Normal variability in the age and first onset of ossification of the triquetral

The age and order of ossification of the triquetral, among the bones of the hand and wrist, were determined from serial radiographs of 108 males and 103 females of the Oxford Child Health Survey. Although the median ages of ossification agreed reasonably well with the mean ages reported by other authors, the distributions suggested that the former is the more appropriate statistic. The distributions of order of ossification were distinctly bimodal for both sexes because of the tendency of the triquetral to appear along with the epiphyseal centers. The triquetral appeared before the lunate in 184 children, after it in eight, and could not be sequentially distinguished in 19. Excluding these 19, triquetral sequence variability in the remaining was achieved, in 190, through alteration of its appearance relative to the epiphyseal centers and, in the other two through an alteration of the lunate appearance. Measures of median or average order of appearance are of very little value for the triquetral.     

De novo methylation in male germ cells of the common marmoset monkey occurs during postnatal development and is maintained in vitro

The timing of de novo DNA methylation in male germ cells during human testicular development is yet unsolved. Apart from that, the stability of established imprinting patterns in vitro is controversially discussed. This study aimed at determining the timing of DNA de novo methylation and at assessing the stability of the methylation status in vitro. We employed the marmoset monkey (Callithrix jacchus) as it is considered the best non-human primate model for human testicular development. We selected neonatal, pre-pubertal, pubertal, and adult animals (n = 3, each) and assessed germ cell global DNA methylation levels by 5-methyl cytosine staining, and Alu elements and gene-specific methylation (H19, LIT1, SNRPN, MEST, OCT4, MAGE-A4, and DDX-4) by pyrosequencing. De novo methylation is progressively established during postnatal primate development and continues until adulthood, a process that is different in most other species. Importantly, once established, methylation patterns remained stable, as demonstrated using in vitro cultures. Thus, the marmoset monkey is a unique model for the study of postnatal DNA methylation mechanisms in germ cells and for the identification of epimutations and their causes.     

Monomorphic Region of the Serotonin Transporter Promoter Gene in New World Monkeys

Genetic variation in the human serotonin system has long been studied because of its functional consequences and links to various neuropsychiatric and behavior‐related disorders. Among non‐human primates, the common marmosets (Callithrix jacchus) and tufted capuchins monkeys (Cebus apella) are becoming increasingly used as models to study the effects of genes, environments, and their interaction on physiology and complex behavior. In order to investigate the independent functions of and potential interactions between serotonin‐related genes, anxiety and neuropsychiatric disorders, we analyzed the presence and variability of the serotonin transporter gene‐linked polymorphic region (5‐HTTLPR) in marmoset and capuchin monkeys. By PCR and using heterologous primers from the human sequence, we amplified and then sequenced the corresponding 5‐HTT region in marmosets and capuchins. The resulting data revealed the presence of a tandem repeat sequence similar to that described in humans, but unlike humans and other Old World primates, no variable length alleles were detected in these New World monkeys, suggesting that if serotonin transporter is involved in modulating behavior in these animals it does so through different molecular mechanisms. Am. J. Primatol. 74:1028‐1034, 2012. © 2012 Wiley Periodicals, Inc.     

Appearance of ossification centers of the lower arm,wrist, lower leg,and ankle in immature orangutans and chimpanzees with an assessment of the relationship of ossification to dental development

This study examines the appearance of the secondary ossification centers in the lower arms, wrists, lower legs, and ankles of a cross-sectional sample of 20 infant orangutans and chimpanzees (15 of known age). The number of tarsal and carpal centers is analyzed relative to the degree of M₁ development and the weight of individual animals. Variation in the appearance of these ossification centers is discussed relative to these variables and others. In addition, a sequence of appearance is established for the carpal and tarsal ossification centers in the orangutan and data is presented on the status of these centers in a fetal and newborn gorilla. Study results indicate that 1) there is variation in the number of secondary epiphyses present in animals of similar ages; 2) tarsal ossification is completed prior to carpal ossification in the orangutan; 3) there are indications of a relationship between weight and the number of ossification centers present in animals of similar age; and 4) there appears to be no evidence of specific relationships between carpal and tarsal development and M₁ development. © 1996 Wiley-Liss, Inc.     

