RPE. Enhancing the success rate by preselecting males for responsiveness may render PVS the sperm collection method of choice in marmoset monkeys.

Animals↗

Relations among birth condition, maternal condition, and postnatal growth in captive common marmoset monkeys (Callithrix jacchus).

The present study characterizes the relations among maternal condition, litter size, birth condition, and growth in body weight for a population of common marmosets. The subjects of the study were marmosets born into a single colony between 1994 and 2001. Three sets of analyses were conducted to answer the following questions: 1) Is there a relationship between litter size, maternal condition, and birth condition? In the study population, maternal body weight, maternal age, litter size, and birth condition were related in a complex fashion. Birth weight and prenatal long-bone growth, as reflected in knee-heel length, were both related to maternal age, with older mothers supporting higher prenatal growth. Age and maternal condition appeared to interact as determinants of long-bone growth, as the combination of older and larger mothers resulted in significantly longer knee-heel lengths in their offspring. 2) Is there a relationship between birth condition or maternal condition and subsequent growth or final adult size? The early growth rate in this population was similar to early growth rates reported for three different marmoset colonies, suggesting that early growth may be relatively inflexible in this species. However, within this population, the variation that did occur in early growth rate was related to birth weight and maternal weight. Later growth and adult weight were related to birth weight and litter size: small twin infants displayed slower later growth rates and were smaller as adults than twins that began life at a higher birth weight, while the birth weight of triplets was not related to adult size. In these marmosets, small infants that were the result of increased litter size differed from small infants whose small birth size resulted from other factors. This reinforces the proposal that the causes of low birth weight will be relevant to the development of the marmoset as a model of prenatal environmental effects.

Age Factors↗

Influence of the mother's reproductive state on the hormonal status of daughters in marmosets (Callithrix kuhlii).

Behavioral and endocrine suppression of reproduction in subordinate females produces the high reproductive skew that characterizes callitrichid primate mating systems. Snowdon et al. [American Journal of Primatology 31:11-21, 1993] reported that the eldest daughters in tamarin families exhibit further endocrinological suppression immediately following the birth of siblings, and suggested that dominant females exert greater control over subordinate endocrinology during this energetically challenging phase of reproduction. We monitored the endocrine status of five Wied's black tufted-ear marmoset daughters before and after their mother delivered infants by measuring concentrations of urinary estradiol (E(2)), pregnanediol glucuronide (PdG), testosterone (T), and cortisol (CORT). Samples were collected from marmoset daughters 4 weeks prior to and 9 weeks following three consecutive sibling-litter births when the daughters were prepubertal (M=6.1 months of age), peripubertal (M=11.9 months), and postpubertal (M=17.6 months). The birth of infants was associated with reduced ovarian steroid excretion only in the prepubertal daughters. In contrast, ovarian steroid levels tended to increase in the postpubertal daughters. Urinary E(2) and T levels in the postpubertal daughters were 73.8% and 37.6% higher, respectively, in the 3 weeks following the birth of infants, relative to prepartum levels. In addition, peak urinary PdG concentrations in peri- and postpubertal daughters were equivalent to luteal phase concentrations in nonpregnant, breeding adult females, and all of the peri- and postpubertal daughters showed clear ovulatory cycles. Cortisol excretion did not change in response to the reproductive status of the mother, nor did the concentrations change across age. Our data suggest that marmoset daughters of potential breeding age are not hormonally suppressed during the mother's peripartum period or her return to fertility. These findings provide an additional example of species diversity in the social regulation of reproduction in callitrichid primates.

Animals↗

Pattern of maternal serum corticotropin-releasing hormone concentration during pregnancy in the common marmoset (Callithrix jacchus).

