Morphometric data on pineal gland of loir (Glis glis) and lerot (Elioyms quercinus) during the annual cycle]

The important involution of the pineal gland of Glis glis and Eliomys quercinus during the months of july and august belongs to a polyglandular involution (anterior lobe of the hypophysis, male and female genital glands) characteristic of estivation.     

Cortical control of mastication in cats. 2. Deficits of masticatory movements following a lesion in the motor cortex

Our previous study suggested that area P in the lateral wall of the presylvian sulcus and MA (masticatory cortex) in the rostral part of the orbital gyrus play an important role in execution of mastication. The aim of this present study is to examine if changes in orofacial behaviors and masticatory movements occur in cats with lesions of area P. First, we explored the locations in area P through the use of single unit recording and ICMS (intracortical microstimulation). Since mastication related neurons (MRNs) with the mechanical receptive field (RF) in facial or intraoral region were intermingled in area P, we performed either a partial or entire lesion in area P by injections of 2 microl or 4 microl of 0.1% kainic acid. Cats with the entire lesion in area P showed a decrease of food intake rates associated with abnormal tongue protrusion and wide jaw-opening, fluctuation of masticatory start, and prolongation of masticatory and food intake periods. Abnormal movements of tongue and jaw did not recover, although their prolongation and fluctuation returned to normal levels in one month. On the other hand, all deficits evoked by cats with the partial lesion recovered in about one month. In cats with the partial and entire lesions, masticatory rhythm remained normal. These findings suggest that area P may regulate accurate and suitable tongue and jaw movements during mastication depending on cortical processing.     

Reduced nucleotide variability at an androgen-binding protein locus (Abpa) in house mice: evidence for positive natural selection.

Previous work has shown that the gene for the alpha subunit of androgen-binding protein, Abpa, may be involved in premating isolation between different subspecies of the house mouse, Mus musculus. We investigated patterns of DNA sequence variation at Abpa within and between species of mice to test several predictions of a model of neutral molecular evolution. Intraspecific variation among 10 Mus musculus domesticus alleles was compared with divergence between M. m. domesticus and M. caroli for Abpa and two X-linked genes, Glra2 and Amg. No variation was observed at Abpa within M. m. domesticus. The ratio of polymorphism to divergence was significantly lower at Abpa than at Glra2 and Amg, despite the fact that all three genes experience similar rates of recombination. Interspecific comparisons among M. m. domesticus, Mus musculus musculus, Mus musculus castaneus, Mus spretus, Mus spicilegus, and Mus caroli revealed that the ratio of nonsynonymous substitutions to synonymous substitutions on a per-site basis (Ka/Ks) was generally greater than one. The combined observations of no variation at Abpa within M. m: domesticus and uniformly high Ka/Ks values between species suggest that positive directional selection has acted recently at this locus.     

Effects of alterations in the immunocompetent status of Mus musculus females on the survival of transferred Mus caroli embryos

The role of the immune system in promoting the midterm death of Mus caroli embryos transferred to the Mus musculus uterus was studied in vivo by transferring M. caroli blastocysts to recipients with altered immune status. Transfers of embryos to chimaeric mothers (Mus musculus in equilibrium Mus caroli), which were expected to be tolerant of species antigens, resulted in survival of M. musculus embryos but death of M. caroli embryos. The preferential survival of M. musculus embryos was explained by showing that M. musculus embryos can survive in the M. caroli uterus. Transfers to T cell-deficient mice of genotype nu/nu and to NK cell-deficient mice of genotype bg/bg as well as treatment of normal transfer recipients with Cyclosporin A or anti-Ia antiserum failed to prolong survival. However, immunization of recipients with M. caroli lymphocytes promoted more rapid and uniform failure of the interspecies pregnancy. Cytotoxic cells were detected in the resorbing embryos on Day 10.5 in immune pregnancies and on Day 12.5 in non-immune pregnancies and these cells were promiscuous in their pattern of lysis, showing equal reactivity against M. caroli, transfer recipient and 3rd party target cells. These experiments show that failure of M. caroli embryos in the M. musculus uterus is complex, but probably does not involve responses by classical cytotoxic T lymphocyte or natural killer cell pathways. Participation of the immune system in the resorption process, however, is confirmed and is associated with generation of promiscuous cytolytic cells.     

