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Type 1 iodothyronine deiodinase in the house musk shrew (Suncus murinus, Insectivora: Soricidae): cloning and characterization of complementary DNA, unique tissue distribution and regulation by T(3).

The house musk shrew Suncus murinus (Insectivora: Soricidae) has been reported as having low thyroxine to 3,3'5-triiodothyronine (T(3)) converting activity in liver and kidney homogenates and was assumed to be type 1 iodothyronine deiodinase (D1)-deficient. To study whether this is due to structural abnormality of shrew D1, we cloned the cDNA and characterized the enzyme. The deduced amino acid sequence of shrew D1 was found to be highly homologous to other known D1s and the enzyme itself to have similar catalytic activity. However, unlike in other species, the D1 activity was detected only in liver. Moreover, the D1 activity in liver of the shrew was less than half of that in rat liver and its expression was not up-regulated by T(3). In contrast, a very high activity of D2 was demonstrated in brain and brown adipose tissue. The present study also revealed that the serum level of T(3) in the shrew was in the same range as these in other mammals. These results suggest that D2 contributes to the production and maintenance of T(3) levels in the house musk shrew.

Adipose Tissue, Brown↗

Fatty liver and hyperlipidemia in IDDM (insulin-dependent diabetes mellitus) of streptozotocin-treated shrews.

Severe IDDM (insulin-dependent diabetes mellitus) was produced in the musk shrew (Suncus murimus, Insectivora) by a high dose (a single intraperitoneal injection of 100 mg/kg Body Weight) of streptozotocin (STZ) injection. All shrews that were administered a high dose of STZ exhibited hyperglycemia (449 +/- 16 mg/dl vs 73 +/- 4 mg/dl in controls) and hypoinsulinemia(0.25 +/- 0.07 ng/ml vs 10.96 +/- 1.97 ng/ml in controls) with ketosuria 10 days after injection. Their livers were enlarged and exhibited ayellowish-brown color with marked triglyceride (TG) accumulation (63.25 +/- 7.10 mg/g Liver vs 2.11 +/- 0.19 mg/g Liver in controls). It is probable that the increased influx of fatty acids into the liver induced by hypoinsulinemia and the low capacity of excretion of lipoprotein secretion from liver in the musk shrew resulting from a deficiency of apolipoprotein B synthesis play important roles in fatty liver formation. Hyperlipidemia was another feature in shrews with severe IDDM. The blood TG level was especially high in these shrews (899 +/- 178 mg/dl vs 23 +/- 5 mg/dl in controls). These results indicate that the IDDM shrew, induced by high doses of STZ, is a unique model characterized by fatty liver and hyperlipidemia and may be useful for studying lipid metabolism of IDDM.

Animals↗

Significance of the localization of large granules in the thyrogonadotrophs of the musk shrew (Suncus murinus).

In order to examine the significance of small round and large granules in the thyrogonadotrophs of the musk shrew, we investigated the differences of numbers, distribution patterns and/or immunoreactivities of the large granules in the thyrogonadotrophs among the newborn, infant and adult musk shrews. As a result, we observed that the thyrogonadotrophs exist in newborn musk shrew, but the large granules were few in number and immunoreactivity on the granules was weak. The ratio of the number of large granule-containing cells to all LH beta positive cells was very low (6.2%) in newborn musk shrew, but high (98.2%) in adult male animals. We assume that thyrogonadotrophs of newborn shrews may not yet synthesize active beta-subunits, that the synthesis of beta-subunits is gradually increased with aging, and excessive products of beta-subunits may be stored in the large granules. It is known that female musk shrews are induced ovulators; therefore, we investigated morphological changes in the large and small granules in the thyrogonadotrophs at the LH surge which occurs after mating. We observed that the small granules were markedly decreased in number, or had almost vanished, 2 h after mating, but that the large granules still remained. Consequently, we assume that the large granules may be storage sites of excessive products of beta-subunits in older animals. On the other hand, small granules (release type granules) may be exhausted by release of a large quantity of gonadotrophin 2 h after mating, and this acute exhaustion of small granules may induce a sharp depression of the LH surge.

Amino Acid Sequence↗

Molecular phylogenetic evidence confirming the Eulipotyphla concept and in support of hedgehogs as the sister group to shrews.

