Original articles:
Expression
of macrophage colony-stimulating factor, scavenger receptors, and macrophage proliferation
in the pregnant mouse uterus(KYAW, Y.)
An
ultrastructural and immunohistochemical study of PC12 cells during apoptosis induced by
serum deprivation with special reference to autophagy and lysosomal cathepsins(OHSAWA, Y.)
Stage-specific
localization of basigin, a member of the immunoglobulin super-family, during mouse
spermatogenesis(MAEKAWA, M.)
Postnatal
development of the rat sublingual glands. A morphometric and radioautographic study(TAGA,
R.)
Immunohistochemical
localization of secretory immunoglobulins in the main excretory duct of the human
submandibular gland(PERRA, M.)
Spermatids
of prepubertal male rats are susceptible to carbendazim during early spermiogenesis(NAKAl,
M.)
Structural
organization of Iymphatics in the monkey esophagus as revealed by
enzyme-histochemistry(SHIMODA, H.)
Immunocytochemical
demonstration of the synovial membrane in experimentally induced arthritis of the rat
temporomandibular joint(NOZAWA-INOUE)
Amelogenin
protein in tooth germs of the snake ElaPhe quadrivirgata, immunohistochemistry, cloning
and cDNA sequence(ISHIYAMA, M.)
Immunohistochemical
localization of pleiotrophin and midkine in the lingual epithelium of the adult
rat(WAKISAKA, S.)
Summary
Title
Expression
of Macrophage colony-stimulating Factor, Scavenger Receptors, and Macrophage Proliferation
in the pregnant Mouse Uterus
Author
Yadanar KYAW, Go HASEGAWAl, Hisakazu TAKATSUKA, Motoko SHIMADA-HIRATSUKAl, Hajime UMEZU,
Masaaki ARAKAWA and Makoto NAITO
Address
Second Department of Pathology and Second Department of Internal Medicine, Niigata
University School of Medicine, Niigata, Japan
Summary
During pregnancy, mouse uterine epithelial cells produce and secrete a large amount of
macrophage colony - stimulating factor ( M-CSF/CSF-1 ). Macro phages accumulate and
proliferate in the undecidualized endometrium of the pregnant uterus. Observations showed
that macrophages expressed scavenger receptor class A (type I and type II) and class C
(macrosialin). Scavenger receptors appeared to be involved in the removal of apoptotic
cells in the degenerated decidual tissue. The expression of class A and class C
scavengerreceptor mRNAs in the uterus of pregnant mice was elevated but the expression of
class B scavenger receptor (CD36) mRNA was similar to that of non-pregnant mice. The
expression of various cytokines and chemokines, including M-CSF, monocyte cheomattractant
protein-1 (MCP-1) and macrophage inflammatory protein 1-alpha (MIP1-alpha), was enhanced
in the uterus of pregnant mice, suggesting that these molecules regulate macrophage
chemotaxis and immunological function in the uterus. These findings imply that the
pregnant uterus provides a microenvironment for the recruitment, differentiation, and
proliferation of macrophages and the regulation of scavenger receptor and cytokine
expression for a successful pregnancy.
Dr. Yadanar KYAW
Second Department of Pathology
Niigata University School of Medicine
Asahimachi-dori 1, Niigata
951-8510 Japan
Tel: +81-25-227-2102
Fax: +81-25-227-0761
E-mail: 2byori@med.niigata-u.ac.jp
Title
An
Ultrastructural and Immunohistochemical Study of PC12 Cells During Apoptosis Induced by
Serum Deprivation with Special Reference to Autophagy and Lysosomal Cathepsins
Author
Yoshiyuki OHSAWA, Kyoko ISAHARA, Shiro KANAMORI, Masahiro SHIBATA, Satoshi KAMETAKA,
Takahiro GOTOW, Tsuyoshi WATANABE, Eiki KOMINAMI and Yasuo UCHIYAMA
Address
Department of Cell Biology and Anatomy, 0saka University Medical School, Suita;
Department of Nutrition, Koshien University, Takarazuka; and Department of
Biochemistry, Juntendo University School of Medicine, Tokyo, Japan
Summary
In addition to the caspase family of proteinases, cathepsin D, a lysosomal aspartic
proteinase, has been suggested to act as a proapoptotic mediator in mammalian cells. To
further understand the roles of cathepsins B and D in apoptosis of the cells, we examined
the precise alteration processes of ultrastructures and immunoreactivity for these enzymes
in PC12 ceils cultured under serum deprivation. Laser scanning microscopy showed
immunoreactivity for cathepsins B and D to be finely distributed in the cytopiasm of PC12
celis at the onset of culture under serum deprivation. At 3 h after the onset of culture,
the immunoreactivity for cathepsin B slightly decreased in the cells, while immunodeposits
for cathepsin D in the cells became more intense and larger in size than those at O h.
