Masato Imada

A Morphological Study of Age Changes in Adult Human Auricular Cartilage With Special Emphasis on Elastic Fibers

Objective It is well known that the size of the human auricle increases after it has finished development. The reason why the size of the human auricle continues to enlarge until advanced age after reaching adulthood was investigated by observation of the ultrastructure of elastic fibers in human auricular cartilage.

Three-dimensional aspects of blood vessels in thyroids from normal, low iodine diet-treated, TSH-treated, and PTU-treated rats

SummaryThree-dimensional images of blood vessels in thyroid glands from normal, low iodine diet-treated, thyroid-stimulating hormone (TSH)-treated and propylthiouracil (PTU)-treated rats were investigated by use of the corrosion-cast method. The vascular casts made by the injection of methacrylate resin were observed with the scanning electron microscope. In normal animals, each follicle is surrounded by a clearly defined basket-like capillary network, which is generally independent of adjacent networks, though a few anastomoses or common capillaries are sometimes seen. In low iodine diet-treated or TSH-treated animals, the capillaries in the basket-like network become markedly dilated and fuse with one another. Though the vascular casts of PTU-treated animals are similar to those of low iodine diet-treated or TSH-treated ones in some aspects, most basket-like networks become distorted and irregular in shape, and the capillaries are heterogeneously dilated and show many buds, branches and anastomoses. We consider that these peculiar changes in the thyroid of the PTU-treated animals are due not only to the elevation of serum TSH but also to other unknown factors. It is clear that the distribution and morphology of the thyroid capillaries are extremely affected and changed by functional states of the gland.

Identification of neuronal cell lineage-specific molecules in the neuronal differentiation of P19 EC cells and mouse central nervous system

P19 embryonic carcinoma (EC) cells are one of the simplest systems for analyzing the neuronal differentiation. To identify the membrane‐associated molecules on the neuronal cells involved in the early neuronal differentiation in mice, we generated two monoclonal antibodies, SKY‐1 and SKY‐2, by immunizing rats with a membrane fraction of the neuronally committed P19 EC cells as an antigen. SKY‐1 and SKY‐2 recognized the carbohydrate moiety of a 90 kDa protein (RANDAM‐1) and the polypeptide core of a 40 kDa protein (RANDAM‐2), respectively. In the P19 EC cells, the expression of RANDAM‐1 was colocalized to a part of Nestin‐positive cells, whereas that of RANDAM‐2 was observed in most Nestin‐positive cells as well as β‐III‐tubulin positive neurons. In the embryonic and adult brain of mice, RANDAM‐1 was expressed at embryonic day 8.5 (E8.5), and the localization of antigen was restricted on the neuroepithelium and choroid plexus. The RANDAM‐2 expression commenced at E6.0, and the antigen was distributed not only on the neuroepithelium of embryonic brain but on the neurons of adult brain. Collectively, it was concluded that RANDAM‐1 is a stage specific antigen to express on the neural stem cells, and RANDAM‐2 is constitutively expressed on both the neural stem cells and differentiated neuronal cells in mouse central nervous system (CNS).

Application of rapid freezing followed by freeze-substitution acrolein fixation for cytochemical studies of the rat stomach

SummaryA method involving rapid freezing followed by substitution fixation was developed, using acrolein as a fixative. This was then applied to several cytochemical stainings, and showed well preserved and clear cell structures. Membranes were apparently negatively stained and the ultrastructure of mitochondria, rough endoplasmic reticulum and Golgi apparatus was clearly discernible. The mitochondrial and cytoplasmic matrices were stained rather densely compared with routine chemically fixed preparations, implying a good preservation of matrix substrances. Periodic acid-thiocarbohydrazide-silver proteinate staining was applied to the present method. The mucous granules of surface covering epithelial cells indicated fine staining of bipartite structure and the Golgi apparatus of mucous cells showed clear staining differences based on polarity. Postembedding lectin-ferritin and immunocytochemical stainings were applicable to the present preparations and stable stainings of secretory granules were obtained. A low temperature embedding material, Lowicryl K4M, was also examined. The cell preservation of these samples was not as good as those embedded in Epon, but the rough endoplasmic reticulum and Golgi apparatus of chief cells were stained with anti-pepsinogen antibody as were the secretory granules. The present method was also applicable to light microscopy.

Rose odor can innately counteract predator odor

When animals smell a predator odor such as 2,5-Dihydro-2,4,5-trimethylthiazoline (TMT), even if it is a novel substance, the hypothalamo-pituitary-adrenal (HPA) axis is activated, causing stress-like behaviors. Although the medial part of the bed nucleus of stria terminalis (mBST) is known to be involved in this process, the mechanism remains unclear. Moreover, it is unknown whether there is any odor that can counteract the predator odor, even when the odorants are novel substances for the animals. In this study, we assessed whether rose odor can counteract by counting the number of activated neurons in mice brain following the presentation of rose odor with or without TMT for 30 min. The number of activated cells in the mBST and in the ventrorostral part of the anterior piriform cortex (APC) was significantly reduced by a mixture of TMT and rose odor; however, no significant differences were noted in the dorsal part of the APC and in the olfactory bulb (OB) following TMT presentation with or without rose odor. The results suggest that rose odor may counteract the TMT-induced stress response in the OB and/or APC and suppress the neural circuit to the mBST. It also indicates that there are some odors that can innately counteract predator odor, even when they have not been experienced before.

Anatomical analysis of the human lacrimal drainage pathway under an operating microscope

Mesoscopic structures of the lacrimal drainage pathway were observed under an operating microscope in six cases of 5 adult cadavers. In all these cases the upper and lower canaliculi fused to form the common canaliculus. Two cases with a large common canaliculus showed large projections of mucosa which reduced the inner space of the common canaliculus. The remaining 4 cases with a small common canaliculus did not show any projections. These findings indicate that the human lacrimal system can function physiologically through a small narrow space of the common canaliculus.

