Deep Learning-Based Spermatogenic Staging in Tissue Sections of Cynomolgus Macaque Testes.

The indirect assessment of adverse effects on fertility in cynomolgus monkeys requires that tissue sections of the testis be microscopically evaluated with awareness of the stage of spermatogenesis that a particular cross-section of a seminiferous tubule is in. This difficult and subjective task could very much benefit from automation. Using digital whole slide images (WSIs) from tissue sections of testis, we have developed a deep learning model that can annotate the stage of each tubule with high sensitivity, precision, and accuracy. The model was validated on six WSI using a six-stage spermatogenic classification system. Whole slide images contained an average number of 4938 seminiferous tubule cross-sections. On average, 78% of these tubules were staged with 29% in stage I-IV, 12% in stage V-VI, 4% in stage VII, 19% in stage VIII-IX, 18% in stage X-XI, and 17% in stage XII. The deep learning model supports pathologists in conducting a stage-aware evaluation of the testis. It also allows derivation of a stage-frequency map. The diagnostic value of this stage-frequency map is still unclear, as further data on its variability and relevance need to be generated for testes with spermatogenic disturbances.

Comparative study of blood pressure and echocardiographic values of male cynomolgus macaques (Macaca fascicularis).

BACKGROUND: Primates represent a unique object for biomedical research, in particular in the field of physiology and pathology of the cardiovascular system. Echocardiography is the most important non-invasive method for the intravital study of the heart structure and function, intracardiac and systemic haemodynamics. The available data on reference values of echocardiographic parameters in primates are limited. METHODS: We determined and described 29 structural and functional parameters in echocardiographic examination using B-mode (two-dimensional scanning), M-mode (one-dimensional scanning) and in various Doppler modes together with blood pressure in 17 male cynomolgus macaques with an average age of 5.7 ± 0.6 years. We compared available literature data on reference values of echocardiography in this species. RESULTS AND CONCLUSIONS: Echocardiographic values in cynomolgus macaques depend on age, sex composition and the anaesthesia method. There is lack of presentation in the published studies of complete list of parameters that can be obtained by echocardiographic examination.

Ultrasound-guided, Transabdominal, Intrauterine Artificial Insemination for Cynomolgus Macaques (Macaca fascicularis) Based on Estimated Timing of Ovulation.

Intrauterine sperm injection for artificial insemination is difficult in cynomolgus macaques (Macaca fascicularis) and rhesus macaques (M. mulatta) due to the complex structure of the cervical canal, which differs from that of humans. Despite the availability of several artificial insemination methods for macaques, pregnancy rates are inconsistent, and details regarding ovulation are unclear, thus warranting more effective methods. Therefore, we developed an effective, ultrasound-guided, transabdominal intrauterine artificial insemination method for cynomolgus macaques that involves timing sperm injection to coincide with the periovulation phase estimated according to rapid hormone measurement. We performed our intrauterine artificial insemination on 6 female macaques; 4 of the 5 animals that were predicted to have ovulated soon after insemination became pregnant, whereas the 1 macaque that was predicted not to have ovulated did not. Furthermore, we saw no evidence of injury, such as a conspicuous needle hole or bleeding on the surface of or inside the uterus, nor did our method result in any abnormalities in the mothers or their offspring. Thus, our ultrasound-guided, transabdominal, intrauterine artificial insemination method is rapid, safe, and effective in cynomolgus macaques.

Bolstering geometric morphometrics sample sizes with damaged and pathologic specimens: Is near enough good enough?

