Daisuke Kondoh

Macrophages are the determinant of resistance to and outcome of nonlethal Babesia microti infection in mice.

ABSTRACT In the present study, we examined the contributions of macrophages to the outcome of infection with Babesia microti, the etiological agent of human and rodent babesiosis, in BALB/c mice. Mice were treated with clodronate liposome at different times during the course of B. microti infection in order to deplete the macrophages. Notably, a depletion of host macrophages at the early and acute phases of infection caused a significant elevation of parasitemia associated with remarkable mortality in the mice. The depletion of macrophages at the resolving and latent phases of infection resulted in an immediate and temporal exacerbation of parasitemia coupled with mortality in mice. Reconstituting clodronate liposome-treated mice at the acute phase of infection with macrophages from naive mice resulted in a slight reduction in parasitemia with improved survival compared to that of mice that received the drug alone. These results indicate that macrophages play a crucial role in the control of and resistance to B. microti infection in mice. Moreover, analyses of host immune responses revealed that macrophage-depleted mice diminished their production of Th1 cell cytokines, including gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α). Furthermore, depletion of macrophages at different times exaggerated the pathogenesis of the infection in deficient IFN-γ−/− and severe combined immunodeficiency (SCID) mice. Collectively, our data provide important clues about the role of macrophages in the resistance and control of B. microti and imply that the severity of the infection in immunocompromised patients might be due to impairment of macrophage function.

Histological features of the vomeronasal organ in the giraffe, Giraffa camelopardalis

The vomeronasal organ (VNO) that preferentially detects species‐specific substances is diverse among animal species, and its morphological properties seem to reflect the ecological features of animals. This histological study of two female reticulated giraffes (Giraffa camelopardalis reticulata) found that the VNO is developed in giraffes. The lateral and medial regions of the vomeronasal lumen were covered with sensory and nonsensory epithelia, respectively. The vomeronasal glands were positive for periodic acid‐Schiff and alcian blue (pH 2.5) stains. The VNO comprises several large veins like others in the order Cetartiodactyla, suggesting that these veins function in a pumping mechanism in this order. In addition, numerous thin‐walled vessels located immediately beneath the epithelia covering the lumen entirely surrounded the vomeronasal lumen. This sponge‐like structure might function as a specific secondary pump in giraffes.

Morphological and histological features of the vomeronasal organ in the brown bear

The vomeronasal organ (VNO) is a peripheral receptor structure that is involved in reproductive behavior and is part of the vomeronasal system. Male bears exhibit flehmen behavior that is regarded as the uptake of pheromones into the VNO to detect estrus in females. However, the morphological and histological features of the VNO in bears have not been comprehensively studied. The present study investigated the properties and degree of development of the VNO of the brown bear by histological, histochemical and ultrastructural methods. The VNO of bears was located at the same position as that of many other mammals, and it opened to the mouth like the VNO of most carnivores. The shape of the vomeronasal cartilages and the histological features of the sensory epithelium in the bear VNO were essentially similar to those of dogs. Receptor cells in the VNO of the bear possessed both cilia and microvilli like those of dogs. The dendritic knobs of receptor cells were positive for anti‐G protein alpha‐i2 subunit (Gαi2) but negative for anti‐G protein alpha‐o subunit, indicating preferential use of the V1R‐Gαi2 pathway in the vomeronasal system of bears, as in other carnivores. The VNO of the bear possessed three types of secretory cells (secretory cells of the vomeronasal gland, multicellular intraepithelial gland cells and goblet cells), and the present findings showed that the secretory granules in these cells also had various properties. The vomeronasal lumen at the middle region of the VNO invaginated toward the ventral region, and this invagination contained tightly packed multicellular intraepithelial gland cells. To our knowledge, this invagination and intraepithelial gland masses in the VNO are unique features of brown bears. The VNO in the brown bear, especially the secretory system, is morphologically well‐developed, suggesting that this organ is significant for information transmission in this species.

