Cytogenetically Elusive Sex Chromosomes in Scincoidean Lizards

The lizards of the species-rich clade Scincoidea including cordylids, gerrhosaurids, skinks, and xantusiids, show an almost cosmopolitan geographical distribution and a remarkable ecological and morphological divergence. However, previous studies revealed limited variability in cytogenetic traits. The sex determination mode was revealed only in a handful of gerrhosaurid, skink, and xantusiid species, which demonstrated either ZZ/ZW or XX/XY sex chromosomes. In this study, we explored the karyotypes of six species of skinks, two species of cordylids, and one gerrhosaurid. We applied conventional and molecular cytogenetic methods, including C-banding, fluorescence in situ hybridization with probes specific for telomeric motifs and rDNA loci, and comparative genomic hybridization. The diploid chromosome numbers are rather conserved among these species, but the chromosome morphology, the presence of interstitial telomeric sequences, and the topology of rDNA loci vary significantly. Notably, XX/XY sex chromosomes were identified only in Tiliqua scincoides, where, in contrast to the X chromosome, the Y chromosome lacks accumulations of rDNA loci. We confirm that within the lizards of the scincoidean clade, sex chromosomes remained in a generally poor stage of differentiation.     

DNA Organization along Pachytene Chromosome Axes and Its Relationship with Crossover Frequencies

During meiosis, the number of crossovers vary in correlation to the length of prophase chromosome axes at the synaptonemal complex stage. It has been proposed that the regular spacing of the DNA loops, along with the close relationship of the recombination complexes and the meiotic axes are at the basis of this covariation. Here, we use a cytogenomic approach to investigate the relationship between the synaptonemal complex length and the DNA content in chicken oocytes during the pachytene stage of the first meiotic prophase. The synaptonemal complex to DNA ratios of specific chromosomes and chromosome segments were compared against the recombination rates obtained by MLH1 focus mapping. The present results show variations in the DNA packing ratios of macro- and microbivalents and also between regions within the same bivalent. Chromosome or chromosome regions with higher crossover rates form comparatively longer synaptonemal complexes than expected based on their DNA content. These observations are compatible with the formation of higher number of shorter DNA loops along meiotic axes in regions with higher recombination levels.     

Use of predator odors as repellents to reduce feeding damage by herbivores

This study investigated the influence of the major anal-gland compounds from the stoat (Mustela erminea) and fecal and urine compounds from the red fox (Vulpes vulpes) in generating an avoidance response by montane voles (Microtus montanus), as well as suppressing feeding by montane and meadow (M. pennsylvanicus) voles on apple trees in orchards. In trap bioassays, a 11 mixture of 2-propylthietane and 3-propyl-1,2-dithiolane significantly reduced vole captures. Other mixtures of stoat compounds reduced the number of new voles captured but not total individuals. 2,5-Dihydro-2,4,5-trimethylthiazoline, a component of fox feces, significantly reduced vole captures in one of two bioassays. Deer mice (Peromyscus maniculatus) did not show a negative response to any predator odor. In overwinter field bioassays, mixtures of 2-propylthietane and 3-propyl-1,2-dithiolane clearly reduced vole feeding on apple trees in four test blocks. 2,5-Dihydro-2,4,5-trimethylthiazoline and a synthetic fox urine mixture also significantly reduced vole attack in respective orchard blocks. Similarly, the intensity of vole feeding, in terms of amount of bark and vascular tissues removed from trees, was reduced by 60% to 97% in predator odor treatments compared with the control. Our study reports the first long-term (four to five months) use of synthetic semiochemicals as area repellents for crop protection from vole feeding damage.     

Comparison of release devices for stoat (Mustela ermined) semiochemicals used as montane vole (Microtus montanus) repellents

Fenced enclosures were used to simulate peak populations of montane voles (Microtus montanus) for field bioassays of a stoat (Mustela erminea) scent mixture in various controlled-release devices. A 1 1 mixture of 2-propylthietane and 3-propyl-1,2-dithiolane was dispensed in capillary tubes, clay pellets (activated alumina), rubber septa, and plastic rope. Release devices were placed near, or attached to, young apple trees planted in blocks in two enclosures containing high populations of voles. The stoat scent mixture in rubber septa and plastic rope significantly reduced vole attack of trees compared with a control, pellets, and capillary tubes. In terms of complete girdling (tree mortality), this odor in septa and rope also significantly reduced feeding compared with the control, and the rubber septa formulation was more effective than either the pellets or capillary tubes. Fenced populations of voles may be used effectively to conduct field bioassays during low years in vole cycles. Variations of release devices (with this stoat scent mixture) based on rubber septa for broadcast application to depress vole population density and survival, and plastic rope attached to individual trees to generate an avoidance response, should be used for forest and agricultural crop protection.     

