Crab-eating Monkeys Research Articles

The enhancer activity of long interspersed nuclear element derived microRNA 625 induced by NF-\u03baB

Transposable elements (TEs) are DNA sequences that cut or introduced into the genome, and they represent a massive portion of the human genome. TEs generate a considerable number of microRNAs (miRNAs) are derived from TEs (MDTEs). Numerous miRNAs are related to cancer, and hsa-miRNA-625 is a well-known oncomiR derived from long interspersed nuclear elements (LINEs). The relative expression of hsa-miRNA-625-5p differs in humans, chimpanzees, crab-eating monkeys, and mice, and four primers were designed against the 3′UTR of GATAD2B to analyze the different quantities of canonical binding sites and the location of miRNA binding sites. Luciferase assay was performed to score for the interaction between hsa-miRNA-625 and the 3′UTR of GATAD2B, while blocking NF-κB. In summary, the different numbers of canonical binding sites and the locations of miRNA binding sites affect gene expression, and NF-κB induces the enhancer activity of hsa-miRNA-625-5p by sharing the binding sites.

Enhancer Function of MicroRNA-3681 Derived from Long Terminal Repeats Represses the Activity of Variable Number Tandem Repeats in the 3' UTR of SHISA7.

microRNAs (miRNAs) are non-coding RNA molecules involved in the regulation of gene expression. miRNAs inhibit gene expression by binding to the 3′ untranslated region (UTR) of their target gene. miRNAs can originate from transposable elements (TEs), which comprise approximately half of the eukaryotic genome and one type of TE, called the long terminal repeat (LTR) is found in class of retrotransposons. Amongst the miRNAs derived from LTR, hsa-miR-3681 was chosen and analyzed using bioinformatics tools and experimental analysis. Studies on hsa-miR-3681 have been scarce and this study provides the relative expression analysis of hsa-miR-3681-5p from humans, chimpanzees, crab-eating monkeys, and mice. Luciferase assay for hsa-miR-3681-5p and its target gene SHISA7 supports our hypothesis that the number of miRNA binding sites affects target gene expression. Especially, the variable number tandem repeat (VNTR) and hsa-miR-3681-5p share the binding sites in the 3’ UTR of SHISA7, which leads the enhancer function of hsa-miR-3681-5p to inhibit the activity of VNTR. In conclusion, hsa-miR-3681-5p acts as a super-enhancer and the enhancer function of hsa-miR-3681-5p acts as a repressor of VNTR activity in the 3′ UTR of SHISA7.

Alzheimer’s disease related genes during primate evolution

During primate evolution, the neuronal and cognition-related genes have evolved rapidly. These genes seem to induce neurological illnesses such as Alzheimer’s disease (AD). In this study, we analyzed genes APOE, TOMM40, and PICALM known as the risk factors of AD. We performed bioinformatics analyses in relation to evolution, phylogeny, and protein structure for those genes in humans, Neanderthals, chimpanzees, bonobos, gorillas, orangutans, crab-eating monkeys, and rhesus monkeys. Cholesterol-related genes showed relatively rapid evolution toward a lower risk of AD. Neanderthals showed relatively higher polymorphism in genes APOE, TOMM40, and PICALM than humans did. Phylogeny indicated different topologies in the trichotomy of humans, chimpanzees, and gorillas in terms of genes APOE, TOMM40, and PICALM. These results provide to hominin-specific patterns in three genes, and give clues to the modern human-specific traits of AD and shed light on further functional research helping to understand AD.

Gain of a New Exon by a Lineage-Specific Alu Element-Integration Event in the BCS1L Gene during Primate Evolution.

BCS1L gene encodes mitochondrial protein and is a member of conserved AAA protein family. This gene is involved in the incorporation of Rieske FeS and Qcr10p into complex III of respiratory chain. In our previous study, AluYRa2-derived alternative transcript in rhesus monkey genome was identified. However, this transcript has not been reported in human genome. In present study, we conducted evolutionary analysis of AluYRa2-exonized transcript with various primate genomic DNAs and cDNAs from humans, rhesus monkeys, and crab-eating monkeys. Remarkably, our results show that AluYRa2 element has only been integrated into genomes of Macaca species. This Macaca lineage-specific integration of AluYRa2 element led to exonization event in the first intron region of BCS1L gene by producing a conserved 3′ splice site. Intriguingly, in rhesus and crab-eating monkeys, more diverse transcript variants by alternative splicing (AS) events, including exon skipping and different 5′ splice sites from humans, were identified. Alignment of amino acid sequences revealed that AluYRa2-exonized transcript has short N-terminal peptides. Therefore, AS events play a major role in the generation of various transcripts and proteins during primate evolution. In particular, lineage-specific integration of Alu elements and species-specific Alu-derived exonization events could be important sources of gene diversification in primates.

