Morphological Characterization of the Developing Greater Cane Rat (Thryonomys swinderianus) Brain.

Developmental mode along the altricial-precocial spectrum is well known to be influenced by brain development and maturation. The greater cane rat (GCR) is an indigenous precocial African rodent with uncommon phenotypes and life traits. This study was therefore designed to characterize and describe distinctive external developmental features in the prenatal GCR brain across the entire gestational length using the emergence and differentiation of external features of the brain vesicles. Four gross morphometric brain parameters (weight, length, width, and height) were evaluated and expressed as mean ± SEM. Relationship between all brain morphometrics and gestation length were analyzed using one-way ANOVA and linear regression. Developmental milestones in the prenatal GCR were then compared with closely related precocial mammals. The earliest time point with gross observable features in the prenatal GCR brain was at gestation day (GD) 60. The period with the most remarkable gross developmental features was noted between GD80 and GD100. Some of these gross features include differentiation of the cerebellar plate into vermis and lateral lobes, emergence of the piriform lobes, mammillary bodies, colliculi bodies, cerebral peduncles, and primordial pons. By GD130, most gross topographic neural features were already established. Cerebellar lobation and patterning at GD130 were the last recognizable gross developmental features noticed in the prenatal GCR brain. This coincided with the time of first eye opening in the GCR fetus. The developmental pattern observed in the prenatal GCR brain is similar to those noted in precocial rodent like the guinea pig. However, the onset of these milestones was delayed, and their duration was relatively shorter in the GCR. This study provides a frame of baseline reference of morphological brain features in the GCR embryos and fetuses that will be useful for fetal age estimation, for home grown neurodevelopmental and eco-toxicological studies, as this rodent is being proposed as a research model.

Patterning and post-patterning modes of evolutionary digit loss in mammals.

A reduction in the number of digits has evolved many times in tetrapods, particularly in cursorial mammals that travel over deserts and plains, yet the underlying developmental mechanisms have remained elusive. Here we show that digit loss can occur both during early limb patterning and at later post-patterning stages of chondrogenesis. In the 'odd-toed' jerboa (Dipus sagitta) and horse and the 'even-toed' camel, extensive cell death sculpts the tissue around the remaining toes. In contrast, digit loss in the pig is orchestrated by earlier limb patterning mechanisms including downregulation of Ptch1 expression but no increase in cell death. Together these data demonstrate remarkable plasticity in the mechanisms of vertebrate limb evolution and shed light on the complexity of morphological convergence, particularly within the artiodactyl lineage.

Periodic patterns in Rodentia: Development and evolution.

Mammalian periodic pigment patterns, such as spots and stripes, have long interested mathematicians and biologists because they arise from non-random developmental processes that are programmed to be spatially constrained, and can therefore be used as a model to understand how organized morphological structures develop. Despite such interest, the developmental and molecular processes underlying their formation remain poorly understood. Here, we argue that Arvicanthines, a clade of African rodents that naturally evolved a remarkable array of coat patterns, represent a tractable model system in which to dissect the mechanistic basis of pigment pattern formation. Indeed, we review recent insights into the process of stripe formation that were obtained using an Arvicanthine species, the African striped mouse (Rhabdomys pumilio), and discuss how these rodents can be used to probe deeply into our understanding of the factors that specify and implement positional information in the skin. By combining naturally evolved pigment pattern variation in rodents with classic and novel experimental approaches, we can substantially advance our understanding of the processes by which spatial patterns of cell differentiation are established during embryogenesis, a fundamental question in developmental biology.

Cell migration in the developing rodent olfactory system.

The components of the nervous system are assembled in development by the process of cell migration. Although the principles of cell migration are conserved throughout the brain, different subsystems may predominantly utilize specific migratory mechanisms, or may display unusual features during migration. Examining these subsystems offers not only the potential for insights into the development of the system, but may also help in understanding disorders arising from aberrant cell migration. The olfactory system is an ancient sensory circuit that is essential for the survival and reproduction of a species. The organization of this circuit displays many evolutionarily conserved features in vertebrates, including molecular mechanisms and complex migratory pathways. In this review, we describe the elaborate migrations that populate each component of the olfactory system in rodents and compare them with those described in the well-studied neocortex. Understanding how the components of the olfactory system are assembled will not only shed light on the etiology of olfactory and sexual disorders, but will also offer insights into how conserved migratory mechanisms may have shaped the evolution of the brain.

