The vomeronasal organ and incisive duct of harbor seals are modified to secrete acidic mucus into the nasal cavity.

Most terrestrial mammals have a vomeronasal system to detect specific chemicals. The peripheral organ of this system is a vomeronasal organ (VNO) opening to the incisive duct, and its primary integrative center is an accessory olfactory bulb (AOB). The VNO in seals is thought to be degenerated like whales and manatees, unlike otariids, because of the absence of the AOB. However, olfaction plays pivotal roles in seals, and thus we conducted a detailed morphological evaluation of the vomeronasal system of three harbor seals (Phoca vitulina). The VNO lumen was not found, and the incisive duct did not open into the oral cavity but was recognized as a fossa on the anteroventral side of the nasal cavity. This fossa is rich in mucous glands that secrete acidic mucopolysaccharides, which might originate from the vomeronasal glands. The olfactory bulb consisted only of a main olfactory bulb that received projections from the olfactory mucosa, but an AOB region was not evident. These findings clarified that harbor seals do not have a VNO to detect some chemicals, but the corresponding region is a specialized secretory organ.

Morphological variation of tail bone among two chicken breeds and their F1 progeny.

Fancy breeds of Japanese indigenous chicken display extensive morphological diversity, particularly in tail feathers. Although marked differences in tail and bone traits have been reported between Tosa-jidori (wild type) and Minohikichabo (rich type) breeds, little is known about the pattern of genetic inheritance in cross experiments. Therefore, this study aimed to investigate the strain and sex effects, and inheritance patterns, in the morphometric variation of pygostyle bones among Tosa-jidori, Minohikichabo, and their F1 hybrids. Five morphological traits, angle of the apex of the pygostyle, pygostyle length, margo cranialis length, tail feather number, and body weight, were evaluated at the adult stage. A significant strain difference was detected in all traits, whereas significant sex differences were observed in only three traits, but not in the angle of the apex of the pygostyle and tail feather number. In F1 hybrids, the angle of the apex of the pygostyle was significantly different to that of Tosa-jidori but not that of Minohikichabo, whereas the pygostyle length and tail number of F1 hybrids were significantly different from those of Minohikichabo but not those of Tosa-jidori. A significant heterosis effect was found in the margo cranialis length and body weight. All five traits showed nonadditive inheritance patterns but varied in each trait between partial dominance (angle of the apex of pygostyle), full dominance (pygostyle length and tail feather number), and over-dominance (margo cranialis length and body weight). Interestingly, different patterns of genetic inheritance in the F1 hybrid were observed at different locations, even within the same pygostyle bone. Using the Japanese indigenous chicken model, these results provide a substantial step toward understanding the genetic architecture of morphology in chickens.

The vomeronasal system in semiaquatic beavers.

In contrast to the main olfactory system that detects volatile chemicals in the nasal air, the vomeronasal system can detect nonvolatile chemicals as well as volatiles. In the vomeronasal system, chemicals are perceived by the vomeronasal organ (VNO) projecting axons to the accessory olfactory bulb (AOB). Beavers (Castor spp.) are semiaquatic mammals that have developed chemical communication. It is possible that the beaver's anal gland secretions, nonvolatile and insoluble substances, may work as a messenger in the water and that beavers may detect the nonvolatile chemicals floating on the water surface via the VNO. The present study aimed to clarify the specificities of the beaver vomeronasal system by histologically and immunohistochemically analyzing the VNO and AOB of 12 Eurasian beavers (C. fiber). The VNO directly opened to the nasal cavity and was independent of a narrow nasopalatine duct connecting the oral and nasal cavities. The VNO comprised soft tissues including sensory and nonsensory epithelium, glands, a venous sinus, an artery, as well as cartilage inner, and bone outer enclosures. The AOB had distinct six layers, and anti-G protein α-i2 and α-o subunits were, respectively, immunoreactive in rostral and caudal glomeruli layers indicating expressions of V1Rs and V2Rs. According to gene repertories analysis, the beavers had 23 and six intact V1R and V2R genes respectively. These findings suggested that beavers recognize volatile odorants and nonvolatile substances using the vomeronasal system. The beaver VNO was developed as well as in other rodents, and it had two specific morphological features, namely, disadvantaged contact with the oral cavity because of a tiny nasopalatine duct, and a double bone and cartilage envelope. Our results highlight the importance of the vomeronasal system in beaver chemical communication and support the possibility that beavers can detect chemicals floating on the water surface via the VNO.