Localization of α1-2 Fucose Glycan in the Mouse Olfactory Pathway

Glycoconjugates in the olfactory system play critical roles in neuronal formation, and α1-2 fucose (α1-2Fuc) glycan mediates neurite outgrowth and synaptic plasticity. Histochemical findings of α1-2Fuc glycan in the mouse olfactory system detected using Ulex europaeus agglutinin-I (UEA-I) vary. This study histochemically assessed the main olfactory and vomeronasal pathways in male and female ICR and C57BL/6J mice aged 3-4 months using UEA-I. Ulex europaeus agglutinin-I reacted with most receptor cells arranged mainly at the basal region of the olfactory epithelium. The olfactory nerve layer and glomerular layer of the main olfactory bulb were speckled with positive UEA-I staining, and positive fibers were scattered from the glomerular to the internal plexiform layer. The lateral olfactory tract and rostral migratory stream were also positive for UEA-I. We identified superficial short-axon cells, interneurons of the external plexiform layer, external, middle and internal tufted cells, mitral cells and granule cells as the origins of the UEA-I-positive fibers in the main olfactory bulb. The anterior olfactory nucleus, anterior piriform cortex and olfactory tubercle were negative for UEA-I. Most receptor cells in the vomeronasal epithelium and most glomeruli of the accessory olfactory bulb were positive for UEA-I. Our findings indicated that α1-2Fuc glycan is located within the primary and secondary, but not the ternary, pathways of the main olfactory system, in local circuits of the main olfactory bulb and within the primary, but not secondary, pathway of the vomeronasal system.

Anchorless cell surface proteins function as laminin-binding adhesins in Lactobacillus rhamnosus FSMM22

Abstract Anchorless cell surface proteins (CSPs) were extracted with 1 M lithium chloride solution from Lactobacillus rhamnosus FSMM22. Loss of the anchorless CSPs resulted in a 2‐fold decrease in FSMM22 cells bound to a constitutive extracellular matrix glycoprotein, laminin, in vitro. DNA‐binding protein HU, glyceraldehyde‐3‐phosphate dehydrogenase, lactate dehydrogenase and 30S ribosomal protein S19 (RpsS) were identified by mass spectrometry in the extract as laminin‐binding adhesins. Among the four proteins, RpsS was immunohistochemically confirmed to exist on the cell surface. Our findings strongly suggest that anchorless CSPs can enhance bacterial adhesion to the host.

Testicular regulation of seasonal change in apocrine glands in the back skin of the brown bear ( Ursus arctos )

Brown bears communicate with other individuals using marking behavior. Bipedal back rubbing has been identified as a common marking posture. Oily substances are secreted via enlarged sebaceous glands in the back skin of male bears during the breeding season. However, whether apocrine gland secretions are associated with seasonal changes remains unknown. The present study aimed to identify histological and histochemical changes in the secretory status and the glycocomposition of the apocrine glands in the back skin of male bears in response to changes in seasons and/or reproductive status. The apocrine glands of intact males during the breeding season were significantly larger and more active than those of castrated males during the breeding season and those of intact males during the non-breeding season. Lectin histochemical analyses revealed a more intense reaction to Vicia villosa agglutinin (VVA) in the cytoplasm, mainly Golgi zones of apocrine cells during the breeding season among castrated, compared with intact males. Positive staining for VVA was quite intense and weak in intact males during the non-breeding and breeding seasons, respectively. Ultrastructural analysis revealed VVA positivity in the Golgi zone, especially around secretory granules in apocrine cells. Changes in lectin binding might reflect a change in the secretory system in the apocrine cells. The present histological and histochemical findings of changes in the secretory status and glycocomposition of the apocrine glands according to the season and reproductive status suggest that these glands are important for chemical communication.

Age-dependent decrease in glomeruli and receptor cells containing α1–2 fucose glycan in the mouse main olfactory system but not in the vomeronasal system

Receptor cells of the olfactory epithelium (OE) and vomeronasal organ (VNO) project axons to glomeruli in the main olfactory bulb (MOB) and accessory olfactory bulb (AOB), respectively and undergo continuous turnover throughout life. Alpha1–2 fucose (α1–2Fuc) glycan mediates neurite outgrowth and synaptic plasticity and plays important roles in the formation of the olfactory system during development. We previously confirmed the localization of α1–2Fuc glycan in the olfactory system of 3- to 4-month-old mice but whether such localization persists throughout life remains unknown. Here, the MOB, AOB, OE and VNO of 1-, 3- and 8-month-old mice were histochemically examined using Ulex europaeus agglutinin-I (UEA-I) that specifically binds to α1–2Fuc glycan. Binding sites for UEA-I in the MOB were similar among all age groups but the ratio of UEA-I-positive glomeruli significantly decreased with aging. The frequency of UEA-I-positive receptor cells in the OE of the two older groups was also significantly lower than that of 1-month-old mice. On the other hand, UEA-I binding in the AOB and VNO did not significantly differ among all three groups. These findings suggest that the primary pathway of the main olfactory system requires the role of α1–2Fuc glycan in young mice rather than old mice, while the vomeronasal pathway equally requires this glycan in both young and old mice.

