Original articles
- Morphological
changes and recovery process in the tenotomized soleus muscles of the rat （E, A, ABOU
SALEM）
- Effects
of pinealectomy and sham-surgery on the area postrema in rats: A quantitative histological
study with special reference to capillaries and neuronal cell nuclei （KUDOU, H.）
- Development
of astrocytes in the mouse hippocampus as tracked by tenascin-C gene expression（YUASA,
S.）
- Immunolocalization
of aquaporin-8 in rat digestive organs and testis （TANI, T.）
- Role
of heme oxygenase-1 and Kupffer cells in the production of bilirubin in the rat liver （HIRANO,
K.）
- Three-dimensional
analysis of nephrogenesis in the neonatal rat kidney: light and scanning electron
microscopic studies （IINO, N.）
- Morphological
characteristics of Schwann cells in the islets of Langerhans of the murine pancreas（SUNAMI,
E.）
- Purkinje
cells in the adult cat cerebellar cortex possess a perineuronal net of proteoglycans （MABUCHI,
M.）
- Effects
of photoperiod and melatonin on the development of growth hormone cells and the
pituitary-adrenal axis in the Djungarian hamster, Phodopus sungorus（HIRA, Y.）
- Asialoglycoprotein
receptors on rat dendritic cells: possible roles for binding with Kupffer cells and
ingesting virus particles （UWATOKU, R.）
Summary
Title
Morphological Changes and Recovery Process in the Tenotomized Soleus
Muscles of the Rat
Author
E, A, ABOU SALEM, Noboru FUJIMAKI, Harunori ISHIKAWA, Tomoko TASHIRO and Yoshiaki KOMIYA
Address
Department of Anatomy and Department of Cellular and Molecular Neurobiology, Gunma
University School of Medicine, Maebashi, Japan
Summary
Tenotomized soleus muscles of adult rats were analyzed morphologically and biochemically
with special reference to the recovery process. Light microscopic observations of
semi-thin sections showed that the characteristic central core lesion was most extensive
at 1 week after tenotomy and began to diminish in extent at 2 weeks until no trace of
lesion could be seen by 6th week, as confirmed by thin-section electron microscopy. Three
phases of changes in the crosssectional area of muscle fibers after tenotomy were
demonstrated by morphometry: phase I designated as the initial increase up to the 3rd day,
phase II as the progressive decrease until the 4th week, and phase III as the recovery to
normal or even hypertrophy. In electron microscopy, the earliest alteration of myofibrils
was recognized at 3 days after tenotomy. The Z discs showed a wavy or zigzag profile with
frequent longitudinal splitting of myofibrils. From the 2nd week on, muscle fibers
underwent a process of recovery, replacing the central core lesion with new myofibrils in
which a reassembly of thick filaments into bundles of thin filaments took place, with Z
discs being aligned adjacent to the peripheral complete myofibrils. In SDSpolyacrylamide
gel electrophoresis, the molar ratio of myosin to actin diminished markedly as the central
core lesion developed and gradually returned to normal with time, correlating well with
the loss and subsequent reassembly of thick filaments.
Dr. E, A, ABOU SALEM
Department of Anatomy
Gunma University School of Medicine
Schowa-machi, Maebashi
371-8511 Japan
Tel: +81-27-220-7912
Fax: +81-27-220-7916
Title
Effects of Pinealectomy and Sham-Surgery on the Area Postrema in
Rats: A Quantitative Histological Study with Special Reference to Capillaries and Neuronal
Cell Nuclei
Author
Hisashi KUDOU, Takashi KACHI, Takao SUZUKI and Yoshiharu SAITO
Address
Department of Obstetrics and Gynecology, and Department of Anatomy, Hirosaki University
School of Medicine, Hirosaki, Japan
Summary
This study aimed to clarify the effects of pinealectomy and sham-surgery on the area
postrema (AP) by quantitative histological methods. Male, Wistar rats of normal (NO),
sham-operated (SX), and pinealectomized (PX) groups were used in the late dark phase at
7weeks of age. Consecutive frontal sections including the AP were stained with hematoxylin
and eosin, and immunostained using PGP 9.5 for neurons, or GFAP or vimentin for glial
cells. Consecutive sections of the AP were separated into five portions starting from the
point of the central canal opening to the fourth ventricle in the caudal direction, and
used for measurements. Mean cross-sectional areas of capillaries showed a lower value in
the SX group than in the other two groups (vs NO, P<0.005; vs PX, P<0.03). In
addition, the frequency distributions of the nuclear diameters of nerve cells showed
different patterns among the three experimental groups (P<0.01), the frequency of large
nuclei being higher in the SX group than in the other two groups. Possible mechanisms of
the effects of sham-pinealectomy and pinealectomy and significance of the pineal-AP
relation are discussed. The results of this study indicate that structural changes in the
AP can be induced by intracranial surgery, suggesting certain pineal involvement in these
changes.
