A Reinke-like inclusion within Leydig cells of the marmoset monkey (Callithrix jacchus)
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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↗
Leukaemia inhibitory factor in the endometrium of the common marmoset Callithrix jacchus: localization, expression and hormonal regulation.
In the present study, changes in the immunohistochemical localization of leukaemia inhibitory factor (LIF) in the endometrium during various phases of ovarian cyclicity of the common marmoset have been reported. LIF was absent during the early and late follicular phases. LIF was observed mainly in the cytoplasm of the endometrial glands during the early luteal phase, reached maximum intensity during the mid-luteal phase and declined again during late luteal phase. In-situ hybridization also showed a similar cyclic pattern in the expression of LIF. Stromal cells only showed signals for LIF during the mid-luteal phase. In ovariectomized marmosets, graded dosages of oestradiol alone failed to induce the appearance of LIF protein. Progesterone treatment following oestradiol priming, however, induced distinct glandular localization of LIF, indicating that LIF is a progesterone-dependent protein. Thus endometrial LIF is under maternal control and is secreted in response to the increased progesterone concentrations in circulation. It is possible that high concentrations of LIF during mid-luteal phase may prepare the endometrium for blastocyst implantation in marmosets.
Animals↗
Luteinizing hormone/chorionic gonadotropin bioactivity in the common marmoset (Callithrix jacchus) is due to a chorionic gonadotropin molecule with a structure intermediate between human chorionic gonadotropin and human luteinizing hormone.
Chorionic gonadotropin (CG), a pregnancy-specific heterodimeric hormone found in primates, is responsible for CL rescue with pregnancy maintenance. Of the primates, the human and baboon gene sequences are the only structures so far determined. In order to study the structure and function of CG in other primates, we have isolated and sequenced the coding regions for the two subunits of marmoset CG (mCG) by the reverse transcription/polymerase chain reaction method. Study of multiple clones confirmed a high degree of homology with the human sequences (88% and 80% for the alpha and beta nucleotide sequences, respectively). Marmoset CG alpha has an extra four amino acids compared to hCG alpha, whereas the mCG beta sequence has a 3-bp deletion that maintains the reading frame and C-terminal amino acid sequence. Most of the differences between hCG beta and mCG beta peptides occur in the C-terminal region, which includes the loss of two of the O-linked glycosylation consensus sequences and the presence of an N-linked glycosylation consensus sequence. When mCG alpha and beta were co-expressed in CHO cells, assembly of biologically active hormone was confirmed by induced steroid secretion by MA10 cells. Partially purified mCG beta was used to raise anti-mCG antibodies. To date, an antibody has been obtained that is capable of detecting recombinant mCG beta, recombinant mCG dimer, and mCG dimer secreted by cultured marmoset trophoblast. Marmoset CG alpha and beta were also detectable at the transcriptional level in cultured trophoblast by in situ hybridization. This suggests that the LH/CG bioactivity reported from marmoset placentae and embryos is due to a molecule with structural features common to hLH (glycosylation pattern) and hCG (CG beta C-terminal structure).
Amino Acid Sequence↗
Patterns of relaxin and steroids in the reproductive cycle of the common marmoset (Callithrix jacchus): effects of prostaglandin F2 alpha on relaxin and progesterone secretion during pregnancy.
