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The protein requirement of adult marmosets: nitrogen balances and net protein utilization of milk proteins, soy protein, and amino acid mixtures.

Nitrogen balance studies were conducted in adult male marmosets (Callithrix jacchus) using purified and semipurified diets with protein levels between 0% and 7%. Daily nitrogen loss in a state of prolonged protein free nutrition was 131 +/- 16 mg/kg body weight0.75. Zero nitrogen balance resulted from mean daily intake of 261 mg nitrogen/kg0.75 when high quality protein sources were used. Very low protein intake or the lack of arginine and histidine in an amino acid mixture induced coprophagy. It is concluded that the protein requirement of adult marmosets is very similar to the protein requirement of adult humans (based on metabolic body weight). About 6-7% high quality protein, based on dry matter, suffice to avoid a negative nitrogen balance in all individuals.

Amino Acids↗

Islet hyperplasia in callitrichids.

Five callitrichids (three common marmosets -Callithrix jacchus -, a black tufted-eared marmoset -C. penicillata-, and a saddle-back tamarin -Saguinus fuscicollis) were diagnosed with islet hyperplasia by histopathology and immunohistochemistry. All were privately-owned, unrelated callitrichids ranging from 2- to 4-year-old. Relevant findings were anorexia (3/5), vomiting (2/5), ptyalism (1/5), polyuria/polydipsia (1/5), respiratory distress (1/5), hyperglycemia (2/3) and glycosuria (1/1); hyperglycemia and glycosuria were associated with pregnancy in a common marmoset and resolved after reducing simple carbohydrates in diet. All five animals died, three of them after few premonitory signs; in two cases, other concurrent diseases unrelated to islet hyperplasia were considered the cause of death. Additional animals from two facilities had high weight (4), physical obesity (3), polyuria/polydipsia/polyphagia/uriposia (1), hyperglycemia (1), and/or glycosuria (2). Pathologic findings in the deceased callitrichids were: islet hyperplasia (5/5); hemosiderosis (5/5); lipomatosis (4/5) of several tissues (atria, 3/5; pancreas, gall bladder, intestine, esophagus, and thyroid, 2/5; liver, 1/5); pancreatic necrosis or steatonecrosis, and/or acute pancreatitis (3/5); and vacuolation of hepatocytes and renal tubular cells most likely consistent with hepatorenal lipidosis (2/5). The islets of Langerhans were more numerous and larger than in a control, and morphologically normal in all cases, except in a common marmoset that had a few cells with a foamy cytoplasm and shrunken hyperchromatic or picknotic nucleus. Insulin (5/5), glucagon (3/5), and somatostatin (3/5) immunohistochemistry revealed that most cells stained positively for insulin diffusely in their cytoplasm (5/5) (staining restricted to the vascular pole of b-cells in the control). These findings suggest that obesity, insulin resistance and/or type II diabetes may be implicated and thus a prospective study on these diseases in callitrichids is necessary to determine their etiopathogenesis.

Animals↗

A reexamination of the phylogenetic position of Callimico (primates) incorporating new mitochondrial DNA sequence data.

The New World monkeys are divided into two main groups, Callitrichidae and Cebidae. Callimico goeldii shares traits with both the Cebidae and the Callitrichidae. Recent morphological phyletic studies generally place Callimico as the most basal member of the Callitrichidae. In contrast, genetic studies (immunological, restriction fragment, and sequence data) have consistently placed Callimico somewhere within the Callitrichidae, not basal to this clade. A DNA sequence data set from the terminal 236 codons of the mitochondrial ND4 gene and the tRNA(His), tRNA(Ser), and tRNA(Leu) genes was generated to clarify the position of Callimico. The sequences of 887 base pairs were analyzed by maximum-parsimony, neighbor-joining, and maximum-likelihood methods. The results of these various methods are generally congruent and place Callimico within the Callitrichidae between the marmosets (Callithrix and Cebuella) and the tamarins (Saguinus and Leontopithecus). Combined analyses of all suitable nuclear and mitochondrial gene sequences confirm the position of Callimico between the marmosets and the tamarins. As available molecular evidence indicates that Callimico is more closely related to the marmosets than to the tamarins, a reconsideration of the morphological evidence in light of the consensus tree from DNA sequence analyses is warranted. The marmosets and tamarins share four morphological characters (loss of the third molar, loss of the hypocone, reduced body size, reproductive twinning). Dwarfism may have evolved repeatedly among the Callitrichidae. It is well-known that the loss of a character can occur many times independently. The reproduction of marmosets and tamarins is extremely specialized and it is difficult to imagine that this complex and unique twinning system evolved separately in marmosets and tamarins. However, it is possible that a secondary reversal to single offspring took place in Callimico.