For more than a century, living insectivore-like mammals have been viewed as little removed from the ancestral mammalian stock based on their retention of numerous primitive characteristics. This circumstance has made "insectivores" a group of special interest in the study of mammalian evolution. included hedgehogs, moles, shrews, solenodons, golden moles, tenrecs, flying lemurs, tree shrews, and elephant shrews in Insectivora. Subsequently, morphologists excluded flying lemurs, tree shrews, and elephant shrews from Insectivora and placed these taxa in the orders Dermoptera, Scandentia, and Macroscelidea, respectively. The remaining insectivores constitute Lipotyphla, which is monophyletic based on morphology. In contrast, molecular data suggest that lipotyphlans are polyphyletic, with golden moles and tenrecs placed in their own order (Afrosoricida) in the superordinal group Afrotheria. Studies based on nuclear genes support the monophyly of the remaining lipotyphlans (=Eulipotyphla) whereas mitochondrial genome studies dissociate hedgehogs from moles and place the former as the first offshoot on the placental tree. One shortcoming of previous molecular studies investigating lipotyphlan relationships is limited taxonomic sampling. Here, we evaluate lipotyphlan relationships using the largest and taxonomically most diverse data set yet assembled for Lipotyphla. Our results provide convincing support for both lipotyphlan diphyly and the monophyly of Eulipotyphla. More surprisingly, we find strong evidence for a sister-group relationship between shrews and hedgehogs to the exclusion of moles.

Animals↗

Taxonomic status and origin of the shrews (Soricidae) from the Canary islands inferred from a mtDNA comparison with the European Crocidura species.

European island shrews are either relicts of the endemic Pleistocene fauna, e.g.,. Crocidura zimmermanni, or were introduced from continental source populations. In order to clarify the taxonomic status and the origin of the two shrew species from the Canary islands, a 981bp fragment of cytochrome b gene was investigated in all European Crocidura species and compared with the Canary shrew (Crocidura canariensis) and the Osorio shrew (Crocidura osorio). The first shares its karyotype with the Sicilian shrew Crocidura sicula (2N=36), the second with the Greater white-toothed shrew Crocidura russula (2N=42), suggesting possible sister species relationships. Results confirm the monophyly of taxa sharing the same karyotype. Genetic distances between C. sicula and C. canariensis suggest a separation since 5 Myr. The first was probably isolated from the North African ancestor after the Messinian desiccation; the second arrived on the Canary islands by natural jump dispersal. Within the 2N=42 cluster, a first split separated an Eastern line (Tunisia) from a western line (Morocco/Europe) of C. russula. C. osorio clusters together with C. russula from Spain, indicating conspecificy. This suggests a recent introduction from Spain by human.

Animals↗

Fiber capillarization relative to mitochondrial volume in diaphragm of shrew.

The objective was to examine fiber capillarization in relation to fiber mitochondrial volume in the highly aerobic diaphragm of the shrew, the smallest mammal. The diaphragms of four common shrews [Sorex araneus; body mass, 8.2 +/- 1.3 (SE) g] and four lesser shrews (Sorex minutus, 2.6 +/- 0.1 g) were perfusion fixed in situ, processed for electron microscopy, and analyzed by morphometry. Capillary length per fiber volume was extremely high, at values of 8,008 +/- 1,054 and 12,332 +/- 625 mm(-2) in S. araneus and S. minutus, respectively (P = 0.012), with no difference in capillary geometry between the two species. Fiber mitochondrial volume density was 28.5 +/- 2.3% (S. araneus) and 36.5 +/- 1.4% (S. minutus; P = 0.025), yielding capillary length per milliliter mitochondria values (S. araneus, 27.8 +/- 1.5 km; S. minutus, 33.9 +/- 2.2 km; P = 0.06) as high as in the flight muscle of the hummingbird and small bats. The size of the capillary-fiber interface (i.e., capillary surface per fiber surface ratio) per fiber mitochondrial volume in shrew diaphragm was also as high as in bird and bat flight muscles, and it was about two times greater than in rat hindlimb muscle. Thus, whereas fiber capillary and mitochondrial volume densities decreased with increased body mass in S. araneus compared with S. minutus Soricinae shrews, fiber capillarization per milliliter mitochondria in both species was much higher than previously reported for shrew diaphragm, and it matched that of the intensely aerobic flight muscles of birds and mammals.

