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Neural distribution of estrogen receptor immunoreactive cells in the female musk shrew.

In this report we describe the neural distribution of estrogen receptor immunoreactivity (ER-IR) in the female musk shrew (Suncus murinus). The highest concentrations of neurons containing ER-IR were found in the preoptic areas, the lateral septum, the anterior arcuate and ventromedial nuclei of the hypothalamus, the medial nuclei of amygdala, and the midbrain central grey. Additional preoptic, hypothalamic, limbic and midbrain nuclei also contained ER-IR cells. The distribution of ER-IR was similar to that described in other mammals and birds, with several important differences. In the female musk shrew, there was little difference in ER-IR intensity or distribution when brains from gonadally intact and ovariectomized musk shrews were compared. In addition, long-term treatment with a supraphysiological dose of estradiol was required to detect a decrease in ER-IR intensity. Finally ER-IR was noted in both nuclear and cytoplasmic regions of cells in ovariectomized and gonadally intact musk shrews. The dense ER-IR noted in intact females as well as the presence of cytoplasmic stain may be due to the unusual relationship between estradiol, ovulation and sexual receptivity in this species.

Animals↗

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--05. 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-alpha-amino acids with aliphatic side chains at both 0.02 and 0.2 M. Increase in the aliphatic side chain length of DL-alpha-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.

Amino Acids↗

Scent marking in the musk shrew (Suncus murinus).

The hormonal regulation of scent marking was examined in male and female musk shrews. In males castration depressed flank, perineal, and throat marking frequencies, and a physiological dose of testosterone restored perineal and throat marking to intact levels. Females exhibited high levels of marking whether tested in a non-pregnant state, late in gestation, or when post-partum. Moreover, ovariectomy had no influence on marking behavior. Adrenalectomy, in addition to ovariectomy, caused a significant decline in two of the three marking behaviors examined, flank and perineal marking. These results demonstrate that these sexually monomorphic scent marking behaviors are under gonadal control in male musk shrews, but not in females. Furthermore, two of the three different types of scent marking examined in female musk shrews appear to require adrenal hormones. Throat marking in the female musk shrew apparently is not regulated by steroidal hormones.

Adrenal Glands↗

The role of nausea in taste avoidance learning in rats and shrews.

When paired with a novel flavoured solution, the injection of an emetic drug, such as lithium chloride, produces avoidance of that solution in both non-emetic rats and in emetic shrews. On the other hand, the pairing of a novel flavour with a drug with rewarding properties results in conditioned taste avoidance in rats, but in conditioned taste preference in shrews. It, therefore, appears that nausea may be necessary for the establishment of conditioned taste avoidance in the emetic shrew, but not in the non-emetic rat. Indeed, pre-treatment with the anti-emetic agents, ondansetron or Delta9-tetrahydrocannabinol, interferes with the establishment of lithium-induced conditioned taste avoidance in the shrew, but does not even attenuate the establishment of lithium-induced conditioned taste avoidance in the rat. The results of a number of studies suggest that the nature of flavour-drug associations varies on the basis of the emetic capacity of the species.

Animals↗

Water balance and kidney function in the least shrew (Cryptotis parva).

We assessed renal function in least shrews (Cryptotis parva, body mass 4.7 g) within the context of overall water balance. The glomerular filtration rate (GFR) of shrews with unlimited food and water was 2.4 ml/h, about 60% of the rate predicted from body mass. Of this, about 3% (0.075 ml/h) was excreted as urine with an osmolality of 1944 mmol/kg, 5.5 times plasma osmolality. Shrews had a total water turnover (5 ml/day) two to three times higher than expected from allometry for a small mammal of this size. Water influx was partitioned among preformed water from food (65%), drinking (16%), and metabolic water (20%). Water efflux was divided among urine flow (35%), fecal water loss (estimated as 23%), and evaporation (by difference, 42%). Least shrews had a high water turnover rate and relatively high urine flow rate (UFR); this likely reflects a combination of factors, including high metabolism, active lifestyle, and wet diet.

Animal Feed↗

Apparent scarcity of glial fibrillary acidic protein expression in the brain of the pygmy shrew Sorex minutus as revealed by immunocytochemistry.