Bark consumption by voles in relation to mineral contents

Recent field experiments with impregnated wooden sticks have demonstrated a pronounced use by small rodents of mineral supplies, especially sodium, and such findings seemed related to vole damage to forestry seedlings. Consumption of the bark of experimentally introduced aspen twigs and of sodium-impregnated sticks by voles (mainly or onlyMicrotus agrestis) correlated significantly on clear-cuts but not on unmanipulated abandoned fields. Such a correlation appeared when abandoned fields were cut continuously in summer. At vole peak densities, bark of pine seedlings experimentally fertilized with sodium was consumed but not bark of seedlings fertilized with calcium or control seedlings. Field pine seedlings attacked by voles had significantly higher levels of calcium, sodium, and phosphorus than the nearest untouched seedling. However, sodium and phosphorus contents correlated strongly. Sodium and calcium supply to voles in laboratory feeding trials did not diminish the moderate interest in pine bark. Such conditions are, however, assumed to mimic a situation of bark sampling in low-density populations. Sodium, and possibly also calcium, requirements are concluded to be partial determinants of the destructive bark consumption by voles at the peaks of their multiannual population cycles.     

Alterations in Tau Protein Level and Phosphorylation State in the Brain of the Autistic-Like Rats Induced by Prenatal Exposure to Valproic Acid

Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by deficient social interaction and communication besides repetitive, stereotyped behaviours. A characteristic feature of ASD is altered dendritic spine density and morphology associated with synaptic plasticity disturbances. Since microtubules (MTs) regulate dendritic spine morphology and play an important role in spine development and plasticity the aim of the present study was to investigate the alterations in the content of neuronal α/β-tubulin and Tau protein level as well as phosphorylation state in the valproic acid (VPA)-induced rat model of autism. Our results indicated that maternal exposure to VPA induces: (1) decrease the level of α/β-tubulin along with Tau accumulation in the hippocampus and cerebral cortex; (2) excessive Tau phosphorylation and activation of Tau-kinases: CDK5, ERK1/2, and p70S6K in the cerebral cortex; (3) up-regulation of mTOR kinase-dependent signalling in the hippocampus and cerebral cortex of adolescent rat offspring. Moreover, immunohistochemical staining showed histopathological changes in neurons (chromatolysis) in both analysed brain structures of rats prenatally exposed to VPA. The observed changes in Tau protein together with an excessive decrease in α/β-tubulin level may suggest destabilization and thus dysfunction of the MT cytoskeleton network, which in consequence may lead to the disturbance in synaptic plasticity and the development of autistic-like behaviours.     

Lespedeza phenolics andPenstemon alkaloids: Effects on digestion efficiencies and growth of voles

Lespedeza cuneata contains high levels of phenolics and is a common food plant of the meadow vole (Microtus pennsylvanicus);Penstemon digitalis contains substantial quantities of alkaloids and is a common food plant of meadow voles and prairie voles (M. ochrogaster). We investigated the palatability of these plants and the effects of their secondary compounds on the digestion efficiencies and growth of both species of voles. Voles ate very little of either plant when alternative food was present. Phenolics and alkaloids were extracted from the plants, incorporated into separate artificial diets, and fed to weanling voles for three weeks.Lespedeza phenolics reduced the growth of meadow voles but not prairie voles throughout the feeding trial. These compounds disrupted digestion, reducing protein digestibility by more than half.Penstemon alkaloids lowered only the initial growth rates of prairie voles and had no significant effect on meadow voles. Prairie voles tended to increase food consumption rates on bothLespedeza andPenstemon diets. This response offset some of the decrease in digested protein intake in the first case, and offset the increased metabolic cost of processing the diet in the latter case. Our results indicate that althoughLespedeza phenolics andPenstemon alkaloids do influence consumption and digestibility of artificial diets, they do not greatly reduce the performance (i.e., growth or survival) of voles when consumed at levels generally observed for wild voles. However, these compounds probably do place an upper limit on the amount of freshLespedeza andPenstemon that can be consumed and contribute to the generalist feeding strategy of voles.     

