Original articles
- Dendritic
cells in the rat pituitary gland evaluated by the use of monoclonal antibodies and
electron microscopy (SATO, T.)
- Lectin
binding patterns in rat nasal-associated lymphoid tissue (NALT) and the influence of
various types of lectin on particle uptake in NALT (TAKATA, S.）
- The
Spatial relationship between the perineuronal proteoglycan network and the synaptic
boutons as visualized by double staining with cationic colloidal iron method and
anti-calbindin-D-28K immunohistochemistry in rat cerebellar nuclei (OHTSUKA, A.)
- Structural
and ultrastructural studies of GH, PRL and SMT cells in goat fetus (Capra hircus) using
immunocytochemical methods (VASQUEZ F. A.)
- Muscular
innervation of the proximal duodenum of the guinea pig (IINO, S.)
- Ultrastructure
and distribution of interstitial glandular cells and associated elements in human fetal
ovaries (NOTTOLA, S. A.)
- Nitric
oxide-containing neurons in the bovine gut, with special reference to their relationship
with VIP and galanin (VITTORIA, A.)
- Time-related
changes in periodontal mechanoreceptors in rat molars after the loss of occlusal stimuli
(MURAMOTO, T.)
- Transient
expression of heat shock protein (Hsp) 25 in the dental pulp and enamel organ during
odontogenesis in the rat incisor (OHSHIMA, H.)
Short communication
- Orcein-picroindigocarmine
- a new multiple stain (STEVEN, P.)
Summary
Title
Dendritic Cells in the Rat Pituitary Gland Evaluated by the Use of
Monoclonal Antibodies and Electron Microscopy
Author
Tetsuji SATO and Kouji INOUE
Address
Department of Anatomy (Division II), School of Dental Medicine, Tsurumi University,
Yokohama, Japan
Summary
A detailed analysis of the difference in the localization and the immunoreactivity for
various surface markers among folliculo-stellate cells, macrophages, and dendritic cells
was performed using immunohistochemistry and electron microscopy of the rat pituitary
gland. The folliculo-stellate cells were selectively labeled by an antiserum against S100
protein. The majority of dendritic cells were immunoreactive for the MHC class II (Ia)
antigen (OX6) and/or the dendritic cell antibodies (OX62). The main population of
macrophages was positive for the macrophage antibodies (ED1, ED2, and/or OX42). The
cellular density of adenohypophyseal macrophages was significantly lower than that of
folliculo-stellate cells and of dendritic cells. All the neurohypophyseal microglial cells
were labeled with OX42, while the mAb OX6 labeled a small population of cells different
from the cells identified by OX42 in the neurohypophysis. Double-immunoperoxidase staining
for ED1 and OX6 revealed that positively stained cells could be classified into ED1+OX6-,
ED1+OX6+, and ED1-OX6+ cells. Double staining with OX62 and OX6 mAbs showed that about 60%
of the OX6+ cells were also immunolabeled with OX62 in the anterior lobe: OX62 detects a
subpopulation of dendritic cells but does not recognize macrophage populations.
Furthermore, double staining for S100 and OX6 resulted in no S100+ OX6+ cells. At the
electron-microscopic level, reaction products for OX6 were confirmed in the cell membrane
and labeled cells were distinguished from macrophages and folliculo-stellate cells by
distinctive short, broad cytoplasmic processes and the rare presence of cytoplasmic
organelles. Such cytological characteristics of the OX6-positive cells in the pituitary
gland are similar to dendritic cells. Our results suggest that resident dendritic cells
and folliculo-stellate cells are two different main components of interstitial cells in
the pituitary gland.
Prof. Tetsuji SATO
Department of Anatomy (Division II)
School of Dental Medicine
Tsurumi University
2-1-3 Tsurumi, Tsurumi-ku
Yokohama, 230-8501 Japan
Tel: +81-45-581-1001
Fax: +81-45-573-9599
E-mail: m00777@simail.ne.jp
Title
Lectin Binding Patterns in Rat Nasal-Associated Lymphoid Tissue
(NALT) and the Influence of Various Types of Lectin on Particle Uptake in NALT
Author
Satoru TAKATA, Osamu OHTANI and Yukio WATANABE
Address
Departments of Anatomy and Otolaryngology, Faculty of Medicine, Toyama Medical and
Pharmaceutical University, Toyama
Summary
We investigated the binding of four types of lectin to follicle-associated epithelium
overlying the nasal-associated lymphoid tissue (NALT) of rats in order to identify M-cell
specific surface markers and to determine the influence of lectin administration to NALT
on the uptake of a particulate antigen. The NALT tissues were incubated with a panel of
four types of lectin conjugated to horseradish peroxidase (HRP). Ulex europaeus-1 (UEA-1)
and Dolichos biflorus (DBA) lectin stained the surface of M-cells and goblet cells.