Positive staining for TUNEL in nuclei of the ceils appeared at 6 h, though fewer in
number. Corresponding to the increase in the number of TUNEL-positive cells at 12 h and 24
h, the immunoreactivity for cathepsin B was drastically diminished in the cells, whereas
that for cathepsin D was significantly augmented, especially in TUNEL-positive cells.
Electron microscopically, autophagic vacuoles/autolysosomes appeared in the cytoplasm of
the cells ah after the onset of culture. A distinct nuclear change showing relatively
condensed chromatin first appeared in the peripherai part of the nuclei at 6 h. The number
of PC12 celis having nuclei with chromatin condensation in. creased especially at 24 h,
while these cells showed shrinkage of both their cytoplasm and nuciei. Dense bodies and
autophagic vacuoles with limiting membranes were seen in these cells. These results
showing the occurrence of autophagy and imbalance of protein amounts between cathepsins B
and D during apoptosis may argue for our hypothesis that these enzymes are, in part,
involved in the cell death cascade for PC12 cells following serum deprivation.
Prof. Yasuo UCHIYAMA
Department of Cell Biology and Anatomy I
Osaka University Medical School(A1)
2-2 Yamadaoka, Suita, Osaka
565-0871 Japan
Tel: +81-6-6879-3124
Fax: +81-6-6879-3129
E-mail: uchiya@anat.1med.osaka-u.ac.jp
Title
Stage-specific
Localization of Basigin, a Member of the Immunoglobulin Superfamily, During Mouse
Spermatogenesis
Author
Mamiko MAEKAWA, Fumie SUZUKI-TOYOTA, Yoshiro TOYAMA, Kenji KADOMATSU, Masako HAGIHARA,
Naohiko KUNO, Takashi MURAMATSU, Kayoko DOHMAE and Shigeki YUASA
Address
Department of Anatomy and Developmental Biology, Chiba University School of Medicine,
Chiba; Department of Biochemistry Nagoya University School of Medicine, Nagoya; and
Department of Science for Laboratory Animal Experimentation, Research; Institute for
Microbial Diseases, Osaka University, Osaka, Japan
Summary
Ablation of the transmembrane glycoprotein basigin leads to azoospermic mice, indicating
that this gene is essential for spermatogenesis. To examine the functions of basigin in
the testis, the precise localization of basigin during spermatogenesis was examined
immunohistochemically. In the adult mouse testis, basigin immunoreactivity appeared on the
cell surface of leptotene spermatocytes and gradually increased in intensity during the
meiotic prophase. Cytoplasmic staining, as well as cell surface staining, was detected in
spermatids. The most conspicuous reactivity was found in the spermatids at steps 9-11 and
in the flagella of spermatids. Immuno-electron microscopic analysis demonstrated that
basigin was localized not only on the plasma membranes of spermatocytes and spermatids,
but also on the plasma membrane of the Sertoli cell processes which contact the
spermatocytes and sper. matids. Basigin immunoreactivity was also detected during
postnatal development in spermatocytes and spermatids but not in spermatogonia.
Experimental cryptorchid testes which contain only spermatogonia and Sertoli cells in the
seminiferous epithelium showed no basigin immunoreactivity. Seven days after surgical
reversal of the cryptorchid testis, spermatocytes reappeared in the tubules, along with
basigin immunoreactivity. Furthermore, in sterile mutant mice, in which neither
spermatocytes nor spermatids were generated, no basigin immunoreactivity was detected in
the seminiferous tubules. These findings indicate that expression of basigin is
concomitant with appearance of spermatocytes in the seminiferous tubule, and suggest that
basigin is involved in the interaction between Sertoli cells and germ cells at specific
stages of spermatogenesis.