Aerobiosis↗

Delayed development of reflexes and hyperactive locomotion in the spontaneous mutant "waltzing" of the musk shrew, Suncus murinus.

The autosomal recessive mutation waltzing (wz), displaying abnormal circling and head-shaking behavior, has previously been reported in the musk shrew (Suncus murinus). Postnatal development of reflexes and locomotor patterns in an open arena were examined in wz/wz mutant shrews. The wz/wz shrews showed extreme developmental delays in surface-righting reflex and negative geotaxis until 10-16 days after birth, but both reflexes eventually recovered to the levels of +/wz normal. Nevertheless, the wz/wz adults exhibited bi-directional circling behavior 59 times, head-tossing behavior 22 times and horizontal head-shaking behavior 6 times more frequent than in the +/wz controls. Although the wz/wz adult shrews were extremely hyperactive with daily spontaneous locomotor activity exceeding 4-7 times control shrew activity, they appeared to have a normal circadian rhythm. This shrew mutant may therefore be useful as a model for hyperactivity syndromes in humans.

Animals↗

Blood and liver lipid concentrations in EDS shrews exhibiting spontaneous non-insulin dependent diabetes mellitus (NIDDM).

The EDS (early-onset diabetes in suncus) colony was developed as a new laboratory colony of the musk shrew and is characterized by a high incidence of early-onset spontaneous non-insulin dependent diabetes mellitus (NIDDM). We examined blood lipid (triglyceride [TG], total cholesterol [TC], phospholipid [PL], free fatty acid [FFA]) and liver lipid (TG, TC, PL) concentrations to investigate the features of lipid metabolism in these animals. All lipid concentrations examined both in blood and liver of the diabetic shrews had a tendency toward higher values than those in non-diabetic shrews. The PL concentration was the only parameter that barely showed a significant difference. Values for all blood lipid concentrations in diabetic shrews at 7-9 months tended to be higher than those of 2-month-old diabetic shrews, although the difference was not significant. These findings indicate that diabetic EDS shrews exhibit a much milder defect of lipid metabolism induced by NIDDM than other rodent models.

Animals↗

Alleles at plasma alpha-amylase locus(Amy-1) of the house musk shrew(Suncus murinus): frequency distribution and new variants in laboratory lines and local Asian populations.

Among nine laboratory shrew lines originating from the Japanese islands (Nag, Tok, TKU, Ize, Tr and OKI lines), West Java (Bog), Bangladesh (BAN) and Sri Lanka (SRI), Nag, Tr and Bog were fixed with Amy-1b and SRI with Amy-1a. The remaining lines were still highly polymorphic with the two alleles. A new electrophoretic band C was found in the BAN line and concluded to be expressed by a codominant allele, Amy-1c, which was carried by a single heterozygous female from among eleven wild shrews of the original breeding stock. In most local populations of the Asian shrews surveyed, Amy-1b was more common than Amy-1a. The Sri Lanka population was clearly distinguishable from the others, being nearly fixed with Amy-1a. The C band was found in eleven (one homozygous) of 86 wild shrews caught in Bangladesh. Of a total of 234 wild shrews collected from four Japanese and two Indonesian islands, Bangladesh, and Sri Lanka, only two showed different AMY-bands from AMY-A, -B and -C, and such bands were not found in the laboratory-bred shrews examined.

Animals↗

[The population dynamics of the water shrew Neomys fodiens (Mammalia, Soricidae) and its helminthes fauna in the Northern Baraba].

The water shrew population was studied in Ust-Urgulca (Novosibirsk Province) in 1978-1990; abundance, sex and age structure, reproduction rate of this animal and its helminth fauna were examined. It was found that the abiotic factors (in particular the water level of habitat areas) influenced on the water shrew populations. The number of water shrews increased when the high water level increased. In these conditions, the population of the water shrew rejuvenated and the numbers of females increased with some increasing their fertility. In the helminthes fauna associated with the water shrew, predominated the parasite species developing in water invertebrates or ones living near water basins. When water habitats dried out, the number of water shrews decreased and individuals retained only in some areas and the population became old.