Predicting the energy and protein requirements of the pregnant grasscutter (Thryonomys swinderianus, Temminck) using the changes in weight and composition of the foetus and associated tissues of pregnancy.

Eighteen (18) gravid uteri collected from hunters' kills during February and March 2008 were used to investigate growth and accretion of energy and protein in the gravid uterus during pregnancy in the grasscutter. Each gravid uterus was separated into foetal, placental, empty uterus and foetal fluids (amniotic and allantoic fluids combined) components. Individual tissues were weighed and analyzed for dry matter (DM), ash, crude protein (CP) and crude fat. Regression equations were fitted to the weights of each component to describe the effects of gestational age. The DM, protein and energy contents of individual uterine tissues, with the exception of the DM of the foetal fluids, increased significantly with advancing gestation age so that the DM of the gravid uterus increased ~20-fold from days 45 to 152 (3.85 v. 76.24 g). However, the average elemental N in the foetus was 8.3 g N/100 g. Percentage DM did not change with advancing gestation age. Fat content of the foetal grasscutter was 2.79 %. Changes in foetal protein and energy fitted a multi-phasic regression that consisted of two linear equations. Similarly, changes in foetal fat and water contents fitted a multi-phasic regression that consisted of two linear equations, indicating that protein and energy growth accelerated after days 95 and 97 of gestation, respectively, with increases in fat and water accelerating after days 127 and 96 of gestation, respectively. Foetal protein and energy accretions were 0.04 g day(-1) and 0.001 MJ day(-1) before days 95 and 97 of gestation, respectively, and increased to 0.31 g day(-1) and 0.009 MJ day(-1) after days 95 and 97 of gestation, respectively. Fat and water accretions were also 0.012 g day(-1) and 0.15 ml day(-1) before days 127 and 96 of gestation, respectively, and increased to 0.104 g day(-1) and 1.29 ml day(-1) after the respective days of gestation. Protein needs for tissue protein gain increased ~8-fold after day 95 of gestation. Results of this study indicate that the growth of the foetus and foetal tissues occur at different rates during gestation and points to a two-phase feeding strategy before and after day 95 of gestation for pregnant grasscutters.

Germ cell differentiation and proliferation in the developing testis of the South American plains viscacha, Lagostomus maximus (Mammalia, Rodentia).

Cell proliferation and cell death are essential processes in the physiology of the developing testis that strongly influence the normal adult spermatogenesis. We analysed in this study the morphometry, the expression of the proliferation cell nuclear antigen (PCNA), cell pluripotency marker OCT-4, germ cell marker VASA and apoptosis in the developing testes of Lagostomus maximus, a rodent in which female germ line develops through abolished apoptosis and unrestricted proliferation. Morphometry revealed an increment in the size of the seminiferous cords with increasing developmental age, arising from a significant increase of PCNA-positive germ cells and a stable proportion of PCNA-positive Sertoli cells. VASA showed a widespread cytoplasmic distribution in a great proportion of proliferating gonocytes that increased significantly at late development. In the somatic compartment, Leydig cells increased at mid-development, whereas peritubular cells showed a stable rate of proliferation. In contrast to other mammals, OCT-4 positive gonocytes increased throughout development reaching 90% of germ cells in late-developing testis, associated with a conspicuous increase in circulating FSH from mid- to late-gestation. TUNEL analysis was remarkable negative, and only a few positive cells were detected in the somatic compartment. These results show that the South American plains viscacha displays a distinctive pattern of testis development characterized by a sustained proliferation of germ cells throughout development, with no signs of apoptosis cell demise, in a peculiar endocrine in utero ambiance that seems to promote the increase of spermatogonial number as a primary direct effect of FSH.

The developing ovary of the South American plains vizcacha, Lagostomus maximus (Mammalia, Rodentia): massive proliferation with no sign of apoptosis-mediated germ cell attrition.

