Embryonic development of the house shrew (Suncus murinus)

Summary The embryonic development of the laboratory house shrew (Suncus murinus) was observed during the period from 13 to 29 pairs of somites and was compared with that of other mammalian species. This period ranged from 10 to 12 days after fertilization. Embryos were allocated to stages 11A–C and 12A–C according to somitic count. These embryos were characterized by the following features. 1. The curvature of the body became gradually enhanced, changing from a parenthesis-like to a bracket-like shape. 2. The swelling of the future hindlimb bud appeared at stage 11 A, while the forelimb bud became apparent at stage 12 A. The latter developed rapidly and had exceeded the former in size by the end of stage 12C. 3. The timing of closure of the cephalic neuropores varied, but the order of closure is constant. The prosencephalic neuropore closed later. The caudal neuropore still existed even at the end of stage 12C. 4. In all of the cranial nerve ganglia except for the auditory one, crest-derived cells seemed to contribute to the initial formation, while placodal cells seemed to be involved later. A different pattern of primordial formation was observed between trigeminal/facial and glossopharyngeal/vagal ganglia. 5. By the end of stage 12C, the eye cup and nasal pit began to form, and the endolymphatic appendage also appeared. 6. In the heart, the sinus valves were well developed, and the atrium and ventricle were being divided by the developing interatrial septum I and interventricular septum, respectively. 7. The thyroid primordium was temporarily everted and projected into the pharynx. The internal structure of this protruding thyroid resembled tuberculum thyroideum observed in the human embryo.Abbreviations AA1-4 1st, 2nd, 3rd, and 4th aortic arches - AC axial condensation - Acc accessory nerve - AlC alimentary canal - AT arterial trunk - At atrium - CE cardiac eminence - Clo cloaca - ClM cloacal membrane - CN caudal neuropore - Co intraembryonic coelom - DA dorsal aorta - Di diencephalon - DMg dorsal mesogastrium - DP dorsal pancreas - DR dorsal pharyngeal ridge - EP1-2 1st and 2nd epipharyngeal placodes - FB forelimb bud - G5 7–10, trigeminal, facial, auditory, glossopharyngeal, and vagal ganglion primordia - Gb gall bladder - HB hindlimb bud - HbS swelling of future hindlimb bud - HC hepatic cord - HD hepatic diverticulum - lAA6 left 6th aortic arch - lLB left lung bud - LP lens placode - lVt left ventricle - Ma maxillary process - MnT mesonephric tubule - Mn mandibular process - NaP nasal placode - ND nephric duct - NF neural fold - Nc notochord - NP neural plate - NT neural tube - NV nephric vesicle - OB omental bursa - OpV optic vesicle - OV otic vesicle - PA1-4 1st, 2nd, 3rd, and 4th pharyngeal arches - PAF interatrial foramen I - PBr primary bronchi - Ph pharynx - PP3 3rd pharyngeal pouch - rAA6 right 6th aortic arch - RCV rostral cardinal vein - RD respiratory diverticulum - ReP retinal plate - rLB right lung bud - RP Rathke's pouch - rVt right ventricle - So somite - Sm stomach - SV sinus venosus - Tel telencephalon - Th thyroid primordium - Thm thymus - VP ventral pancreas - Vt ventricle - VV vitelline vein     

Neurofilament expression in lens cells of the house shrew, Suncus murinus

Immunohistochemical analysis demonstrated that lens cells of the laboratory house shrew, Suncus murinus, expressed many intracellular filaments that immunoreacted with pooled monoclonal antibodies against 70-kDa, 160-kDa, and 210-kDa neurofilament triplet proteins. Immunopositive filaments in lens cells first appeared in day 13 embryos, while the invaginating lens placode was thickening, and this immunoreactivity was still present in immature lens fiber cells of the adult animal. Western blot analysis showed that the immunopositive molecule was a low-molecular-weight neurofilament protein that appeared in the neural tissues of this animal. The immunoreactive pattern of lens cells was quite similar to that of neurons, although there were some peculiar aspects. When the cells of the lens vesicle differentiated into the lens epithelium and fibers, immunoreactivity occurred in both, suggesting that the neurofilaments in the lens cells do not directly relate to lens fiber elongation nor to a determinant of the fiber caliber. The strong immunoreactivity in the embryonic lens and weak expression of this protein in the immature lens fiber cells of the adult animal suggest that low-molecular-weight neurofilament protein is transiently expressed in the differentiating lens cells. This may be a common feature of placodederived cells.     

