Original articles:
- Immunocytochemical
localization of chromogranin A and secretogranin II in female rat gonadotropes. (Watanabe
T; Arch Histol Cytol, 1998 May)
- Direct
observation of t-butyl alcohol frozen and sublimated samples using low-vacuum scanning
electron microscopy. (Hashizume H; Arch Histol Cytol, 1998 May)
- Embryonic
development of the inner ear and otolith of the rainbow trout Oncorhynchus mykiss. (Salem
MA; Arch Histol Cytol, 1998 May)
- Novel
S100 proteins in human esophageal epithelial cells: CAAF1 expression is associated with
cell growth arrest. (Hitomi J; Arch Histol Cytol, 1998 May)
- Immunohistochemical
identification of type B intercalated cells in the rat kidney by a monoclonal antibody.
(Yanase H; Arch Histol Cytol, 1998 May)
- Development
of diaphragmatic lymphatics: the process of their direct connection to the peritoneal
cavity. (Shao XJ; Arch Histol Cytol, 1998 May)
- Molecular
weight-dependent effects of hyaluronate on the arthritic synovium. (Asari A; Arch Histol
Cytol, 1998 May)
- Evidence
for polymorphism of Merkel cells in the adult human oral mucosa. (Tachibana T; Arch Histol
Cytol, 1998 May)
Summary
Title
Immunocytochemical localization of chromogranin A and secretogranin II in
female rat gonadotropes.
Author
Watanabe T; Azuma T; Banno T; Jeziorowski T; Ohsawa Y; Waguri S; Grube D; Uchiyama Y
Address
Department of Cell Biology and Anatomy I, Osaka University Medical School, Suita-City,
Japan.
Summary
Ultrastructures of pituitary gonadotropes are known to show a prominent sex-related
difference: typical male rat gonadotropes contain both large- and small-sized granules,
whereas typical female rat gonadotropes appear to exhibit uniformly small-sized granules.
Our preceding studies have demonstrated that two representative granins, chromogranin A
(CgA) and secretogranin II (SgII), are separately localized to each type of granule in
male rat gonadotropes. To clarify whether or not there is a certain relationship between
granin proteins and characteristic features of secretory granules in female rat
gonadotropes, we examined the expression levels and immunocytochemical localizations of
CgA and SgII in the cells. Northern blot and immunoblot analyses demonstrated that both
CgA and SgII were synthesized and stored in the female pituitary, although the amount of
CgA was much lower in the female than that in the male pituitary. Immunocytochemical
observations clarified that gonadotropes in the female pituitary possessed intermediate
secretory granules containing both CgA and SgII, in addition to solely CgA-positive and
SgII-positive ones. However, secretory granules containing CgA in the female gonadotropes
were much smaller in size and appeared less frequently than those in the male cells,
whereas no sexual difference was discerned in SgII-positive granules. Moreover, the size
and appearance of CgA-positive secretory granules varied depending on stages of the
estrous cycle. These findings suggest that the size and appearance of secretory granules
containing CgA are closely associated with the expression and storage levels of CgA in the
pituitary.
Title
Direct
observation of t-butyl alcohol frozen and sublimated samples using low-vacuum scanning
electron microscopy.
Author
Hashizume H; Itoh S; Tanaka K; Ushiki T
Address
Department of Anatomy, Niigata University School of Medicine, Japan.