We measured the concentrations of relaxin (Rlx), progesterone, and estradiol-17 beta in serum samples obtained twice or three times weekly from marmosets during the estrous cycle and pregnancy. The cyclic patterns and concentrations of progesterone and estradiol-17 beta were similar to those reported by previous investigators. Rlx was not detected in individual serum samples ( < 0.62-1.25 ng/ml) obtained from nonpregnant marmosets. However, pooling of luteal serum from all animals permitted assay of much larger volumes of serum (0.4 ml vs. 0.1 ml), and a concentration of about 1 ng/ml was detected. Rlx was first detected in serum in the second or third week of the 21-wk marmoset pregnancy, rose to a peak during Weeks 10-14, and then declined slowly as the time of parturition approached. The pattern of Rlx was unlike that observed during pregnancy in Old World monkeys, chimpanzees, or women, and resembled, instead, that seen in rodents, carnivores, and equids. Progesterone and estradiol-17 beta likewise increased throughout pregnancy, and their patterns were similar to those previously described for marmosets by other investigators. The concentrations of the steroids and Rlx in serum of pregnant marmosets was 10-fold or more higher than those found in Old World monkeys, baboons, chimpanzees, or women. Spontaneous abortions in two of the marmosets were accompanied by precipitous falls in serum levels of progesterone, estradiol-17 beta, and Rlx. Following s.c. injection of the luteolytic agent prostaglandin F2 alpha (PGF2 alpha) into two marmosets at midpregnancy, serum progesterone and Rlx fell to low levels. These animals received a progestin, 17 alpha-ethyl-19-nortesterone, to preclude abortion. Serum progesterone rose again, but serum Rlx remained low for the duration of pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)
Abortion, Veterinary↗
Metabolism of reproductive steroids during the ovarian cycle in two species of callitrichids, Saguinus oedipus and Callithrix jacchus, and estimation of the ovulatory period from fecal steroids.
Gonadal steroids were measured in daily fecal samples providing comparative data on steroid metabolism in two genera of New World primates. Circulating bioactive LH and progesterone concentrations and fecal progesterone, pregnanediol, estradiol, and estrone concentrations were measured by collecting blood and daily fecal samples from four captive common marmoset females and four cotton-top tamarin females for 30 days. High recoveries (> 80%) of labeled steroids that were added directly to the feces before extraction were recovered from feces of both species. Because of the presence of complex steroid conjugates, only one fifth the amount of estradiol was measured without solvolysis as compared to the amount measured with solvolysis. In tamarins, steroids were metabolized rapidly, with all postovulatory increases occurring within two days after the circulating LH peak (an increase of 2 SD higher than mean follicular levels). In marmosets, steroid excretion was slower; increased steroid levels occurred 2-4 days after the LH peak except in the case of estrone, which did not consistently increase after the LH peak. Circulating estrone and estradiol both contributed to the high excretion of estradiol in the feces from both species. The timing in the delay in excretion of fecal steroids was used to accurately determine the ovulatory period to within a 2-day window. This degree of accuracy is possible when the duration of the delay to the LH peak is known for a given species. Additionally, steroid concentrations were highly correlated between frozen and lyophilized fecal samples (0.81 +/- 0.07 SEM), indicating that fluid removal from the feces did not effectively alter steroid profiles.
Animals↗
A local oxytocin system is part of the luteinization process in the preovulatory follicle of the marmoset monkey (Callithrix jacchus).
Luteinization is a complex differentiation process involving the interaction of extrinsic and intraovarian factors. The aim of this study was to examine the components of an intraovarian oxytocin (OT) system during the periovulatory period in the marmoset monkey, as well as the possible relationship of these components to other factors involved in the luteinization process, using immunohistochemistry and cell culture techniques. Ovaries were collected on Day 7 of the follicular phase (before the endogenous LH surge) and on Day 8 (22 h after an exogenous hCG application, but before ovulation). Before the endogenous LH increase, OT immunoreactivity was detectable at low levels in most antral follicles, where its presence was confined to antral granulosa cell (GC) layers. In contrast, immunoreactivity for the OT receptor (OTR) was localized primarily in the basal GC layer. After application of exogenous hCG, there was a marked enhancement in both the staining intensity and the number of cells positive for OT and the OTR in all GC layers of antral follicles, especially in the preovulatory follicle. Progesterone receptor and 3beta-hydroxysteroid dehydrogenase activity in GC were clearly present only when follicles were obtained after gonadotropin stimulation. Secretion of authentic OT was demonstrated from cultured GC obtained before the LH surge, with highest amounts in cells cultured from preovulatory as opposed to smaller antral follicles. OT production could be stimulated by the application of hCG to the GC cultured from preovulatory follicles, whereas the gonadotropin was without effect on GC from small follicles. FSH had no effect on OT production by GC from either follicle type. Application of OT to the cultures caused an increase in progesterone production by GC from large preovulatory follicles but was without effect on steroidogenesis by cells from small antral follicles. These results describing the presence and distribution of OT and OTR and their modulation by hCG, as well as the luteotrophic effect of OT in cultured GC from preovulatory follicles, implicate OT as a paracrine mediator in the luteinization process in the primate ovary.