We examined astroglial cells in the brain of the pygmy shrew Sorex minutus (Insectivora). For that purpose we labeled glial fibrillary acidic protein (GFAP) immunohistochemically in brain sections with a polyclonal antibody. Antigen retrieval experiments were performed to counteract formaldehyde fixation masking of GFAP epitopes. Our results showed remarkable paucity of GFAP-immunoreactive cells and fibers in the cerebral cortex and nuclei, as well as in the majority of the diencephalic and mesencephalic structures. In the forebrain, significant numbers of GFAP-containing astrocytes were found only in the ependyma and subventricular zones, superficial part of layer I of the cerebral cortex, and the majority of white matter structures. In the diencephalon, habenular nuclei were rich in GFAP-immunopositive astrocytes and labeled radial fibers were extended between median eminence and the third ventricle. A considerably higher density of labeled astrocytes was detected in the caudal brainstem and cerebellum. In contrast, in the mouse brain, immunoreactive astrocytes were present in large quantities in various structures. Staining of sections of the shrew brain against glutamine synthetase revealed abundance of immunofluorescent astrocytes in many areas, especially in the shrew cerebral cortex. It seems probable that in the shrew brain only a limited fraction of astroglia expresses GFAP, while other astroglial cells can be detected with different markers. It is possible that the rodent type of astroglial GFAP expression might not be common to insectivores and probably to some other mammalian orders.

Animals↗

Placenta and fetal membranes of the four-toed elephant shrew, Petrodromus tetradactylus.

The four-toed elephant shrew is the only extant member of the genus Petrodromus. Some features of its placentation are found in other elephant shrews and may be important in consolidating the elephant shrews in the order Macroscelidea. These include implantation in an embryo chamber at the caudal end of the uterine horn; a large yolk sac; a large, lobulated allantoic sac; a distinct layer of proliferative trophoblastic cells between the labyrinthine and spongy zones of the placenta; an accessory epitheliochorial placenta; and the formation of a mesoplacenta by the decidua basalis. Of particular note are the large granulated cells that occupy the walls of decidual, myometrial and mesometrial arteries. They appear to be uterine natural killer cells. The persistence of a relatively large yolk sac and a large lobulated allantoic sac is a feature which Petrodromus and other elephant shrews share with other members of the superordinal clade Afrotheria.

Animals↗

Sex-specific regulation of marking behavior by sex hormones and conspecifics scent in tree shrews (Tupaia belangeri).

In tree shrews (Tupaia belangeri), both males and females mark their surroundings with urine and skin gland secretions. In this study we examined in standard tests the modulation of marking behavior of male and female tree shrews by hormones and chemical signals of conspecifics. Males mark more frequently than females in the absence of conspecific scent. Female scent elicits only low marking responses in both sexes. Male scent stimulates marking of both sexes, with a much higher response in females. Increasing amounts of male scent result in a corresponding increase in marking by females. In contrast, male marking is maximally stimulated by low scent concentrations of male conspecifics, whereas higher scent concentrations inhibit male marking. Gonadectomy diminishes male marking activities under all scent conditions, whereas in females only the response to male scent is reduced. In unscented cages, the low marking activities of castrated males and intact females are stimulated by treatment with testosterone or estradiol. In the presence of male scent, however, only testosterone, but not estradiol, stimulates marking of castrated males. Marking of intact males or females is not increased by either hormone. As this study shows, external (scent of conspecifics) and internal factors (sex hormone levels) interact to regulate tree shrews' marking behavior in a sex-specific manner. These results are discussed with respect to the sex-specific contexts and functions of tree shrews' marking behavior.

Animal Communication↗

Liver apoptosis after dimethylnitrosamine administration in shrews.