Uniform staining by Triticum vulgaris (WGA) was detected in the M-cells, ciliated cells
and goblet cells. In contrast, staining of Griffonia simplicifolia I isolectin-B4 (GSI-B4)
was almost exclusively M-cell specific. The administration of M-cell specific lectin (GS
I-B4) to NALT suppressed the uptake of baker's yeast particles administered later, whereas
the non-specific one (UEA-1) had no influence on the uptake. These results indicate that
GS I-B4 is a useful marker for the identification of rat NALT M-cells and that such a
specific expression of surface glycoconjugates by M-cells may permit the targeting of
vaccines and drugs to the antigen sampling sites of the nose. It also appears possible to
block the uptake of pathogens by an administration of M-cell specific lectin to NALT.
Dr. Satoru TAKATA
Department of Otolaryngology
Faculty of Medicine
Toyama Medical and
Pharmaceutical University
Sugitani, Toyama
930-0194 Japan
Title
The Spatial Relationship between the Perineuronal Proteoglycan
Network and the Synaptic Boutons as Visualized by Double Staining with Cationic Colloidal
Iron Method and Anti-Calbindin-D-28K Immunohistochemistry in Rat Cerebellar Nuclei
Author
Aiji OHTSUKA, Takehito TAGUCHI, Ramadan SAYED and Takuro MURAKAMI
Address
Department of Anatomy, Faculty of Medicine, Okayama University Medical School, Okayama,
Japan; and Department of Anatomy and Histology, Faculty of Veterinary Medicine, Assiut
University, Assiut, Egypt
Summary
The present study demonstrated the precise spatial relationship between meshes in the
perineuronal proteoglycan network and the terminal boutons of synaptically associated
axons. Sections from the rat cerebellum were stained with cationic colloidal iron (pH
1.0-1.5), and successively immunostained with anti-calbindin-D-28K monoclonal antibody.
Cationic iron stained sulfated proteoglycans around the nerve cell of the medial
cerebellar nucleus, whereas the anti-calbindin antibody labeled the Purkinje cells
including their axons terminating on large neurons in the cerebellar nucleus. It was found
that each synaptic bouton fits into a mesh of the perineuronal network. The individual
meshes appeared to be divided by partitions faintly stained with the colloidal iron.
Electron microscopy of cationic colloidal iron-stained ultrathin sections revealed that
the synaptic boutons were separated from each other by the proteoglycan matrix and that
each of them was further divided into two or more contact areas of presynaptic membrane by
the same matrix. This suggests that individual synapses are protected against the effects
of adjacent synaptic transmission, and that each of them may be subdivided by this manner
of partitioning, like pads of a cat's paw.
Dr. Aiji OHTSUKA
Department of Anatomy
Faculty of Medicine
Okayama University Medical School
2-5-1 Shikata-cho, Okayama
700-8558 Japan
Tel: +81-86-235-7089
Fax: +81-86-235-7095
E-mail: aiji@med.okayama-u.ac.jp
Title
Structural and Ultrastructural Studies of GH, PRL and SMT Cells in
Goat Fetus (Capra hircus) Using Immunocytochemical Methods
Author
F. A. VASQUEZ, M. A. GOMEZ, J. SERRANO and A. BERNABE
Address
Histologia y Embriologial Facultad de Ciencias Veterinaria, Universidad Nacional de
Rosario, Argentina; Histologia y Anatomia Patologica; and Biologia, Facultad de
Veterinaria, Universidad de Murcia, Murcia, Spain
Summary
The first data based on immunolabeling techniques of goat fetus adenohypophysis show that
the structure and ultrastructure of growth hormone (GH)-, prolactin (PRL)-, and GH- plus
PRL-secreting cells (SMT cells) in fetuses aged 100 days differ from those in the adult.