Histological findings of sperm storage in green turtle (Chelonia mydas) oviduct.

Green turtles (Chelonia mydas) are seasonal breeders with a time lag between mating and nesting periods. We therefore investigated whether female turtles store sperm like some other animals by histologically and ultrastructurally analyzing oviducts collected from three mature female free-ranging green turtles during the breeding season in the Ogasawara Islands, Japan. The oviduct comprised an infundibulum, magnum, isthmus, uterus, and vagina. Sperm was found in the isthmus of all turtles examined. Some spermatozoa were found in the duct and acini of glands in the isthmus of two turtles with oviducts containing eggs, and a few were also located in the transition area between the uterus and vagina of one of the turtles. On the other hand, we also found abundant spermatozoa on the luminal surface of the isthmus of one turtle captured during mating. In most reptiles, fertilization occurs in the infundibulum or albumen region, and thus the isthmus near those areas might be suitable for storing sperm in female turtles.

Contraceptive effect of a gonadotropin-releasing hormone vaccine on a captive female African Lion (Panthera leo): a case study.

Lions (Panthera leo) breed well under captivity, so contraception has been commonly conducted for population management, leading to a demand for a less invasive and reversible contraceptive approach in lions. In this study, we examined the efficacy of a commercial gonadotropin-releasing hormone vaccine as a method of suppressing reproductive activity in a sexually matured female lion. Under behavioral restraint, the vaccine was injected twice (days 0 and 109). After the initial vaccination, ovarian activity is still observed. After the second vaccination, contraceptive effect was confirmed for 246 days until restart of estrous cycles. We confirmed only a slight swelling around the injection site after the second vaccination. This study may suggest an alternative option for a contraceptive method in lions.

Associations between plasma testosterone levels and season, nutritional status, age, and body size in free-ranging male Asian black bears (Ursus thibetanus) in central Honshu, Japan.

Testosterone is an important regulator of male reproduction in terms of spermatogenesis and physiological changes in the accessory reproductive organs. Despite the important role of the testosterone level in male reproduction, testosterone levels vary among male bears even during the breeding season, and the causative underlying factors remain unknown. We postulated that testosterone levels are higher during the breeding season than during other seasons in free-ranging male bears, and that testosterone levels increase with advancing age, larger body size, and better nutritional status. We assessed potential factors (season, time of sampling, body condition index, head circumference, and age) associated with the testosterone level in 80 blood samples collected from free-ranging Asian black bears (Ursus thibetanus) captured in barrel traps in the Ashio-Nikko Mountains and Okutama Mountains in central Honshu Island, Japan. The plasma testosterone level was higher during the breeding season (May-July) than during the non-breeding season (August-November). The body condition index was significantly and positively associated with the plasma testosterone level. None of the other factors were significantly associated with the plasma testosterone level. Therefore, the body condition index may be essential for maintaining high plasma testosterone levels. These findings imply that testosterone secretion might be activated in free-ranging males during the breeding season, and animals with good nutritional status may be able to maintain high plasma testosterone levels. This study uncovered an association between nutritional conditions and reproductive activity in male bears.

Seasonal ultrastructural changes in apocrine gland cells in back skin of male brown bears (Ursus arctos).