Obtaining coordinate data for geometric morphometric studies often involves the sampling of dry skeletal specimens from museum collections. But many specimens exhibit damage and/or pathologic conditions. Such specimens can be considered inadequate for the analyses of shape and are excluded from study. However, the influences that damaged specimens may have on the assessment of normal shape variation have only been explored in two-dimensional coordinate data and no studies have addressed the inclusion of pathological specimens to date. We collected three-dimensional coordinate data from the cranium and mandible of 100 crab-eating macaques (Macaca fascicularis). Tests typically employed to analyze shape variation were performed on five datasets that included specimens with varying degrees of damage/pathology. We hypothesized that the inclusion of these specimens into larger datasets would strengthen statistical support for dominant biological predictors of shape, such as sex and size. However, we also anticipated that the analysis of only the most questionable specimens may confound statistical outputs. We then analyzed a small sample of good quality specimens bolstered by specimens that would generally be excluded due to damage or pathologic morphology and compared the results with previous analyses. The inclusion of damaged/pathologic specimens in a larger dataset resulted in increased variation linked to allometry, sexual dimorphism, and covariation, supporting our initial hypothesis. We found that analyzing the most questionable specimens alone gave consistent results for the most dominant aspects of shape but could affect outputs for less influential principal components and predictors. The small dataset bolstered with damaged/pathologic specimens provided an adequate assessment of the major components of shape, but finer scale differences were also identified. We suggest that normal and repeatable variation contributed by specimens exhibiting damage and/or pathology emphasize the dominant components and shape predictors in larger datasets, however, the various unique conditions may be more influential for limited sample sizes. Furthermore, we find that exclusion of damaged/pathologic specimens can, in some cases, omit important demographic-specific shape variation of groups of individuals more likely to exhibit these conditions. These findings provide a strong case for inclusion of these specimens into studies that focus on the dominant aspects of intraspecific shape variation. However, they may present issues when testing hypotheses relating to more fine-scale aspects of morphology.

Variability in the number of infraorbital foramina in rhesus macaques (Macaca mulatta) and cynomolgus macaques (Macaca fascicularis).

This study aimed to determine the number of infraorbital foramina in monkeys of the Papionini tribe. The authors performed a µCT analysis of the morphology of the infraorbital foramina. A total number of 52 simian skulls belonged to two macaque species: Macaca mulatta and Macaca fascicularis were used in the study. The number of infraorbital foramina was counted macroscopically and with the use of a magnifying glass. Next, the skull representing the most common morphological type was selected and scanned by micro-computed tomography (µCT). The Shapiro-Wilk normality test was used in the study. To compare the differences in the number of infraorbital foramen between species, sex and sides, the Mann-Whitney U test was applied. Three infraorbital foramina were present in most individuals from the test group. The Mann-Whitney test revealed no statistically significant difference between the number of foramina on the right- and left-hand side. Likewise, no statistically significant differences between the numbers of infraorbital foramina across sexes were observed. Volumetric reconstructions revealed the presence of separate infraorbital canals for each infraorbital foramen. Craniofacial innervation in macaques is formed by complex branching patterns of cranial nerves. Variability in the number of infraorbital foramina suggests a variable maxillary innervation pattern in these animals. Based on the analysis of volumetric projections, the presence of two labial branches and a single nasal branch of the infraorbital nerve is suggested. Detailed descriptions are supported by quantitative data and µCT evidence.

Comparison of the anatomical morphology of cervical vertebrae between humans and macaques: related to a spinal cord injury model.

Non-human primates are most suitable for generating cervical experimental models, and it is necessary to study the anatomy of the cervical spine in non-human primates when generating the models. The purpose of this study was to provide the anatomical parameters of the cervical spine and spinal cord in long-tailed macaques (Macaca fascicularis) as a basis for cervical spine-related experimental studies. Cervical spine specimens from 8 male adult subjects were scanned by micro-computed tomography, and an additional 10 live male subjects were scanned by magnetic resonance imaging. The measurements and parameters from them were compared to those of 12 male adult human subjects. Additionally, 10 live male subjects were scanned by magnetic resonance imaging, and the width and depth of the spinal cord and spinal canal and the thickness of the anterior and posterior cerebrospinal fluid were measured and compared to the relevant parameters of 10 male adult human subjects. The tendency of cervical parameters to change with segmental changes was similar between species. The vertebral body, spinal canal, and spinal cord were significantly flatter in the human subjects than in the long-tailed macaques. The cerebrospinal fluid space in the long-tailed macaques was smaller than that in the human subjects. The anatomical features of the cervical vertebrae of long-tailed macaques provide a reference for establishing a preclinical model of cervical spinal cord injury.

