A new viewpoint on antlers reveals the evolutionary history of deer (Cervidae, Mammalia).

Recent molecular phylogeny of deer revealed that the characters of antlers previously focused on are homoplasious, and antlers tend to be considered problematic for classification. However, we think antlers are important tools and reconsidered and analysed the characters and structures to use them for classification. This study developed a method to describe the branching structure of antlers by using antler grooves, which are formed on the antlers by growth, and then projecting the position of the branching directions of tines on the burr circumference. By making diagrams, comparing the branching structure interspecifically, homologous elements (tines, beams, and processes) of the antlers of 25 species of 16 genera were determined. Subsequently, ancestral state reconstruction was performed on the fixed molecular phylogenetic tree. It was revealed that Capreolinae and Cervini gained respective three-pointed antlers independently, and their subclades gained synapomorphous tines. We found new homologous and synapomorphous characters, as the antler of Eld's deer, which has been classified in Rucervus, is structurally close to that of Elaphurus rather than that of Rucervus, consistent with molecular phylogeny. The methods of this study will contribute to the understanding of the branching structure and phylogeny of fossil species and uncover the evolutionary history of Cervidae.

Chromosome observation by scanning electron microscopy using ionic liquid.

Electron microscopy has been used to visualize chromosome since it has high resolution and magnification. However, biological samples need to be dehydrated and coated with metal or carbon before observation. Ionic liquid is a class of ionic solvent that possesses advantageous properties of current interest in a variety of interdisciplinary areas of science. By using ionic liquid, biological samples need not be dehydrated or metal-coated, because ionic liquid behaves as the electronically conducting material for electron microscopy. The authors have investigated chromosome using ionic liquid in conjunction with electron microscopy and evaluated the factors that affect chromosome visualization. Experimental conditions used in the previous studies were further optimized. As a result, prewarmed, well-mixed, and low concentration (0.5∼1.0%) ionic liquid provides well-contrasted images, especially when the more hydrophilic and the higher purity ionic liquid is used. Image contrast and resolution are enhanced by the combination of ionic liquid and platinum blue staining, the use of an indium tin oxide membrane, osmium tetroxide-coated coverslip, or aluminum foil as substrate, and the adjustment of electron acceleration voltage. The authors conclude that the ionic-liquid method is useful for the visualization of chromosome by scanning electron microscopy without dehydration or metal coating.

Immunohistochemical study of endocrine cells in the gastrointestinal tract of the barking deer, Muntiacus muntjak.

The morphology, distribution and relative frequencies of endocrine cells in the gastrointestinal tract of the barking deer (Muntiacus muntjak) were investigated. The immunohistochemical method employed used seven types of antisera against chromogranin, serotonin, gastrin, cholecystokinin, somatostatin, glucagon and insulin. Chromogranin and serotonin immunoreactive (IR) cells were found throughout the gastrointestinal tract. Conversely, gastrin-IR cells were only found in the oxyntic gland, pyloric gland and duodenum, while cholecystokinin-IR and somatostatin-IR cells were detected in the oxyntic gland, pyloric gland and small intestines. Somatostatin-IR cells were also seen in the caecum. Glucagon-IR cells were found in all parts of the gastrointestinal tract apart from the colon and rectum. No insulin-IR cells were found in the gastrointestinal tract of this species. The cells in the small intestine were generally spindle shaped with long cytoplasmic processes ending in the lumen (open type), while in the stomach and large intestine, they were occasionally round or spherical in shape (closed type). An uncommon distribution pattern of endocrine cells in the gastrointestinal tract of the barking deer was noted for cholecystokinin- and glucagon-IR cells.

Morphological study of the lingual papillae in the barking deer, Muntiacus muntjak.

The morphology of the tongue of the adult barking deer, Muntiacus muntjak, was examined by light and scanning electron microscopy. The result showed that the tongue of the barking deer was elongated with a rounded apex. Four types of lingual papillae were observed: filiform, fungiform, vallate and large conical papillae. The filiform papillae represented the most numerous types of lingual papillae. The fungiform papillae were distributed among the filiform papillae on the rostral and the body portions of the tongue. Ten to thirteen vallate papillae were distributed on both sides of the lingual prominence among the large conical papillae. Histologically, both the fungiform and vallate papillae contain taste buds in the epithelial layer. The distribution and types of lingual papillae found in the barking deer are similar to those in the other species that belong to the family Cervidae.