Aging↗

Short-tailed shrews as reservoirs of the agents of Lyme disease and human babesiosis.

To determine whether short-tailed shrews (Blarina brevicauda) serve as reservoir hosts for the Lyme disease spirochete (Borrelia burgdorferi) and the agent of human babesiosis (Babesia microti), we examined nymphal ticks that had fed as larvae on shrews collected from 3 enzootic sites in coastal Massachusetts for evidence of infection by either or both of these agents. Xenodiagnosis indicated that 11 of 14 shrews were infected by B. burgdorferi. One of 3 piroplasm-infected shrews also infected ticks with B. microti. In a site where the piroplasm is endemic, 11 of 17 shrews showed patent parasitemias by thin blood smears. Of these, 4 had parasitemias exceeding 40%. Few immature ticks infested shrews, however, suggesting that B. brevicauda, although abundant in some endemic sites and serving as a competent reservoir, would contribute minimally to the population of infected nymphs.

Animals↗

Preservation of hippocampal neuron numbers and hippocampal subfield volumes in behaviorally characterized aged tree shrews.

Aging is associated with a decreased ability to store and retrieve information. The hippocampal formation plays a critical role in such memory processes, and its integrity is affected during normal aging. We used tree shrews (Tupaia belangeri) as an animal model of aging, because in many characteristics, tree shrews are closer to primates than they are to rodents. Young and aged male tree shrews performed a holeboard spatial memory task, which permits assessment of reference and working memory. Upon completion of the behavioral measurements, we carried out modified stereological analyses of neuronal numbers in various subdivisions of the hippocampus and used the Cavalieri method to calculate the volumes of these subfields. Results showed that the working memory of aged tree shrews was significantly impaired compared with that of young animals, whereas the hippocampus-dependent reference memory remained unchanged by aging. Estimation of the number of neurons revealed preserved neuron numbers in the subiculum, in the subregions CA1, CA2, CA3, and in the hilus of the dentate gyrus. Volume measurements showed no aging-related changes in the volume of any of these hippocampal subregions, or in the molecular and granule cell layers of the dentate gyrus of tree shrews. We conclude that the observed changes in memory performance in aging tree shrews are not accompanied by observable reductions of hippocampal neuron numbers or hippocampal volume, rather, the changes in memory performance are more likely the result of modified subcellular mechanisms that are affected by the aging process.

Aging↗

Alpha 2-adrenergic binding sites in the medulla oblongata of tree shrews demonstrated by in vitro autoradiography: species related differences in comparison to the rat.

Alpha 2-adrenergic binding sites in the medulla oblongata of tree shrews and rats were detected and quantified by in vitro-autoradiography with the alpha 2-antagonist 3H-rauwolscine (3H-RAUW). The autoradiographic pattern of the radioligand binding in the tree shrew medulla oblongata resembles that which has been described by others for the human myelencephalon. This pattern coincides well with the occurrence of catecholaminergic structures detected by immunocytochemistry with antibodies against phenylethanolamine-N-methyltransferase and tyrosine hydroxylase. In contrast to the rat, where only the nucleus tractus solitarii and the nucleus dorsalis nervi vagi were labeled, five discrete nuclei specifically bound 3H-RAUW in tree shrews. The highest number of binding sites was detected in the nucleus dorsalis nervi vagi (nX; Bmax: 333 fmoles/mg) and the nucleus tractus solitarii (NTS; 311 fmoles/mg), followed by the nucleus nervi hypoglossi (nXII; 297 fmoles/mg), the nucleus reticularis parvocellularis (FRS; 230 fmoles/mg), and the area of the catecholamine cell groups A1 and C1 (area C1; 202 fmoles/mg). Maximal binding in the two labeled nuclei of the rat was 158 fmoles/mg. The discrete nuclei of the two species also showed different affinities for 3H-RAUW with Kd ranging from 0.17 to 0.83 nM in tree shrews and 1.80 to 1.95 nM in rats. Competition experiments revealed that the radioligand bound specifically to alpha 2-binding sites. In the tree shrew, nX, nXII and the area C1, also have a relatively high affinity for the alpha 1-antagonist prazosin which is a quality of the adrenoceptor subtype alpha 2B. Furthermore, in the area C1, 3H-RAUW binding was inhibited by the dopamine antagonist haloperidol. There are thus species related as well as regional differences with respect to the number, the affinity, and the pharmacological properties of alpha 2-binding sites in the medulla oblongata. In tree shrews, alpha 2-adrenoceptors can be autoradiographically quantified in regions which are not labeled in the rat, although former data predicted the existence of such receptors, e.g., in the area of the adrenaline cell group C1.