Both cell number and cell size are smaller in the fetus, and the percentage of dark cells
decreases with development. The data do not support the hypothesis that SMT cells
represent the common origin of GH- and PRL-cells.
Prof. Dr. A. BERNABE
Histologia y Anatomia Patologica
Facultad de Veterinaria
Universidad de Murcia
Apdp. Correos 4021
E-30071 Murcia Spain
Tel: +34-968-364705
Fax: +34-968-364147
E-mail: abernabe@fcu.urn.es
Title
Muscular Innervation of the Proximal Duodenum of the Guinea Pig
Author
Satoshi IINO
Address
Department of Anatomy, Fukui Medical University, Matsuoka, Fukui, Japan
Summary
We investigated the muscular structure and innervation of the gastroduodenal junction in
the guinea pig. In the gastroduodenal junction, the innermost layer of the circular muscle
contained numerous nerve fibers and terminals. Since this nerve network continued onto the
deep muscular plexus (DMP) of the duodenum, we surmised that the numerous nerve fibers in
the gastroduodenal junction were specialized DMP in the most proximal part of the
duodenum. The innermost layer containing many nerve fibers was about 1,000 microm in
length and 100 microm in thickness in the proximal duodenum. This layer contained numerous
connective tissue fibers composed of collagen and elastic fibers. Five to 30 smooth muscle
cells lay in contact with each other and were surrounded by fine connective tissue. The
nerve fibers in the proximal duodenum contained nerve terminals immunoreactive for choline
acetyltransferase, dynorphin, enkephalin, galanin, gastrin-releasing peptide, nitric oxide
synthase, substance P, and vasoactive intestinal polypeptide. Adrenergic fibers which
contained tyrosine hydroxylase immunoreactivity were rare in the proximal duodenum. In the
innermost layer of the proximal duodenum, there were numerous c-Kit immunopositive cells
that were in contact with nerve terminals. This study allowed us to clarify the specific
architecture of the most proximal portion of the duodenum. The functional significance of
the proximal duodenum in relation to the electrical connection and neural cooperation of
the musculature between the antrum and the duodenum is also discussed.
Dr. Satoshi IINO
Department of Anatomy
Fukui Medical University
Matsuoka, Fukui
910-1193 Japan
Tel: +81-776-61-8302
Fax: +81-776-61-8132
E-mail: iinosa@fmsrsa.fukui-med.ac.jp
Title
Ultrastructure and Distribution of Interstitial Glandular Cells and
Associated Elements in Human Fetal Ovaries
Author
Stefania A. N0TTOLA, Sayoko MAKABE, Tiziana STALLONE, Guido MACCHIARELLI, Silvia CORRER
and Pietro M. M0TTA
Address
Department of Anatomy, University of Rome "La Sapienza", Rome, Italy; and
Department of Obstetrics and Gynecology, Toho University, Tokyo, Japan
Summary
In order to understand the fine structure and distribution of the interstitial glandular
cells (IGCs) and associated elements in the human fetal ovary, we studied human fetal
ovaries at 16 weeks post fertilization (p. f.) by transmission electron microscopy.
Semithin sections revealed voluminous typical IGCs usually grouped in clusters, located in
the interstitium among the ovigerous cords. Isolated primordial follicles were seen in the
cords located close to the interstitium in which IGCs were present. Besides the main
ultrastructural characteristics of steroid secreting cells, the IGCs showed lipofuscin
granules and stacks of annulate lamellae in their cytoplasm. Fibrocytes, macrophages and
mast cells were detected close to the IGCs. In particular, the fibrocytes were located
around the IGCs, with which they occasionally formed focal cell contacts. Fibrocytes
issued numerous long projections, which, together with collagen fibers, surrounded the
clusters of IGCs and small vessels (mainly capillaries), often extending into the
intercellular spaces among IGCs. These data indicated that, already at the initiation of
folliculogenesis, the IGCs are present numerously in a close association with the
ovigerous cords. The morphological aspects of IGCs were comparable to that of fetal testis
interstitial (Leydig) cells and hilar cells in adult ovary, and suggest that fetal IGCs
may be source of adult ovary hilar cells. In addition, we have here demonstrated for the
first time that IGCs are associated with stromal cells whose distribution seems to support
IGCs microtopography. Fetal ovarian fibrocytes revealed a structural arrangement similar
to that of the "compartmentalizing cells" previously described in the adult
testis. Macrophages and mast cells presumably have a role as local modulators of steroid
synthesis. Mast cells may also affect fibrocyte organization and vascular permeability. We
thus suggest that IGCs and associated cells may form a glandular unit in the human fetal
ovary similar to that in the adult testis, and this structure is likely involved in early
steroid secretion during gonadal differentiation.