Taste preferences for amino acids in the house musk shrew, Suncus murinus

Taste preferences in house musk shrews for amino acids as well as NaCl, sucrose, quinine hydrochloride, HCl and saccharin Na were studied by employing the two-bottle preference technique. Shrews showed a preference for 0.2–0.5 M sucrose but a moderate rejection to NaCl and a strong rejection to quinine, HCl and saccharin. They exhibited a marked preference for many naturally occurring L-α-amino acids with aliphatic side chains at both 0.02 and 0.2 M. Increase in the aliphatic side chain length of DL-α-amino acids resulted in both lowering of the preference threshold and increase in the preference magnitude. Amino acids with side chains containing sulfur atoms, basic groups and Phe at 0.02 M were preferred to water, but Cys and Arg at 0.2 M was rejected. Shrews showed neither preference nor rejection to Trp, Asn, Gln and monosodium glutamate at 0.02 M, but rejected strongly Asp and Glu. D-Met from 0.001 to 0.1 M was preferred as well as L-Met, while D-Phe was more preferred than L-Phe. Such preferences for a wide variety of amino acids in shrews could be attributed to their food habit of predating on various kinds of insects and worms.     

The median and lateral substantia gelatinosa in the cervical cord of the musk shrew (Suncus murinus) and its synaptic composition

Summary The extent and laminar arrangement of the substantia gelatinosa (SG) were examined in the cervical spinal cord of the shrew. Between C1 and C6 or C7 the three dorsal layers on either side of the gray matter were confluent at the commissura posterior grisea in shrews of both sexes. Lamina I was thin with no large marginal cells. Lamina II comprised the major part of the SG, consisting of outer cellular and inner neuropil layers. Lamina III was composed of a meshwork of axon terminals, dendritic profiles and myelinated fibers. Unlike the situation in other mammals, in shrew the incidence of axon terminals with round vesicles was similar in the three uppermost layers, but the occurrence of terminal profiles with flat vesicles was significantly greater in deeper laminae. Lamina IV was restricted to the dorsal horn and did not extend through the midline.     

Ultrastructural study of postnatal development of the tonsillar crypt epithelium of the musk shrew, Suncus murinus

Summary An ultrastructural study was made of the postnatal development of the tonsillar crypt epithelium in the musk shrew, Suncus murinus. On day 3 after birth, a particular kind of large lymphoid cell was first seen to move through the basement membrane into the epithelium. The next migration was that of lymphocytes, which passed through holes in the basement membrane. On days 5 to 7, the lymphocytes formed clusters, and pale epithelial cells of low electron density appeared. The cell clusters and pale epithelial cells fused on day 10. By day 14, these epithelial cells extended cytoplasmic projections to the surface of the epithelium, which had many heterophagic vacuoles and some microvilli-like structures. These findings suggest that the lymphoepithelial relationship is important for the organization of the immunological microenvironment in tonsillar crypt epithelium of the neonatal musk shrew.     

A stereological approach to the study of neural organogenesis in Xenopus laevis

Summary Stereological methods are applied to the study of structural changes undergone, during neurulation, by the neuroepithelium of early embryos of Xenopus laevis. In light microscopy, we evaluate: the section area of the neuroepithelium, the nucleocytoplasmic ratio, the volume fraction of the intercellular spaces, of the nuclei and of the cytoplasm. In electron microscopy, the volumetric density and the surface ratio of mitochondria as well as the surface density of the endoplasmic reticulum are measured. Occasionally, the data found for Xenopus are compared to those found earlier in a similar study of the chicken embryo.     