Female house musk shrews (Suncus murinus, Insectivora) were given a single i.p. dose of 30 mg/kg dimethylnitrosamine (DMN) at 8 weeks of age which was lethal 36 to 48 hrs after dosing. Liver tissues were collected from shrews killed 3, 6, 16, 24 and 36 hrs after treatment, and the sequential development of the lesions was characterized. DMN induced acute centrilobular cell injury. In 6 hrs, a few cells became apoptotic in the centrilobular area; the number increased at 16 hrs and 24 hrs, and was prominent at 36 hrs. There was no inflammatory reaction or necrosis and hemorrhage was not obvious. These apoptotic cells as well as normal appearing cells in the centrilobular area were labeled by the TUNEL method. In both hepatocytes and endothelial cells, apoptosis was confirmed electron microscopically as nuclear chromatin condensation at the periphery with no mitochondria swelling. When an i.p. dose of 10 mg/kg DMN was given twice at 8 and 9 weeks of age, no acute toxicity was induced, and the liver of shrews surviving for 50 weeks of age was normal with no tumor formation. These findings indicate that a single i.p. administration of 30 mg/kg DMN induced severe and fatal toxicity on liver tissues in shrews due to apoptosis, whereas 2 x 10 mg/kg DMN had no carcinogenic effect.

Animals↗

Rod bipolar cells in the cone-dominated retina of the tree shrew Tupaia belangeri.

The tree shrew has a cone-dominated retina with a rod proportion of 5%, in contrast to the common mammalian pattern of rod-dominated retinae. As a first step to elucidate the rod pathway in the tree shrew retina, we have demonstrated the presence of rod bipolar cells and studied their morphology and distribution by light and electron microscopy. Rod bipolar cells were labeled with an antiserum against the protein kinase C (PKC), a phosphorylating enzyme. Intense PKC immunoreactivity was found in perikarya, axons, and dendrites of rod bipolar cells. The cell bodies are located in the sclerad part of the inner nuclear layer, the dendrites ascend to the outer plexiform layer where they are postsynaptic to rod spherules, and an axon descends towards the inner plexiform layer (IPL). The axons branch, and terminate in the vitread third of the IPL where mammalian rod bipolar cells are known to terminate. Two amacrine cell processes are always seen as the postsynaptic elements (dyads). Dendritic and axonal arbors of rod bipolar cells are rather large, up to 100 microns in diameter. The topographical distribution of the rod bipolar cells was analyzed quantitatively in tangential sections. Their density ranges from 300 cells/mm2 in peripheral retina to 900 cells/mm2 more centrally. The distribution is rather flat with no local extremes. Consistent with the low rod proportion in tree shrew, the rod bipolar cell density is low compared to the rod-dominated cat retina for example (36,000-47,000 rod bipolar cells/mm2). Rod-to-rod bipolar cell ratios in the tree shrew retina range from smaller than 1 to about 7, and thus are also lower than in cat.

Animals↗

Alkoxyresorufin-O-deethylase activities and polychlorinated biphenyl patterns in shrews as biomarkers in environmental risk assessments: sensitivity and specificity.

Alkoxyresorufin-O-deethylase (AROD) biomarkers are useful indicators of the exposure of organisms to dioxin-like compounds. In the current study, an in vivo validation of the use of such biomarkers in shrews was conducted. Furthermore, the use of changes in polychlorinated biphenyl (PCB) patterns as an animal-friendly alternative to AROD biomarkers was evaluated. Two experiments and a field study were conducted in which dose-response relations were established between levels of sigma-PCBs in shrews on one hand and their AROD activities and changes in PCB patterns on the other. We demonstrate that the changes in PCB patterns are as sensitive as the classic AROD biomarkers. The experiments also showed a substrate-specific induction of AROD biomarkers and a related PCB congener-specific metabolism. This implies that congener-specific analysis of PCBs can reveal activities of specific AROD biomarkers. Gender-specific induction of AROD activities in shrews was shown in the field study, whereas the relationship between exposure and changes in PCB patterns did not differ between genders. It is concluded that (i) AROD biomarkers are useful biomarkers to assess exposure of shrews to specific organochlorines and that (ii) changes in PCB patterns can be used as an animal-friendly alternative to these AROD biomarkers.

Animals↗

Ondansetron and Delta-9-tetrahydrocannabinol interfere with the establishment of lithium-induced conditioned taste avoidance in the house musk shrew (Suncus murinus).