Histological study of the parotid and mandibular glands of barking deer (Muntiacus muntjak) with special reference to the distribution of carbohydrate content.

We investigated the histology and carbohydrate content of the parotid and mandibular glands of the barking deer (Muntiacus muntjak). Three adult males were used. Paraffin wax sections of the glands were stained with haematoxylin and eosin (HE), alcian blue (AB), pH 2.5 and periodic acid Schiff (PAS). The acinar cells of the parotid gland were serous, whereas those of the mandibular gland were of the mixed type. The acini of the mandibular gland comprised serous and mucous cells with the mucous type predominating. AB and PAS staining showed high concentrations of acidic and neutral carbohydrates in the mucous cells, but not in the serous cells of the mandibular gland. These carbohydrates were also found in moderate-to-high concentrations in the secreted material in the mandibular duct lumen. However, these carbohydrates were not found in acinar cells of the parotid gland or in the serous cells of the mandibular gland. Thus, carbohydrates in the saliva of the barking deer appear to be produced mainly by the mucous cells of the mandibular glands.

One gland, two lobes: organogenesis of the "Harderian" and "nictitans" glands of the Chinese muntjac (Muntiacus reevesi) and fallow deer (Dama dama).

The nictitans and Harderian glands are enigmatic glands situated in the anterior aspect of the orbit. Traditionally, the nictitans and Harderian glands of mammals have been considered to be two fundamentally distinct glands. However, a consistent, unambiguous distinction between these two glands has remained elusive due to conflicting anatomical and histochemical definitions. The Harderian gland was originally described, and first distinguished from the nictitans gland, in adult deer. We examined the organogenesis and histochemistry of the anterior orbital glandular mass in two species of deer (Muntiacus reevesi and Dama dama) to determine whether it comprises two distinct glands or one bilobed gland. The anterior orbital regions of 30 fetal specimens of both species, along with some adult material, were examined histologically. Four stages of glandular organogenesis were observed. Most notably, both glandular portions developed from the same inception point, but the deep lobe developed faster than the superficial lobe. The common inception point and the relationship of the collecting ducts clearly shows that this is a single glandular mass that differentiates into two lobes rather than two distinct glands. Moreover, although the histochemical profiles of the two lobes differ slightly, both lobes produce lipids, which is further indication that these are not profoundly different glands but part of a single, heterogeneously developed gland. Thus, it is proposed that the terms nictitans and Harderian glands, as separate entities, be discontinued and that the entire gland be referred to as the anterior orbital gland (glandula orbitalis anterior), with superficial and deep lobes (pars superficialis and pars profundus, respectively).

Comparative morphological study on the lingual papillae and their connective tissue cores (CTC) in reeves' muntjac deer (Muntiacus reevesi).

The lingual papillae and their connective tissue cores (CTC) from Reeves' muntjac deers (herbivorous artiodactyla) were studied using light and scanning electron microscopy and then compared to those of other mammalian species. At the posterior portion of the tongue, the Reeves' muntjac has a lingual prominence on which large conical papillae are distributed. On the dorsal surface of the anterior tongue, numerous filiform papillae were found. Externally, each filiform papilla consists of a rod-shaped main process and several small accessory processes. Their CTCs consist of 10 or more rod-shaped processes arranged in a horseshoe pattern and several posterior processes forming a small circular pattern. This structure is a common characteristic of artiodactyla, through which Reeves' muntjac deer can be categorized in a position in the artiodactyla class lying between the bighorn sheep and the East African bongo. Fungiform papillae are distributed among the filiform papillae on the anterior portion of the tongue. Large fungiform papillae are also sparsely distributed on the lingual prominence and have several taste buds in the epithelium on the surface. Ten or more vallate papillae are distributed at the postero-lateral area of the lingual prominence and numerous taste buds are distributed in the epithelium of their side.