A study of postnatal appendicular skeletal maturation in captive-born squirrel monkeys (Saimiri boliviensis)

This study is based on a mixed longitudinal radiographic sampling of appendicular bones in 82 captive-born squirrel monkeys (Saimiri boliviensis). All appendicular ossification centers had appeared radiographically by 17 weeks of age, and epiphyseal fusion was complete by 47-53 months of age. No statistically significant differences were found between the sexes in comparisons carried out at birth and at 6 and 8 months of age. Valid sex comparisons could not be made at other ages owing to the small size of the sample. The sequence of appearance of the ossification centers (Spearman correlation coefficient of Saimiri vs. Callithrix = 0.68) shows greater interspecies differences than the sequence of fusion (Saimiri vs. Callithrix = 0.88).     

Micropuncture and microanalytical studies of rhesus monkey and baboon epididymis and the human ductus deferens

Using micropuncture and microanalytical techniques, we studied the microenvironment surrounding the maturing spermatozoa in different regions along the epididymis of the rhesus monkey (Macaca mulatta; 3 animals) and the baboon (Papio cynocephalus; 1 animal) and in the human ductus deferens. In the monkey and baboon, samples of luminal contents (luminal fluid and spermatozoa) of approximately 50 to 300 nanolitres were collected from several epididymal sites and the luminal fluid analyzed for inositol. Similarly, a sample of approximately 0.5 to 1.0 μl of luminal contents was collected from each human ductus deferens and the luminal fluid analyzed for sodium, potassium, chloride, inositol, carnitine, glycerophosphocholine (GPC), phosphocholine and total phosphate. Each analysis required the modification of standard methods to accommodate the very small sample volumes collected. We show that the microenvironment in the monkey, baboon, and human is different from those in other species with respect to the concentration of compounds estimated. In the luminal fluid of the human ductus deferens, the majority of the osmoticallyactive compounds are the inorganic ions which is in direct contrast to the rat, hamster, rabbit, ram and boar. In these species, organic compounds contribute significantly more to the osmolarity of the luminal fluid than do inorganic ions. Although the significance of these findings is unclear, a relationship seems to exist between the appearance of carnitine in the luminal fluid of the proximal caput epididymidis of the rat and the point where spermatozoa develop the potential for motility. These investigations also raise the question of which species most closely reflects the physiology of the reproductive system of man.     

全身一体化成像技术在恒河猴动物实验中的应用

目的探讨并比较CT与核磁共振成像（magnetic resonance imaging,MRI）在恒河猴全身成像研究中的应用范围,以及全身一体化技术（total imaging Matrix,TIM）配合增强扫描对恒河猴骨骼、循环、神经系统成像,总结其影像学表现中与人相应系统解剖的差异。方法应用64层螺旋CT的表面遮盖成像重建技术（3D-SSD）对恒河猴全身骨骼系统进行全身平扫及重建,3．0TMRI的TIM技术与最大密度投影重建（MIP）对恒河猴循环系统、神经系统进行全身平扫及增强扫描并重建。结果64层螺旋CT骨骼系统成像显示恒河猴腰椎骨6块、骶椎骨8块、尾椎骨11块、真肋骨9根,数量多于人类;颈椎棘突未见分叉,与人类不同。MRI动态增强技术与一体化成像技术联用显示恒河猴的动脉系统与人类相近,端脑矢状位最长径与冠状位最长径比值比人类略低,脊髓膨大位置比人类略低。结论多层螺旋CT与MRI的TIM技术可应用于对大型实验动物进行无创性的解剖学分析,是一种无创、快速、可以在体研究并动态连续观察其结构的科学有效方法。     

Cynomolgus Monkey (Macaca fascicularis) Cathepsin K: Cloning, Expression, Purification, and Activation