Animals↗

Development of the lateral geniculate nucleus: interactions between retinal afferent, cytoarchitectonic, and glial cell process lamination in ferrets and tree shrews.

We have studied the relationship of retinal afferents, glial cell processes, and neuronal cytoarchitectonics in the lateral geniculate nucleus (LGN) of two species: tree shrews (Tupaia belangeri) and ferrets (Mustela putoris). Both species are relatively immature at birth, allowing the development of these features to be studied in the perinatal period. Retinal afferents, visualized by intraocular injection of a wheat germ agglutinin/horseradish peroxidase conjugate (WGA-HRP), are apparently the first elements of the developing LGN to exhibit a characteristic layered pattern in tree shrews and ferrets. Some radial glia still remain in the LGN of both species as the retinal afferents are in the process of segregating. Glial cell processes were visualized immunohistochemically with antibodies to glial fibrillary acidic protein (GFAP) or vimentin. In both the ferret and tree shrew, layering of glial cell processes is first seen as the overlap of retinal terminal fields diminishes. In the tree shrew LGN, these bands of dense glial cell staining are seen in apparent future cellular layers, whereas in the ferret, glial cell banding appears in interlaminar zones. If one or both eyes are removed at birth in tree shrews (before LGN cell layers are formed), the glial cell pattern seen 1 week later is in accord with the distribution of surviving nerve cells. The glial processes do not appear to invade regions left by degenerating retinal terminals or dying LGN cells. Several days after the appearance of layered glial cell processes (in the tree shrew) or at about the same time as glial layering (in the ferret), the first interlaminar spaces develop between neuronal cells, marking the beginning of cytoarchitectonic lamination, with its distinctive alternating cell-rich and cell-poor zones. Over the next several weeks, LGN neurons in both species continue to segregate into characteristic layers until the final, adult pattern of neuronal lamination is evident; as this process is completed, glial cell lamination disappears. These observations suggest that glial cells may be involved in establishing the neuronal layers that characterize the mature LGN of many species.

Afferent Pathways↗

Human hepatitis B virus and hepatocellular carcinoma. I. Experimental infection of tree shrews with hepatitis B virus.

Tree shrews (Tupaia belangeri chinenesis) can be experimentally infected with human hepatitis B virus (HBV) by inoculation with human serum positive for HBV, the experimental infection rate being 55.21%. Successive infections have been passed through five generations among the tree shrews inoculated with HBV-positive sera from the infected animals, the average infection rate being 94.0%. The experimental infection of tree shrews with HBV may be prevented by immunization with hepatitis B vaccine, the protection rate being 88.89%. Standard serum containing HBV at 10(8) CID (chimpanzee infection dose)/ml, was diluted 10(-6), 10(-7), 10(-8), 10(-9), and 10(-10) and produced infection rates of 80.0%, 88.8%, 66.7%, 55.6% and 42.9% respectively. Thus the CID50 in tree shrews may reach a dilution of 10(-9), which shows that tree shrews are sensitive to HBV infection. These results successfully establish tree shrews as a reliable and useful animal model for research on HBV infection and its relation to hepatocarcinogenesis.

Animals↗

Hippocampal glucocorticoid receptor expression in the tree shrew: regulation by psychosocial conflict.