Postnatal development of 3H-rauwolscine binding sites in the dorsal lateral geniculate nucleus and the striate cortex of the tree shrew (Tupaia belangeri)

Summary Noradrenaline has been shown to play an important role within the visual system of the brain. To analyze the postnatal development of alpha₂-noradrenergic receptors in the visual system of tree shrews, we localized and quantified binding sites for the antagonist [³H]-rauwolscine by in vitro-autoradiography in the dorsal lateral geniculate nucleus and the striate cortex at different postnatal ages. At birth, the dorsal lateral geniculate nucleus is only slightly labeled by [³H]-rauwolscine. During the postnatal period, the number of binding sites increases to reach a maximum around postnatal day 20. Since the young tree shrews open their eyes at approximately day 19, it appears that this high concentration of alpha₂-adrenoceptors is related to eye opening. In the adult animal, [³H]-rauwolscine labeling shows a laminated pattern in the dorsal lateral geniculate nucleus. Laminae 1, 2, and 3 are more strongly labeled than laminae 4, 5, and 6. In the striate cortex, the pattern of [³H]-rauwolscine-binding sites changes dramatically during the early postnatal period. Immediately after birth, there is only one layer, located within the subplate zone, which is labeled. From postnatal day 5 onwards, all cortical layers which can be distinguished on histologically stained sections reveal [³H]-rauwolscine-binding sites, but in layer IV, which is known to receive major inputs from the dorsal lateral geniculate nucleus, there is very little labeling during the first two postnatal weeks. In this layer, a large number of [³H]-rauwolscine-binding sites occurs between postnatal day 15 and 20, that is slightly before and around the time of eye opening. From this time onwards, the pattern of [³H]-rauwolscine binding in the striate cortex is very similar to that in the adult, where all layers are labeled although to different degrees. Since around postnatal day 20, maximal numbers of [³H]-rauwolscine binding sites are present in the dorsal lateral geniculate nucleus and a large number of these binding sites also emerges in layer IV of the striate cortex, alpha₂-noradrenergic receptors are probably important for processes related to the opening of the eye and/or for the visual system to function.     

The FAAH inhibitor URB-597 interferes with cisplatin- and nicotine-induced vomiting in the Suncus murinus (house musk shrew)

Considerable evidence implicates the endocannabinoid system as a neuromodulator of nausea and vomiting. The action of anandamide (AEA) can be prolonged by inhibiting its degradation, through the use of URB597 (URB), a Fatty Acid Amide Hydrolase (FAAH) enzyme inhibitor. Here we present evidence that the FAAH inhibitor, URB, interferes with cisplatin- and nicotine-induced vomiting in the Suncus murinus. In Experiment 1, shrews were injected with URB (0.9 mg/kg) or vehicle 120 min prior to the behavioral testing. They received a second injection of AEA (5 mg/kg) or vehicle 15 min prior to being injected with cisplatin (20 mg/kg) or saline and the number of vomiting episodes were counted for 60 min. In Experiment 2, shrews were injected with vehicle or URB (0.9 mg/kg) 120 min prior to receiving an injection of nicotine (5 mg/kg) or saline and the number of vomiting episodes were counted for 15 min. Experiment 3 evaluated the potential of the CB₁ antagonist, SR141716, to reverse the effect of URB on nicotine-induced vomiting. URB attenuated vomiting produced by cisplatin and nicotine and the combination of URB + AEA suppressed vomiting produced by cisplatin. The effect of URB on nicotine-induced vomiting was reversed by SR141716. These data suggest that the EC system plays a tonic role in the regulation of toxin-induced vomiting.     

Differential activity of drugs to induce emesis and pica behavior in Suncus murinus (house musk shrew) and rats

We have previously reported that emetic stimuli induce kaolin ingestion behavior (pica behavior) in rats and mice (i.e., species that do not have the emetic reflex) and that the behavior may be analogous to gastrointestinal discomfort, such as nausea and emesis. We hypothesized that pica behavior may also occur in species capable of vomiting and that it may serve as an additional index of discomfort relevant to antiemetic drug development. The present experiments were conduced using Suncus murinus and rats and kaolin consumption was measured at 24 h after the administration of nicotine (1.25-5 mg/kg, s.c.), copper sulfate (10-120 mg/kg, p.o.), lithium chloride (50-200 mg/kg, i.p.) and cisplatin (1-30 mg/kg, i.p.). In S. murinus, all treatments, excepting lithium chloride, were emetic but none induce kaolin consumption. Conversely, all treatments induced kaolin consumption in rats without inducing emesis. The results indicate that pica behavior is not likely to be useful to assess gastrointestinal discomfort in S. murinus.     