Summary
Frozen biological specimens in t-butyl alcohol were examined under a low-vacuum
environment in a "wet SEM" or "variable pressure SEM (scanning electron
microscope)" equipped with a cooling stage and highly sensitive backscattered
detector of the YAG type. After fixation with glutaraldehyde and osmium tetroxide, rat
tissue blocks (tracheae and kidneys), and cultured human carcinoma cells were dehydrated
with a graded series of t-butyl alcohol. The specimens were directly frozen on the cooling
stage at -10 degrees C, evacuated to 20 Pa in the specimen chamber, and observed by
detecting backscattered electrons at accelerating voltages of 5-6 kV. The images became
clearer 20 min after the vacuum reached 20 Pa and revealed had good quality by 30 min,
probably because t-butyl alcohol was sublimated during the time. The cilia of tracheal
ciliated cells, end-feet of the podocytes of the renal glomerulus, and processes of
cultured cells were clearly observed without any serious preparation artifacts. Since the
low-vacuum SEM of t-butyl alcohol frozen samples is both simple and provides high imaging
quality, it is expected to be useful in a variety of biological fields such as the rapid
pathological diagnosis.
Title
Embryonic
development of the inner ear and otolith of the rainbow trout Oncorhynchus
mykiss.
Author
Salem MA; Omura Y
Address
Laboratory of Animal Information Biology, Graduate School of Bioagricultural Sciences,
Nagoya University, Chikusa, Japan.
Summary
The embryonic development of the inner ear, especially the sensory epithelia and otoliths
in the rainbow trout Oncorhynchus mykiss, was studied by light and electron microscopy.
Light microscopically, the auditory vesicle, saccular otolith and statoacoustic ganglion
were first observed by 12 days after fertilization, while the utricular otolith appeared
at 15 days after fertilization. Both the saccular and utricular maculae were more
developed at 22 days after fertilization, and well developed by 27 days. The crista
ampullaris of the horizontal canal was also developed at 27 days after fertilization,
while the other cristae were not yet distinguished. Electron microscopically, vesicular
structures and short microvilli were found on the sensory epithelia of the maculae by 15
days after fertilization. At 22 days after fertilization, the saccular otolith possessed 7
incremental layers, and developing cilia, microvilli, and aggregates of secretory
materials also appeared on the apical surface of the sensory epithelia. At 27 days after
fertilization, the apical surface of each hair cell was covered with a hair bundle
consisting of a single kinocilium and a bundle of stereocilia. These findings are
discussed with special regard to the environmental factors on early development in fishes.
Title
Novel
S100 proteins in human esophageal epithelial cells: CAAF1 expression is associated with
cell growth arrest.
Author
Hitomi J; Kimura T; Kusumi E; Nakagawa S; Kuwabara S; Hatakeyama K; Yamaguchi K
Address
Department of Anatomy, Niigata University School of Medicine, Japan.
Summary
CAAF1 and CAAF2, newly identified calcium-binding proteins from bovine amniotic fluid,
have been revealed to be members of the S100 protein family preferentially produced by
fetal squamous epithelial cells, including epidermal keratinocytes. Having previously
cloned the cDNA of human CAAF1 protein from the esophageal epithelium, we report here on
the characteristic expression pattern of CAAF1 and related S100 proteins in human
esophageal epithelial cells. Normal cells of the human esophageal epithelium expressed
CAAF1, and also expressed the homologous novel S100 proteins including CAAF2, MRP8, and
MRP14, but not S100alpha. An immunohistochemical study with specific monoclonal antibodies
against CAAF1 proteins demonstrated that CAAF1 proteins were produced by the esophageal
epithelial cells in the process of cell differentiation. The immature proliferating cells
in the epithelium did not produce CAAF1 proteins, but the differentiated cells expressed
CAAF1, which overlay the immature cells and were stratifying in the epithelium. These CAA
1-producing cells did not show any proliferating activities. Esophageal carcinoma cells
did not express CAAF1, except for the keratinized cells with no proliferating activity. In
addition, the forced expression of CAAF1 proteins in the carcinoma cells resulted in a
marked decrease in DNA synthesis. These findings indicate that human esophageal epithelial
cells express the multiple genes of S100 proteins including CAAF proteins, and that CAAF1
is closely associated with the terminal differentiation of these cells. CAAF1 expression
also might play some role in cell growth.
Title
Immunohistochemical
identification of type B intercalated cells in the rat kidney by a monoclonal antibody.