実験動物サルのMHCクラスI多様性

旧世界ザルに属するアカゲザル（Macaca mulatta）やカニクイザル（Macaca fascicularis）は，ヒトに最も近い重要な実験動物として，エイズワクチン開発や薬剤の安全性確認をはじめとする各種の医学研究に欠かせない役割を担っている。このような動物実験，なかでも免疫に関わる実験の結果を正しく評価するためには，各個体の免疫応答に関わる遺伝的背景，特にMHC情報を把握することが重要である。筆者らの研究室では，以前よりエイズワクチン開発モデルであるサル免疫不全症候群ウイルス（simian immunodeficiency virus: SIV）に対するワクチン開発実験に用いられているアカゲザルやカニクイザルについて，RSCA（reference strand-mediated conformation analysis）法やcDNAクローニングによりMhcクラスI遺伝子群の解析を行い，多数の新規アリルを同定する一方，家系解析などから旧世界ザルに特徴的なMHC ハプロタイプ構造を見出している。本稿では，旧世界ザルMhcクラスI解析に関する研究の現状を紹介する。

Phenotypical Analysis after Administration of Anti-HLA ClassII-DR Humanized Monoclonal Antibody, IMMU-114 in Non-Human Primate

Background: Anti-HLA classII-DR humanized monoclonal antibody, IMMU-114 depletes HLA-DR positive cells by inducing apoptosis. To apply IMMU-114 for human trial, studies in non-human primate model is mandatory. The phenotypical changes of peripheral blood mononuclear cells (PBMCs) by IMMU-114 administration were investigated by flowcytemtery in monkey. Methods: IMMU-114 at a dose of 3 mg/kg was given intravenously, to two crab-eating monkeys, weighing 3-4 kg. PBMCs were obtained before (Pre) and 7 days (D7) after IMMU-114 administration, and analyzed by flowcytemtry. Results were expressed as the mean frequency of antigen-positive cells, in order of Pre vs. D7. Results: HLA-DR+ cells were 14.2 vs. 14.7%. CD3+CD25+ T cells were 2.5 vs. 6.2%. CD4+CD25+ T cells were 5.0 vs. 8.9%. CD8+CD25+ T cells were 8.8 vs. 9.6%. CD20+ B cells were 18.7 vs. 16.8%, and CD20+HLA-DR+ B cells were 14.8 vs. 13.0%. CD16+ NK cells were 17.9 vs. 11.6% and CD16+ HLA-DR+ NK cells were 5.4% vs. 2.35%. CD14+ macrophages were 12.8 vs. 8.7%, and CD14+ HLA-DR+ macrophages were 8.5% vs. 3.5%. Conclusions: IMMU-114, at a dose of 3 mg/kg had mild effects on decreasing the frequency of HLA-DR-expressing B cells, NK cells, and macrophages in mokey. However, the deletion of HLA-DR-expressing cells was mild, increasing the dose or repeated administration of IMMU-114 are required in non-human primates model.

Food Preference of Semi-Provisioned Macaques Based on Feeding Duration and Foraging Party Size

The long-tailed macaques ( Macaca fascicularis ; also called crab-eating monkeys) have broad geographic distribution in continent and archipelago of Southeast Asia. They have wide ecological plasticity to adapt to various environments. Due to disturbance of habitat and intensive contact with human, long-tailed macaques change their feeding behavior. Here we present food preferences of long-tailed macaques that live in Cikakak Monkey Park in Central Java. By recording the number of individuals who fed on a food patch and the duration of eating the food, we found that proportion of their food from natural resources is greater than those from human sources. They shifted to omnivory feeding mode to adapt to the changing environment. At many times, this omnivorous feeding brought forth crop-raiding which were not in natural behavioral repertory. Conservation effort of long-tailed macaques, and primate in general, should consider the aspect of human-modulated behavior in feeding ecology if we wish to be successful.