Gross anatomical study of the sympathetic cardiac nerves in the house musk shrew (Suncus murinus)

The sympathetic cardiac nerves originating from the cervical and upper thoracic sympathetic ganglia in the house musk shrew (Suncus murinus) were examined using macroscopic and whole‐mount immunohistochemical methods. Based on the results, the nerves were macroscopically classified into the following three groups: nerves innervating the cervical sympathetic ganglia mainly to the arterial porta of the heart; nerves supplying the stellate and thoracic sympathetic ganglia at the level of T2–T5 or T6 for both the arterial and venous portae of the heart; and nerves innervating the thoracic sympathetic ganglia at the level of T4–T9 to the esophagus and lung and then the heart via the blood vessels within the mediastinal pleura. These findings in the house musk shrew suggest a possible primitive morphological pattern of the cervical and thoracic sympathetic nervous system that may be related to those in other mammals, including humans. Anat Rec, 2007. © 2007 Wiley‐Liss, Inc.     

The distribution of crossed and uncrossed optic fibers in the different layers of the lateral geniculate nucleus in the tree shrew (Tupaia glis)

Summary The laminar distribution of crossed and uncrossed optic fibers was studied in the lateral geniculate nucleus (LGN) in the tree shrew (Tupaia glis) following unilateral enucleation. For the investigation of the termination of optic fibers the transneuronal degeneration method and experimental EM were employed. By using formvar film-coated slot grids, all six layers of the LGN could be studied in a single ultrathin section. Degeneration of crossed optic fibers was observed in layers 1, 3, 4 and 5 of the contralateral LGN. The uncrossed retinofugal fibers supply layers 2 and 6 of the LGN. The degeneration in layer 4 was less pronounced than that in the other layers. Ipsilateral and contralateral optic fibers were well separated. Filamentous as well as dark types of degeneration were found in the LGN after enucleation. The optic terminals (RL boutons) were seen only in the synaptic glomeruli.     

Callosal projections between areas 17 in the adult tree shrew (Tupaia belangeri)

Summary In the primary visual cortex (area 17) of the tree shrew (Tupaia belangeri) neurons projecting to the contralateral area 17 via the corpus callosum were identified by horseradish peroxidase histochemistry (HRP, WGA-HRP). The distribution of homotopic and heterotopic connections was studied. We found that a narrow stripe of area 17 close to the dorsal area 17/18 border — which corresponds to the visual field along the vertical meridian — is connected via homotopic callosal projections. The adjacent dorsal part of area 17, which largely corresponds to the binocular visual field, is connected via homotopic as well as heterotopic projections. Heterotopic projections originate in the cortical stripe along the area 17/18 border and their contralateral targets are displaced medially. Callosal neurons are located mostly in supragranular but also occur in infragranular layers. The supragranular neurons in general are pyramidal cells. In addition to these findings, we confirmed earlier reports on ipsilateral projections of the primary visual area to the dLGN, the claustrum, area 18 and other visual areas.The authors wish to dedicate this paper to Prof. W. Lierse in honour of his 60th birthday     

Deceleration and acceleration in the rate of posterior neuropore closure during neurulation in the curly tail (ct) mouse embryo