Author
Yanase H; Orikasa M; Shimizu F; Kawachi H; Iwanaga T
Address
Laboratory of Anatomy, Graduate School of Veterinary Medicine, Hokkaido University,
Sapporo, Japan.
Summary
We recently produced monoclonal antibodies using macrophagic cells derived from cultured
rat glomeruli as the antigen. One of the antibodies, named OS-3, was found to detect a
cell population scattered in collecting ducts of the rat kidney as well as macrophages in
various tissues. The present study deals with the cellular and subcellular localization of
immunoreactivities with OS-3 in the kidney and other organs of rats. Double immunostaining
using OS-3 and an anti-serum against either calbindin or epidermal cytokeratin showed that
OS-3-immunoreactive cells exist exclusively in both the connecting segment and cortical
collecting duct, and differ from calbindin- or cytokeratin-positive epithelial cells.
Ultrastructurally, OS-3-immunoreactive cells appeared spherical in shape with few
cytoplasmic microprojections on the narrow apical surface. Their relatively dark cytoplasm
contained numerous mitochondria and a developed tubulo-vesicular system. The intense
immunoreactivity was selectively localized in the basolateral membrane exhibiting shallow
but complicated infoldings. Distribution and ultrastructural properties of the
OS-3-immunoreactive cells showed that they were type B intercalated cells, which are
engaged in the regulation of the acid-base balance mainly by secreting HCO3-. Another
positive staining with OS-3 was found in the macula densa and some epithelial cells of
Bowman's capsule, the former monitoring Cl- concentrations in the urine. Immunoblotting of
extracts from the rat kidney demonstrated a protein band immunoreactive to OS-3 at a
molecular weight of 43 kDa. Aside from the kidney, a specific and intense immunoreactivity
with OS-3 was also found in the epithelial cells of the pancreatic excretory duct and in
the secretory cells of the salivary, pyloric and duodenal glands, all of which are HCO3-
-secreting cells. These immunohistochemical findings imply that OS-3 is useful for the
detection of type B intercalated cells and recognizes a functional molecule involved in
the production/secretion of HCO3- or transport of Cl-.
Title
Development
of diaphragmatic lymphatics: the process of their direct connection to the peritoneal
cavity.
Author
Shao XJ; Ohtani O; Saitoh M; Ohtani Y
Address
Department of Anatomy, Toyama Medical and Pharmaceutical University,
Sugitani, Japan.
Summary
The development of the lymphatic system in the rat diaphragm was studied from embryonic
day 16 to 25 weeks after birth by histochemistry for 5'-nucleotidase, scanning electron
microscopy of KOH-treated or intact tissues, and transmission electron microscopy of thin
sections. On embryonic day 16, distinct lymphatics were noted in the subpleural space of
the diaphragm periphery. The endothelial cells at this stage contained an abundance of
rough endoplasmic reticulum, a developed Golgi apparatus and mitochondria, and fewer
pinocytotic vesicles than those in adults. The subpleural lymphatics subsequently
increased and formed a polygonal network. They possessed many valves, and by postnatal
week 6, some thick collecting lymphatics became endowed with smooth muscle cells. On
embryonic day 19, some lymphatics appeared in the subperitoneal space. They extended
centripetally and had many lateral projections that subsequently became elongated and
connected with those from adjacent lymphatics, thus forming a lattice-like network. During
the early postnatal days, the subperitoneal lymphatics projected many bulges that
subsequently became elongated, and came into contact with the pores among the mesothelial
cells, thus forming lymphatic stomata connecting the lymphatic lacunae to the peritoneal
cavity. The lymphatic stomata increased until postnatal week 10. The results show that
lymphatics appear as early as embryonic day 16 in the subpleural space of the diaphragm
periphery, and develop with age by sprouting to form networks in both the subpleural and
the subperitoneal spaces, and that the direct connection of the lymphatic lacunae to the
peritoneal cavity is formed after birth.