Dr. Hisashi KUDOU
Department of Obstetrics and Gynecology
Hirosaki University School of Medicine
Zaifuchou 5, Hirosaki
036-8562 Japan
E-mail (to Prof. T. KACHI): kachitak@cc.hirosaki-u.ac.jp
Title
Development of Astrocytes in the Mouse Hippocampus as Tracked by
Tenascin-C Gene Expression
Author
Shigeki YUASA
Address
Laboratory of Neural Information, National Institute for Physiological Sciences, Okazaki,
Japan
Summary
Tenascin-C (TN-C) is an astroglia-derived extracllular matrix protein that has been shown
to be an early marker for astroglial precursors in the embryonic mouse brain. This study
examined astroglial generation, migration, and differentiation in the developing mouse
hippocampus by in situ hybridization histochemistry for TN-C mRNA. Special reference was
given to the difference in the mode of astroglial development between the two cortical
structures of the hippocamups: the dentate gyrus and Ammon's horn. TN-C-positive cells
were found in the ventricular germinative zone of the hippocampus as early as the 15th
gestational day, and the labeled cells in the zone apposed to the fimbria migrated
tangentially through the subpial area towards the forming dentate gyrus. The TN-C-positive
cells aligned in the dentate gyrus exhibited the characteristic morphology of unipolar
astrocytes as revealed by double labeling with glial fibrillary acidic protein
(GFAP)-immunohistochemistry. On the other hand, the TN-C-positive cells ranging over a
wide area of the ventricular germinative zone facing the forming Ammon's horn migrated
radially towards the cortex, with most of them aligned in the Ammon's horn exhibiting a
GFAP-positive stellate morphology. The onset of migration towards the dentate gyrus was
two days earlier than that towards the Ammon's horn. TN-C-positive cells in both cortical
structures exhibited a DNA-replicating activity after settlement in the early postnatal
stage and were considered to further generate astrocytes. On the other hand, TN-C-positive
cells with DNA-replicating activity were also found in the subpial migratory stream moving
towards the dentate gyrus and were considered to form the subpial matrix for the
generation of the dentate astrocytes. Migratory TN-C-positive cells directed towards both
the dentate gyrus and Ammon's horn were apposed to radial glial processes and were
believed to be guided by contact with these processes in a manner similar to migratory
immature neurons. These findings indicate that TN-C-positive cells for the dentate gyrus
and those for the Ammon's horn have different migratory patterns and undergo different
morphological differentiations depending on their site of origin at the early stage of
astrogliogenesis and corresponding to the different modes of neurogenesis in the two
cortical structures.
Prof. Shigeki YUASA
Department of Anatomy
and Developmental Biology
Chiba University School of Medicine
Chiba, 260-8670 Japan
Fax: +8l-43-226-2021
E-mail: yuasa@med.m.chiba-u.ac.jp
Title
Immunolocalization of Aquaporin-8 in Rat Digestive Organs and Testis
Author
Tatsuo TANI, Yu KOYAMA, Kouei NIHEI, Satoru HATAKEYAMA, Kazufumi OHSHIRO, Yutaka YOSHIDA,
Eishin YAOITA, Yasuo SAKAI, Katsuyoshi HATAKEYAMA and Tadashi YAMAMOTO
Address
Department of Structural Pathology, Institute of Nephrology and lst Department of Surgery,
Faculty of Medicine, Niigata University, Niigata, Japan
Summary
Expression of aquaporin-8 mRNA has previously been shown in hepatocytes, pancreatic acinar
cells, colon epithelial cells and seminiferous tubules of the testis in the rat by in situ
hybridization technique. However, immunolocalization of this water channel has not yet
been demonstrated. In the present study, the localization of immunoreactive aquaporin-8
and expression of the mRNA were examined in rat organs (cerebrum, cerebellum, eye,
salivary gland, heart, lung, liver, pancreas, esophagus, stomach, jejunum, ileum, colon,
testis, ovary, kidney, spleen and lymphnode) by immunohistochemistry using an antibody
against aquaporin-8 and ribonuclease protection assay. Aquaporin-8 was distinctly
immunolocalized on the apical membranes of pancreatic acinar cells and mucosal epithelium
of the colon and jejunum. In the liver, the bile canalicular membrane of hepatocytes was
immunostained. In the testis, immunoreactive aquaporin-8 was demonstrated on the luminal
side of the seminiferous tubules. At high magnification, the peroxidase reaction products
appeared on the ramified cytoplasmic membrane of Sertoli cells surrounding the residual
bodies or spermatogenic cells. Specificity of the antibody was verified by Western blot
analysis showing a minor ～28kDa band (deduced deglycosylated form of aquaporin-8) and a
major ～30kDa band (glycosylated form) in these organs. The intensity of aquaporin-8
immunoreactivity was approximately comparable to that of aquaporin-8 mRNA expression in
the liver, pancreas, colon, jejunum and testis. The aquaporin-8 mRNA expression in the
hepatocytes was presumed to be closely associated with the structure of bile canaliculi
since the message was detected in hepatocytes immediately after isolation from the liver
but not in cells following cultivation for three days.