Molecular cloning and long terminal repeat sequences of intracisternal A-particle genes in Mus caroli

We isolated DNA clones of intracisternal A-particle (IAP) genes from the genome of an Asian wild mouse, Mus caroli. A typical M. caroli IAP gene was 6.5 kilobase pairs in length and had long terminal repeat (LTR) sequences at both ends. The size of the LTR was 345 base pairs in clone L20, and two LTRs at both ends of this clone were linked to directly repeating cellular sequences of 6 base pairs. Each LTR possessed most of the structural features commonly associated with the retrovirus LTR. The restriction map of the M. caroli IAP gene resembled that of Mus musculus, although the M. caroli IAP gene was 0.4 kilobase pairs shorter than the M. musculus IAP gene in two regions. Sequence homology between the M. caroli and M. musculus IAP LTRs was calculated as about 80%, whereas the LTR sequence of the Syrian hamster IAP gene was about 60% homologous to the M. caroli LTR. The reiteration frequency of the M. caroli IAP genes was estimated as 200 to 400 copies per haploid genome, which is at least 10 times the reported value. These results suggest that the IAP genes observed in the genus Mus are present in multiple copies with structures closely resembling the integrated retrovirus gene.     

Development and validation of a mastication simulator

More and more research are being done on food bolus formation during mastication. However, the process of bolus formation in the mouth is difficult to observe. A mastication simulator, the Artificial Masticatory Advanced Machine (AM2) was developed to overcome this difficulty and is described here. Different variables can be set such as the number of masticatory cycles, the amplitude of the mechanical movements simulating the vertical and lateral movements of the human lower jaw, the masticatory force, the temperature of the mastication chamber and the injection and the composition of saliva. The median sizes of the particles collected from the food boluses made by the AM2 were compared with those of human boluses obtained with peanuts and carrots as test foods. Our results showed that AM2 mimicked human masticatory behavior, producing a food bolus with similar granulometric characteristics.     

Trends in the Actions of Mammalian Masticatory Muscles

The actions of the masticatory muscles of a variety of mammalsin which feeding behavior and the configuration of the masticatoryapparatus differ have been reported. The most common approachused in these studies involves (1) obtaining a good anatomicalperception of the musculature, (2) deriving a theoretical modelof the actions of these muscles during jaw movement, and (3)testing this model by recording muscle activity and jaw movementssimultaneously. A catalogue of the activity patterns in eleven species of mammalsduring food reduction reveals certain trends in the actionsof the masticatory muscles. Horizontal jaw movements are generatedprimarily by differential activities of the deep temporalis,superficial masseter, and medial pterygoid. Vertical movementsand the maintenance of tooth to food contact apparently areproduced by action of the superficial temporalis, deep masseter,and zygomaticomandibularis. Thus, horizontal movements are seeminglygenerated by muscles having fibers arranged in marked anteroposteriordirection, whereas vertical movements are generated by muscleshaving more or less vertically arranged fibers. The asymmetry of jaw movement and the muscular activity generatingit suggest that mastication involves an interactionbetween anunbalanced and flexible functional unit (muscles) and a balancedand stable structural unit (skull and teeth). Thus, any unbalancingof the structural unit results in a further unbalancing of themasticatory process.     

Immune presensitization and local intrauterine defences as determinants of success or failure of murine interspecies pregnancies

Putatively immuno-incompetent Mus musculus females exhibited failure to support pregnancy of Mus caroli embryos. These results for M. musculus females (i.e. treated by cyclosporine A, of the nu/nu genotype, and as an interspecies chimaera) can be explained in immunological terms. Mus musculus females possessed pre-sensitized cytotoxic T cells against Mus caroli antigen. Nu/nu mice possessed activated NK cells and macrophages, and selectively discriminated against Mus caroli embryos early in pregnancy unlike normal +/+ females; the requirement for T cells to activate non-specific cytotoxic effector mechanisms was bypassed in nu/nu mice. Mus caroli are not inbred, and interspecies chimaeras which are tolerant of the antigens on the Mus musculus donor strain were not tolerant of cells from unrelated Mus caroli. Interspecies chimaeras also behaved as if they were pre-sensitized to Mus caroli. Our results show that Mus caroli embryos recruit fewer active suppressor cells even when gestating in Mus caroli decidua as compared to Mus musculus embryos in Mus musculus decidua and that the ability of Mus caroli placental cells to directly inhibit cytotoxic effector cell killing was inherently less than the inhibitory activity of placental cells from Mus musculus. Mus caroli embryos therefore appear to be less well defended against maternal immune attack even when gestating in a uterus possessing compatible Mus caroli decidual tissue.     