Corticotropin-releasing hormone (CRH), a potent neuropeptide, is produced by the placenta of anthropoid primates. No other mammals, including prosimian primates, are known to produce placental CRH. In humans, placental CRH appears to play an important role in the progression of pregnancy to parturition. Maternal circulating CRH begins to rise early in pregnancy and increases until parturition. Gorillas and chimpanzees share this pattern of increasing maternal CRH during pregnancy with humans. In humans, chimpanzees, and gorillas, maternal CRH and estradiol concentrations are correlated, consistent with the hypothesis that CRH is involved in the biosynthetic pathway for placental estrogen production. In contrast, in baboons, maternal circulating CRH rises precipitously early in pregnancy and then declines, though CRH is detectable until birth. This research was designed to investigate the pattern of maternal circulating CRH in the common marmoset during pregnancy. Blood samples were taken across gestation from nine subjects over 11 pregnancies, and the plasma was assayed for CRH. The pattern of maternal circulating CRH in the common marmoset was similar to that of the baboon, with a rapid rise starting at about 50 days postconception and a peak at approximately 70 days postconception. By 110 days postconception, CRH concentration had plateaued at a significantly lower value. The peak and mean values for CRH were associated with fetal number (e.g., females gestating triplets had higher values than females gestating twins). Urinary estradiol showed no association with plasma CRH concentration. Marmosets appear to differ from the great apes in this regard, and to share a pattern of maternal CRH during pregnancy with the baboon, indicating that the baboon and marmoset pattern may be ancestral. The function of the early rapid rise of CRH in baboons and marmosets, and the significance of this difference between monkeys and apes, are not known.

Animals↗

Bipolar cell diversity in the primate retina: morphologic and immunocytochemical analysis of a new world monkey, the marmoset Callithrix jacchus.

The aim of this study was to identify the bipolar cell types in the retina of a New World monkey, the common marmoset, and compare them with those found in the Old World macaque monkey. Retinal whole-mounts, sections, or both, were stained by using DiI labeling and immunohistochemical methods. Semithin sections were analyzed by using quantitative methods. We show that the same morphologic types of bipolar cell as described for the Old World macaque monkey by Boycott and Wässle (Boycott and Wässle [1991] Eur. J. Neurosci. 3:1069-1088) are present in marmoset retina: two types of midget bipolar cells, six type of diffuse bipolar cells, a blue cone bipolar cell, and one type of rod bipolar cell. The pattern of staining with different immunohistochemical markers ("fingerprint") of each bipolar cell type in marmoset was also the same as described for macaque, with one exception: the flat midget bipolar cell (FMB) class is labeled by antibodies to recoverin in macaque but is labeled by antibodies to CD15 in marmoset. The labeled FMB cells in marmoset make contact with multiple cone photoreceptors throughout most of the extrafoveal retina. The spatial density of bipolar cells in marmoset is shown to be sufficient to support one-to-one connectivity of midget bipolar and ganglion cells in the fovea and to allow for parallel pathways to ganglion cells throughout the retina. Quantitative differences in the morphology and receptor connectivity between marmoset and macaque can be related to differences in cone and rod photoreceptor density between the species. We conclude that bipolar cell diversity is a preserved feature of the primate retina.

ATP-Binding Cassette Transporters↗

Cytoarchitectonic subdivisions of the dorsolateral frontal cortex of the marmoset monkey (Callithrix jacchus), and their projections to dorsal visual areas.

We describe the organization of the dorsolateral frontal areas in marmoset monkeys using a combination of architectural methods (Nissl, cytochrome oxidase, and myelin stains) and injections of fluorescent tracers in extrastriate areas (the second visual area [V2], the dorsomedial and dorsoanterior areas [DM, DA], the middle temporal area and middle temporal crescent [MT, MTc], and the posterior parietal cortex [area 7]). Cytoarchitectural field 8 comprises three subdivisions: 8Av, 8Ad, and 8B. The ventrolateral subdivision, 8Av, forms the principal source of frontal projections to the "dorsal stream," having connections with each of the injected visual areas. The cytoarchitectural characteristics of 8Av suggest that this subdivision corresponds to the marmoset's frontal eye field. The intermediate subdivision of area 8 (8Ad) has efferent projections to area 7, while the dorsomedial subdivision (8B) has few or no connections with extrastriate cortex. Area 46, located rostrolateral to area 8Av, has substantial connections with the medial extrastriate areas (DM, DA, and area 7) and with MT, while the cortex lateral to 8Av (area 12/45) projects primarily to MT and to the MTc. The rostromedial prefrontal (area 9) and frontopolar (area 10) regions have very few extrastriate projections. Finally, cells in dorsal area 6 (6d) have sparse projections to DM, MT, and the MTc, as well as strong projections to DA and to area 7. These results illuminate aspects of the evolutionary development of the primate frontal cortex, and serve as a basis for further research into cognitive functions using a marmoset model.