Environmental Contributions to Cranio-facial Variation in Crab-eating Monkeys (Macaca fascicularis)

The purposes of the present study were to investigate the difference in craniofacial size of M. fascicularis (crab-eating monkey) living in wild and captive environments and to analyze the allometry between craniofacial size and body weight. Total of 111 M. fascicularis female skulls, included 82 wild, and 29 captive or bred in artificial environment in Japan Monkey Centre were examined. Thirty two cranial landmarks, body weight, humeral length and femoral length were measured using a digital caliper. Also, cranial index, difference of the measurement (D) and ratio of the difference to wild (R) were used. Allometric analysis was performed among those measurements in two groups. It could be seen that all measured dimensions were larger in the captive group than the wild. These differences in craniofacial size seemed to be related to the high level of nutrition. The measurements (D) and (R) of the facial region tended to be higher than those of neurocranial region and limbs. Slopes of all measurements were low against ideal values, isometric baseline or references. These implied that increase in body weight in captive environments were excessive against the other measurements.

Novel cynomolgus macaque MHC‐DPB1 polymorphisms in three South‐East Asian populations*

Cynomolgus macaques (Macaca fascicularis, Mafa), alias the crab-eating monkeys or long-tailed macaques, live across a vast range of South-East Asia. These non-human primates have emerged as important animal models in infectious and chronic diseases and transplantation studies, necessitating a more extensive characterization of their major histocompatibility complex polymorphic regions. The current information on the polymorphic variation or diversity of the Mafa-DPB1 locus is largely limited in comparison with the more commonly studied rhesus macaque DPB1 locus. In this article, to better elucidate the degree and types of polymorphisms and genetic differences of Mafa-DPB1 locus among three South-East Asian populations and to investigate how the allele differences between macaques and humans might affect their respective immune responses, we identified 40 alleles within exon 2 of the Mafa-DPB1 locus by DNA sequencing using 217 individuals. We also performed evolutionary and population analyses using these sequences to reveal some population-specific alleles and trans-species allelic conservation between the cynomolgus macaques and the rhesus macaques. Of the 40 new alleles, eight belong to a newly identified lineage group not previously found in the rhesus macaque species. This allele information will be useful for medical researchers using the cynomolgus macaques in disease and immunological studies.

Variations in long- and middle-wavelength-sensitive opsin gene loci in crab-eating monkeys

We analyzed variations in long (L)- and middle (M)-wavelength-sensitive opsin gene loci in crab-eating monkeys. Unlike humans, most monkeys have a single L and a single M gene. Two variant genotypes, one with only one opsin gene (dichromatic) and one with tandemly arrayed multiple genes, were also found in the monkeys. However, the frequency of the former was 0.47%, and that of the latter was 5% in the monkeys, while 2% and 66%, respectively, in Caucasian males. The two variants were found only in Java Island, Indonesia, and South Thailand, respectively. The data suggest that the frequency of each genotype is different among Old World primates.

Noninstrumented enzyme‐linked immunosorbant assay for detection of early pregnancy in macaques

A practical, noninstrumented enzyme-linked immunosorbant assay (NELISA) for the measurement of urinary monkey chorionic gonadotropin (mCG) has been developed for the detection of early pregnancy in macaque monkeys for use in both the laboratory and the field. Five rhesus monkeys (Macaca mulatta) and six crab-eating monkeys (Macaca fascicularis) were tested for the presence of mCG in urine on gestational days (GDs) 12 to 35. The mCG NELISA detected pregnancy as early as GD 14, with an average earliest detection at GD 16.5 +/- 1.4 (n = 11). Out of 90 tests, 27 false-negative and zero false-positive tests were obtained, for an accuracy of 70.0%. Without the aid of a spectrophotometer, the presence of mCG in pregnant monkey samples was indicated by a dark green color change. Nonpregnant monkey urine samples, on the other hand, exhibited no color change. These findings suggest that the simple, economical, and reliable urinary mCG NELISA may be useful for diagnosing early pregnancy in these and related species. Because capture and restraint are unnecessary for collecting urine samples, the mCG NELISA has widespread potential for confined and free-ranging animals.

Concentrations of C-reactive protein in normal monkeys (Macaca irus) and in monkeys inoculated with Bordetella bronchiseptica R-5 and measles virus.