3-Hydroxysteroid Dehydrogenases↗
Differential splicing and expression of the relaxin-like factor gene in reproductive tissues of the marmoset monkey (Callithrix jacchus).
The relaxin-like factor (RLF) is a novel member of the insulin/relaxin/insulin-like growth factor family of growth factors and hormones that is expressed predominantly in the reproductive system, with highest expression in the Leydig cells of the testis. Using a combination of molecular and immunological techniques, we have characterized the structure and expression of the RLF gene from a primate model, the marmoset monkey, with the intention of comparing this with recent results on the closely related hormone relaxin in this species. As in other species, including the human, RLF gene products can be detected maximally in Leydig cells and in the follicular theca interna cells and corpora lutea of the ovary. Reverse transcription-polymerase chain reaction analysis confirmed this expression and showed that in the corpus luteum, testis, and epididymis, a second, alternative RLF gene transcript was present that is expressed at low levels and that appears to be derived by differential splicing of a novel exon. Analysis of genomic DNA from the marmoset showed that in this species, the single-copy gene contains a longer intronic region separating the two exons described for the human. Alternative splicing introduces a novel exon 1A between exons 1 and 2, which leads to an altered open reading frame, with a new stop codon, such that if translated, the novel transcript will encode a truncated polypeptide comprising a C-terminally extended B-domain.
Alternative Splicing↗
Persistent Epstein-Barr virus infection in the common marmoset (Callithrix jacchus).
Epstein-Barr virus (EBV) infection of the common marmoset causes long-term infection, with production of antibodies to virus-induced antigens, without clinical illness. Attempts to show the presence of EBV DNA in saliva of infected animals by PCR were initially unsuccessful, although slot-blot hybridization analysis demonstrated that viral DNA was present. Further investigations showed that most samples of pilocarpine-induced saliva, and 33% of the samples of whole mouth fluids (WMF) tested, were inhibitory to PCR. Similar results were found using human WMF. A method of assessing samples of marmoset WMF for the presence of EBV, by PCR using an EBV BamHI W probe, and removing inhibition with Chelex 100, is described. A total of 202 samples from 21 EBV infected, and seven non-infected animals was tested. Five seropositive animals shed virus on every occasion, and 15 intermittently. Two marmosets, infected as neonates, showed progressively increasing humoral responses to viral antigens, and shed virus on every occasion tested over 3 years. When mated with uninfected animals, the latter seroconverted 4 and 6 weeks later, respectively, and later shed virus into their WMF. The naturally infected animals were paired with naive marmosets, and were able to pass on infection. These results establish that long-term, permissive EBV infection occurs in the common marmoset, and demonstrate again the similarities in the response to EBV between marmoset and man.
Animals↗
Diurnal and annual variations in plasma androgen levels in the adult male marmoset (callithrix jacchus).
Diurnal and annual variations in plasma androgen levels were studied in adult male marmosets. A distinct diurnal pattern, characterized by low androgen levels during the lights-on period and high levels during the lights-off period was observed (n = 7). In contrast to this, no definite circannual pattern in androgen levels was observed (n = 6).
Androgens↗
Continuous light environment has no effect on the circadian testosterone rhythm, spermatogenesis or fertility of the marmoset (Callithrix jacchus).
Maintenance of adult male marmosets in continuous light for 60 days had no effect on the circadian rhythmicity of plasma testosterone levels, spermatogenesis or fertility. The results indicate that photoperiodicity is not the environmental determinant that regulates reproduction in male marmosets.
Animals↗
Maturation of sperm motility in the epididymis of the common marmoset (Callithrix jacchus) and the cynomolgus monkey (Macaca fascicularis).