The Meiosis-Specific Crs1 Cyclin Is Required for Efficient S-Phase Progression and Stable Nuclear Architecture

Cyclins and CDKs (Cyclin Dependent Kinases) are key players in the biology of eukaryotic cells, representing hubs for the orchestration of physiological conditions with cell cycle progression. Furthermore, as in the case of meiosis, cyclins and CDKs have acquired novel functions unrelated to this primal role in driving the division cycle. Meiosis is a specialized developmental program that ensures proper propagation of the genetic information to the next generation by the production of gametes with accurate chromosome content, and meiosis-specific cyclins are widespread in evolution. We have explored the diversification of CDK functions studying the meiosis-specific Crs1 cyclin in fission yeast. In addition to the reported role in DSB (Double Strand Break) formation, this cyclin is required for meiotic S-phase progression, a canonical role, and to maintain the architecture of the meiotic chromosomes. Crs1 localizes at the SPB (Spindle Pole Body) and is required to stabilize the cluster of telomeres at this location (bouquet configuration), as well as for normal SPB motion. In addition, Crs1 exhibits CDK(Cdc2)-dependent kinase activity in a biphasic manner during meiosis, in contrast to a single wave of protein expression, suggesting a post-translational control of its activity. Thus, Crs1 displays multiple functions, acting both in cell cycle progression and in several key meiosis-specific events.     

Searching for Major Urinary Proteins (MUPs) as Chemosignals in Urine of Subterranean Rodents

Chemosensory information mediates behavior in many rodent genera. Major Urinary Proteins (MUPs) facilitate chemical communication in some species of mice. We sought to demonstrate the importance of MUPs in chemosignaling across a range of rodent genera that live in different habitats and social structures. We analyzed urine from three subterranean rodent genera from different continents, and with diverse social systems: eusocial Zambian mole-rats (Fukomys), solitary Israeli blind mole rats (Spalax), and social Chilean coruros (Spalacopus). 2D gel electrophoresis revealed low levels of protein, with sequences similar to aphrodisin, in Fukomys mole-rat urine, but no MUPs in urine of any of the studied species. Previous research demonstrated that subjects from the tested genera responded differentially to odors indicating transmission of individuality, family/colony or population, species, and reproductive state in secretions and excretions of conspecifics. This extends, to subterranean rodents, the evidence that rodent species can successfully transmit and receive chemosignals without the necessity of MUPs.     

Lichen Compounds Restrain Lichen Feeding by Bank Voles <Emphasis Type="Italic">(Myodes glareolus</Emphasis>)

Some lichen compounds are known to deter feeding by invertebrate herbivores. We attempted to quantify the deterring efficiency of lichen compounds against a generalist vertebrate, the bank vole (Myodes glareolus). In two separate experiments, caged bank voles had the choice to feed on lichens with natural or reduced concentrations of secondary compounds. We rinsed air-dry intact lichens in 100% acetone to remove extracellular compounds non-destructively. In the first experiment, pairs of control and rinsed lichen thalli were hydrated and offered to the bank voles. Because the lichens desiccated fast, we ran a second experiment with pairs of ground control and compound-deficient thalli, each mixed with water to porridge. Eight and six lichen species were tested in the first and second experiment, respectively. In the first, bank voles preferred compound-deficient thalli of Cladonia stellaris and Lobaria pulmonaria, but did not discriminate between the other thallus pairs. This was likely a result of deterring levels of usnic and stictic acid in the control thalli. When lichens were served as porridge, significant preference was found for acetone-rinsed pieces of Cladonia arbuscula, C. rangiferina, Platismatia glauca, and Evernia prunastri. The increased preference was caused mainly by lower consumption of control thalli. Grinding and mixing of thallus structures prevented bank voles from selecting thallus parts with lower concentration of secondary compounds and/or strengthened their deterring capacity. We conclude that some lichen secondary compounds deter feeding by bank voles.     

Variants of STAR,AMH and ZFPM2/FOG2 May Contribute towards the Broad Phenotype Observed in 46,XY DSD Patients with Heterozygous Variants of NR5A1

Variants of NR5A1 are often found in individuals with 46,XY disorders of sex development (DSD) and manifest with a very broad spectrum of clinical characteristics and variable sex hormone levels. Such complex phenotypic expression can be due to the inheritance of additional genetic hits in DSD-associated genes that modify sex determination, differentiation and organ function in patients with heterozygous NR5A1 variants. Here we describe the clinical, biochemical and genetic features of a series of seven patients harboring monoallelic variants in the NR5A1 gene. We tested the transactivation activity of novel NR5A1 variants. We additionally included six of these patients in a targeted diagnostic gene panel for DSD and identified a second genetic hit in known DSD-causing genes STAR, AMH and ZFPM2/FOG2 in three individuals. Our study increases the number of NR5A1 variants related to 46,XY DSD and supports the hypothesis that a digenic mode of inheritance may contribute towards the broad spectrum of phenotypes observed in individuals with a heterozygous NR5A1 variation.     