The concentrations of C-reactive protein (CRP) in serum from normal crab-eating monkeys (Macaca irus) were measured by means of a monkey-specific turbidimetric immunoassay (TIA), and the changes in the serum CRP concentrations in crab-eating monkeys inoculated with Bordetella bronchiseptica R-5 and measles virus (Ichinose or NK 3 strain) were also examined. The CRP concentrations in sera from 54 normal crab-eating monkeys ranged from 0 to 8.3 microg/ml (mean 2.2 +/- 1.9). No significant difference was found in the CRP concentrations between males and females (p > 0.05). The concentrations of CRP in the sera from four crab-eating monkeys inoculated intrabronchially with 10(9) live B. bronchiseptica increased gradually to a peak at 2 days after inoculation. The peak concentrations of CRP were from 102.4 to 313.2 microg/ml, 54-96 times the preinoculative values of 1.9-5.6 microg/ml. When the same four crab-eating monkeys were inoculated intrabronchially with measles virus 34 days after inoculation of B. bronchiseptica, the serum CRP concentrations did not increase. Monitoring of CRP is useful for assessing monkeys with acute B. bronchiseptica infection and will probably be of value in the diagnosis of other bacterial infections.

The antithrombotic effects of recombinant human soluble thrombomodulin (rhsTM) on tissue factor-induced disseminated intravascular coagulation in crab-eating monkeys (Macaca fascicularis).

We evaluated the antithrombotic effects of recombinant human soluble thrombomodulin (rhsTM) in plasma and in a monkey model. rhsTM dose-dependently prolonged activated partial thromboplastin time (APTT) in the following order: humans > monkeys > rats >> rabbits. The prolongation of APTT by rhsTM was also observed in protein C-deficient plasma. rhsTM activated protein C and inactivated factor Va in human and monkey plasma, but not in rat plasma. These findings suggest that the antithrombotic activities of rhsTM are fully expressed in human and monkey. Therefore, to evaluate the whole activity of rhsTM in a clinical model, tissue factor (TF) was intravenously infused into crab-eating monkeys to induce disseminated intravascular coagulation (DIC). Pretreatment with rhsTM reduced fall in fibrinogen with a biphasic and moderate dose-dependency curve, and reduced thrombin-antithrombin III (TAT) levels with a flat linear dose-dependency, while heparin prevented fall in fibrinogen with a steep linear dose-dependency curve without reducing TAT levels. Further evidence suggesting that rhsTM activates protein C in vivo was also obtained. Taken together, the data indicate that rhsTM fully expresses its antithrombotic activities in human and monkey but not in rat and rabbit, and rhsTM prevents TF-induced DIC in monkeys by suppressing thrombin generation.

Conflict and reconciliation in two groups of crab-eating monkeys differing in social status by birth

Two groups of captive macaques (M. fascicularis) were studied at Kassel University, Germany. One included animals whose mothers were high-ranking, another, those whose mothers were low-ranking. The first group was a despotic community in which conflicts were severe and occurred mainly between single individuals; the reconciliation tendency was weak, the male leader was the controlling animal, and the affiliative preferences were marked. The second group was an egalitarian community split into two mutually hostile conalitions; the conflicts were less severe, the tendency for reconciliation was strong, the male leader could control only his own bloc and had no strong affiliative ties with other group members.

Response of plasma tissue factor pathway inhibitor to diet-induced hypercholesterolemia in crab-eating monkeys.

Tissue factor pathway inhibitor (TFPI), a protease inhibitor associated with lipoproteins in plasma and endothelial cells, can inhibit the initial reactions of the tissue factor-mediated coagulation pathway. A positive relationship between TFPI and cholesterol has been demonstrated in human plasma. To investigate this relation in more detail, in the present study we measured TFPI in the plasma of monkeys on a high-cholesterol diet. After diet treatment, cholesterol levels and TFPI activity were increased 3- and 1.5-fold, respectively. Three forms of TFPI, low-density lipoprotein (LDL)/very-low-density lipoprotein (VLDL)-associated TFPI, high-density lipoprotein (HDL)-associated TFPI, and free TFPI, were measured after gel filtration of plasma. In hypercholesterolemic monkeys, levels of TFPI activity and antigen in the LDL/VLDL fraction were increased to about three times those of normal monkeys. Changes in HDL-associated TFPI and free TFPI were not significant compared with the change in LDL/VLDL-associated TFPI. After the monkeys received heparin infusions TFPI was increased about fivefold, but there was no significant difference in these increases between normal and hypercholesterolemic monkeys. The increase in TFPI after heparin infusion is discussed in terms of the relationship between lipoprotein-associated TFPI in plasma and endothelial cell-associated TFPI.

The Bronchial Tree and Lobular Division of the Crab-Eating Monkey (Macaca fascicularis) Lung.

The lungs of 70 crab-eating monkeys were examined. The crab-eating monkey has the dorsal, lateral, ventral and medial bronchiole systems on both sides. According to the bronchial ramification, the right lung consists of the upper, middle, lower, and accessory lobes, whereas the left lung consists of a middle lobe which is bilobed and a lower lobe. These findings were based on the comparative anatomy of the bronchial tree and blood vessels of the mammalian lung.