Morphological demonstration of the failure of Mus caroli trophoblast in the Mus musculus uterus

A histological study of Mus caroli embryos gestating in the Mus musculus uterus was undertaken at Day 8.5 of gestation, 1 day after such embryos are reported to be normal and 1 day before the earliest events associated with death of the xenogeneic embryos. In comparison to control M. caroli embryos recovered from M. caroli and to control M. musculus embryos recovered from M. musculus, the xenogeneically transferred embryos showed intrauterine growth retardation that was associated with trophoblastic insufficiency. Trophoblast cell degeneration was observed, in the absence of lymphocytic infiltration. Therefore, loss of trophoblast cell function rather than lymphocyte-mediated destruction of trophoblast appears to underlie the death of M. caroli embryos in the M. musculus uterus.     

An evolutionary switch in tissue-specific gene expression. Abundant expression of alpha 1-antitrypsin in the kidney of a wild mouse species

alpha 1-Antitrypsin (AT), one of the major proteinase inhibitors in mammalian serum, is generally considered to be synthesized exclusively in the liver. We have found that a wild-derived Mus species, Mus caroli, expresses AT mRNA in kidney at levels approaching that in liver; no other mouse, inbred or wild-derived, exhibits this striking property. Liver and kidney mRNAs from M. caroli encode very similar AT polypeptides that are distinct from that encoded by Mus musculus liver mRNA. In vivo, liver AT is secreted into the bloodstream, while kidney AT, which is processed differently from the liver protein, is excreted into the urine. Analysis of RNA from a hybrid between M. musculus and M. caroli indicates that a cis-acting genetic element may be responsible for the difference in AT expression. Restriction enzyme digestion patterns of AT genomic sequences in M. caroli DNA are considerably different from those in M. musculus; in addition, these sequences are undermethylated in liver DNA from M. musculus and in liver and kidney DNA from M. caroli, reflecting the respective patterns of expression. Further studies of the altered tissue specificity of AT expression that is apparent in these two related species should lead to new insights into the nature and evolution of genetic determinants of tissue-specific phenotypes.     

Tempo and mode of concerted evolution in the L1 repeat family of mice

A 300-bp DNA sequence has been determined for 30 (10 from each of three species of mice) random isolates of a subset of the long interspersed repeat family L1. From these data we conclude that members of the L1 family are evolving in concert at the DNA sequence level in Mus domesticus, Mus caroli, and Mus platythrix. The mechanism responsible for this phenomenon may be either duplicative transposition, gene conversion, or a combination of the two. The amount of intraspecies divergence averages 4.4%, although between species base substitutions accumulate at the rate of approximately 0.85%/Myr to a maximum divergence of 9.1% between M. platythrix and both M. domesticus and M. caroli. Parsimony analysis reveals that the M. platythrix L1 family has evolved into a distinct clade in the 10-12 Myr since M. platythrix last shared a common ancestor with M. domesticus and M. caroli. The parsimony tree also provides a means to derive the average half-life of L1 sequences in the genome. The rates of gain and loss of individual copies of L1 were estimated to be approximately equal, such that approximately one-half of them turn over every 3.3 Myr.      