Quantitative assessment of amygdala in Macaca fascicularis monkeys.

The amygdaloid complex (AC) is involved in very relevant cognitive and emotional pathways and exhibits changes in aging and in some neurological and psychiatric disorders. The quantitative estimators of AC could be useful to understand the impact of amygdaloid pathology in these processes, both globally and for each nucleus in particular, and their neurons. The present study analyzes morphometric and stereological estimators in the whole AC and its three main nuclei (lateral [La], basal [Ba], and accessory basal [AB]) in six Macaca fascicularis monkeys. All the brains were fixed and sectioned in the coronal plane; Nissl-stained sections were used for estimation of size and form parameters in both, the AC, and the La, Ba, and AB nuclei separately. The study includes stereological estimates of the volume and surface area of the AC; also, volume of the neurons in the amygdaloid nuclei was estimated using the point-sampled intercepts method. Our results show that the AB nucleus is smaller than both the La and Ba nuclei in both morphometric and stereological estimators. Brain hemispheric side had not significant influence on any of quantitative estimates. The neuron volume was higher in the AB nucleus relative to LA and Ba of the nuclei. These data describe some quantitative parameters of the amygdaloid complex and their main nuclei that could help us to detect small changes in neurodegenerative and other pathological processes.

Ontogenetic changes in magnitudes of integration in the macaque skull.

OBJECTIVES: Magnitudes of morphological integration may constrain or facilitate craniofacial shape variation. The aim of this study was to analyze how the magnitude of integration in the skull of Macaca fascicularis changes throughout ontogeny in relation to developmental and/or functional modules. MATERIALS AND METHODS: Geometric morphometric methods were used to analyze the magnitude of integration in the macaque cranium and mandible in 80 juvenile and 40 adult M. fascicularis specimens. Integration scores in skull modules were calculated using integration coefficient of variation (ICV) of eigenvalues based on a resampling procedure. Resultant ICV scores between the skull as a whole, and developmental and/or functional modules were compared using Mann-Whitney U tests. RESULTS: Results showed that most skull modules were more tightly integrated than the skull as a whole, with the exception of the chondrocranium in juveniles without canines, the chondrocranium/face complex and the mandibular corpus in adults, and the mandibular ramus in all juveniles. The chondrocranium/face and face/mandibular corpus complexes were more tightly integrated in juveniles than adults, possibly reflecting the influences of early brain growth/development, and the changing functional demands of infant suckling and later masticatory loading. This is also supported by the much higher integration of the mandibular ramus in adults compared with juveniles. DISCUSSION: Magnitudes of integration in skull modules reflect developmental/functional mechanisms in M. fascicularis. However, the relationship between "evolutionary flexibility" and developmental/functional mechanisms was not direct or simple, likely because of the complex morphology, multifunctionality, and various ossification origins of the skull.

Nonpyramidal neurons in the primate basolateral amygdala: A Golgi study in the baboon (Papio cynocephalus) and long-tailed macaque (Macaca fascicularis).

Nonpyramidal GABAergic interneurons in the basolateral nuclear complex (BNC) of the amygdala are critical for the regulation of emotion. Remarkably, there have been no Golgi studies of these neurons in nonhuman primates. Therefore, in the present study we investigated the morphology of nonpyramidal neurons (NPNs) in the BNC of the baboon and monkey using the Golgi technique. NPNs were scattered throughout all nuclei of the BNC and had aspiny or spine-sparse dendrites. NPNs were morphologically heterogeneous and could be divided into small, medium, large, and giant types based on the size of their somata. NPNs could be further divided on the basis of their somatodendritic morphology into four types: multipolar, bitufted, bipolar, and irregular. NPN axons, when stained, formed dense local arborizations that overlapped their dendritic fields to varying extents. These axons always exhibited varying numbers of varicosities representing axon terminals. Three specialized NPN subtypes were recognized because of their unique anatomical features: axo-axonic cells, neurogliaform cells, and giant cells. The axons of axo-axonic cells formed "axonal cartridges," with clustered varicosities that contacted the axon initial segments of pyramidal neurons (PNs). Neurogliaform cells had small somata and numerous short dendrites that formed a dense dendritic arborization; they also exhibited a very dense axonal arborization that overlapped the dendritic field. Giant cells had very large irregular somata and long, thick dendrites; their distal dendrites often branched extensively and had long appendages. In general, the NPNs of the baboon and monkey BNC, including the specialized subtypes, were similar to those of rodents.