Animals↗

Identification of evolutionarily invariant sequences in the protein C gene promoter.

Recent studies on human protein C gene expression have revealed the presence of three transcription factor binding sites in close proximity to the transcription start site. Binding sites for the liver-enriched hepatocyte nuclear factors 1 and 3 (HNF-1 and HNF-3, respectively) are located immediately upstream of the transcription start site, whereas just downstream of the start site a presently unidentified transcription factor may bind. To identify other candidate transcription factor binding sites in the protein C promoter, we studied the promoter sequence identity in a number of evolutionarily close and more distant species: Gorilla gorilla, Pongo pygmaeus, Pan troglodytes, Homo sapiens, Cebus apella, Macaca mulatta, Callithrix jacchus, Papio hamadryas, Macaca fascicularis, and Rattus norvegicus. This analysis showed that a high degree of identity (78%) exists among the different primates. Comparison of the primate consensus sequence with the Rattus norvegicus protein C promoter sequence revealed the presence of seven identical regions (I to VII). Two of these regions overlap with established regulatory sequences for HNF-3 and HNF-1 (region VI) and for PCE-1 (region VII), respectively. The functional importance and the transcription factors that may bind to the other five identical regions are now to be determined.

Animals↗

SRY evolution in Cebidae (Platyrrhini: Primates).

Sex determination in mammals is dependent on the presence of SRY, which codes for a protein with a DNA binding motif (the HMG-box domain). Here we analyze the evolution of SRY among seven genera of New World monkeys belonging to the family Cebidae. Estimates of the number of synonymous and nonsynonymous substitutions indicated the absence of positive selection acting on SRY evolution. The presence of indels at the C-terminus coding region in different genera and species maintained an open reading frame, indicating a selective pressure constraining the evolution of this coding region. Available data on the fertility of natural and captive interspecific hybrids failed to show any relationship between SRY evolution and speciation for the genera herein studied. Our phylogenetic arrangement for Cebidae genera was similar to previous topologies based on mitochondrial and autosomal DNA sequences. This arrangement also corroborated the division of Cebus into two species groups. However, for Callithrix the differences among SRY topology and those derived from autosomal and mitochondrial genes suggested a Y-chromosome ancestral polymorphism.

Amino Acid Sequence↗

Molecular evolution of prolactin in primates.

Pituitary prolactin, like growth hormone (GH) and several other protein hormones, shows an episodic pattern of molecular evolution in which sustained bursts of rapid change contrast with long periods of slow evolution. A period of rapid change occurred in the evolution of prolactin in primates, leading to marked sequence differences between human prolactin and that of nonprimate mammals. We have defined this burst more precisely by sequencing the coding regions of prolactin genes for a prosimian, the slow loris (Nycticebus pygmaeus), and a New World monkey, the marmoset (Callithrix jacchus). Slow loris prolactin is very similar in sequence to pig prolactin, so the episode of rapid change occurred during primate evolution, after the separation of lines leading to prosimians and higher primates. Marmoset prolactin is similar in sequence to human prolactin, so the accelerated evolution occurred before divergence of New World monkeys and Old World monkeys/apes. The burst of change was confined largely to coding sequence (nonsynonymous sites) for mature prolactin and is not marked in other components of the gene sequence. This and the observations that (1) there was no apparent loss of function during the episode of rapid evolution, (2) the rate of evolution slowed toward the basal rate after this burst, and (3) the distribution of substitutions in the prolactin molecule is very uneven support the idea that this episode of rapid change was due to positive adaptive selection. In the slow loris and marmoset there is no evidence for duplication of the prolactin gene, and evidence from another New World monkey (Cebus albifrons) and from the chimpanzee and human genome sequences, suggests that this is the general position in primates, contrasting with the situation for GH genes. The chimpanzee prolactin sequence differs from that of human at two residues and comparison of human and chimpanzee prolactin gene sequences suggests that noncoding regions associated with regulating expression may be evolving differently from other noncoding regions.

Amino Acid Sequence↗

Extensive Mhc-DQB variation in humans and non-human primate species.