Insulin secretion and IP levels in two distant lineages of the genus Mus: comparisons with rat islets

Islet responses of two different Mus geni, the laboratory mouse (Mus musculus) and a phylogenetically more ancient species (Mus caroli), were measured and compared with the responses of islets from rats (Rattus norvegicus). A minimal and flat second-phase response to 20 mM glucose was evoked from M. musculus islets, whereas a large rising second-phase response characterized rat islets. M. caroli responses were intermediate between these two extremes; a modest rising second-phase response to 20 mM glucose was observed. Prior, brief stimulation of rat islets with 20 mM glucose results in an amplified insulin secretory response to a subsequent 20 mM glucose challenge. No such potentiation or priming was observed from M. musculus islets. In contrast, M. caroli islets displayed a modest twofold potentiated first-phase response upon subsequent restimulation with 20 mM glucose. Inositol phosphate (IP) accumulation in response to 20 mM glucose stimulation in [(3)H]inositol-prelabeled rat or mouse islets paralleled the insulin secretory responses. The divergence in 20 mM glucose-induced insulin release between these species may be attributable to differences in phospholipase C-mediated IP accumulation in islets.     

Ultrastructural changes in the renal filtration barrier during hibernation of the common dormouse (Eliomys quercinus L.)

We have studied kidney samples of 16 garden dormouses (Eliomys quercinus L.) divided into two groups, 8 hibernating and 8 non-hibernating. Hibernation produces structural modifications in the glomerular ultrafilter: (1) the endothelial pores decrease in number and size; (2) the podocytic food processes increase in number and the slit pores decrease in size; (3) in the basement membrane there are no structural morphological modifications. In short, we could say that the permeability of the glomerular ultrafilter decreases during hibernation. This fact helps to understand the decrease in the rate of urine formation that takes place in the presence of a low body temperature and a metabolic depression.     

Specificity of androgen resistance inMus caroli kidney

Androgen controls the expression of beta-glucuronidase and several other proteins in the kidney of the standard laboratory mouse, Mus musculus. Other species within the genus Mus exhibit a variety of response patterns for kidney beta-glucuronidase and other markers of androgen action. We have investigated the mechanism of androgen action in M. caroli, a Mus species that does not produce beta-glucuronidase in response to testosterone. The failure of testosterone to induce beta-glucuronidase in M. caroli females cannot be overcome by treatment with dihydrotestosterone, with pharmacological doses of testosterone propionate or dihydrotestosterone propionate, or with a variety of potent androgen analogues. All of these compounds induce kidney beta-glucuronidase in M. musculus females and kidney ornithine decarboxylase, submandibular gland renin, and submandibular gland epidermal growth factor in both M. caroli and M. musculus females. Furthermore, kidney androgen receptor proteins from M. caroli and M. musculus animals have the same sedimentation characteristics on sucrose density gradients. These data indicate that androgen resistance in M. caroli is not due to deficient 5 alpha-reductase or aberrant hormone metabolism producing suboptimal levels of functional androgen and is not caused by a defective androgen receptor. They suggest that the resistance of beta-glucuronidase in M. caroli kidney to induction by androgen occurs at the level of the beta-glucuronidase gene.     

CENP-B binds a novel centromeric sequence in the Asian mouse Mus caroli.

Minor satellite DNA, found at Mus musculus centromeres, is not present in the genome of the Asian mouse Mus caroli. This repetitive sequence family is speculated to have a role in centromere function by providing an array of binding sites for the centromere-associated protein CENP-B. The apparent absence of CENP-B binding sites in the M. caroli genome poses a major challenge to this hypothesis. Here we describe two abundant satellite DNA sequences present at M. caroli centromeres. These satellites are organized as tandem repeat arrays, over 1 Mb in size, of either 60- or 79-bp monomers. All autosomes carry both satellites and small amounts of a sequence related to the M. musculus major satellite. The Y chromosome contains small amounts of both major satellite and the 60-bp satellite, whereas the X chromosome carries only major satellite sequences. M. caroli chromosomes segregate in M. caroli x M. musculus interspecific hybrid cell lines, indicating that the two sets of chromosomes can interact with the same mitotic spindle. Using a polyclonal CENP-B antiserum, we demonstrate that M. caroli centromeres can bind murine CENP-B in such an interspecific cell line, despite the absence of canonical 17-bp CENP-B binding sites in the M. caroli genome. Sequence analysis of the 79-bp M. caroli satellite reveals a 17-bp motif that contains all nine bases previously shown to be necessary for in vitro binding of CENP-B. This M. caroli motif binds CENP-B from HeLa cell nuclear extract in vitro, as indicated by gel mobility shift analysis. We therefore suggest that this motif also causes CENP-B to associate with M. caroli centromeres in vivo. Despite the sequence differences, M. caroli presents a third, novel mammalian centromeric sequence producing an array of binding sites for CENP-B.     