Dr. Mamiko MAEKAWA
Department of Anatomy and Developmental Biology
Chiba University School of Medicine
Inohana, Chuo-ku, Chiba 260-8670 Japan
Tel: +81-43-226-2020
Fax: +81-43-226-2021
E-mail: maekawa@med.m.chiba-u.ac.jp
Title
Postnatal
Development of the Rat Sublingual Glands. A Morphometric and Radioautographic Study
Author
R. TAGA and A. SESSO
Address
Department of Morphology, Faculty of Dentistry of Bauru, Sao Paulo University; Laboratory
of Molecular Pathology, Department of pathology, Faculty of Medicine of Sao Paulo, Sao
Paulo University, SP, Brazil
Summary
The postnatal development of rat sublingual glands was analyzed by morphometric and
radioautographic studies. The absolute number of each cell type was evaluated by the
Aherne II morphometric method for cell counting and labeling indices of these cell types
were determined in radioautographs from animals injected with 3H,thymidine The
quantitative cell population kinetic studies were accompanied by morphologic analysis of
the modifications in each gland structure. The data concerning evolution of number of each
cell type were submitted to analysis by least squares fitexponential curve. The
exponential equations duplication times for the acinar, serous demilune, intercalated
duct, striated duct and stroma cells from 2 to 30 days of age were 7.5, 9.0, 10.8 and 9.5
days, respectively. On the other hand, the mean labeling indices for the same cell types
during the same period were 9.5%, 5.8%, 7.2%, 3.3% and 4.3%, respectively. Thus, the
intercalated duct cells exhibited the second highest labeling index and the slowest growth
rate, while the striated duct cells showed the lowest labeling index and the third highest
duplication time. The fact that the striated duct cell labeling index does not explain the
relatively short duplication time of these cells, suggests that cells from other
neighboring morphologic compartments, probably from intercalated duct, migrate and
dilrerentiate into striated ducts cells.
Prof. Rumio TAGA
Departamento de Morfologia-Histologia
Faculdade de Odontologia de Bauru-USP
Al. Octavio Pinheiro Brisola, 9-75, C.P.73
CEP: 17043-101, Bauru, SP Brazil
Tel: 55014-235-8259
Fax: 55014-223-0415
Title
Immunohistochemical
Localization of Secretory Immunoglobulins in the Main Excretory Duct of the Human
Submandibular Gland
Author
Maria Teresa PERRA, Roberto PUXEDDU, Cristina MAXIA and Paola SIRIGU
Address
Department of Cytomorphology, Department of Surgical Sciences and Organ
Transplantation-Section of Otolaryngology, University of Cagliari, Cagliari, Italy
Summary
The localization of IgA, IgG, and IgM was investigated immunohistochemically in the
mucosal surface of the main excretory duct of the human submandibular gland in order to
verify the possible antimicrobial properties of this duct. Only secretory
IgA-immunoreactivity was recognized in the epithelial cells of the duct. An intense
immunoreactivity was observed in the cytoplasm of some cells and at the lumina! surface of
most of the cells. Clusters of IgA-positive immunocompetent cells were also recognizable
in the subepithelial layers. No reactivity for IgG and IgM was noticed. The results
suggest that the ductal epithelium may actively be involved in the release of secretory
IgA, which could play a prominent role in the local defense mechanism of the duct.
Prof. Dr. Paola SIRIGU
Dipartimento di Citomorfologia
Cittadella Universitaria SS. 554-Bivio per Sestu
09042 Monserrato (CA) Italy
Tel: +39-070-6754075
Fax: +39-070-6754003
E-mail: psirigu@vaxca1.unica.it
Title
Spermatids
of Prepubertal Male Rats are Susceptible to Carbendazim During Early Spermiogenesis
Author
Masaaki NAKAI, Kiyotaka TOSHIMORI, Kazuya YOSHINAGA, Tetsuo NASU and Rex A. HESS
Address
Department of Veterinary Anatomy Faculty of Agriculture, Miyazaki University, Miyazaki;
Department of Anatomy Miyazaki Medical College, Kiyotake, Miyazaki; and Department of
Veterinary Biosciences, University of Illinois at Urbana-Champaign, Urbana, Ill. USA
Summary
The fungicide carbendazim, a male reproductive system toxicant, affects the adult testis,
while prepubertal animals are assumed to be unsusceptible to the chemical. The present
study was conducted to re-evaluate the susceptibility of rat prepubertal testis (25-30
days old) to carbendazim based on the incidence of spermatid abnormalities, including
nuclear enlargement (megaspermatids) and binucleate round spermatids. In control
prepubertal rats treated orally with corn oil vehicle alone, seminiferous tubules
containing magno- and/or binucleate round spermatids were often observed in the groups at
stages II-III, IV-V and VI-VII. At 3.0days after treatment with carbendazim (100 mg/kg),
seminiferous tubules containing spermatids with identical abnormalities were significantly
increased, including groups at all stages. Significant increases were also observed at
stages IV-V and VI-VII at day 4.5, and VI-VII at day 7.5. In contrast, the frequency of
spematids with these abnormalities was reduced in the groups at stages II-III and IV-V at
day 7.5. These findings confirm that the prepubertal rat testis is susceptible to
carbendazim during early spermiogenesis.