Susceptibility to vole attacks due to bark phenols and terpenes inPinus contorta provenances introduced into Sweden

Seedlings of North AmericanPinus contorta introduced to Sweden and Finland are severely gnawed by voles, e.g.,Microtus agrestis. The level of damage varies between provenances. Chemical analyses of various phenolic compounds, monoterpenes, and resin acids of different provenances and of damaged and undamaged stems showed that some phenolic substances in the bark increased after damage without deterring the animals, that monoterpene differences between provenances were not related to vole damage, and that certain resin acids occurred in larger amounts in lightly than in severely damaged provenances. Levopimaric and neoabietic acid, and possibly abietic acid, may be important for a partial resistance to vole browsing.     

Molecular diagnosis of 5α-reductase type II deficiency in Brazilian siblings with 46,XY disorder of sex development

The steroid 5α-reductase type II enzyme catalyzes the conversion of testosterone (T) to dihydrotestosterone (DHT), and its deficiency leads to undervirilization in 46,XY individuals, due to an impairment of this conversion in genital tissues. Molecular analysis in the steroid 5α-reductase type II gene (SRD5A2) was performed in two 46,XY female siblings. SRD5A2 gene sequencing revealed that the patients were homozygous for p.Gln126Arg missense mutation, which results from the CGA > CAA nucleotide substitution. The molecular result confirmed clinical diagnosis of 46,XY disorder of sex development (DSD) for the older sister and directed the investigation to other family members. Studies on SRD5A2 protein structure showed severe changes at NADPH binding region indicating that structural modeling analysis can be useful to evaluate the deleterious role of a mutation as causing 5α-reductase type II enzyme deficiency.     

The Cdc14 Phosphatase Controls Resolution of Recombination Intermediates and Crossover Formation during Meiosis

Meiotic defects derived from incorrect DNA repair during gametogenesis can lead to mutations, aneuploidies and infertility. The coordinated resolution of meiotic recombination intermediates is required for crossover formation, ultimately necessary for the accurate completion of both rounds of chromosome segregation. Numerous master kinases orchestrate the correct assembly and activity of the repair machinery. Although much less is known, the reversal of phosphorylation events in meiosis must also be key to coordinate the timing and functionality of repair enzymes. Cdc14 is a crucial phosphatase required for the dephosphorylation of multiple CDK1 targets in many eukaryotes. Mutations that inactivate this phosphatase lead to meiotic failure, but until now it was unknown if Cdc14 plays a direct role in meiotic recombination. Here, we show that the elimination of Cdc14 leads to severe defects in the processing and resolution of recombination intermediates, causing a drastic depletion in crossovers when other repair pathways are compromised. We also show that Cdc14 is required for the correct activity and localization of the Holliday Junction resolvase Yen1/GEN1. We reveal that Cdc14 regulates Yen1 activity from meiosis I onwards, and this function is essential for crossover resolution in the absence of other repair pathways. We also demonstrate that Cdc14 and Yen1 are required to safeguard sister chromatid segregation during the second meiotic division, a late action that is independent of the earlier role in crossover formation. Thus, this work uncovers previously undescribed functions of the evolutionary conserved Cdc14 phosphatase in the regulation of meiotic recombination.     

DYRK1A Negatively Regulates CDK5-SOX2 Pathway and Self-Renewal of Glioblastoma Stem Cells

Glioblastoma display vast cellular heterogeneity, with glioblastoma stem cells (GSCs) at the apex. The critical role of GSCs in tumour growth and resistance to therapy highlights the need to delineate mechanisms that control stemness and differentiation potential of GSC. Dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) regulates neural progenitor cell differentiation, but its role in cancer stem cell differentiation is largely unknown. Herein, we demonstrate that DYRK1A kinase is crucial for the differentiation commitment of glioblastoma stem cells. DYRK1A inhibition insulates the self-renewing population of GSCs from potent differentiation-inducing signals. Mechanistically, we show that DYRK1A promotes differentiation and limits stemness acquisition via deactivation of CDK5, an unconventional kinase recently described as an oncogene. DYRK1A-dependent inactivation of CDK5 results in decreased expression of the stemness gene SOX2 and promotes the commitment of GSC to differentiate. Our investigations of the novel DYRK1A-CDK5-SOX2 pathway provide further insights into the mechanisms underlying glioblastoma stem cell maintenance.     