Methodology for the production of recombinant active cynomolgus monkey (Macaca fascicularis) cathepsin K (EC 3.4.22.38) was elucidated. The cDNA encoding the cathepsin K was cloned from femaleM. cynomolgusmonkey mRNA. The deduced amino acid sequence ofM. cynomolguspreprocathepsin K from the cDNA sequence showed 94.2% identity to human preprocathepsin K. Sequence differences occurred only in the prepro- domains; the mature domains were identical. The recombinantM. cynomolguscathepsin K was expressed as a secreted proenzyme using baculovirus-infected SF21 insect cells having the predicted N-terminus (LYPEEILDTH … ), indicating proper cleavage of the secretion sequence. Purified monkey procathepsin K was activated under autocatalytic conditions at pH 4.0. The mature enzyme was composed of mixture of enzymes having N-termini of Gly¹¹³and Arg¹¹⁴. The molecular weight was determined to be 23,668.3 Da by MALDI-TOF-MS which is consistent with the absence of carbohydrate on the mature enzyme. These results indicate that monkey procathepsin K is able to autoactivate and produces a mature enzyme which is identical to that of human cathepsin K. Since the sequence of monkey and human mature cathepsin K are identical and thein vitroactivation mechanisms appear to be indistinguishable, monkeys are predicted to be a good animal model for evaluating cathepsin K inhibitorsin vivoas therapeutic agents for diseases characterized by excessive bone loss, such as osteoporosis.     

Assessment of the nucleotide sequence variability in the bovine T-cell receptor alpha delta joining gene region

The sequence of 2,193 nucleotides from the bovine T-cell receptor alpha/delta joining gene region (TCRADJ) was determined and compared with the corresponding human and murine sequences. The identity was 75.3% for the comparison of the Bos taurus vs. the Homo sapiens sequence and 63.8% for the Bos taurus vs. the Mus musculus sequence. This comparison permitted the identification of the putatively functional elements within the bovine sequence. Direct sequencing of 2,110 nucleotides in nine animals revealed 12 variable sites. Estimates, based on direct sequencing in three Holstein Friesian animals, for the two measures of sequence variability, nucleotide polymorphism (u) and nucleotide diversity (p), were 0.00050 (60.00036) and 0.00077 (60.00056), respectively. The test statistic, Tajima's D, for the comparison of the two measures indicates that the difference between u and p is close to significance (P < 0.05), suggesting the possibility of selective forces acting on the studied genomic region. Allelic variation at 5 of the 12 variable sites was analysed in 359 animals (48 Anatolian Black, 56 Braunvieh, 115 Fleckvieh, 47 Holstein Friesian, 50 Simmental and 43 Pinzgauer) using the oligonucleotide ligation assay (OLA) in combination with the enzyme linked immunoabsorbant assay (ELISA). Nine unambiguous haplotypes could be derived based on animals with a maximum of one heterozygous site. Four to seven haplotypes were present in the different breeds. When taking into account the frequencies of the haplotypes in the different breeds, especially in Anatolian Black, an ancestral cattle population, we could establish the likely phylogenetic relationships of the haplotypes. Such haplotype trees are the basis for cladistic candidate gene analysis. Our study demonstrates that the systematic search of single nucleotide polymorphisms (SNPs) is useful for analysing all aspects of variability of a given genomic region.     

Comparative ossification and development of the skull in palaeognathous birds (Aves: Palaeognathae)

Ratites and tinamous are a morphologically diverse group of flightless and weakly flighted birds. As one of the most basal clades of extant birds, they are frequently used as an outgroup for studies discussing character evolution within other avian orders. Their skeletal development is not well known in spite of their important phylogenetic position, and studies have historically been plagued with small sample sizes and limited anatomical and temporal scope. Here, I describe the ossification of the skull in the emu (Dromaius novaehollandiae), ostrich (Struthio camelus), greater rhea (Rhea americana), and elegant crested‐tinamou (Eudromia elegans). Skeletal development is remarkably consistent within palaeognaths, in spite of large differences in absolute size and incubation period. Adult morphology appears to play a role in interordinal differences in the sequence and timing of ossification of certain bones. Neither the timing of cranial ossification events relative to stage nor the sequence of ossification events provides any evidence in support of a paedomorphic origin of the palaeognathous palate. This study provides an important first look at the timing and sequence of skull development in palaeognathous birds, providing data that can be compared to better‐studied avian systems in order to polarize ontogenetic characters. © 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 156 , 184–200.     