Apoptosis-dependent massive germ cell death is considered a constitutive trait of the developing mammalian ovary that eliminates 65-85% of the germinal tissue depending on the species. After birth and during adult lifetime, apoptotic activity moves from the germ cell proper to the somatic compartment, decimating germ cells through follicular atresia until the oocyte reserve is exhausted. In contrast, the South American rodent Lagostomus maximus shows suppressed apoptosis-dependent follicular atresia in the adult ovary, with continuous folliculogenesis and massive polyovulation, which finally exhausts the oocyte pool. The absence of follicular atresia in adult L. maximus might arise from a failure to move apoptosis from the germinal stratum to the somatic compartment after birth or being a constitutive trait of the ovarian tissue with no massive germ cell degeneration in the developing ovary. We tested these possibilities by analysing oogenesis, expression of germ cell-specific VASA protein, apoptotic proteins BCL2 and BAX, and DNA fragmentation by TUNEL assay in the developing ovary of L. maximus. Immunolabelling for VASA revealed a massive and widespread colonisation of the ovary and proliferation of germ cells organised in nests that disappeared at late development when folliculogenesis began. No sign of germ cell attrition was found at any time point. BCL2 remained positive throughout oogenesis, whereas BAX was slightly detected in early development. TUNEL assay was conspicuously negative throughout the development. These results advocate for an unrestricted proliferation of germ cells, without apoptosis-driven elimination, as a constitutive trait of L. maximus ovary as opposed to what is normally found in the developing mammalian ovary.

Investigating gradients of gene expression involved in early human cortical development.

The division of the neocortex into functional areas (the cortical map) differs little between individuals, although brain lesions in development can lead to substantial re-organization of regional identity. We are studying how the cortical map is established in the human brain as a first step towards understanding this plasticity. Previous work on rodent development has identified certain transcription factors (e.g. Pax6, Emx2) expressed in gradients across the neocortex that appear to control regional expression of cell adhesion molecules and organization of area-specific thalamocortical afferent projections. Although mechanisms may be shared, the human neocortex is composed of different and more complex local area identities. Using Affymetrix gene chips of human foetal brain tissue from 8 to 12.5 post-conceptional weeks [PCW, equivalent to Carnegie stage (CS) 23, to Foetal stage (F) 4], human material obtained from the MRC-Wellcome Trust Human Developmental Biology Resource (http://www.hdbr.org), we have identified a number of genes that exhibit gradients along the anterior-posterior axis of the neocortex. Gene probe sets that were found to be upregulated posteriorally compared to anteriorally, included EMX2, COUPTFI and FGF receptor 3, and those upregulated anteriorally included cell adhesion molecules such as cadherins and protocadherins, as well as potential motor cortex markers and frontal markers (e.g. CNTNAP2, PCDH17, ROBO1, and CTIP2). Confirmation of graded expression for a subset of these genes was carried out using real-time PCR. Furthermore, we have established a dissociation cell culture model utilizing tissue dissected from anteriorally or posteriorally derived developing human neocortex that exhibits similar gradients of expression of these genes for at least 72 h in culture.

A Comparative Approach of Cellular Reprogramming in the Rodentia Order.

Cellular reprogramming mainly involves induction of reactivation of genes responsible for nuclear plasticity, a process that can be performed in vitro through production of cloned embryos by somatic cell nuclear transfer or by induction of cells into the pluripotent state through exogenous transcription factor expression. While these techniques are already well known and utilized in mice and rats, their application in other rodent species would be greatly beneficial, especially for conservation purposes. Within the diverse Rodentia order, wild species stand out as they play an important role in balancing the ecosystem by facilitating seed diversion, soil aeration, and consequently, reforestation. Many of these species are currently approaching extinction, and application of techniques, such as nuclear reprogramming, aimed at species conservation and multiplication and to produce stem cells is of interest. Thus, in this review, we aimed to present the evolution and success of nuclear reprogramming, mainly highlighting its potential application for the conservation of wild rodents.

Early and natural embryonic death in Lagostomus maximus: Association with the uterine glands, vasculature, and musculature.