Prof. Pietro M. MOTTA
Department of Anatomy
Faculty of Medicine
University of Rome ''La Sapienza"
Via A. Borelli 50
I-00161 Rome, Italy
Phone: +39-064462623
Fax: +39-064452349
E-mail: pietro.motta@mliromal.it
Title
Nitric Oxide-containing Neurons in the Bovine Gut, with Special
Reference to Their Relationship with VIP and Galanin
Author
A. VITTORIA, A. COSTAGLIOLA, E. CARRESE, B. MAYER and A. CECIO
Address
Department of Biological Structures, Functions and Technology University of Naples
"Federico II", Naples, Italy; and Institute of pharmacology and Toxicology,
Karl-Franzens University, Graz, Austria
Summary
The presence and distribution of nicotinamide dinucleotide phosphate diaphorase
(NADPH-d)-containing neurons have been studied by means of NADPH-d histochemistry in
different regions of the adult cow gut, from the esophagus to the rectum. NADPH-d and
nitric oxide synthase (NOS) were constantly recognized to be colocalized in the same
neuron. The colocalization of vasoactive intestinal polypeptide (VIP) and galanin in such
nitrergic neurons was also studied by means of combined histochemical and
immunofluorescence techniques. NADPH-d-positive neurons were present along the myenteric
plexus of the entire gut, and in the submucous plexus from the abomasum to the rectum.
Notably, they formed two types of nerve networks in the submucous connective tissue of the
jejunum-ileum. NADPH-d-positive innervation of the muscle layers occurred throughout the
tract, and sometimes a clear correspondence was noted between the number of reactive
fibres and the thickness of the muscle. Nitrergic fibres also occurred in the mucosa and
often were in relation to glands and blood vessels. The nitrergic neurons varied in size,
shape, and intensity of staining, and often their terminals were seen to surround
unstained perikarya. Various types of neurons were recognized on the basis of their
chemical content; one of them contained galanin, VIP and NOS simultaneously. The present
results suggest that the nitrergic neurons of the bovine gastrointestinal tract play roles
presumably for controlling the motility of the gut and the conduction of interneuronal
impulses.
Dr. Anna COSTAGLIOLA
Department of Biological Structures,
Functions and Technology
Naples University "Federico II"
Faculty of Veterinary Medicine
Via Delpino, l
80137 Napoli Italy
Phone: +39-081-5644226
Fax: +39-081-5644230
E-mail: costagli@unina.it
Title
Time-Related Changes in Periodontal Mechanoreceptors in Rat Molars
after the Loss of Occlusal Stimuli
Author
Takeshi MURAMOTO, Yoshiro TAKANO and Kunimichi SOMA
Address
First Department of Orthodontics and Second Department of Oral Anatomy, Faculty of
Dentistry, Tokyo Medical and Dental University, Tokyo, Japan
Summary
The effect of a loss of occlusal stimuli upon the distribution and structure of the
periodontal mechanoreceptors of the rat mandibular molar was examined after extracting
opposing molars. The hypofunctional periodontal ligament narrowed significantly two weeks
after tooth extraction, associated with an altered morphology of the Ruffini endings that
showed typical dendritic profiles in normal controls. At four weeks and later periods
after extraction, the Ruffini endings-including those without light microscopic changes
demonstrated unusual ultrastructural features such as the eccentric localization of
mitochondria along the axonal membrane and loss of other cell organelles, unusual
elongation of axonal microprojections, or a deep invagination of the Schwann sheath into
the axoplasm. Immunoreactivity for the growth-associated protein-43 (GAP-43) in the
Ruffini endings was restricted to the Schwann element in both the normal and
hypofunctional periodontal ligament, but the reaction was weaker and even negligible in
some cases in the latter ligament. The present results suggest that occlusal stimuli are
essential for maintaining the structural integrity of the periodontal ligament, including
that of periodontal mechanoreceptors. A decreased immunoreactivity for GAP-43 in the
Schwann sheaths supports the notion of a possible functional alteration in the Ruffini
endings that showed no structural abnormality.