The localization of immunoreactive aquaporin-8 indicated functions for this water channel
in the secretion of bile and pancreatic juice, and the secretion or absorption of water in
the colon and jejunum, and the maturation or liberation of spermatogenic cells in the
testis.
Prof. Tadashi YAMAMOTO, M.D. Ph.D.
Department of Structural Pathology
Institute of Nephrology
Faculty of Medicine Niigata University
1-757 Asahimachi-dori, Niigata
951-8510 Japan
Tel: +81-25-227-2151
Fax: +81-25-227-0768
E-mail: tdsymmt@med.niigata-u.ac.jp
Title
Role of Heme Oxygenase-1 and Kupffer Cells in the Production of
Bilirubin in the Rat Liver
Author
Ken-ichiro HIRANO, Takashi KOBAYASHI, Takaoki WATANABE, Takashi YAMAMOTO, Go HASEGAWA,
Katsuyoshi HATAKEYAMA, Makoto SUEMATSU, and Makoto NAITO
Address
Second Department of Pathology and First Department of Surgery, Niigata University School
of Medicine, Niigata; and Department of Biochemistry, School of Medicine, Keio University,
Tokyo, Japan
Summary
Heme oxygenase (HO)-1, the heme-degrading enzyme in macrophages, plays a key role in
bilirubin metabolism. HO-1 is expressed in various tissue macrophages, especially Kupffer
cells. This study aimed to examine the roles of macrophages and HO-1 in the modulation of
heme catabolism in rat livers. Rats treated with or without liposome-encapsulated
dichloromethylene diphosphonate, a macrophage-depleting reagent, were administered with
heat-denatured red blood cells (h-RBC), and the time course of the biliary output of
bilirubin and the expressions of HO-1 mRNA and protein were monitored.
Immunohistochemistry in the control rat liver revealed that Kupffer cells constitute a
major cellular component expressing HO-1, while hepatocytes exhibited little expression.
The levels of HO-1 expression in Kupffer cells were elevated immediately after injection
of h-RBC. In Kupffer cell-depleted livers, however, HO-1 expressing cells were not
detected even after h-RBC administration. HO-1 mRNA levels were elevated at 2h after
administration of h-RBC in control rat livers, while they were very low in Kupffer
cell-depleted rat livers. The control and Kupffer cell-depleted groups exhibited distinct
time courses of biliary bilirubin excretion. In the untreated control rats, total
bilirubin excretion increased about two-fold at 5h after h-RBC administration. In
contrast, the Kupffer cell-depleting treatment decreased the level of bilirubin
production; administration of h-RBC to Kupffer cell-depleted rats did not accelerate the
generation of bilirubin. These results suggest that Kupffer cells serve both as a sensor
for scenesent RBC clearance and an effector that upregulates heme-degrading capacity and
bilirubin production.
Prof. Makoto NAITO, M. D.