The uterus is an organ with great plasticity due to the morphological and physiological changes it experiences during the estrous cycle and pregnancy. In mammals, pregnancy requires diverse sex hormones, growth factors and cytokines, among others, for promoting uterine remodeling to favor implantation, placentation, and embryo/fetus survival and growth. The hystricognathi rodent Lagostomus maximus (plains viscacha) has a high rate of embryonic resorption. The cranial and middle implants are reabsorbed 25-35 days after intercourse while the caudal embryos continue with their development until two precocial offspring are born. So far, no uterine studies of non-pregnant L. maximus females were performed to determine the possible existence of variations in the organ that could be related to the differential survival of the implants. We used ultrasonography, as well as morphological, morphometric, histochemical, lectinhistochemical, and immunohistochemical methods to study differences in the uterine glands (area), vasculature (area), and musculature (thickness) along the uterine horns in non-pregnant females. Along the uterus, all these structures were in more advanced developmental condition in the caudal region as compared to more anterior positions. These regional variations could be decisive in explaining the reason why only caudal implantations come to term. In contrast, no differences in the in the luminal and glandular epithelial cells, nor in the degree of cell proliferation and apoptosis, and hormonal receptor staining were found. These parameters could be related to implantation along the uterine horns, but not to the differential survival of the implants.

Reduced water intake: Implications for rodent developmental and reproductive toxicity studies.

In developmental and reproductive toxicity studies, drinking water is a common means of delivering the test agent. Reduced consumption of toxicant-containing water raises questions about indirect effects of reduced maternal fluid consumption resulting from unpalatability, versus direct effects of the test compound. Issues to consider include: objective assessment of dehydration and thirst, the relative contributions of innate and learned behaviors to drinking behavior and flavor preference, and the objective assessment of physiologic stress. Not only do lab animals under ad lib conditions consume more water than the minimum required to maintain fluid balance, animals faced with water restriction have substantial physiologic capacity for protection of metabolic processes. Measures of blood biochemistry can provide quantifiable, objective indications of fluid balance, but changes in these parameters could result from other causes such as effects of a test toxicant. Consummatory behaviors in response to perceived need are highly influenced by learning. Hence, the drinking behavior, water intake, and flavor acceptance/preference of animals used in toxicology experiments could be subject to learning experiences with the test compound. Physiological symptoms of stress produced by water deprivation may be distinguishable from the symptoms associated with other generalized stressors, such as food deprivation, but doing so may be beyond the scope of most developmental or reproductive toxicity studies. Use of concurrent controls, paired to test groups for water consumption, could help distinguish between the direct effects of a test toxicant as opposed to effects of reduced water consumption alone.

Developmental horizons in the pre-natal development of the Greater cane rat (Thryonomys swinderianus).

The Greater cane rat (GCR, Thyronomys swinderianus) is a precocial rodent predominantly found within Africa. Economic and scientific interests have led to several research efforts towards the domestication and better understanding of the biology and development of this rodent. Despite these efforts, information on the pre-natal development of this rodent is currently lacking. This study characterises distinct developmental milestones including skin pigmentation, emergence and distributions of hairs, calvarium consistency, teeth eruption, development of appendages, sensory organs and external genitalia in the pre-natal GCR and assesses quantitative body parameters, that is body weight, body and crown-rump lengths across its entire gestation length (gestation days [GDs] 10-140). Using these external features, we provide baseline reference ontogenetic scales for GCR embryos and fetuses, employable for stage, age and sex estimation of the pre-natal GCR in future studies. We observed that the first evidence of an embryo was not seen before the end of the first trimester (GD50) and that the late second trimester (GD80-GD100) marks the transition from embryogenesis to fetogenesis in the GCR. As both events occur at a much later developmental time point when compared to precocial non-rodents including human, sheep and pig and slightly later when compared to other precocial rodents such as guinea pig, our data provide first indication that the pre-natal GCR development might be associated with a reproductive delay. Together, this study expands our knowledge of the development and biology of the GCR, which will improve reproductive and breeding management, and native species conservation of this hystricomorph mammal.

Diverse developmental strategies of X chromosome dosage compensation in eutherian mammals.

In eutherian mammals, dosage compensation arose to balance X-linked gene expression between sexes and relatively to autosomal gene expression in the evolution of sex chromosomes. Dosage compensation occurs in early mammalian development and comprises X chromosome upregulation and inactivation that are tightly coordinated epigenetic processes. Despite a uniform principle of dosage compensation, mechanisms of X chromosome inactivation and upregulation demonstrate a significant variability depending on sex, developmental stage, cell type, individual, and mammalian species. The review focuses on relationships between X chromosome inactivation and upregulation in mammalian early development.