Complementary DNA cloning and characterization of RANDAM-2, a type I membrane molecule specifically expressed on glutamatergic neuronal cells in the mouse cerebrum

A membrane‐surface glycoprotein, RANDAM‐2, is one of the neuronal cell lineage‐specific antigens involved in the neuronal differentiation of P19 embryonic carcinoma (EC) cells and the mouse central nervous system (CNS). Complementary DNA cloning of RANDAM‐2 indicated that its nucleotide sequence completely matched that of PA2.26 antigen, a sialomucin‐like transmembrane glycoprotein previously found on tumorigenic keratinocytes. RANDAM‐2 transcripts were detectable from the embryonic stage of 6.5 days, and then the expression continued throughout the remaining embryonic stages and adulthood, with a localization restricted to the CNS. In growth factor‐induced neurospheres and adult cerebrum, RANDAM‐2‐expressing cells coincided well not only with nestin‐positive cells but also with glutamate‐positive neurons, but not with γ‐aminobutyric acid‐positive ones. These results indicate that RANDAM‐2 is one of the type I membrane surface antigens constitutively expressed on undifferentiated neuronal cells and the glutamatergic neuronal cells during mouse neurogenesis.

Scanning electron microscopy of vascular architecture in the gastric mucosa of the golden hamster

SummaryThe three-dimensional architectures and the regional differences of the vascular system in the mucosa of the hamster stomach were revealed by scanning electron microscopy of corrosion casts. In the forestomach, the vascular network spreads two-dimensionally in a thin lamina propria. In the corpus and the antrum, the capillaries in the thick lamina propria are well developed, extending three-dimensionally along the gastric pits and glands. In the corpus, the submucosal arteries enter the lamina propria to become ascending capillaries, which project toward the top of the lamina propria and anastomose to create a capillary network beneath the mucosal epithelium. A subepithelial capillary is much wider in diameter than an ascending capillary and is, therefore, a sinusoid capillary. Subepithelial capillaries join descending venules, which are less numerous than the ascending capillaries. Near the gastric lumen, the capillaries in the corpus can be classified into two types: arched type in the cephalic (upper) region and honeycomb type in the caudal (lower) region. In the antrum, the submucosal arterial plexus is less well developed than that in the corpus. The mucosal aspect of the corrosion cast shows many clumps, formed by a unit of capillary network. Functional significances of different vascular architectures in the gastric mucosa of the forestomach, corpus, and antrum are discussed.

The prevalence of and factors related to calcium pyrophosphate dihydrate crystal deposition in the knee joint.

OBJECTIVES The purpose of this study was to reveal the accurate prevalence and related factors to the presence of calcium pyrophosphate dihydrate (CPPD) crystal deposition in cadaveric knee joints. DESIGN Controlled laboratory study. METHODS Six hundred and eight knees from 304 cadavers (332 male knees and 276 female knees, formalin fixed, Japanese anatomical specimens) were included in this study. The average age of the cadavers was 78.3 ± 10.7 years. Knees were macroscopically evaluated for the existence of CPPD, and the depth of cartilage degeneration of the femoro-tibial joint following the Outerbridges classification. CPPD crystal was confirmed under Fourier transform infrared spectroscopy (FTIR) analysis using light microscopy. Statistical analysis was performed to reveal the correlation between the occurrence of CPPD deposition in the knee joint and gender, age, and the depth of cartilage degeneration of the femoro-tibial joint. RESULTS The prevalence of grossly visible CPPD crystal was 13% (79 knees). In all of these knees, CPPD crystal was confirmed under FTIR analysis. Statistical analysis showed significant correlation between the occurrence of CPPD deposition and gender (P < 0.001), and depth of cartilage degeneration in the femoro-tibial joint (P < 0.001). In the cartilage degeneration positive knees (Over grade 3 in Outerbridges classification), average age of CPPD deposition knee was significantly higher than CPPD negative knees. CONCLUSIONS In this study, the prevalence of CPPD deposition disease was evaluated in a relatively large sample size of cadaveric knees. The prevalence of CPPD deposition disease was 13%, and was significantly correlated with the subjects age, gender, and severity of cartilage degeneration in the femoro-tibial joint.

Stress-related activities induced by predator odor may become indistinguishable by hinokitiol odor.

Predator odors, such as 2,5-dihydro-2,4,5-trimethylthiazoline (TMT), induce a stress-like behavior in some rodents, and there is activation of a complex mix of brain regions including the anterior piriform cortex (APC) and the bed nucleus of stria terminalis (BST). In contrast, rose odor can counteract TMT-induced activation of the ventrorostral part of APC and the medial part of BST. In the present study, two novel odors, woody (hinokitiol) and caraway [S(+)-carvone] odors, were evaluated to determine whether they have an antistress effect. Plasma adrenocorticotropic hormone levels, a marker of stress, and the number of c-Fos-immunopositive cells were determined in APC and BST. Plasma adrenocorticotropic hormone levels were increased by TMT alone and in combination with S(+)-carvone; however, hinokitiol with or without TMT did not have an effect. The number of activated cells in the medial part of BST was increased by TMT alone and in combination with S(+)-carvone or hinokitiol. Although TMT alone activated the medial part of BST, a mixture of TMT and hinokitiol activated both the medial and the lateral part of BST. These data suggest that the selective responses to TMT in the medial part of BST were obscured by activation of more odor-related regions by hinokitiol with TMT. In addition, the ratio of medial to lateral BST activation may be critical in stress-related behavior. In conclusion, hinokitiol can alleviate TMT-induced stress; however, the underlying mechanism appears to be different from that of the rose odor, as found in our previous study.