The acquisition by spermatozoa of the capacity for movement upon liberation from the epididymal duct into buffer is one of the obvious changes associated with sperm maturation and, hence, studies of sperm kinematics might provide good indices for sperm maturation. Profiles of development in the movement pattern of spermatozoa taken from six regions of the epididymis were established in marmoset and cynomolgus monkeys using computer-assisted sperm analysis. Motion of spermatozoa from initial regions was sluggish and irregular, then developed in the more distal region into erratic movement effected by an increase in the vigour of flagellation as reflected by curvilinear velocity. The tendency of immature spermatozoa to display circular motion was more common in the marmoset than in the cynomolgus monkey. In the marmoset the most drastic changes in percentage motility and kinematic parameters of spermatozoa occurred between the distal caput and the proximal corpus; maximal values for these parameters were found around the distal corpus epididymis. For the cynomolgus spermatozoa the sharpest gain in percentage motility was also achieved between the caput and proximal corpus epididymis, but with full kinematic development in the proximal cauda epididymis. These maturational changes were not synchronous among spermatozoa from the same site as they moved along the epididymis, as reflected in the heterogeneity of each sample in the kinematic properties of individual spermatozoa, especially those from the proximal corpus epididymis. The maturational profiles in the monkeys are comparable to the situation in the human, for which they may provide a useful model.
Animals↗
Temporal contrast sensitivity in the lateral geniculate nucleus of a New World monkey, the marmoset Callithrix jacchus.
1. The temporal contrast sensitivity of koniocellular, parvocellular and magnocellular cells in the lateral geniculate nucleus (LGN) of nine adult marmosets was measured. The receptive fields of the cells were between 0.3 and 70 deg from the fovea. The stimulus was a large spatially uniform field which was modulated in luminance at temporal frequencies between 0.98 and 64 Hz. 2. For each cell group there was a gradual increase in modulation sensitivity, especially for temporal frequencies below 8 Hz, with increasing distance from the fovea. At any given eccentricity, magnocellular cells had the greatest sensitivity. In central visual field, the sensitivity of koniocellular cells lay between that of parvocellular and magnocellular cells. In peripheral visual field (above 10 deg eccentricity) koniocellular and parvocellular cells had similar sensitivity. 3. The contrast sensitivity of each cell class was dependent on the anaesthetic used. Cells from animals anaesthetized with isoflurane were less sensitive than cells from animals anaesthetized with sufentanil. This effect was more marked for temporal frequencies below 4 Hz. 4. These results are incompatible with the notion that the koniocellular pathway is functionally homologous to a sluggish, W-like pathway in other mammals. At least in terms of their temporal transfer properties, many koniocellular cells are more like parvocellular cells.
Animals↗
Spatial properties of koniocellular cells in the lateral geniculate nucleus of the marmoset Callithrix jacchus.
1. The receptive field dimensions, contrast sensitivity and linearity of spatial summation of koniocellular (KC), parvocellular (PC) and magnocellular (MC) cells in the lateral geniculate nucleus (LGN) of 11 adult marmosets were measured using achromatic sinusoidal gratings. 2. The receptive field centre diameter of cells in each (PC, KC and MC) class increases with distance from the fovea. There is substantial overlap in centre size between the three cell classes at any eccentricity, but the PC cells have, on average, the smallest centres and the KC cells have the largest. Some PC and KC cells did not respond at all to the grating stimulus. 3. The contrast sensitivity of the receptive field centre mechanism in KC cells decreases in proportion to the centre area. A similar trend was seen for the surround mechanism. These characteristics are common to PC and MC cells, suggesting that they originate at an early stage of visual processing in the retina. 4. The KC cells showed, in general, lower peak evoked discharge rates than PC or MC cells. The spontaneous discharge rate of KC cells was lower than that of PC cells and similar to that of MC cells. 5. The majority of cells in all divisions of the LGN show linear spatial summation. A few cells did show non-linear spatial summation; these cells were predominantly located in the MC and ventral KC layers. 6. The ventral KC layers below and between the MC layers contain cells with larger and more transiently responding receptive fields than cells in the more dorsal KC layers. 7. We conclude that many of the contrast-dependent spatial properties of cells in the marmoset LGN are common to PC, MC and KC cells. The main difference between KC cells and the other two classes is that there is more variability in their response properties, and they are less responsive to high spatial frequencies.
Animals↗
Patterns of prenatal survival in the common marmoset (Callithrix jacchus).