Summary Curly tail (ct) is a mouse mutant producing spinal neural tube defects as a result of delayed closure of the posterior neuropore (PNP). The purpose of the present study was to determine in ct/ct embryos the time of onset of the delay in PNP closure, and the pattern of this closure, as well as to study the possibility that reopening of the neural tube occurs. Normal spinal neurulation was studied in non-mutant Swiss (Sw) embryos. In the latter, the average PNP length diminished steadily between the 7- and 25-somite stages, and then decreased more rapidly, indicating an acceleration of closure rate, until the 30- to 32-somite stage, when all PNPs closed. PNP width decreased steadily between the stages of 7 and 30 somites. In ct/ct embryos the average PNP length showed a slight increase between the stage of 23 to 28 somites, indicating a temporary deceleration of closure rate, and the range of PNP sizes increased markedly. This was followed by a decrease in PNP length until the 37-somite stage, indicating an acceleration of closure rate. From the stage of 32 somites onwards, the proportion of embryos with closed PNPs gradually increased to 90%. The population of ct/ct embryos was subdivided. Embryos with large PNPs showed a marked deceleration of closure rate during a period of 11 somite stages, followed by a brief but very high acceleration of closure rate. This resulted in closure of the PNP in a proportion of these embryos, while in the remainder of the embryos the deceleration phase had been too enhanced to allow complete catch up of closure during the acceleration phase; these embryos would develop spina bifida. Embryos with relative small PNPs also showed a deceleration of closure rate, but only during a period of four somite stages. This was followed by an acceleration, resulting in closure of all PNPs at the stage of 32 to 33 somites. The enlargement of the PNP in ct/ct embryos was not due to re-opening of a closed neural tube, but resulted from a sharp decline in the rate of PNP closure combined with a normal rate of caudal elongation of the embryo. It is concluded that the ct strain forms a homogeneous population, with a large variation of its specific phenotype: deceleration of PNP closure during a restricted period. The disturbance of spinal neurulation in ct/ct embryos takes the form of a deceleration/acceleration pattern, resulting in a net delay of closure. It is suggested that, due to the ct mutation, forces are generated in the embryonic axis which oppose a normal neurulation process at a specific stage of development.     

Stages in the prenatal development of the Chinese hamster (Cricetulus griseus)

Summary A system of staging embryos is described for the Chinese hamster (Cricetulus griseus). This system of staging, based on Streeter's developmental horizons in human embryos, comprises three sets of criteria: 1) data on postconceptional age, size and number of somites, 2) external characteristics and 3) internal characteristics. A comparison has been made with data in mouse (Theiler, 1972) as well as in primates. It seems that the order of organogenesis, i.e. the sequence in which individual organs are formed, is basically similar in all mammals studied so far.     

Potential energetic implications of emesis in the house musk shrew (Suncus murinus)

During the course of studies investigating novel anti-emetic therapies we serendipitously observed a previously unreported behaviour related to emesis in the house musk shrew. This behaviour consisted of spontaneous ingestion of vomit in about half of the animals (males and females) in which emesis was induced by either nicotine (4 mg kg-1 sc.) or horizontal motion (1 Hz, 4 cm, 10 min). Analysis of vomit samples and gastric contents revealed that in a "typical" individual the gastric contents would be voided by as few as 3 vomits. Energetic calculations of the metabolisable energy of food, gastric contents, vomit and field metabolic rate (FMR) predict that a male weighing 60 g would lose 17.3% of its hourly energy requirement for FMR if it vomited once. A 40 g female, however, would experience an hourly energy loss of approximately 22.8%. The possible energetic consequences and resulting ecological implications of this unusual behaviour are discussed.     

c-Fos expression in the visual system of the tree shrew (Tupaia belangeri)

c-Fos is a nuclear phosphoprotein coded by the proto-oncogen c-fos which can be detected immunohistochemically after both physiological and pathological stimuli. This property is of great importance, because it offers a valuable tool for morphofunctional identification of activated neurons. We have studied the neuronal activity in the visual pathway of Tupaia belangeri within the following anatomical structures: retina, superior colliculus (SC), dorsal lateral geniculate nucleus (dLGN), pulvinar (Pu), parabigeminal (PBG) nucleus and primary visual cortex (V1) analyzing the c-Fos expression after exposing the tree shrews to different light stimuli (white light –control positive group–, green light, blue light and darkness conditions –control negative group–). Our findings suggest that in the retina, the ganglion cells and the cells of the inner nuclear layer respond better to blue and green light stimuli, when comparing the c-Fos expression between white, green, blue lights and darkness conditions. However, in the SC, dLGN, Pu, PBG nucleus and V1 another pattern of c-Fos expression is observed: a maximum expression for the control positive group, a minimum expression for the control negative group and intermediate expressions within the blue and green light groups. Conclusion: the expression levels of c-Fos protein are able to show significant differences between distinct light stimuli in all anatomical structures studied (retina, SC, dLGN, Pu, PBG and V1) of T. belangeri.