Determining overweight and underweight with a new weight-for-height index in captive group-housed macaques.

Housing primates in naturalistic groups provides social benefits relative to solitary housing. However, food intake may vary across individuals, possibly resulting in overweight and underweight individuals. Information on relative adiposity (the amount of fat tissue relative to body weight) is needed to monitor overweight and underweight of group-housed individuals. However, the upper and lower relative adiposity boundaries are currently only known for macaques living solitarily in small cages. We determined the best measure of relative adiposity and explored the boundaries of overweight and underweight to investigate their incidence in group-housed adult male and female rhesus macaques and long-tailed macaques living in spacious enclosures at the Biomedical Primate Research Centre (BPRC), the Netherlands. During yearly health checks different relative adiposity measures were obtained. For long-tailed macaques, comparable data on founder and wild animals were also available. Weight-for-height indices (WHI) with height to the power of 3.0 (WHI3.0) for rhesus macaques and 2.7 (WHI2.7) for long-tailed macaques were optimally independent of height and were highly correlated with other relative adiposity measures. The boundary for overweight was similar in group-housed and solitary-housed macaques. A lower boundary for underweight, based on 2% body fat similar to wild primates, gave a better estimate for underweight in group-housed macaques. We propose that for captive group-housed rhesus macaques relative adiposity should range between 42 and 67 (WHI3.0) and for long-tailed macaques between 39 and 62 (WHI2.7). The majority of group-housed macaques in this facility have a normal relative adiposity, a considerable proportion (17-23%) is overweight, and a few (0-3%) are underweight.

Tridimensional mapping of Phox2b expressing neurons in the brainstem of adult Macaca fascicularis and identification of the retrotrapezoid nucleus.

Chemosensitivity is a key mechanism for the regulation of breathing in vertebrates. The retrotrapezoid nucleus is a crucial hub for respiratory chemoreception within the brainstem. It integrates chemosensory information that are both peripheral from the carotid bodies (via the nucleus of the solitary tract) and central through the direct sensing of extracellular protons. To date, the location of a genetically defined RTN has only been ascertained in rodents. We first demonstrated that Phox2b, a key determinant for the development of the visceral nervous system and branchiomotor nuclei in the brainstem including the RTN, had a similar distribution in the brainstem of adult macaques compared to adult rats. Second, based on previous description of a specific molecular signature for the RTN in rats, and on an innovative technique for duplex in situ hybridization, we identified parafacial neurons which coexpressed Phox2b and ppGal mRNAs. They were located ventrally to the nucleus of the facial nerve and extended from the caudal part of the nucleus of the superior olive to the rostral tip of the inferior olive. Using the previously described blockface technique, deformations were corrected to allow the proper alignment and stacking of digitized sections, hence providing for the first time a 3D reconstruction of the macaque brainstem, Phox2b distribution and the primate retrotrapezoid nucleus. This description should help bridging the gap between rodents and humans for the description of key respiratory structures in the brainstem.

Variability of Spleen and Mesenteric Lymph Node in Control Cynomolgus Monkeys ( Macaca fascicularis) from Nonclinical Safety Studies: A Retrospective Assessment.

We assessed the variability of spleen and mesenteric lymph node (MLN) microscopic observations and the correlations of these observations with other study data from 478 control cynomolgus monkeys from 53 routine nonclinical safety studies. Spleen weight parameters (absolute and relative to body or brain weights) were highly variable both within a control group on an individual study (up to 5.11-fold) and among animals with the same light microscopic observation. Grades for microscopic observations were also highly variable. The most frequent microscopic observations for spleen were changes in the size and number of germinal centers (58%), acidophilic (hyaline) material in lymphoid follicles (52%), and compound lymphoid follicles (20%). The most frequent microscopic observations in the MLN were eosinophil infiltrates (90%), changes in size and number of germinal centers (42%), and brown pigment (21%). The only meaningful relationships ( r2 > 0.3) were positive correlations between reticuloendothelial hyperplasia and malarial pigment in the spleen and between each of these observations and spleen weight parameters. We conclude that determination of test article-related effects on the immune system in routine monkey toxicology studies requires careful consideration and a weight-of-evidence approach due to the low numbers of animals/group, the inherent variability in spleen and MLN parameters, and the infrequent correlation among immune system-related end points.