Considerable evidence suggests that rats can learn to avoid a taste in the absence of nausea. The current experiments evaluated the potential of the antiemetic agents, ondansetron (OND) and delta-9-tetrahydrocannabinol (THC), to interfere with lithium chloride (LiCl)-induced taste avoidance in the house musk shrew, Suncus murinus, an insectivore that, unlike rats, is capable of vomiting. At a dose that did not modify saccharin (Experiment 1) or sucrose (Experiment 2) intake, OND prevented the establishment of LiCl-induced taste avoidance in the shrew. A low dose of THC (1 mg/kg), which did not modify sucrose intake during conditioning, also prevented the establishment of LiCl-induced taste avoidance in the shrew. Higher doses of THC were also effective, but they also suppressed sucrose consumption during conditioning. These results suggest that nausea is a necessary component of the unconditioned stimulus for the establishment of conditioned taste avoidance in the shrew, unlike the rat, which does not vomit when injected with a toxin.

Analgesics, Non-Narcotic↗

Distribution pattern of HNF-3beta proteins in developing embryos of two mammalian species, the house shrew and the mouse.

The pattern of expression of HNF-3beta in organizing centers and axial structures during early vertebrate development suggests an important role for this protein in the establishment of the vertebrate body plan. To establish whether the pattern of expression during embryogenesis is species specific, a comparative immunohistochemical study of two mammalian species, the house shrew, insectivore, and the mouse was carried out; it is difficult to obtain accurate morphological differences from the study of remotely related animals. The earliest expression of HNF-3beta appeared in the node and hypoblast (or endoderm) in both species, where the presumptive foregut endoderm showed intense immunoreactivity prior to the formation of the axial mesoderm, suggesting a role different from that in axial formation. The anterior extension of immunopositive axial mesoderm and the median band of the neural plate varied between the two species, and was delayed in the house shrew. HNF-3beta in the anterior end of the foregut disappeared transiently in the house shrew but persisted in the mouse embryo. An asymmetric pattern of distribution in the primitive streak was also observed in the mouse but not in the house shrew. The present immunohistochemical study elucidated that the distribution of HNF-3beta is conserved initially but soon manifests species specificities in development even between mammalian species.

Animals↗

Olfactory mucosa ultrastructure in the short-tailed shrews, Blarina brevicauda and Blarina carolinensis.

The olfactory mucosae of the northern short-tailed shrew, Blarina brevicauda, and the southern short-tailed shrew, Blarina carolinensis, were examined by light and electron microscopy. A well-developed olfactory epithelium was observed that included all of the cells necessary to provide for a sensitive olfactory system, suggesting that olfaction plays a major role in the behavior of these animals. There were no significant differences between the olfactory mucosae of these two species. The general features of the olfactory epithelium in these shrews were similar to those reported for several other macrosmatic mammals. A new type of supporting cell, called the light supporting cell, was observed in these shrews. The light supporting cell cytoplasm exhibited very little staining by light microscopy and had low electron density by transmission electron microscopy compared to that of the more common dark supporting cell. The light supporting cell had a convex apical surface with microvilli and lacked the large amounts of smooth endoplasmic reticulum (SER) typical of the apical cytoplasm of the dark supporting cell. In the lamina propria of the mucosa, the Bowman's glands consisted of two cell types, one with electron-lucent, alcian blue-positive granules, and the other with electron-dense PAS-positive granules. The cell with electron-lucent granules contained large amounts of SER and small clumps of rough ER. The cells with electron-dense granules had large amounts of RER and little SER.

Animals↗

Cytochrome P-450-dependent fatty acid hydroxylase system in Japanese house musk shrew, Suncus murinus.