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
Three-dimensional Analysis of Nephrogenesis in the Neonatal Rat
Kidney:
Light and Scanning Electron Microscopic Studies
Author
Noriaki IINO, Fumitake GEJYO, Masaaki ARAKAWA and Tatsuo USHIKI
Address
Department of Anatomy and Histology, and Department of Internal Medicine, Faculty of
Medicine, Niigata University, Niigata, Japan
Summary
In order to clarify the process of renal development more precisely than previously, the
present study observed the rat neonatal kidney by scanning electron microscopy (SEM) of
KOH digested tissue as well as by light microscopy of plastic sections. In the subcapsular
region, aggregation of the mesenchymal cells was closely associated with the upper side of
the ureteric duct ampulla. These mesenchymal cells projected a number of fine irregular
processes at the basal portion facing the ureteric duct. A spherical cluster transformed
from the mesenchymal cell aggregation was found on the lower side of the terminal ampulla,
and was differentiated into the renal vesicle. Some cells at the top of the renal vesicle
formed a cone-shaped projection and invaded the ureteric duct ampulla, forming a
connection with it. In the advanced stage, a shallow transverse cleft appeared on the
outer lateral side of the renal vesicle, and a second cleft was formed on the opposite
side close to the junction between the renal vesicle and the ampulla. As the two clefts
deepened, the vesicle assumed the well-known S-shaped body. In the advanced S-shaped body,
the lower limb became cup-shaped, while the segment between the middle and lower limbs of
the "S" elongated to form a tubular structure (i.e., the prospective proximal
tubule and Henle's loop). The upper limb of the "S" also increased its length to
form a distal tubule. The middle limb of the "S", however, was attached firmly
to the cup-shaped lower limb (i.e., the prospective renal corpuscle) and was considered to
become the macula densa of the mature nephron. In the maturing renal corpuscle,
irregularly shaped cells were observed as a sheet-like aggregation at its vascular pole
and were continuous with the vascular smooth muscle cells. These findings will help toward
a better understanding of the morphological complexities of nephrogenesis.
Dr. Noriaki IINO
Department of Internal Medicine
Faculty of Medicine, Niigata University
1-757 Asahimachi-dori, Niigata
951-8510 Japan
Tel: +81-25-227-2193
Fax: +81-25-227-0775
E-mail: noriaki@med.niigata-u.ac.jp
Title
Morphological Characteristics of Schwann Cells in the Islets of
Langerhans of the Murine Pancreas
Author
Eiji SUNAMI, Hiroaki KANAZAWA, Hiroya HASHIZUME, Masaei TAKEDA, Katsuyoshi HATAKEYAMA and
Tatsuo USHIKI
Address
Department of Anatomy and Histology, and Department of Surgery Faculty of Medicine,
Niigata University, Niigata; and Department of Radiological Technology, Nagoya University
School of Health Sciences, Nagoya, Japan
Summary
The present study demonstrated the three-dimensional architecture of periinsular nerve
plexuses in the murine pancreas by the combined use of light microscopy of S-100
immunostained sections, transmission electron microscopy (TEM) of thin sections, and
scanning electron microscopy (SEM) of KOH digested tissues. By light microscopy of thin
sections immunostained with anti-S-100 antibody, Schwann cells were often found on the
margin of the islets as if delimiting the islet and exocrine parenchyma. In thick
sections, Schwann cells of the islet connected their thin and slender processes with each
other to form a delicate network on the surface of the islet. By TEM, Schwann cells were
observed as an attenuated sheet that invested the surface of the islet. Axon terminals
were usually found on the outer surface of these membranous Schwann cells. SEM of KOH
digested tissues revealed that nerves reaching the islet spread on the insular surface.
Schwann cells in this portion extended their thin membranous processes, which directly
covered the basal part of several endocrine cells as a whole. Numerous axons with
varicosities were usually found on the surface of these membranous Schwann cells, but
sometimes crept beneath them.
These findings indicate that ''the interstitial cells" described by light
microscopists are peculiar-shaped Schwann cells present in the islets. The functional
significance of the rich innervation of the islets is also briefly discussed in the
present study.
Prof. Tatsuo USHIKI
Department of Anatomy and Histology Faculty of Medicine, Niigata University
Asahimachi-dori 1, Niigata
951-8510 Japan
Phone: +81-25-227-2058
Fax: +81-25-224-1767
E-mail: t-ushiki@med.niigata-u.ac.jp
Title
Purkinje Cells in the Adult Cat Cerebellar Cortex Possess a
Perineuronal Net of Proteoglycans
Author
Masaru MABUCHI, Shinichiro MURAKAMI, Takehito TAGUCHI, Aiji OHTSUKA and Takuro MURAKAMI
Address
Department of Anatomy, Faculty of Medicine; and Department of Radiological Technology,
Faculty of Health Sciences, Okayama University Medical School, Okayama, Japan
Summary
The Purkinje cells in the adult cat cerebellar cortex were found to possess perineuronal
proteoglycans which could be stained with our fine cationic iron colloid and Fujita's
highly concentrated aldehyde fuchsin, and digested by chondroitinase
ABC/keratanase/heparitinase and hyaluronidase. The Purkinje cells are surrounded by some
collagenous elements which are stained with Gomori's ammoniacal silver and digested by
collagenase. The Purkinje cells also express nerve cell surface glycoproteins which are
labeled with lectin Vicia villosa agglutinin and digested by a double treatment with
collagenase and endo-alpha-N-acetylgalactosaminidase.Sole digestion by
endo-alpha-N-acetylgalactosaminidase never erased the lectin labeling of the nerve cell
surface glycoproteins. These findings suggest that the collagenous elements mediate the
linkage of the perineuronal proteoglycans to the nerve cell surface glycoproteins. It is
presumed that in mice and rats, the perineuronal nets of proteoglycans and nerve cell
surface glycoproteins of the Purkinje cells are so thin or coarse that they can not be
sufficiently visualized under the light microscope.