Title
Molecular
weight-dependent effects of hyaluronate on the arthritic synovium.
Author
Asari A; Miyauchi S; Matsuzaka S; Ito T; Kominami E; Uchiyama Y
Address
Tokyo Research Institute, Seikagaku Corporation, Higashiyamato, Japan.
Summary
Intra-articular injection of hyaluronate (HA) is widely used in the treatment of
arthropathies. However, the mechanism of the effects of HA preparations on the arthritic
synovium and the relationship between their effects and molecular weights (MW) remains
unknown. The objectives of this study were to compare the effects of two hyaluronate
preparations, HA84 (MW: 84 X 10(4)) and HA230 (MW: 230 X 10(4)), on the synovium of an
arthritis model and to examine the mechanism of the effects of HA. The HA preparations
were intra-articularly injected in a model of canine arthritis induced by anterior
cruciate ligament transection for a total trial of 5 weeks. To define the accessibility of
HA preparations to the synovial lining layers, fluorescein-labeled HA84 or HA230 was
injected at the last administration. Pathological changes analyzed included increases in
volumes and prostaglandin E2 concentrations in synovial fluids, thickening of the synovial
lining layers, vacuolar alterations in the lining cells, and stainability of HA in the
synovium. Expression levels of Heat shock protein 72 (Hsp72) were immunohistochemically
detected in the tissues to investigate the ability of the cells to survive the
degeneration. The pathological changes described above were more significantly suppressed
in the HA84-treated than in the HA230-treated groups. In most cases of the HA84-treated
group (five cases out of six), fluorescein particles were intensely distributed in the
synovial lining layers, but only two cases in the HA230-treated group showed a weak
distribution of fluorescein particles in the layers, indicating a certain difference in
the accessibility of HA preparations to the lining cells between the two HA molecules.
Moreover, the immunoreactivity for Hsp72 in the lining cells as more intense in the
HA84-treated than in the HA230-treated groups. The difference in the accessibility of HA
molecules corresponded well with that in the inducibility of Hsp72 in the lining cells.
These results suggest that the up-regulation of Hsp72 may offer a new concept concerning
mechanism of the effects of HA preparations on the arthritic synovium.
Title
Evidence
for polymorphism of Merkel cells in the adult human oral mucosa.
Author
Tachibana T; Kamegai T; Takahashi N; Nawa T
Address
Department of Oral Anatomy, Iwate Medical University School of Dentistry, Morioka, Japan.
Summary
We recently reported that Merkel cells in the normal palatine mucosa of adult rodents are
highly polymorphic. In order to ascertain whether or not this polymorphism is also evident
in the human oral mucosa, palatine mucosae from cadavers without oral diseases and
perilesional palatine mucosae of patients with pleomorphic adenoma were examined by
immunohistochemistry using an antibody against cytokeratin 20. Findings showed that Merkel
cells in the human normal palatine mucosa were polymorphic, and a number of
irregular-shaped Merkel cells (dendritic Merkel cells) with apparent cytoplasmic
projections were present among typical oval to round Merkel cells. The mucosa usually
contained a small number of oval to round Merkel cells residing in ectopic places such as
prickle and granular cell layers. On the other hand, the slightly inflamed perilesional
palatine mucosa contained an increased incidence of dendritic Merkel cells. Ectopic Merkel
cells were rare in the perilesional palatine mucosa. Characteristics of dendritic Merkel
cells were examined using specimens from perilesional palatine mucosae by means of
immunohistochemistry and electron microscopy. It was shown that every dendritic Merkel
cell and most roundish Merkel cells in the perilesional mucosa lacked innervation.
Electron microscopy suggested that dendritic Merkel cells release secretory granules from
the tip of the cytoplasmic process and the basal cytoplasm towards the lamina propria
mucosae, in a manner resembling the case of similar cells in rodents.