Oily secretions from the back skin are involved in the marking behavior of male brown bears (Ursus arctos), and apocrine glands in back skin are activated during the breeding season. Here, we investigated seasonal changes in the intracellular organelles of apocrine gland cells in the back skin of male brown bears using transmission electron microscopy (TEM) and osmium-maceration scanning electron microscopy (OM-SEM). The morphological features of mitochondria and intracellular granules, and secretory mechanisms obviously differed between breeding and non-breeding seasons. The TEM findings showed that contents of low-density granules were released into the glandular lumen by frequent exocytosis, and sausage-shaped mitochondria were located in the perinuclear region during the non-breeding season. In contrast, high-density granules appeared in the apical region and in projections during the breeding season, and swollen mitochondria and lysosome-like organelles separating into high-density granules were located in the perinuclear region. The OM-SEM findings revealed swollen mitochondria with only a few partially developed cristae, and small mitochondria with cristae shaped like those in swollen mitochondria in the apical regions during the breeding season. These findings indicated that the small mitochondria corresponded to the high-density granules identified by TEM. These findings suggested that mitochondria in apocrine gland cells swell, degenerate, fracture into small pieces, and are finally released by apocrine secretions during the breeding season. Small mitochondria released in this secretory manner might function as the source of chemical signals in the oily secretions of brown bears during the breeding season.

Behavioral effects of scents from male mature Rathke glands on juvenile green sea turtles (Chelonia mydas).

Sea turtles can detect airborne and waterborne odors, but whether they recognize scents from the same species and if so, how they affect their behavior remains unknown. The present study evaluated the behavioral effects of odorants on juvenile green sea turtles (Chelonia mydas). The odorants were derived from Rathke glands (external scent glands) of mature male green sea turtles, and from two types of food. The activity of the juveniles increased when exposed to food scents, and significantly decreased compared with controls when exposed to scents from Rathke glands. These findings indicated that scents from the same species affect behavior, and that chemical communication via olfaction has important outcomes for sea turtles.

Comparative histological studies on properties of polysaccharides secreted by vomeronasal glands of eight Laurasiatheria species.

Most mammalian species have a vomeronasal organ that detects specific chemical substances, such as pheromones. Mucous fluid covering the vomeronasal sensory epithelium is secreted by vomeronasal glands, and the properties of these fluids have been suggested to be involved in chemical detection. Histological studies using periodic acid-Schiff (PAS) and Alcian blue pH 2.5 (AB) stains, which respectively detect natural and acidic polysaccharides, have suggested variations in the nature of the vomeronasal glands among species. Here, we investigated the responsivity of the vomeronasal glands to PAS and AB stains in eight Laurasiatheria species. All species studied herein possessed vomeronasal glands that stained positive for PAS, like other many reported species. The vomeronasal glands of dogs and minks - like rodents, were AB-negative, whereas those of cows, goats, sika deer, musk shrews and two bat species were positive. Considering the present findings and previous reports, the vomeronasal glands in most of Laurasiatheria species appear to be fundamentally abundant in acidic polysaccharides, whereas those in carnivores essentially contains neutral polysaccharides.

Ultrastructural changes in colonic epithelial cells in a rat model of inflammatory bowel disease.

Inflammatory bowel disease (IBD) is a global, chronic intractable disease. The functions of drugs and food components have been evaluated in models of IBD induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS). Here, we used transmission (TEM) and osmium-maceration scanning (SEM) electron microscopy to evaluate the ultrastructure of colonic epithelial cells in rat models of IBD induced by TNBS. Histological evaluation revealed that the intestinal crypts in the most regions of the IBD-model colons were deformed and we classified them as having high cell migration rates (HMIG). The remaining regions in the intestinal crypts retained a relatively normal structure and we classified them as having low cell migration rates (LMIG). Osmium-maceration SEM revealed the mucosal fluid flowing in spaces without secretory granules in crypt goblet cells of both HMIG and LMIG regions, indicating the depletion of goblet cell mucin that is found in patients with IBD. The Golgi apparatus in absorptive cells was stacked and curled in both regions. Osmium-maceration SEM showed membrane network structures resembling endoplasmic reticulum that were large and expanded in absorptive cells with HMIG rather than with LMIG regions in IBD-model colons. These findings indicated that endoplasmic reticulum stress is associated with susceptibility to IBD and that the effects of various agents can be evaluated according to endoplasmic reticulum stress revealed by using electron microscopy in models of IBD induced by TNBS.

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.

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.

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.