Prof. Takuro MURAKAMI, M.D., Ph.D.
Department of Anatomy
Faculty of Medicine Okayama University Medical School
2-5-l Shikata-cho, Okayama
700-8558 Japan
Tel: +81-086-235-7088
Fax: +81-086-235-7095
E-mail: em2kai@md.okayama-u.ac.jp
Title
Effects of Photoperiod and Melatonin on the Development of Growth
Hormone Cells and the Pituitary-Adrenal Axis in the Djungarian Hamster, Phodopus sungorus
Author
Yoshiki HIRA, Yuko SAKAI and Shoji MATSUSHIMA
Address
Department of Anatomy, Asahikawa Medical College, Asahikawa, Japan
Summary
The development of GH cells and the pituitary-adrenal axis was morphologically examined in
male Djungarian hamsters (Phodopus sungorus) exposed to short days and those kept under
long days and receiving daily afternoon injections of melatonin, from the time of weaning
(20 days) until 100 days of age. The postnatal increase in area of ACTH cells under long
days was inhibited in short-day-exposed or melatonin-treated animals. It was suggested
that a short photo-period may suppress, via melatonin, the development of ACTH cells. GH
cells were not affected by age, photo-period or exogenous melatonin. Under long days, the
zona fasciculata decreased in volume with age, while the zona reticularis increased. Such
changes in the volumes of these adrenocortical zones were depressed under short days. In
addition, the volumes of the zona fasciculata and zona reticularis in long-day-housed
animals became respectively larger and smaller subsequent to orchidectomy and melatonin
administration. These results suggest that fasciculata cells in deeper levels become
progressively dilrerentiated into reticularis cells, that short photoperiod inhibits
development of both zonae, and that such an inhibition is caused mainly by the decreased
secretion of androgens.
Dr. Yoshiki HIRA
Department of Anatomy
Asahikawa Medical College
Midorigaoka-higashi 2-1-1-1
Asahikawa, 078-8510 Japan
Tel: +80-166-68-2312
Fax: +81-166-68-2319
Title
Asialoglycoprotein Receptors on Rat Dendritic Cells: Possible Roles
for Binding with Kupffer Cells and Ingesting Virus Particles
Author
Ryosuke UWATOKU, Kazuaki AKAIKE, Kazuhito YAMAGUCHI, Tosisuke KAWASAKI, Masayuki ANDO and
Kenjiro MATSUNO
Address
Departments of AnatomyⅡ, Internal MedicineⅠ and Microbiology, Kumamoto University
School of Medicine, Kumamoto; Institute of Laboratory Animals, Yamaguchi University School
of Medicine, Ube; and Department of Biological Chemistry, Kyoto University Graduate School
of Pharmaceutical Sciences, Kyoto, Japan
Summary
Rat dendritic cells selectively bind to Kupffer cells in vitro. The present study aimed to
reveal adhesion molecules on dendritic cells and their roles in the host defense system.
The in situ binding assay to examine the effects of pretreatment of dendritic cells with
various kinds of monosaccharides suggested that N-acetylgalactosamine was necessary for
the binding of dendritic cells to Kupffer cells. This binding was also attenuated when
dendritic cells were injected into an ex vivo liver perfusion circuit together with
N-acetylgalactosamine. It was further shown that the majority of rat lymph dendritic cells
and some interdigitating dendritic cells in the lymph nodes possessed asialoglycoprotein
receptors specific for N-acetylgalactosamine/galactose as detected by immunostaining.
Lymph dendritic cells could ingest virus particles in vitro, even though these cells
showed no phagocytic activity for latex particles. The results indicate that rat dendritic
cells possess asialoglycoprotein receptors which are probably utilized to recognize
Kupffer cells for their recruitment to the liver and possibly to recognize virus particles
prior to phagocytosis.
Prof. Kenjiro MATSUNO
Department of Anatomy I
Dokkyo University School of Medicine
Mibu, Tochigi
321-0293 Japan
Tel: +81-282-87-2123 (direct)
Fax: +81-282-86-6229 (direct)
E-mail address: keniiro@dokkyomed.ac.jp