Dr. Msaaki NAKAI
Department of Veterinary Anatomy
Faculty of Agriculture Miyasaki University
Gakuen Konohanadai, Miyazaki 889-2192 Japan
Tel: +81-985-58-2811
Fax: +81-985-58-2884
Title
Structural
Organization of Lymphatics in the Monkey Esophagus as Revealed by Enzyme-Histochemistry
Author
Hiroshi SHIMODA
Address
Department of Anatomy, Oita Medical University, Oita, Japan
Summary
The fine structure and distribution of lymphatic vessels in the monkey esophageal wall
were studied by light and electron microscopy using an enzyme.histochemical method.
Identification of lymphatics was made by 5'-nucleotidase staining, whereas blood vessels
were visualized by alkaline phosphatase staining. This technique revealed intramural
lymphatic networks in the mucosa, submucosa, and myenteric layer of the esophagus. The
organization of lymphatics in the esophagus basically conformed to that of the small
intestine, although they showed certain distribu. tion patterns peculiar to the esophagus.
The lamina propria mucosae exhibited a double.layered lymphatic network, and lymphatic
capillaries extended into its papillae. Despite their forming blind ends and being closed
by endothelial cells, the lymphatics in the mucosal papillae were found to contain
lymphocytes in their lumen. This suggests that free cells might pene. trate the
endothelium to enter these initial portions of the lymphatics.
Dr. Hiroshi SHIMODA
Department of Anatomy
Oita Medical University
1-1 Idaigaoka, Hasama-machi, Oita 879-5593 Japan
Tel/Fax: +81-97-586-5623
Title
Immunocytochemical
Demonstration of the Synovial Membrane in Experimentally Induced Arthritis of the Rat
Temporomandibular Joint
Author
Kayoko NOZAWA-INOUE, Ritsuo TAKAGI, Tatsuaki KOBAYASHI, Yasushi OHASHI and Takeyasu MAEDA
Address
2nd Department of Oral amd Maxillofacial Surgery, and 2nd Department of Oral Anatomy,
Niigata University School of Dentistry, Niigata, Japan
Summary
The present study is first to report an experimental model of adjuvant-induced arthritis
in the rat temporomandibular joint (TMJ). Arthritis was induced by simultaneous
intradermal administrations of Freund's complete adjuvant, one at the parietal scalp and
the other at the base of the tail. In this model, we demonstrated responses of the
synovial membrane by immunocytochemistry using antibodies to OX6 and ED1 which recognize
Ia antigen in MHC class II antigen. expressing cells and the macrophage/monocyte lineage,
respectively. Three weeks after administration, no remarkable signs of inflammation were
macroscopically recognizable in the TMJ, but microscopically the synovial membrane in the
TMJ revealed marked changes such as enhanced vascularization and hemostasis in the
sublining layer and a thickening in the synovial lining cell layer. Intense
OX6-immunoreactivity was found in the synovial lining cells at lesions in the experimental
group but not in the control group. Immunoelectron microscopy revealed that these
OX6-immunopositive synovial lining cells developed dense cytoplasmic processes and
numerous vacuoles and vesicles, resembling type A cells. Part of the type A cells also
showed ED1-immunoreactivity. The expression of OX6 or ED1 immunoreactivity in the synovial
lining cells might be involved in the initial immune responses in this arthritis model
because the synovial membranes are exposed to the synovial fluids which have been believed
to contain antigenic substances.