Progressive loss of PAX6, TBR2, NEUROD and TBR1 mRNA gradients correlates with translocation of EMX2 to the cortical plate during human cortical development.

The transcription factors Emx2 and Pax6 are expressed in the proliferating zones of the developing rodent neocortex, and gradients of expression interact in specifying caudal and rostral identities. Pax6 is also involved in corticoneurogenesis, being expressed by radial glial progenitors that give rise to cells that also sequentially express Tbr2, NeuroD and Tbr1, genes temporally downstream of Pax6. In this study, using in situ hybridization, we analysed the expression of EMX2, PAX6, TBR2, NEUROD and TBR1 mRNA in the developing human cortex between 8 and 12 postconceptional weeks (PCW). EMX2 mRNA was expressed in the ventricular (VZ) and subventricular zones (SVZ), but also in the cortical plate, unlike in the rodent. However, gradients of expression were similar to that of the rodent at all ages studied. PAX6 mRNA expression was limited to the VZ and SVZ. At 8 PCW, PAX6 was highly expressed rostrally but less so caudally, as has been seen in the rodent, however this gradient disappeared early in corticogenesis, by 9 PCW. There was less restricted compartment-specific expression of TBR2, NEUROD and TBR1 mRNA than in the rodent, where the gradients of expression were similar to that of PAX6 prior to 9 PCW. The gradient disappeared for TBR2 by 10 PCW, and for NEUROD and TBR1 by 12 PCW. These data support recent reports that EMX2 but not PAX6 is more directly involved in arealization, highlighting that analysis of human development allows better spatio-temporal resolution than studies in rodents.

Chorioallantoic and yolk sac placentation in the dassie rat Petromus typicus and its significance for the evolution of hystricognath rodents.

Placental characters are most important in understanding the evolutionary history of hystricognath rodents of which some act as animal models for human pregnancy. The data available deal mostly with species native to South America, but the current paper presents novel findings on chorioallantoic and yolk sac placentation in an Old World hystricognath and discusses its significance for the evolution of the group. Several hystricognath stem species characters are verified for Petromus, such as the unique trophoblast growth pattern within the chorioallantoic placenta. Subsequently, a novel set of characters belonging to the visceral yolk sac is added to the stem species pattern of the group. The nourishment of the embryo is facilitated by an inverted visceral yolk sac placenta from early pregnancy onward, later complemented by the chorioallantoic placenta. About mid term, the visceral yolk sac becomes partly folded and attached to the parietal yolk sac cover of the chorioallantoic placenta, suggesting a functional shift to the transfer of substances between the two placental types. Thus, the chorioallantoic and yolk sac placenta collaborate in nurturing the embryo. This apparently represents an evolutionary transformation along the stem lineage of hystricognaths.

Identification of rCop-1, a new member of the CCN protein family, as a negative regulator for cell transformation.

By using a model system for cell transformation mediated by the cooperation of the activated H-ras oncogene and the inactivated p53 tumor suppressor gene, rCop-1 was identified by mRNA differential display as a gene whose expression became lost after cell transformation. Homology analysis indicates that rCop-1 belongs to an emerging cysteine-rich growth regulator family called CCN, which includes connective-tissue growth factor, CYR61, CEF10 (v-src inducible), and the product of the nov proto-oncogene. Unlike the other members of the CCN gene family, rCop-1 is not an immediate-early gene, it lacks the conserved C-terminal domain which was shown to confer both growth-stimulating and heparin-binding activities, and its expression is lost in cells transformed by a variety of mechanisms. Ectopic expression of rCop-1 by retroviral gene transfers led to cell death in a transformation-specific manner. These results suggest that rCop-1 represents a new class of CCN family proteins that have functions opposing those of the previously identified members.

The development of mastication in rodents: from neurons to behaviors.