1. This study was conducted to determine whether chronic psychosocial conflict alters the expression of glucocorticoid receptor (GR) mRNA in the hippocampus of male tree shrews (Tupaia belangeri). 2. To generate probes for the in situ hybridization, the tree shrew GR gene was partly cloned. There was a 90% homology between the deduced amino acid sequence of the cloned tree shrew GR and that of the corresponding human GR sequence. 35S-Labeled riboprobes which had been transcribed from the tree shrew GR clone hybridized to pyramidal neurons in all subregions of the tree shrew hippocampal formation and to granule neurons in the dentate gyrus. 3. After in situ hybridization, the expression of GR mRNA was semiquantitatively determined by counting silver grains over single neurons of the hippocampal formation of psychosocially stressed tree shrews and control animals. After 12 days of social conflict, the number of silver grains in the CA1 and CA3 pyramidal neurons of stressed animals was significantly lower than in controls. No statistically significant differences in mRNA expression were observed in the pyramidal neurons of the subiculum and in the granule neurons of the dentate gyrus. 4. The present results suggest that psychosocial stress leads to a site-specific down-regulation of hippocampal GR via modification of mRNA expression.

Amino Acid Sequence↗

Localization of dopamine receptors in the tree shrew brain using [3H]-SCH23390 and [125I]-epidepride.

The tree shrew is a mammalian species, which is phylogenetically related to insectivores and primates. The aim of the present study was to investigate the distribution of dopamine receptor D1- and D2-like binding sites in the brain of this non-rodent, non-primate mammal. Using in vitro autoradiography and employing the radioligands [3H]-SCH23390 and [125I]-epidepride, dopamine receptors were mapped and quantified. Significant findings with regard to the D1-like binding pattern include the presence of a "patchy" binding in the striatum. In the cortex, D1-like binding sites were observed in both the superficial and the deep layers. In the hippocampal formation, D1-like binding sites were seen primarily in the CAI region and not in the dentate gyrus. These characteristics of the D1 pattern in the tree shrew brain are shared by cat and monkey and human brain, but not by rodent brain. Significant findings with regard to the D2-like binding pattern include the presence of D2-like binding in the claustrum. In addition, the striatum demonstrated "patchy" D2-like binding. These characteristics of the D2 pattern in the tree shrew brain are shared by cat and monkey and human brain, but not by rodent brain. On the other hand, the significant densities of D2-like binding sites in the glomerular layer of the tree shrew olfactory bulb is a finding that discriminates tree shrews from higher evolutionary species who lack such binding. Overall, the evidence coincides with the view that tree shrews are phylogenetically related to primates.

Animals↗

Molecular phylogenetic relationships of moles, shrew moles, and desmans from the new and old worlds.

A Rich variety of anatomical and physiological specializations has enabled members of the family Talpidae (moles, shrew moles, and desmans) to exploit a diverse range of habitats: terrestrial, semi-aquatic, aquatic/fossorial, semi-fossorial, and fossorial. While numerous morphological and biochemical studies pertaining to the origin and radiation of the Talpidae have been completed, phylogenetic hypotheses remain controversial. To address this shortcoming we sequenced the mitochondrial DNA cytochrome b gene (1140bp) from 29 individuals spanning 12 talpid species. Phylogenetic trees incorporating 12 New and Old World genera (18 species; all 3 extant subfamilies) were then constructed using NJ, MP, ML, and NJ-ML (NJ with ML parameters) methods. Our results provide molecular support for a mononphyletic Talpidae, and suggest that the 12 genera are clustered into seven major clades; (1) Asiatic shrew-like moles (Uropsilus), (2) North American aquatic/fossorial moles (Condylura), (3) North American fossorial moles (Parascalops, Scalopus, and Scapanus), (4) North American semi-fossorial shrew moles (Neurotrichus), (5) Japanese semi-fossorial shrew moles (Dymecodon and Urotrichus), (6) European semi-aquatic desmans (Desmana), and (7) Eurasian fossorial moles (Euroscaptor, Mogera, and Talpa). None of these groupings comprised mole species from both continents. In fact, North American moles and shrew moles do not appear to have specific affinities with Asian moles and shrew moles, respectively. Although low bootstrap support was generally found for evolutionary nodes uniting the major talpid clades, all gene trees constructed identified fossorial North American and Eurasian mole lineages as nonmonophyletic groups, suggesting subterranean specializations arose independently at least twice during the evolution of the Talpidae. Additionally, our data set provides molecular support for a basal divergence and long independent history of Uropsilus from the main talpid line, and refutes the traditional taxonomic status and secondarily basal phylogenetic placement of the subfamily Desmaninae within the Talpidae.

Animals↗