Pharyngeal mastication and food transport in the carp (Cyprinus carpio L.): A cineradiographic and electromyographic study

Cyprinids constitute the largest fish family and are characterized by their pharyngeal teeth. The masticatory mechanism is still poorly understood. The complex of structures that determine the movements of pharyngeal teeth and chewing pad in the carp (Cyprinus carpio L.) is analyzed. Activities in 16 head muscles of a free-swimming carp were recorded. X-ray cinerecordings, synchronized with electromyograms, were made of the intake, transport, mastication, and deglutition of radiopaque food pellets. Metal markers allowed a detailed movement analysis. Masticatory cycles are bilaterally synchronous and show distinct crushing and grinding patterns. Direct masticatory muscles that suspend and connect the pharyngeal bones steer and stabilize the masticatory movements. Baudelot's ligament, between skull and pectoral girdle, is applied as fulcrum, effects a crucial shift of the rotation axis of the pharyngeal jaw, and transforms crushing into grinding; simultaneous abduction lengthens the grinding stroke. Body muscles supply indirectly the power for mastication; they also appear to be regulated more distantly. The epaxial muscles lift the skull and thereby the levators of the pharyngeal bones, thus transmitting high forces to the teeth. They also stretch the levator of the bone as soon as occlusion is reached and thus optimize its production of forces during grinding. The hypaxial muscles retract the pharyngeal bones indirectly during grinding and power the teeth in sliding. The chewing pad, previously assumed to be motionless, rotates rostroventrad with the skull and intensifies grinding. Respiration and mastication are mutually related. The extensive movements of the pharyngeal bones are permitted only by the simultaneous expansion of the buccopharynx and a slide-coupling in the branchial floor. Muscular pads that line the pharynx are shown to transport food toward the teeth. The constrictor pharyngis effects deglutition. Natural food, intestinal contents, and feces of the carp were analyzed with respect to the capacity for distinct masticatory operations. During the experiments pellets, barley, and worms were fed. The carp is specialized for polyphagy and this appears to be based on the profiles of the heterodont teeth rather than on drastic changes in the two preprogrammed activity patterns. Comparison of the pharyngeal jaw system in the carp and higher teleosts emphasizes the structural design for the application of large forces in this cyprinid.     

A cinefluorographic study of mandibular movement during feeding in the rat (Rattus norvegicus)

The anatomy of the masticatory apparatus, and particularly of the mandibular joints, has led to the view that mandibular movement in the Rodentia is predominantly propalinal, or forwards and backwards in direction. As part of an investigation into the mechanism of function of the mandibular joints in these animals, the feeding behaviour of "August" strain and "Wistar" rats was examined by cinephotography and cinefluorography. The rats were trained to feed on barium sulphate impregnated biscuit and animal cake and to drink radio-opaque liquids. Cinefluorographic recordings of ingestion, mastication, deglutition and of drinking were taken in both the lateral and dorso-ventral projections.
Analysis of the recordings has shown a fundamental separation of ingestive and masticatory activity in the rat, which can be attributed to the morphology of the jaws and particularly to the disparity in the lengths of the mandibular and maxillary diastemas. To bring the incisor teeth into occlusion for ingestion, the mandible is brought forward through the rest position and the condyle into articulation with the anterior part of the fossa. In mastication the condyle is moved backwards to bring the molar teeth into occlusion and the condyle into articulation with the posterior articular facet on the fossa. Once the mandible has been moved into the appropriate position for either ingestion or mastication and deglutition, the movements involved in the separation or chewing of the food are cyclical with combined horizontal and transverse movements as well as the fundamental vertical movement acting to open and close the mouth. The basic movement of ingestion carries the mandibular incisors upwards and forwards across the lingual surfaces of the maxillary incisors, so separating the bite. The grinding stroke of mastication is a horizontal movement carrying the mandibular molars anteriorly across the maxillary.     