Validation of Sysmex XT-2000iV analyzer-generated quantitative bone marrow differential counts in cynomolgus monkeys, Beagle dogs, and CD-1 mice.

BACKGROUND: In a previous study, the validation of rat bone marrow (BM) collection, processing, and analysis using the Sysmex XT-2000iV (Sysmex Corporation, Kobe, Japan) hematology analyzer showed that the Sysmex hematology analyzer produced BM differential counts that were comparable to those obtained with microscopic differential counts. OBJECTIVE: This study was conducted to expand the validation of the Sysmex TNCC (total nucleated cell count) and 5-part BM differential in cynomolgus monkeys, Beagle dogs, and CD-1 mice, which are alternate species that are also frequently used in preclinical safety studies. METHODS: The Sysmex 5-part BM differential counts were generated with a two-step process, whereby proliferating and maturing erythroid and myeloid cells were determined by preset gating and lymphocytes were determined using species-specific B- and T-lymphocyte antibodies and a magnetic cell-sorting method (MACS). Agreement with microscopic myelograms with 500-cell differential counts was determined from BM suspensions of 62 cynomolgus monkeys, 47 Beagle dogs, and 44 CD-1 mice. RESULTS: The correlation coefficients between methods for myeloid to erythroid (M:E) ratios in all three species was > 0.928. The Bland-Altman differences between methods were approximately ± 0.3 units for the M:E ratio in dogs and mice, and +0.6 and -0.4 in monkeys. The upper limits of agreement for all three species were ≤7% for maturing myeloid cells, ≤6% for maturing erythroid cells, and ≤4% for proliferating myeloid cells, proliferating erythroid cells, and lymphocytes. CONCLUSIONS: The Sysmex XT-2000iV produces an automated M:E ratio and a 5-part differential count equivalent to microscopic differential counts in cynomolgus monkeys, Beagle dogs, and CD-1 mice.

Ontogenetic changes to muscle architectural properties within the jaw-adductor musculature of Macaca fascicularis.

OBJECTIVES: Changes to soft- and hard-tissue components of the masticatory complex during development can impact functional performance by altering muscle excursion potential, maximum muscle forces, and the efficiency of force transfer to specific bitepoints. Within Macaca fascicularis, older individuals exploit larger, more mechanically resistant food items and more frequently utilize wide-gape jaw postures. We therefore predict that key architectural and biomechanical variables will scale during ontogeny to maximize bite force and gape potential within older, larger-bodied individuals. MATERIALS AND METHODS: We analyzed 26 specimens of M. fascicularis, representing a full developmental spectrum. The temporalis, superficial masseter, and deep masseter were dissected to determine muscle mass, fiber length, and physiologic cross-sectional area (PCSA). Lever-arm lengths were also measured for each muscle, alongside the height of the temporomandibular joint (TMJ) and basicranial length. These variables were scaled against two biomechanical variables (jaw length and condyle-molar length) to determine relative developmental changes within these parameters. RESULTS: During ontogeny, muscle mass, fiber length, and PCSA scaled with positive allometry relative to jaw length and condyle-molar length within all muscles. TMJ height also scaled with positive allometry, while muscle lever arms scaled with isometry relative to jaw length and with positive allometry (temporalis) or isometry (superficial and deep masseter) relative to condyle-molar length. CONCLUSION: Larger individuals demonstrate adaptations during development towards maximizing gape potential and bite force potential at both an anterior and posterior bitepoint. These data provide anatomical evidence to support field observations of dietary and behavioral differences between juvenile and adult M. fascicularis.