Non-human primates are often used in biomedical research, and the application of these animals as a model in immune-related diseases necessitates the characterisation of their MHC system. In particular, the MHC class II regions of the chimpanzee (Pan troglodytes), the rhesus macaque (Macaca mulatta)and the common marmoset (Callithrix jacchus) have been subject of molecular biological studies in recent years. In this study the emphasis was on MHC class II genes of another macaque species, Macaca fascicularis(crab eating macaque or cynomolgous monkey). The exon 2 of the Mhc-DQB gene (Mafa-DQB) was sequenced in each of a random panel of 60 non-pedigreed cynomolgous monkeys. This resulted in the detection of 23 Mafa-DQB1alleles that had not previously been published. In addition, unreported alleles were found in chimpanzees, rhesus macaques, orang-utans (Pongo pygmaeus) and stump-tailed macaques (Macaca arctoides), of which a few individuals were included in this study. Phylogenetic analyses confirm the trans-species model of evolution of the MHC-DQBlineages, in which a group of major alleles is passed on in the phylogeny, and has led to the sharing of allelic lineages by different species of non-human primates. The sharing of alleles is observed only for closely related macaque species. Furthermore, this manuscript provides an overview of all published, and whenever necessary corrected, non-human primate Mhc-DQB exon 2 alleles.

Alleles↗

Evolutionary history of chromosome 10 in primates.

We have tracked the evolutionary history of chromosomes homologous to HSA10 (PHYL-10) in primates using appropriate panels of PCP, YAC, and BAC probes. This approach allowed us to delineate more precisely the PHYL-10 constitution in the ancestor of catarrhine, platyrrhine, and prosimians. The results suggest that (i) in the ancestor of prosimians PHYL-10 was organized in two separate PHYL-10p and PHYL-10q chromosomes; (ii) in the progenitor of New World monkeys PHYL-10p was a separate chromosome, while PHYL-10q was associated with a chromosome homologous to HSA16; (iii) in the ancestor of Old World monkeys PHYL-10 was a unique chromosome with a marker order corresponding to the orang form. We have also analyzed the cat, chosen as an outgroup for its very conserved karyotype. In agreement with published data our experiments show that the PHYL-10 in cat is structured in two blocks, PHYL-10p and PHYL-10q, both as part of larger chromosomes. The overall data indicate that, contrary to common opinion, PHYL-10p and PHYL-10q were distinct chromosomes in the primate ancestor. Analysis of the Saimiri sciureus (SSC) PHYL-10q marker order showed that it was isosequential with the Callithrix jacchus PHYL-10q, as well as with the PHYL-10q platyrrhine ancestral form. The SSC centromere, nevertheless, was located in a different chromosomal region, therefore suggesting that a centromeric repositioning event occurred in this species.

Animals↗

Can molecular data place each neotropical monkey in its own branch?

Four different DNA datasets, representative of all extant neotropical primate genera, were tandemly aligned, comprising some 6,763 base pairs (bp) with 2,086 variable characters and 674 informative sites. Maximum Parsimony, Maximum Likelihood and Neighbor-Joining analyses suggested three monophyletic families (Atelidae, Pitheciidae and Cebidae) that emerged almost at the same time during primate radiation. Combined molecular data showed congruent branching inside the atelid clade, placing Alouatta as the most basal lineage followed by Ateles and a more derived branch including Brachyteles and Lagothrix as sister groups. In the Pitheciidae, Callicebus was the most basal lineage with respect to Pithecia and to the more derived sister groups (Cacajao and Chiropotes). Conjoint analysis strongly supported the monophyly of the Cebidae, grouping Aotus, Cebus and Saimiri with the small callitrichines. Within callitrichines, Cebuella merged with Callithrix, Callimico appeared as a sister group of Callithrix/Cebuella, Leontopitecus as a sister group of the previous clade, and Saguinus was the earliest callitrichine offshoot. Two major points remained to be clarified in platyrrhine phylogeny: (i) the exact branching pattern of Aotus, Cebus, Saimiri and the callitrichines, and (ii), which two of these three families (Atelidae, Pitheciidae and Cebidae) are more closely related to one another.

Animals↗

Progesterone antagonist lilopristone: a potent abortifacient in the common marmoset.