Visible and "cryptic" segregation of parental chromosomes in embryonic stem hybrid cells

Chromosome segregation of the parental chromosomes was studied in 20 interspecific hybrid clones obtained by fusion of Mus musculus embryonic stem cells with Mus caroli splenocytes. FISH analysis with labeled species specific probes and microsatellite markers was used for identification of the parental chromosomes. Cytogenetic analysis has shown significant intra- and interclonal variability in chromosome numbers and ratios of the parental chromosomes in the hybrid cells: six clones contained all M. caroli chromosomes, nine clones showed moderate segregation of M. caroli chromosomes (from 1 to 7), and five clones showed extensive loss of M. caroli chromosomes (from 12 to complete loss of all M. caroli autosomes). Both methods demonstrated "cryptic" segregation of the somatic partner chromosomes. For instance, five clones with near-tetraploid chromosome sets contained only few M. caroli chromosomes (from 1 to 8). The data obtained suggest that the tetraploid chromosome set per se is not a sufficient criterion for conclusion on the absence of chromosome loss in the hybrid cells. Note that "cryptic" chromosome segregation occurred at a high frequency in the examined hybrid clones. Thus, "cryptic" segregation should be borne in mind for assessing pluripotency and genome reprogramming of embryonic stem hybrid cells.     

Expression of glucose phosphate isomerase in interspecific hybrid (Mus musculus x Mus caroli) mouse embryos.

Hybrid Mus musculus x Mus caroli embryos were produced by inseminating M. musculus (C57BL/OlaWs) females with M. caroli sperm. Control M. caroli embryos developed more rapidly than did control M. musculus embryos and implanted approximately 1 day earlier. At 1 1/2 days, both the hybrid embryos and those of the maternal species (M. musculus) had cleaved to the 2-cell stage. By 2 1/2 days some of the hybrids were retarded compared to M. musculus, and by 3 1/2 days most were lagging behind. This is consistent with the idea that the rate of development of hybrid embryos declines once it becomes dependent on embryo-coded gene products. We have used this difference in rate of preimplantation development, between hybrid and M. musculus embryos, to try to determine whether the activation of embryonic Gpi-1s genes, that encode glucose phosphate isomerase (GPI-1), is age-related or stage-related. In control M. musculus embryos (both mated and Al groups), the GPI-1AB and GPI-1A allozyme, indicative of paternal gene expression, were detected in 7 of 9 samples of 3 1/2-day compacted morula stage embryos and were seen in all 19 samples of 3 1/2-day blastocysts. In hybrid embryos, these allozymes were detected 1 day later. They were not detected in any 3 1/2-day samples (12 samples of compacted morulae) but were consistently detected at 4 1/2 days (4 samples of blastocysts and 2 samples of uncompacted morulae). Our interpretation of the results is that gene activation in hybrid embryos is stage-specific, rather than age-specific, and probably begins around the 8-cell stage, with detectable levels of enzyme accumulating later. Analysis of GPI-1 electrophoresis indicated that both the paternal (M. caroli) and maternal (M. musculus) Gpi-1s alleles were equally expressed in hybrid embryos and that the paternally derived allele was not activated before the maternally derived allele.     

Characterization of food physical properties by the mastication parameters measured by electromyography of the jaw-closing muscles and mandibular kinematics in young adults

The relationship between the physical properties of solid food and the masticatory parameters is clarified. Eight solid foods of varying physical properties were chosen. Electromyography of the jaw-closing muscles and mandibular kinematics in eleven young subjects were recorded. The masticatory parameters were derived from the recorded data for the entire mastication process, for the first bite, and in the early, middle, and late stages of mastication. After calculating values relative to the mean value for each subject, nine parameters representing each group were chosen through a cluster analysis. Three principal components were extracted, each of them related to the masticatory time and cycle, minimum jaw opening at the early stage of mastication, and masticatory force. The principal component scores for each food were different, except for one combination in which the physical properties under large and extra-large deformations were similar, despite different breaking properties or small deformation properties. The masticatory parameters did not correlate with the physical properties of food measured for small deformation.