Comparison of cellular localisation of the Ca2+ -binding proteins calbindin, calretinin and parvalbumin in the retina of four different Macaca species.

Ca2+ -binding proteins are differentially expressed in the nervous system; their functional role often remains unclear. This immunohistochemical study aimed at characterising and comparing the expression pattern of the Ca2+ -binding proteins calbindin (Calb), calretinin (Calr) and parvalbumin (Parv) in the retina of four species of macaque monkeys: Macaca fascicularis (cynomolgus macaque), M. mulatta (rhesus macaque), M. thibetana (Tibetan macaque) and M. fuscata (Japanese macaque). Calb was found in cone photoreceptors and in a subset of bipolar cells. Calr was expressed in a subpopulation of amacrine cells. Parv was present in horizontal and ganglion cells. In addition, Müller cells were stained using antibodies against the specific marker cellular retinaldehyde-binding protein (CRALBP). Immunostainings were used for calculation of the density of different cell populations. The expression pattern was similar between the examined species and between retinal regions.

Morphological integration of anatomical, developmental, and functional postcranial modules in the crab-eating macaque (Macaca fascicularis).

OBJECTIVES: Integration and modularity reflect the coordinated action of past evolutionary processes and, in turn, constrain or facilitate phenotypic evolvability. Here, we analyze magnitudes of integration in the macaque postcranium to test whether 20 a priori defined modules are (1) more tightly integrated than random sets of postcranial traits, and (2) are differentiated based on mode of definition, with developmental modules expected to be more integrated than functional or anatomical modules. MATERIALS AND METHODS: The 3D morphometric data collected for eight limb and girdle bones for 60 macaques were collated into anatomical, developmental, and functional modules. A resampling technique was used to create random samples of integration values for each module for statistical comparison. RESULTS: Our results found that not all a priori defined modules were more strongly integrated than random samples of postcranial traits and that specific types of modules did not present consistent patterns of integration. Rather, girdle and joint modules were consistently less integrated than limb modules, and forelimb elements were less integrated than hindlimbs. DISCUSSION: The results suggest that morphometrically complex modules tend to be less integrated than simple limb bones, irrespective of the number of available traits. However, differences in integration of the fore- and hindlimb more likely reflects the multitude of locomotory, feeding, and social functions involved. It remains to be tested whether patterns of integration identified here are primate universals, and to what extent they vary depending on phylogenetic or functional factors.

Non-Destructive Determination of Muscle Architectural Variables Through the Use of DiceCT.

The fascicular architecture of skeletal muscle dictates functional parameters such as force production and contractile velocity. Muscle microarchitecture is typically determined by means of manual dissection, a technique that is inherently destructive to specimens. Furthermore, fascicle lengths and pennation angles are commonly assessed at only a limited number of sampling sites per muscle. We present the results of a digital technique to non-destructively assess muscle architectural variables for three jaw-adductor muscles within a specimen of the cercopithecine primate Macaca fascicularis (crab-eating macaque). The specimen is first subjected to a contrast-enhanced staining protocol to increase the density of internal soft tissues. High-resolution µCT scans are then collected and segmented to isolate individual muscles. A textural orientation algorithm is then applied to each muscle volume to reconstruct constituent muscle fascicles in three dimensions. Using this technique, we report muscle volume, fascicle length, angle of pennation, and physiological cross-sectional area (PCSA) for each muscle. These data are compared to results collected using traditional dissection of the contralateral muscles. Reconstructions of muscle volume and pennation angle closely correspond to the dissection results. The degree of similarity between measurements of fascicle length and PCSA varies between muscles, with temporalis demonstrating the greatest disparity between techniques; likely reflecting the complex geometry and fascicular arrangement of this muscle. The described technique samples a much larger number of fascicles than had previously been possible and non-destructively investigates the internal architecture of preserved specimens. We conclude that this approach demonstrates great potential for quantifying muscle internal architecture. Anat Rec, 301:363-377, 2018. © 2018 Wiley Periodicals, Inc.

Thalamo-cortical projections to the macaque superior parietal lobule areas PEc and PE.