Amidines↗

Chemoarchitecture of the middle temporal visual area in the marmoset monkey (Callithrix jacchus): laminar distribution of calcium-binding proteins (calbindin, parvalbumin) and nonphosphorylated neurofilament.

We studied the distributions of interneurons containing the calcium-binding proteins parvalbumin and calbindin D-28k, as well as that of pyramidal neurons containing nonphosphorylated neurofilament (NNF), in the middle temporal visual area (MT) of marmoset monkeys. The distributions of these classes of cells in MT are distinct from those found in adjacent areas. Similar to the primary visual area (V1), in MT, calbindin-immunopositive neurons can be objectively classified into "dark" and "light" subtypes based on optical density of stained cell bodies. Calbindin-positive dark neurons are particularly concentrated in layers 2 and 3, whereas light neurons have a more widespread distribution. In addition, a subcategory of calbindin-positive dark neuron, characterized by a "halo" of stained processes surrounding the cell body, is found within and around layer 4 of MT and V1. These cells are rare in most other visual areas. In comparison, parvalbumin-immunopositive cells in area MT have a relatively homogeneous distribution, although with a trend toward higher spatial density in lower layer 3, and are relatively uniform in terms of density of staining. Finally, MT shows a characteristic trilaminar distribution of NNF-immunopositive pyramidal cells, with stained cell bodies evident in layers 3, 5, and 6. Although the laminar distribution of cells stained for the three markers overlap to some extent, these subcategories can be readily distinguished in terms of morphology, including cell body size. Chemoarchitectural parallels observed between MT and V1 suggest comparable physiological requirements and neuronal circuitry.

Animals↗

Interhemispheric connections of visual cortex of owl monkeys (Aotus trivirgatus), marmosets (Callithrix jacchus), and galagos (Galago crassicaudatus).

Interhemispheric connections of visual cortex were studied in owl monkeys, marmosets, and galagos after multiple injections of horseradish peroxidase into one cerebral hemisphere. Areal patterns of connections were revealed in sections of cortex that was flattened and cut parallel to the surface. Results were related to the locations of known visual areas, especially in owl monkeys, in which more visual areas have been established. The connection patterns in owl monkeys and marmosets are very similar, suggesting that the organization of visual cortex differs little in these two New World simians. Galagos have a basically similar pattern, but the connections are more widespread. In all three primates, connections are not restricted to cortex representing the line of decussation of the retina, and even striate cortex has connections displaced from the border. These connections extend up to 2 mm into area 17 in owl monkeys, and they are most extensive in galagos, where they form foci that are coextensive with regions of high cytochrome oxidase activity. Connections are concentrated in the caudal half of area 18, but protrusions of connections cross of the width of the field. The middle temporal visual area (MT) has unevenly distributed connections throughout, with some increase in density along the border. The dorsomedial visual area (DM) of owl monkeys has connections restricted to the rostral border, and a similar region of sparse connections identifies the probable location of DM in marmosets and galagos. Caudal parts of the dorsolateral visual area (DL) of owl monkeys have dense interhemispheric connections. Other visual areas are characterized by unevenly distributed clumps of connections, suggesting that functions are not uniformly distributed, and that semiregular processing modules exist. The results indicate that most extrastriate visual neurons are subject to interhemispheric influences and support the conclusion that callosal connections are functionally heterogeneous.

Animals↗