Serum Estradiol 17β, Progesterone,and Relative Luteinizing Hormone Levels in Saimiri sciureus: Cyclic Variations and the Effect of Laparoscopy and Follicular Aspiration

With increasing use of the squirrel monkey (Saimiri sciureus) in reproductive studies, a precise knowledge of the hormonal variations within the cycle becomes essential for timing of ovulation. The frequent utilization of laparoscopy for follicle aspiration and oocyte recovery warrants analysis of hormonal alterations.     

Individual differences and variability in the timing of motor activity during walking in insects

The uniformity of the neural physiology of an animal population is a fundamental, rarely tested assumption in most neurophysiological work. In this study, the variability of the timing between the movements of pairs of legs during free walking in cockroaches was assessed. Phases (a measure of timing) of motor bursts in muscles of legs in the American cockroach, Periplaneta americana, were calculated for insects walking straight over a flat, level surface. Student's t, Wallraff, Mann Whitney and Watson U² two-sample tests were used to compare the phases of motor bursts of the same pairs of legs in different insects. The comparisons showed that in spite of the homogeneity both of the animal population and of the conditions under which the insects walked, most of the inter-leg phases of the animals that were compared were significantly different statistically. Further testing of greater numbers of insects using analysis of variance to test for population uniformity confirmed that the insects we tested were not members of a single statistical population with respect to the timing of motor bursts of the legs during walking. We infer that this unexpectedly large variability in a population thought to be relatively homogeneous reflects subtle but biologically significant differences between animals. The possible sources of these differences and their consequences for the study of behavior and its physiological basis are discussed.     

Variation and timing of the cranial ossification sequence of the oriental fire-bellied toad,Bombina orientalis (Amphibia,Discoglossidae)

The sequence of appearance of the 17 different skull bones in the oriental fire-bellied toad, Bombina orientalis, is described. Data are based primarily on samples of ten or 11 laboratory-reared specimens of each of 11 Gosner developmental stages (36–46) representing middle through late metamorphosis. Ossification commences as early as stage 37 (hind limb with all five toes distinct), but the full complement of adult bones is not attained until stage 46 (metamorphosis complete). Number of bones present at intermediate stages is poorly correlated with external morphology. As many as four Gosner developmental stages elapse before a given bone is present in all specimens following the stage at which it may first appear. The modal ossification sequence is frontoparietal, exoccipital, parasphenoid, septomaxilla, premaxilla, vomer, nasal, maxilla, angulosplenial, dentary, squamosal, quadratojugal, pterygoid, prootic, interfrontal, sphenethmoid, and mentomeckelian. Most specimens are consistent with this sequence, despite the poor correlation between cranial ossification and external development as assayed by Gosner stage. The timing of cranial ossification in Bombina orientalis differs in many respects from that described for two other, distantly related anurans, the leopard frog (Rana pipiens) and the western toad (Bufo boreas). These include the total number and sequence of appearance of bones, and the timing of ossification relative to the development of external morphology. Interspecific variation may reflect differences in the timing of the tissue interactions known to underlie skeletal differentiation and evolution.     

Primate Language and the Playback Experiment,in 1890 and 1980

The playback experiment – the playing back of recorded animal sounds to the animals in order to observe their responses – has twice become central to celebrated researches on non-human primates. First, in the years around 1890, Richard Garner, an amateur scientist and evolutionary enthusiast, used the new wax cylinder phonograph to record and reproduce monkey utterances with the aim of translating them. Second, in the years around 1980, the ethologists Peter Marler, Robert Seyfarth, and Dorothy Cheney used tape recorders in a broadly similar way to test whether the different predator calls of one monkey species, vervet monkeys, warn about different kinds of predator. This paper explores the circumstances leading to the ca. 1890 invention and the ca. 1980 reinvention of the primate playback experiment. In both instances, I show, the experiment served as a riposte to those arguing, on scientific grounds, that an unbridgeable gap divides human language from animal communication. I also consider how far progress in technology explains the timing of invention and reinvention. I conclude with some reflections on sifting contingent from inevitable aspects of the history of the primate playback experiment, and of scientific achievements more generally.     