The substrate specificity and other properties of a fatty acid hydroxylase system in liver microsomes of the Japanese house musk shrew, Suncus murinus, were examined. Shrew liver microsomes catalyzed the hydroxylation of various saturated fatty acids (C8-C18) to the corresponding omega- and (omega-1)-hydroxy derivatives. The relative activities of the hydroxylase with these substrates were as follows: C12 (100, actual conversion: 7.05 nmol/mg of microsomal protein/min), C10 (90), C14 (87), C16 (23), C8 (19), and C18 (11). The specific activity of the fatty acid hydroxylase in shrew liver was much higher than that in other species. The omega/omega-1-hydroxylation ratio decreased with increasing chain length of fatty acid substrates (C10-C18), but it was 0 for the C8 fatty acid. Both NADPH and O2 were required for hydroxylase activity, and NADH had little effect. The apparent Km value for laurate was 1.6 X 10(-5) M. The hydroxylase activity was 92% inhibited by CO at a CO-O2 ratio of 9. p-Chloromercuribenzoate (0.1 mM) inhibited hydroxylation by 94% whereas iodoacetate (0.1 mM) inhibited it by only 8%. SKF 525-A (1 mM) and menadione (0.01 mM), respectively, caused 41% and 29% inhibition of the activity. It is concluded that the hydroxylase catalyzing fatty acid hydroxylation in shrew liver microsomes is a typical cytochrome P-450-linked monooxygenase.

Animals↗

Cream, a new coat-color mutant in the musk shrew.

A coat-color mutant was found in the wild musk shrew (Suncus murinus, Insectivora). Five musk shrews with gray pelage, the common coat color of this species, were captured in the village of Tambum near Jakarta, Indonesia. Two males and two females were transported to Japan and mated. Matings between one male and two females segregated several cream-colored offspring, a color that had never been seen before in this species. From the pedigree record and data on mating experiments, it was confirmed that this mutant coat color was expressed in the homozygote by an autosomal recessive gene designated cr, and at least three of the four wild shrews examined were carriers of this gene. The cr gene was associated with failure of normal pigmentation in the pelage and skin. The mutant shrews also showed some behavioral abnormalities.

Animals↗

Brevinema andersonii gen. nov., sp. nov., an infectious spirochete isolated from the short-tailed shrew (Blarina brevicauda) and the white-footed mouse (Peromyscus leucopus).

A spirochete which infects short-tailed shrews (Blarina brevicauda) and white-footed mice (Peromyscus leucopus) has been shown previously to be ultrastructurally and serologically distinct from other spirochetes. Two of the original isolates from Connecticut and Minnesota and 16 new isolates obtained from shrews captured in Minnesota were characterized phenotypically and genetically in this study. A comparative analysis of the 16S rRNA sequences of two shrew isolates and one mouse isolate and the 16S rRNA sequences of 16 other spirochetes and Escherichia coli revealed that these organisms exhibited low levels of similarity (range of similarity values, 73.9 to 77.8%; average level of similarity, 74.7%). The shrew and mouse isolates which we examined formed a deeply branching subgroup that was clearly distinct from the other genera of spirochetes examined. These and other results indicated that the new spirochetes represent a unique taxon in the order Spirochaetales. Accordingly, we propose that they should be classified as members of a new genus, Brevinema. The three strains of Brevinema which we examined had 16S rRNA sequences that were nearly identical. We also compared these isolates by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, fatty acid and enzyme analyses, restriction endonuclease analysis, and Southern hybridization and found that the levels of genetic and phenotypic homogeneity among the strains were very high. We concluded that the isolates which we examined were members of a single species, for which we propose the name Brevinema andersonii. The type strain of Brevinema andersonii is CT11616 (= ATCC 43811).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Morphogenesis of esophageal carcinoma induced by N-methyl-N'-nitro-N-nitrosoguanidine in the house musk shrew, Suncus murinus (Insectivora).

The histological changes occurring in the esophageal mucosa of shrews (Suncus murinus) after N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) treatment were investigated sequentially. Six-week-old female shrews were given a 50 micrograms/ml MNNG solution as drinking water for 30 weeks, and 5 selected at random were killed at 10 and 20 weeks of age, and thereafter at 5-week intervals until 45 weeks of age. Controls were killed at 45 weeks of age. The MNNG-induced esophageal lesion in shrews began from basal cell hyperplasia at 20 weeks of age, followed by dysplasia occurring at 25 weeks of age, then progressed toward intraepithelial carcinoma to invasive squamous cell carcinoma at 35 weeks of age. Apparent sequential dysplasia-carcinoma transition was seen. Papillomas were seen from 25 weeks of age but there was no evidence of papilloma-carcinoma sequence. Five MNNG-untreated shrews killed at the end of the experiment were free of esophageal tumors.

Animals↗