The exposed surface of the superior parietal lobule in macaque brain contains two architectonically defined areas named PEc and PE. The aim of the present study is the characterization of thalamic afferents of these two areas. For this purpose, retrograde neuronal tracers were injected, or placed in crystal form, in areas PEc and PE. We found that the two areas show a similar pattern of thalamic inputs, mainly originating from Lateral Posterior (LP), Pulvinar (Pul), Ventral Posterior Lateral (VPL), and Ventral Lateral (VL) nuclei, all structures known to be involved in visual, somatosensory, and/or sensorimotor processing. Minor afferents were observed from the Centromedian/Parafascicular complex (CM/PF), Central Lateral (CL), Ventral Anterior (VA), and Medial Dorsal (MD) nuclei. LP and VL were more strongly connected to PEc than to PE, while the other main thalamic inputs to the two areas showed slight differences in strength. The part of the Pul mostly connected with areas PEc and PE was the Medial Pul. No labeled cells were found in the retinotopically organized Lateral and Inferior Pul. In the somatotopically organized VPL and VL nuclei, labeled neurons were mainly found in regions likely to correspond to the trunk and limb representations (in particular the legs). These findings are in line with the sensory-motor nature of areas PEc and PE, and with their putative functional roles, being them suggested to be involved in the preparation and control of limb interaction with the environment, and in locomotion.

Glucocerebrosidase expression patterns in the non-human primate brain.

Glucocerebrosidase (GCase) is a lysosomal enzyme encoded by the GBA1 gene. Mutations in GBA1 gene lead to Gaucher's disease, the most prevalent lysosomal storage disorder. GBA1 mutations reduce GCase activity, therefore promoting the aggregation of alpha-synuclein, a common neuropathological finding underlying Parkinson's disease (PD) and dementia with Lewy bodies. However, it is also worth noting that a direct link between GBA1 mutations and alpha-synuclein aggregation indicating cause and effect is still lacking, with limited experimental evidence to date. Bearing in mind that a number of strategies increasing GCase expression for the treatment of PD are currently under development, here we sought to analyze the baseline expression of GCase in the brain of Macaca fascicularis, which has often been considered as the gold-standard animal model of PD. Although as with other lysosomal enzymes, GCase is expected to be ubiquitously expressed, here a number of regional variations have been consistently found, together with several specific neurochemical phenotypes expressing very high levels of GCase. In this regard, the most enriched expression of GCase was constantly found in cholinergic neurons from the nucleus basalis of Meynert, dopaminergic cells in the substantia nigra pars compacta, serotoninergic neurons from the raphe nuclei, as well as in noradrenergic neurons located in the locus ceruleus. Moreover, it is also worth noting that moderate levels of expression were also found in a number of areas within the paleocortex and archicortex, such as the entorhinal cortex and the hippocampal formation, respectively.

Testicular Fibrous Hypoplasia in Cynomolgus Monkeys ( Macaca fascicularis): An Incidental, Congenital Lesion.

Testicular fibrous hypoplasia is an incidental lesion characterized by replacement of the testicular parenchyma by mature collagen. A retrospective survey of hematoxylin and eosin-stained testicular sections from 722 purpose-bred Asian and 90 Mauritian cynomolgus monkeys from 56 safety assessment studies conducted between 1999 and 2011 was performed. The incidence of the lesion increased markedly over time. No cases occurred between 1999 and 2004. Between 2005 and 2009, the incidence ranged between 8.1% and 11.0% of the monkeys examined and then rose to 26.1% in 2010 and 30.9% in 2011. Overall, the lesion was identified in 10.94% of Asian monkeys with the highest incidence in animals originating from China and Vietnam; severity ranged from minimal to severe and it occurred unilaterally (38.5%) and bilaterally (61.5%). In Mauritian monkeys, the lesion was predominantly minimal in severity, bilateral in distribution, and affected 6.6% of the animals examined. The lesion occurred regardless of sexual maturation status but when present in mature monkeys was often associated with cystic tubular atrophy of the seminiferous epithelium. Based on the morphological characteristics of the lesion and the unilateral/bilateral distribution, the lesion is considered to be a congenital or developmental abnormality.