Variability in life history parameters of four serotinous plants in the Namib Desert

Plants growing in extreme habitats, such as deserts, are expected to show high plasticity (variability) in life history parameters to compensate for the unpredictability of their environment. In this study variability in life history parameters and possible sources of variability were investigated in four serotinous (= seed retaining) plant species from the Namib Desert.Blepharis grossa, Geigeria alata, Geigeria ornativa andPetalidium setosum were studied in a mosaic of spatio-temporal environmental conditions represented by three sites and three rain seasons. The coefficient of variation and whether or not significant differences occurred between environmental conditions were used to determine the most variable species and parameters. The most variable species was the strictly annualB. grossa, which might be related to its life form strategy and poor competitive ability as a pioneer species. Life history parameters which responded most frequently to varying environmental conditions were source pool, flowering time and seed production. Seedling density was influenced by the source pool and intensity of rainfall, seedling establishment by timing and intensity of rain events, as well as by seed mass. Although it was expected that high within population variability in life history parameters would also result in a response to differing environmental conditions, there was no direct correlation. Variable responses in life history parameters in most species were caused by differences in site condition as well as by differences in seasons, especially with regard to timing and intensity of rain events.     

Determining genetic background in captive stocks of cynomolgus macaques (Macaca fascicularis)

Background As in other model organisms, genetic background in the non‐human primates Macaca mulatta and Macaca fascicularis is an experimental variable that affects the response of other study variables. Genetic background in model organisms is manipulated by breeding schemes but is generally pre‐determined by the source population used to found captive stocks. In M. fascicularis three such sources have been distinguished, however, these are not routinely taken into consideration when designing research. Methods We exemplify a mitochondrial DNA (mtDNA)‐based strategy to trace the maternal geographic origins of M. fascicularis animals of unspecified origins. Results Macaca fascicularis of unspecified origins kept at primate research centers carry mtDNA haplotypes representing all three major genetic subdivisions. Conclusions We suggest that the genetic background of study animals could be better specified in the future using an mtDNA‐based approach, which would enable informed selection of study animals and help reduce variation within and among studies.     

Purification and Characterization of Cynomolgus Monkey Tryptase

Cynomolgus monkey tryptase was purified to homogeneity from lung tissue. Reducing SDS-PAGE analysis of the monkey enzyme produced a doublet at 30–32 kDa, which reacted with antibodies against human lung tryptase on a Western blot. N-terminal sequence analysis of the monkey enzyme yielded a sequence that was identical to human tryptase out to 15 residues. Gel filtration chromatography either in the presence or absence of heparin indicated that the monkey enzyme had a molecular mass of approximately 250 kDa and 140 kDa, respectively, consistent with the formation of a tetramer that can bind heparin. Other similarities between human and monkey tryptase included the ability to degrade vasoactive intestinal peptide and a resistance to inhibition by biological serine protease inhibitors. However, the two enzymes displayed markedly different pH stability profiles. Monkey tryptase was unstable at pH values over 7.0, even in the presence of heparin, displaying a half-life of 10.9 min at pH 8.0. In addition, the stabilizing effect of heparin was pH dependent, being most prevalent at lower pH values. Therefore, the biological activity of monkey tryptase may be controlled by both pH and the availability of heparin.     

Postnatal skeletal ontogeny in Callisaurus draconoides and Uta stansburiana (Iguania: Phrynosomatidae).

The mineralization of the skeleton from hatching to near maximum size in two phrynosomatid lizards, Callisaurus draconoides and Uta stansburiana, is described in detail. Observed patterns in the appearance of epiphyseal secondary centers, ossification centers, apophyseal ossifications, and calcifications, the distribution of sesamoids, and the timing of fusions, are compared and contrasted with observations of other squamates available in the literature. Overall, Callisaurus and Uta exhibit an advanced state of ossification in the hatchling relative to other squamate neonates and share a similar sequence of braincase fusions and appearance of secondary centers. Preliminary observations suggest that patterns of postnatal skeletal development are highly conserved and independent of patterns of prenatal morphogenesis, and thus a potentially rich source of character data for systematic investigations.