The effects of a progesterone antagonist ZK 98.734 (lilopristone) on implantation, early pregnancy, and midpregnancy were studied in the common marmoset, Callithrix jacchus jacchus. Treatment (5 mg/da intramuscularly for 3 consecutive days) on day (n = 8) after the midcycle peak in estradiol levels in mated animals induced a premature drop in plasma progesterone levels and shortened the ovarian cycle length. Treatment on day 20 (n = 5) or day 40 (n = 5) induced a drop in progesterone levels and decidual collapse. In three animals treated on day 40, vaginal bleeding was observed within 46 hours of the initiation of treatment. Treatment on day 80 resulted in expulsion of the fetuses with a mean induction abortion interval of 39 hours (range, 20 to 48 hours). The progesterone antagonistic effects of ZK 98.734 could be a result of the decrease in progesterone synthesis by the corpus luteum and/or placenta in addition to the interference with the progesterone binding to its cellular receptors in the target organ. Our study suggests that ZK 98.734 has potential for fertility regulation. Clinical trials for postcoital contraception, induction of menstruation, and early abortifacient effects are warranted.

Abortifacient Agents↗

Learning impairment following lesion of the basal nucleus of Meynert in the marmoset: modification by cholinergic drugs.

Five common marmosets (Callithrix jacchus) received unilateral ibotenic acid lesions of the basal nucleus of Meynert (nBM). Seven days later, choline acetyltransferase activity was significantly reduced by 50% in the frontal and temporal neocortex, 40% in the amygdala, and approximately 30% in the motor, parietal and occipital cortex in the ipsilateral hemisphere. Four marmosets receiving equivalent bilateral ibotenic acid lesions were severely impaired on new visual object discrimination learning and on relearning an object discrimination learnt prior to surgery when compared with operated controls. New learning in lesioned animals was substantially improved by i.m. administration of the cholinergic agonist arecoline. Lesioned animals' learning ability improved with time but these animals were then differentially sensitive to the disruptive effect of scopolamine on discrimination learning. These results show that lesions of the nBM which destroy the rising cholinergic pathways impair learning ability but that this ability can be substantially restored by administration of a cholinergic agonist.

Acetylcholine↗

Shortening of luteal phase in common marmosets by sheep ovarian follicular fluid peptide.

A low molecular weight peptide has been partially purified from sheep follicular fluid. It inhibited FSH binding to granulosa cells from ovarian follicles of common marmosets (Callithrix jacchus). When injected into cycling marmosets during the follicular phase, it reduced the area under the curve (AUC) of circulating progesterone. The peptide also shortened the luteal phase in all marmosets during the treatment cycle compared to the pretreatment control cycle. These results indicate that the ovarian follicular fluid peptide inhibited FSH binding to granulosa cells thereby probably resulting in decreased progesterone secretion (AUC) from these cells and subsequently inducing luteal insufficiency.

Animals↗

Tissue activities of enzymes of diagnostic interest in the marmoset and rat.

The activities of enzymes of diagnostic interest were investigated in the liver, heart, kidney and muscle of the marmoset (Callithrix jacchus) and the rat. Methods of tissue extraction which gave maximal enzyme activity were used and comparison between the species showed some major differences. AST, LDH and GDH showed a similar distribution in both species but ICDH activity was much higher in the rat heart than in any other rat or marmoset organ. ALP, LAP and GGT were present in much higher activities in the rat kidney than in the marmoset kidney, a finding which was reversed in the liver of these animals. The major ALT-containing organ in the rat was the liver but, in the marmoset, this enzyme was found in relatively large quantities in the heart and muscle also. These differences can be of importance when plasma enzyme activities are measured following tissue damage.

Alanine Transaminase↗

An involvement of acetylcholine in object discrimination learning and memory in the marmoset.

Five marmosets (Callithrix jacchus) were tested, using a Wisconsin General Test Apparatus, on a series of junk object visual discrimination tasks, including new learning, 24-hr reversal and 24-hr retention. The effects of administering the cholinergic receptor blocking agent, scopolamine either just before or immediately after the new learning task, or just before the 24-hr reversal and retention tasks, were assessed. Results suggest that scopolamine impairs new learning and impairs the encoding of new information in long term memory. Some evidence of a mild retrieval deficit under scopolamine was also seen, while state-dependent effects were not apparent.

Acetylcholine↗

Comparative studies on nafenopin-induced hepatic peroxisome proliferation in the rat, Syrian hamster, guinea pig, and marmoset.