Mobility of the forearm in the raccoon (Procyon lotor), raccoon dog (Nyctereutes procyonoides) and red panda (Ailurus fulgens).

The ranges of pronation/supination of forearms in raccoons, raccoon dogs and red pandas were nondestructively examined. Three carcasses of each species were used for CT analysis, and the left forearms were scanned with a CT scanner in two positions: maximal supination and maximal pronation. Scanning data were reconstructed into three-dimensional images, cross-sectional images were extracted at the position that shows the largest area in the distal part of ulna, and then, the centroids of each cross section of the radius and ulna were detected. CT images of two positions were superimposed, by overlapping the outlines of each ulna, and then, the centroids were connected by lines to measure the angle of rotation, as an index of range of mobility. The measurements in each animal were analyzed, using the Tukey–Kramer method. The average angle of rotation was largest in raccoons and smallest in raccoon dogs, and the difference was significant. In the maximally pronated forearm of all species, the posture was almost equal to the usual grounding position with palms touching the ground. Therefore, the present results demonstrate that the forearms of raccoons can supinate to a greater degree from the grounding position with palms on the ground, as compared with those of raccoon dogs and red pandas.

Decrease in an Inwardly Rectifying Potassium Conductance in Mouse Mammary Secretory Cells after Forced Weaning

Mammary glands are physiologically active in female mammals only during nursing. Immediately after weaning, most lactation-related genes are downregulated and milk production ceases. In our previous study, we have detected an inwardly rectifying potassium channel (Kir) 2.1-like current in mammary secretory (MS) cells freshly isolated from lactating mice. This current is highly sensitive to external Ba2+. The potassium permeability of the Kir channels may contribute to the secretion and/or preservation of ions in milk. We hypothesized that the functions of the Kir channels in MS cells are regulated after weaning. To test this hypothesis, we examined the effect of forced weaning on the Ba2+-sensitive Kir current and Kir2.1 expression in the mouse mammary glands. Twenty-four hours after weaning, the lumina of mammary acini were histologically enlarged by milk accumulation. The whole-cell patch-clamp analyses showed that the Ba2+-sensitive Kir current in the post-weaning MS cells was smaller than in the lactating MS cells. The inward conductances of the current in the lactating and post-weaning cells were 4.25 ± 0.77 and 0.93 ± 0.34 nS, respectively. Furthermore, real-time PCR and Western blot analyses showed that Kir2.1 mRNA and protein expression decreased in the post-weaning mammary gland (mRNA, 90% reduction; protein, 47% reduction). Moreover, the local milk accumulation caused by teat sealing decreased Kir conductance in MS cells (2.74 ± 0.45 and 0.36 ± 0.27 nS for control and sealed mammary glands, respectively). This was concomitant with the reduction in the Kir2.1 mRNA expression. Our results suggest that milk stasis after weaning immediately decreases the Kir conductance in MS cells. This decrease in the Kir conductance may be partly caused by the reduction in the Kir2.1 mRNA and protein expression. These alterations during the post-weaning period may be involved in the cessation of ion secretion and/or preservation in the milk.

Lectin histochemical studies on the olfactory gland and two types of gland in vomeronasal organ of the brown bear

Olfaction is mediated by the vomeronasal and main olfactory systems, and the peripheral vomeronasal organ (VNO) processes species-specific chemicals that are associated with various behaviors in mammals. Sensory epithelial surfaces of the olfactory mucosa and VNO are covered by mucosal fluid that contains secretory products derived from associated glands, and glycoconjugates in the mucosal fluid are involved in odorant reception. The VNO of brown bears contains two types of glands; submucosal vomeronasal glands (VNG) and multicellular intraepithelial glands (MIG). The present study determined the labelling profiles of 21 lectins in the olfactory glands (OG), VNG and MIG of young male brown bears. The OG reacted with 12 lectins, and the VNG and MIG were positive for seven and eight lectins, respectively. Six lectins bound only to the OG, while four reacted with both or either of the VNG and MIG, but not the OG. The differences of lectin labelling pattern between the OG and glands in the VNO suggest that glycans in covering mucosal fluids differ between the olfactory mucosa and VNO. In addition, Bandeiraea simplicifolia lectin-I, Sophora japonica agglutinin and Jacalin reacted with the MIG but not the VNG, whereas Datura stramonium lectin and concanavalin A bound to the VNG, but not the MIG. These findings indicate that the properties of secretory substances differ between the two types of glands in the bear VNO, and that the various secretions from these two types of glands may function in the lumen of VNO together.