Accumulation of cardiac lipofuscin in crab-eating monkeys ( Macaca fasicularis): The same rate of lipofuscin accumulation in several species of primates

Previously, we have reported that the aging process begins at sexual maturation (Nakano, M. et al., Mech. Ageing Dev., 52 (1990) 93–106). In this paper, we reported the cardiac lipofuscin accumulation of crab-eating monkeys. The first appearance of cardiac lipofuscin was around sexual maturation, and the rate of accumulation in crab-eating monkey was 0.45. Several primates which have different life spans show the same rate of lipofuscin accumulation in the life stage. Namely, even in a different life span, the amount of lipofuscin accumulation in a given period of life such as puberty, middle age, old age was the same. From these results, it is suggested that the amount of lipofuscin accumulation is the same in the life span of primates having different life spans.

Hazard rates and causes of death in a captive group of crab-eating monkeys (Macaca fascicularis).

In an attempt to examine possible associations between stages of agespecific mortality and various causes of death, vital records of 159 male and 192 female crab-eating monkeys (Macaca fascicularis), housed as a single group, were analyzed. Survival and hazard rates associated with each of five distinct categories of causes of death were estimated for males and females, using the nonparametric kernel method. The obtained overall survival and hazard functions were similar to those reported previously for rhesus monkeys. Among two stages identified in age-specific mortality, the first stage, characterized by rapidly decreasing hazard rates up to about 1.5-2 years of age, was discriminated by the occurrence of deaths due to unfitness for postnatal life. The second stage lasted up to the age of 10-15 years and was largely characterized by a high incidence of violent deaths. The respective hazard rates in males and females attained peaks during the early reproductive period of life and markedly decreased thereafter. This pattern was interpreted to indicate that second stage mortality is unlikely related to senescence, but rather, seems to depend on extrinsic environmental factors. Thus, when considering overall hazard rates in Macaca fascicularis, the onset of senescence, as a result of the specific aspects of simian reproduction, may be hidden from view, and mortality due to aging may only be appreciable after 10 to 15 years of age. © 1993 Wiley-Liss, Inc.

Bone induction in monkeys by bone morphogenetic protein. A trans-filter technique

We investigated bone induction by bone morphogenetic protein in primates, comparing it with that seen in rodents. Twelve Millipore diffusion chambers containing 5 mg of semipurified bone morphogenetic protein were implanted into the dorsal muscles of 12 young, adult crab-eating monkeys (Macaca fascicularis) and were retrieved six weeks later. In six of nine unbroken chambers, new bone with haematopoietic marrow had been induced on the host-side surface of the filter. The incidence of trans-filter bone induction in the monkeys was almost equal to that observed in mice, and the new bone yield was approximately half as much as in mice. Our results show that bone morphogenetic protein can induce new bone formation in primates within six weeks, and support the hope that it will be useful as a substitute for bone graft in man.

Molecular cloning of monkey liver cytochrome P-450 cDNAs : Similarity of the primary sequences to human cytochromes P-450

Three cDNAs coding for monkey cytochrome P-450 (P450) 2C, 2E and 3A (MKmp13, MKj1 and MKnf2, respectively) were isolated from a λgt11 cDNA library of a liver from a 3-methylcholanthrene (3MC)-treated crab-eating monkey, using cDNA fragments for human P450 2C, 2E and 3A as respective probes. MKmp13 and MKnf2 were 1901 and 2032 bp long, containing entire coding regions for polypeptides of 490 and 503 residues, respectively. The deduced N-terminal amino acid sequences of MKmp13 and MKnf2 were identical with those of P450-MK1 and P450-MK2, which had been purified from liver microsomes of untreated and polychlorinated biphenyl (PCB)-treated crab-eating monkeys, respectively. MKj1 was 1508 bp long, encoding a polypeptide of 449 residues, which is presumed to lack N-terminal 45 residues as compared with the sequence for human P450 2E1. Northern blot analysis indicated that monkey P450 2C, 2E and 3A mRNAs were expressed constitutively in monkey livers. P450 2E and 3A mRNAs were induced by both 3MC and PCB, while P450 2C mRNA was induced only by PCB. The deduced amino acid sequences of four monkey cytochrome P-450 cDNAs, including P450 1A1 (MKah1) which we isolated previously, were more than 92% identical with those of corresponding human cytochrome P-450 cDNAs