Nafenopin was administered orally for 21 days to male Sprague-Dawley rats (0.5-50 mg/kg/day), Syrian hamsters (5-250 mg/kg/day), Dunkin-Hartley guinea pigs (50 and 250 mg/kg/day), and marmosets (Callithrix jacchus, 50 and 250 mg/kg/day). With the rat, and to a lesser extent in the hamster, nafenopin treatment produced dose-related increases in liver size and induction of peroxisomal (palmitoyl-CoA oxidation) and microsomal (lauric acid 12-hydroxylase) fatty acid oxidizing enzyme activities. In contrast, in the guinea pig and marmoset, there was no effect on liver size and only comparatively small changes were observed in these enzyme activities. Ultrastructural examination of liver sections from nafenopin-treated rats and hamsters revealed increased numbers of peroxisomes many of which lacked the characteristic crystalline nucleoid. While nafenopin had little effect on peroxisome numbers in either the guinea pig or marmoset, increases in microsomal cytochrome P450 content and mixed function oxidase activities were observed in these species. These results demonstrate marked species differences in nafenopin-induced hepatic peroxisome proliferation with the Syrian hamster being less responsive than the rat and the guinea pig and marmoset being only weakly responsive. As nafenopin is a known hepatocarcinogen in the rat, comparative long-term studies in poorly responsive species, such as the guinea pig and marmoset, may help clarify the role of organelle proliferation in the hepatocarcinogenicity of certain peroxisome proliferators.

Animals↗

A new approach to the role of noradrenaline in learning: problem-solving in the marmoset after alpha-noradrenergic receptor blockade.

Nine marmosets (Callithrix jacchus) were tested on a variety of visual discrimination learning tasks in a Wisconsin General Test Apparatus with or without alpha-noradrenergic receptor blockade achieved by the administration of aceperone. After aceperone, animals were found to be severely and consistently impaired at learning the first task of each test session and to be impaired on new and repeated reversal learning. They were, however, unimpaired on learning another similar task in each test session and on performance of a well-learnt task. Results were interpreted as evidence for defective association formation which can be compensated for by suitable priming or practice.

Adrenergic alpha-Antagonists↗

Further consideration of the learning impairment after aceperone in the marmoset: effects of the drug on shape and colour discrimination and on an alternation task.

Ten marmosets (Callithrix jacchus) learned to discriminate between pairs of small grey objects differing only in shape or small plain plaques differing only in colour, in a Wisconsin General Test Apparatus. Each day, each animal was presented with three consecutive visual discrimination problems in the order shape-colour-shape or colour-shape-colour. After aceperone, an alpha-noradrenergic antagonist, animals were impaired at learning the first but not the subsequent tasks of each trio. These results suggest that the previously observed impairment [10] on the first of a pair of object discrimination tasks after aceperone is a consequence of disruption of a mechanism common to both shape and colour discrimination learning. The fact that there is no impairment on task 2 in a dimension differing from task 1 suggests that the deficit is not one of attending to, or switching attention to, the appropriate visual dimension. Three further marmosets were trained to perform an alternation task and tested under aceperone. No impairment in performance was seen, suggesting that a variety of cognitive skills other than stimulus-reward association were intact. We conclude that the impairment following aceperone is a dysfunction of processes involved in association formation, but that it is one which is manifest only when the animal is faced with a type of task which has not recently been performed and that it can be overcome with persistence even the animal encounters novel stimuli.

Animals↗

Quantitative changes in morphological parameters in the developing visual cortex of the marmoset monkey.

Several quantitative morphological parameters were measured during postnatal development in area 17 of the marmoset monkey (Callithrix jacchus). In a series of 14 animals, at ages from birth to adulthood, we studied changes in the thickness, surface area and volume of area 17, as well as the neuronal and glial numerical densities, and total numbers. We found evidence for a rapid increase in thickness, area and volume, culminating between 6 weeks and 6 months postnatally, and then decreasing. The adult values are close to those observed in one-month-old animals. The overshoot in thickness and volume is greatest in layers II, III, IVa and IVc. The neuronal density shows a trend which is opposite to that of volume, and therefore the total number of neurons is constant postnatally, ca. 38 million neurons in area 17 of one hemisphere. The number of glial cells approximately doubles during the first postnatal month and remains stable afterwards, so that in the adult, there is one glial cell for two neurons. Morphological development of area 17 in this New World monkey is similar to that reported in Old World monkeys, as are the adult values for neuronal and glial densities.

Animals↗