The development of suckling behavior is a fundamental characteristic of mammalian development. The occurrence of this behavior across mammals allows us to extrapolate information from animal models to better understand normal and abnormal masticatory development in infants. This review focuses on prenatal cell, molecular, and morphological changes in rat and/or mouse masticatory muscles, trigeminal motoneurons (Mo5) and mesencephalic trigeminal neurons (Me5) that accompany the development of suckling behavior. A special emphasis is placed on N-methyl-d-aspartate (NMDA) receptor subunit expression because of the important role that NMDA receptors play in the production of rhythmical jaw movements and neuronal development. Prenatally the timing of NMDA subunit changes follows neuromuscular junction formation in masticatory muscles, and is coincident with the emergence of rhythmical jaw movements and in vitro rhythmical trigeminal activity. Our data suggest that NMDA receptor subunit changes in Mo5 and Me5 are synchronized with the emergence of rhythmical jaw movements and trigeminal motor activity.

The patterning and onset of opsin expression in vertebrate retinae.

A fascinating area of current research for developmental biologists concerns the patterning of complex tissues. The distribution of photoreceptors across the vertebrate retina is an excellent example of patterning in a tractable model system. Recent studies defining photoreceptor distribution in developing and mature tissue have set the stage for mechanistic studies of the control of patterning.

Prenatal development of hypothalamic neuropeptide systems in the nonhuman primate.

In the rodent, arcuate nucleus of the hypothalamus (ARH)-derived neuropeptide Y (NPY) and proopiomelanocortin (POMC) neurons have efferent projections throughout the hypothalamus that do not fully mature until the second and third postnatal weeks. Since this process is likely completed by birth in primates we characterized the ontogeny of NPY and melanocortin systems in the fetal Japanese macaque during the late second (G100), early third (G130) and late third trimesters (G170). NPY mRNA was expressed in the ARH, paraventricular nucleus (PVH), and dorsomedial nucleus of the hypothalamus (DMH) as early as G100. ARH-derived NPY projections to the PVH were initiated at G100 but were limited and variable; however, there was a modest increase in density and number by G130. ARH-NPY/agouti-related peptide (AgRP) fiber projections to efferent target sites were completely developed by G170, but the density continued to increase in the postnatal period. In contrast to NPY/AgRP projections, alphaMSH fibers were minimal at G100 and G130 but were moderate at G170. This study also revealed several significant species differences between rodent and the nonhuman primate (NHP). There were few NPY/catecholamine projections to the PVH and ARH prior to birth, while projections were increased in the adult. A substantial proportion of the catecholamine fibers did not coexpress NPY. In addition, cocaine and amphetamine-related transcript (CART) and alpha-melanocyte stimulating hormone (alphaMSH) were not colocalized in fibers or cell bodies. As a consequence of the prenatal development of these neuropeptide systems in the NHP, the maternal environment may critically influence these circuits. Additionally, because differences exist in the neuroanatomy of NPY and melanocortin circuitry the regulation of these systems may be different in primates than in rodents.

The subplacenta of the red-rumped agouti (Dasyprocta leporina L).

BACKGROUND: Hystricognath rodents have a lobed placenta, comprising labyrinthine exchange areas and interlobular trophoblast. These correspond to the labyrinthine and spongy zones of other rodent placentae. Beneath them, however, is a structure unique to hystricognath rodents called the subplacenta. We here describe the subplacenta of the red-rumped agouti and examine the possible functional correlates of this structure. METHODS: Placentae were collected from early in midgestation to near term of pregnancy and examined by standard histological techniques, immunohistochemistry and transmission electron microscopy. In addition, to study the microvasculature of the subplacenta, vessel casts were inspected by scanning electron microscopy. RESULTS: In the subplacenta, lamellae of connective tissue support a layer of mononuclear cytotrophoblast cells. Beneath this is found syncytiotrophoblast. Clusters of multinuclear giant cells occur in the transition zone between the subplacenta and decidua. There are prominent intercellular spaces between the cytotrophoblast cells. The basal membrane of these cells is often close to fetal blood vessels. The syncytiotrophoblast surrounds an extensive system of lacunae. Microvilli project into these lacunae from the plasma membrane of the syncytiotrophoblast. The syncytial cytoplasm contains electron-dense granules. This is probably the amylase-resistant PAS-positive material identified by histochemistry. The subplacenta is supplied entirely from the fetal circulation. Within it the vessels pursue a tortuous course with sinusoidal dilatations and constrictions. CONCLUSION: The functions that have been attributed to the subplacenta include hormone production. Our findings are consistent with this interpretation, but suggest that hormone secretion is directed towards the fetal circulation rather than the maternal tissues.