Fifty common marmoset pregnancies were monitored using ultrasound. The objective was to ascertain if fetal mortality was related to litter size. Prior to analysis we determined abortion rate and litter size were not influenced by repeated ultrasound. Mortality was unrelated to litter size and occurred fairly late in gestation. All singletons born in this study began gestation as twins. It is hypothesized that marmosets may be able to adjust litter size late in pregnancy in response to proximate environmental factors.
Abortion, Veterinary↗
Efficacy and effects of short- and medium-term contraception in the common marmoset (Callithrix jacchus) using melengestrol acetate implants.
This study examines the effect of melengestrol acetate (MGA) implants on reproductive function and various biochemical parameters, ovarian activity, and uterine morphology in ten female common marmosets implanted for either 6-8 or 19-21 months. Measures of body weight, concentrations of urinary glucose and blood liver enzymes were taken. Ovarian activity was assessed by analysis of urinary progestin levels and ultrasound examinations of the ovaries. Ultrasonography was also used to evaluate uterine morphology. MGA was highly effective in preventing pregnancies in the study animals. No changes in biochemical parameters were found; however, seven females developed a substantial weight gain during the study. Follicular development was not suppressed, as indicated by the presence of antral follicles, luteinized structures, and elevated urinary progestin levels. The uteri of the MGA-treated subjects were moderately enlarged with a thickened endometrium that showed a marked change in structural appearance indicative of hypertrophy and decidualization. After implant removal these changes quickly disappeared and all females ovulated within 3 weeks and conceived within 4 months post-treatment. MGA appears to be an acceptable contraceptive in the marmoset, although non-steroidal methods should be evaluated as possible potential alternatives.
Animals↗
Unrearable litters and prenatal reduction of litter size in the common marmoset (Callithrix jacchus).
It is widely believed that common marmosets (Callithrx jacchus) typically give birth to twins under natural conditions. In captivity, however, births of triplets or even larger litters are common, although parents rarely succeed in rearing more than two offspring. The traditional interpretation is that captive conditions, notably the ready availability of food, have led to increased reproductive output, perhaps involving a higher ovulation rate. The present paper provides evidence, combined from ultrasound examinations between ovulation and birth and hysterotomies conducted during the late embryonic and early fetal phase, that the litter size can be progressively reduced during pregnancy without spontaneous abortion. There is an unusually long lag phase prior to the onset of embryonic growth in common marmosets; the fetal stage does not begin until day 80 of the 144-day pregnancy. Reduction in litter size occurs during embryonic stages (up to day 80), and continues into the fetal stages. These results indicate that the common marmoset is adapted for flexible modification of litter size between ovulation and birth. The high incidence of triplet births in captive colonies may therefore be an expression of an adapted natural developmental process under artificial circumstances.
Abortion, Veterinary↗
Immunohistochemical localization of collagenous components in healthy periodontal tissues of the rat and marmoset (Callithrix jacchus). II. Distribution of collagen types IV, V and VI.
The immunohistochemical distribution of collagen types IV, V and VI has been demonstrated in healthy periodontal tissues of rats and marmosets following decalcification of the maxillae and mandibulae in 0.2 N HCl. An intense fluorescence with anti-collagen type IV antibodies was demonstrated in the basement membranes of the epithelium and of the blood vessels and nerves. In the alveolar bone stroma and in the periodontal ligament (PL) collagen type IV was present only in the basal membranes of the blood vessels and nerves. In comparison, collagen type V was observed in a fibrillar pattern in the gingival connective tissue, as well as the PL. In the PL, type V collagenous fibers demonstrated a parallel distribution with stronger fluorescence near the cementum surface. Collagen type VI could be demonstrated in fine fibers present in the gingival connective tissue and the PL. Blood vessels and nerves were not stained in the marmoset, but were in the rat, where a localization of collagen type VI was demonstrated in these areas. Alveolar bone and cementum, as well as the Sharpey's fibers embedded in these tissues, were not stained with antibodies against collagen type V and type VI, but a pericellular localization of these collagenous components could be observed. Collectively, these results provide basic information on the relative distribution of different collagen types in normal tissues of rats and marmosets that will be required for future studies on the effects of pathological, reparative and regenerative processes.
Animals↗