The effects of chronic tannic acid intake on prairie vole (Microtus ochrogaster) reproduction

The hypothesis was tested that the reproductive performance of voles would be reduced when fed diets containing tannins either because of increased metabolic rates, decreased intake, or decreased digestive efficiency. We fed a ration containing 4% tannic acid (TA) (dry mass basis) to 24 pair of prairie voles (Microtus ochrogaster) and compared reproductive performance (litter size, birth weights, body mass of the young until weaning, and mass changes in the dams) to that of 24 pair of prairie vole fed a control ration. We also compared the intake rates, digestive efficiency, and metabolic rates [as measured by Vo₂ consumption (cm³/hr)] of dams and young fed both rations. Voles consuming 4% TA diets produced litters of similar size and mass as did voles fed the control ration. Furthermore, the mass of the young of dams fed the tannic acid ration were similar to the young of dams consuming the control ration through day 19 postpartum. However, dams consuming the tannic acid ration lost mass throughout lactation while the control dams maintained mass. Because the Vo₂ rates of both treated and control dams and litters were similar, we conclude increased metabolic costs were not the reason for the observed mass loss but, rather, reduced digestive efficiency, reduced intake of digestible dry mass, and apparent digestible nitrogen.     

Attractiveness of Opposite-Sex Odor and Responses to It Vary with Age and Sex in Meadow Voles (Microtus pennsylvanicus)

Current models suggest that among short-lived mammals, such as rodents, older adults are less attractive to the opposite sex and spend less time associated with the opposite sex than do younger adults. The objective of this study was to test two hypotheses in three different age groups of meadow voles. The first hypothesis is that 3- to 5 month old voles produce scents that are more attractive to opposite sex conspecifics than those of 7- to 12 month old and 14- to 18 month old voles. The second hypothesis is that 3- to 5 month old voles spend more time than either 7- to 12 month old or 14- to 18 month old voles investigating the scents of an opposite sex conspecific. The first experiment shows that when choosing between two conspecifics, females prefer the odor of the older male within each pair and that males prefer the odor of the 7- to 12 month old females to those of either 3- to 5 month old or 14- to 18 month old females. Thus, the data did not support the first hypothesis. The second experiment shows that the 14- to 18 month old males spent more time investigating female odors than did either the 3- to 5 month old or 7- to 12 month old males and that 7- to 12 month old females spent more time investigating male odors than did the 3- to 5 month old and 12- to 18 month old females. These data did not support the second hypothesis. Overall, older adult male meadow voles are more interested in and attractive to females than are younger adult males. The present data raise questions as to whether current models predict the age-related effects on the behavior of short-lived mammals.     

Differences in Vole Preference, Secondary Chemistry and Nutrient Levels Between Naturally Regenerated and Planted Norway Spruce Seedlings

Field voles (Microtus agrestis) cause severe damage to young Norway spruce (Picea abies) plantations during wintertime in Fennoscandia. We experimentally investigated vole preference for winter-dormant, naturally regenerated seedlings; spring-planted seedlings; or autumn-planted seedlings; and how preference corresponds with seedling chemistry. Voles showed the highest preference for autumn-planted seedlings and the second highest for spring-planted seedlings, while naturally regenerated seedlings were avoided. The stems of the autumn-planted seedlings contained higher concentrations of nitrogen and piperidine alkaloids and lower concentrations of stilbenes than did the other groups. In addition to differences between naturally regenerated and planted seedlings, we investigated seasonal differences in naturally regenerated P. abies needle and bark secondary chemistry. While piperidine alkaloid concentrations did not vary with season, the soluble non-tannin phenolics of needles and the condensed tannins of bark were lower in May than in November or January. At the time of planting, the concentration of bark piperidine alkaloids was higher in autumn-planted than in spring-planted seedlings. We detected two alkaloids not previously found in P. abies, 2-methyl-6-propyl-1,6-piperideine and a tentatively identified pinidine-isomer. Our results demonstrate that vole choice of spruce seedlings is promoted by high nitrogen and low stilbene content, both associated with seedlings planted late in the season. As vole damage is linked to seedling chemistry, damage potentially could be mitigated by advancing planting or by manipulating plant chemistry in nurseries.