Dr. Takeshi MURAMOTO
First Department of Orthodontics
Faculty of Dentistry
Tokyo Medical and Dental University
1-5-45 Yushima, Bunkyo-ku, Tokyo
113-8549 Japan
E-mail: muraortl@dent.tmd.ac.jp
Title
Transient Expression of Heat Shock Protein (Hsp) 25 in the Dental
Pulp and Enamel organ during Odontogenesis in the Rat Incisor
Author
Hayato OHSHIMA, Hisao AJIMA, Yoshiro KAWANO, Kayoko NOZAWA-INOUE, Satoshi WAKISAKA and
Takeyasu MAEDA
Address
Departments of Oral Anatomy and Oral Maxillofacial Surgery, Faculty of Dentistry, Niigata
University, Niigata; and Department of Oral Anatomy and Developmental Biology, Osaka
University Faculty of Dentistry, Osaka, Japan
Summary
The expression of heat shock protein (Hsp) 25 during odontogenesis in the dental pulp and
enamel organ of rat incisors was investigated by immunocytochemistry and confocal
microscopy. In the process of dentin formation, immature odontoblasts first exhibited Hsp
25-immunoreactivity, and increased in immunointensity with the advance of their
differentiation. In the dental pulp, in contrast, intense immunoreaction in the
mesenchymal cells became weak or negative in parallel with the progress of cell
differentiation. The immunoreaction for Hsp 25 in the enamel organ revealed a
characteristic stage-related alteration during amelogenesis. In secretory ameloblasts, the
immunoreaction for Hsp 25 was found throughout their cell bodies, intense reactivity being
located near the proximal and distal terminal webs. At the maturation stage, ruffle-ended
ameloblasts (RA) consistently showed Hsp 25-immunoreactivity throughout the cell bodies,
whereas smooth-ended ameloblasts (SA) lacking a ruffled border were weak in immunoreaction
at the distal cytoplasm. Other cellular elements of the enamel organ were negative. The
subcellular localization of Hsp 25-immunoreactivity in this study appeared essentially
identical to that of actin filaments as demonstrated by confocal microscopy using
rhodamine-labeled phalloidin. These immunocytochemical data suggest that the Hsp 25
molecule is involved in reinforcement of the cell layer following cell movement during
odontogenesis and in the formation and maintenance of the ruffled border of RA.
Dr. Hayato OHSIIIMA
2nd Department of Oral Anatomy
Faculty of Dentistry, Niigata University
2-5254 Gakkocho-dori, Niigata
951-8514 Japan
Tel: +81-25-227-2816
Fax: +81-25-223-6499
E-mail: histoman@dent.niigata-u.ac.jp
Title
Orcein-Picroindigocarmine－a new multiple stain
Author
Philipp STEVEN, Friedrich PAULSEN and Bernhard TILLMANN
Address
Department of Anatomy, Christian Albrecht University, Kiel, Germany
Summary
A new "orcein-picroindigocarmine staining", a colour combination of orcein,
indigo carmine, and picric acid, was developed for histological applications. The new
technique was tested on different human tissues. Colours ranging from red to brown,
yellow, green and blue were observed in paraffine sections of tissues stained by this
method. Nuclear structures in all tissues were stained dark brown to dark blue. Squamous
epithelium was stained light brown with varying shades of blue in upper horny layers,
whereas the ciliated epithelium was tinged blue grey. When connective tissue was stained,
collagen fibrils appeared strongly blue next to elastic fibres, which took on a rust brown
tinge; cellular components were all coloured brown. The matrix of hyaline cartilage was
stained in different shades of blue, with the chondrocytes rust brown. Sections of bone
components appeared dark blue to dark green. Skeletal muscle cells were coloured yellow
and green with blue collagenous septa. The new staining is useful for distinguishing
connective tissue components such as elastic fibres and collagen fibrils. It also
demonstrates chondrocytes in favourable contrast to the cartilage matrix. The technique
produces aesthetic staining colouring that could supplement histological investigations
and provide an alternative to other staining materials.
Philipp STEVEN
Department of Anatomy
Christian Albrecht University
Olshausenstraβe 40
D-24098 Kiel, Germany
Tel: +49 431 880 2597
Fax: +49 431 880 1557
E-mail: psteven@gmx.de