Dr. Kayoko NOZAWA- INOUE
Department of Oral Anatomy
Niigata University School of Dentistry 2-5274, Gakkocho-dori, Niigata 951-8514 Japan
Tel: +81-25-227-2817
Fax: +81-25-223-6499
E-mail: nozawa@dent.niigata-u.ac.jp
Title
Amelogenin
Protein in Tooth Germs of the Snake Elaphe quadrivirgata, Immunohistochemistry, Cloning
and cDNA Sequence
Author
Mikio ISHIYAMA, Masato MIKAMI, Hitoyata SHIMOKAWA and Shinichiro OIDA
Address
Department of Histology and Oral Microbiology, The Nippon Dental University School of
Dentistry at Niigata, Niigata; and Department of Oral Biochemistry, School of Dentistry,
Tokyo Medical and Dental University, Tokyo, Japan
Summary
In the snake, Elaphe quadrivirgata, the occurrence of amelogenin was immunohistochemically
demonstrated in the enamel of developing tooth germs. Teeth of the snake are covered with
a thin true enamel layer about l-2 micro meters in thickness. Light and electron
microscopic immunohistochemistry indicated an intense amelogenin immunoreactivity
occurring in the enamel layer during the secretory stage of tooth development. cloning and
cDNA sequence of snake amelogenin was performed by RT-PCR. The amino acid sequence of the
snake amelogenin cDNA-in its portion corresponding to the area from exon 5 to exon 7 of
human X189 amelogenin gene-showed 45% homology with humans. Regions of both the N-terminus
and C-terminus were well conserved. Furthermore, the positions of prolin in the amino acid
alignment of the snake amelogenin corresponded well with those of human amelogenin. It is
suggested that prolin is an essential constituent of amelogenin and therefore its
positions in the molecule have been conserved after the evolutionary divergence of
reptiles and mammals. This study using reptiles is the first detection of specific
amelogenin immunoreactivity by high resolutional immunoelectron microscopy and the first
cloning of amelogenin cDNA in a non-mammalian animal.
Dr. Mikio ISHIYAMA
Department of Histology
The Nippon Dental University School of Dentistry at Niigata 1-8 Hamaura-cho, Niigata
951-8580 Japan
Tel: +81-25-267-1500
Fax: +81-25-267-1134
E-mail: ishiyama@ngt.ndu.ac.jp
Title
Immunohistochemical
Localization of Pleiotrophin and Midkine in the Lingual Epithelium of the Adult Rat
Author
Satoshi WAKISAKA, Makoto J. TABATA, Takeyasu MAEDA, Kazumasa MATSUMOTO, Akio WANAKA,
Hisako MURAMATSU, Takash MURAMATSU and Kojiro KURISU
Address
Department of Oral Anatomy and Developmental Biology Osaka University Faculty of
Dentistry, Osaka; Second Department of Oral Anatomy Niigata University School of
Dentistry, Niigata; Division of Structural Cell Biology, Nara Institute of Science and
Technology (NAIST), Nara; Department of Cell Science, Institute of Biomedical Sciences,
Fukushima Medical University School of Medicine, Fukushima; and Department of
Biochemistry, Nagoya University School of Medicine, Nagoya, Japan
Summary
Distribution of two heparin-binding molecules, pleiotrophin (PTN) and midkine (MK), was
investigated by immunohistochemistry in the lingual epithelium of the adult rat. In the
lingual epithelium, both PTN- and MK-like immunoreactivities were observed in its basal
cell layers, showing a mesh-like appearance. These molecules were also found along the
surface of the taste bud cells; an intense immunoreaction was detected at the base of the
taste buds in the circumvallate and foliate papillae. At the electron microscope level,
the immunoreactive products were localized on the cell surface and basement membrane at
the base of the taste buds. The immunoreactivity for PTN was distributed more diffusely
than that for MK. It was suggested that these molecules may be involved in the
digerentiation and maintenance of taste bud cells, possibly via their trophic effect upon
approaching nerves.
Dr. Satoshi WAKISAKA
Department of Oral Anatomy and Developmental Biology
Osaka University Faculty of Dentistry
1-8, Yamadaoka, Suita, Osaka 565-0871 Japan
Tel: +81-6-6879-2872
Fax: +81-6-6879-2875
E-mail: wakisaka@dent.osaka-u.ac.jp
