[SEM study on the dorsal lingual surface of the nutria, Myocastor coypus].

The dorsal lingual surface of prenatal and postnatal nutria was examined by scanning electron microscopy (SEM). Filiform (Fi), fungiform (Fu), foliate (Fo) and vallate papillae (Va) were observed. The Fi differed in morphology by their location on the tongue and could be classified into 3 types: 1) conical, 2) long, and 3) short filiform papillae. The Fu were scattered among the Fi. Many Fo were observed on the posterolateral regions of the tongue. There were 2 Va separating the anterior 2/3 from the posterior 1/3 of the tongue. The rudiments of Fu, Fo and Va were visible earlier than those of the Fi.

A novel germline mutation of the MEN1 gene, L259del, in a patient with sporadic multiple endocrine neoplasia type 1 (MEN1).

A Japanese woman was treated for insulinoma when she was 29 years old. Ten years later, heperparathyroidism and non-functioning adrenal tumor were found and she was diagnosed as having multiple endocrine neoplasia type 1 (MEN1). No other family members have developed MEN-related lesion(s). Genomic DNA of the patient was analyzed by sequencing for the MEN1 gene and a novel, three-base in-frame deletion resulting in deletion of an amino acid Leu259 was identified. Her two children showed a wild-type sequence at this codon.

Developing germ cells in mouse testis express pheromone receptors.

Pheromone receptors are expressed in the accessory olfactory system, which is vital for non-specific chemical communication and for sexual behavior. Under the hypothesis that some of the pheromone molecules released from female reproductive organs might regulate sperm chemotaxis or chemokinesis, we examined whether the V1R type pheromone receptor mRNAs are expressed in developing germ cells. By a reverse transcription-PCR method, we obtained nine kinds of cDNA fragments belonging to the receptor family. In situ hybridization analysis in testicular sections using probes of testicular pheromone receptors (TVRs) revealed that TVR mRNAs were expressed by spermatids. TVRs were also expressed in the accessory olfactory organ. In the testis, hybridization signals were localized in subsets of the seminiferous tubules, suggesting that TVRs were expressed by selective subsets of the spermatids. In situ hybridization study suggests also that each sperm expresses multiple pheromone receptors. The testicular pheromone receptors might have an important role in the maturation and/or migration of sperm.

Morphology of the dorsal lingual papillae in the barbary sheep, Ammotragus lervia.

The dorsal lingual surface of a barbary sheep (Ammotragus lervia) was examined by scanning electron microscopy (SEM). The tongue was about 20 cm in length. There were about 30 vallate papillae on both sides. Filiform, conical, fungiform and vallate papillae were found. The filiform papillae were distributed over the entire dorsal surface of the tongue, excepted for the lingual torus where conical papillae were present. The fungiform papillae were present rounded bodies, and more densely distributed as compared to that of the lingual body on the tip and ventral surface of lingual apex. No foliate papillae were seen on the dorsal surface. The vallate papillae were located on both sides of the midline in the caudal part. Each papilla was surrounded by a groove. These findings indicate that the tongue of the barbary sheep is similar to that of the formosan serow, japanese serow and blackbuck.

Diminished function and expression of the cardiac Na+-Ca2+ exchanger in diabetic rats: implication in Ca2+ overload.

1. The present work was carried out in order to determine whether a decrease in cardiac Na+-Ca2+ exchanger (NCX) activity observed in diabetes is caused by a reduction in NCX protein and mRNA levels and to elucidate the significance of this decrease in alterations in [Ca2+]i homeostasis in diabetic cardiomyocytes. 2. The NCX current was significantly reduced in ventricular myocytes freshly isolated from streptozotocin-induced diabetic rat hearts, and its current density was about 55 % of age-matched controls. 3. Diabetes resulted in a 30 % decrease in cardiac protein and mRNA levels of NCX1, a NCX isoform which is expressed at high levels in the heart. 4. The reduced NCX current and the decreased protein and mRNA levels of NCX1 in diabetes were prevented by insulin therapy. 5. Although both diastolic and peak systolic [Ca2+]i were not different between the two groups of myocytes, increasing external Ca2+ concentration to high levels greatly elevated diastolic [Ca2+]i in diabetic myocytes. Inhibition of NCX by reduction in extracellular Na+ by 50 % could produce a marked rise in diastolic [Ca2+]i in control myocytes in response to high Ca2+, as seen in diabetic myocytes. However, cyclopiazonic acid, an inhibitor of sarcoplasmic reticulum Ca2+ pump ATPase, did not modify the high Ca2+-induced changes in diastolic [Ca2+]i in either control or diabetic myocytes. 6. Only in papillary muscles from diabetic rats did the addition of high Ca2+ cause a marked rise in resting tension signifying a partial contracture that was possibly due to an increase in diastolic [Ca2+]i. 7. In conclusion, the diminished NCX function in diabetic myocytes shown in this study results in part from the decreased levels of cardiac NCX protein and mRNA. We suggest that this impaired NCX function may play an important role in alterations in Ca2+ handling when [Ca2+]i rises to pathological levels.

Effects of ethanol on the ultrastructure of the hamster thyroid C-cell.

The morphology of the thyroid C-cells in golden hamsters after short- and long-term treatment with ethanol was studied. Immunohistochemistry was applied to examine the distribution of the C-cells in the thyroid gland. In the short-term experimental animals, the Golgi complexes and the granular endoplasmic reticulum were well developed and the number of the secretory granules was decreased as compared with those of the control animals. These findings suggest that the cellular activity of the thyroid C-cell is stimulated after short-term treatment with ethanol. The morphology of the thyroid C-cells of the long-term experimental animals was similar to that of the controls. It is conceivable that long-term treatment with ethanol does not affect the function of the C-cell.

Morphology of the dorsal lingual papillae in the black rhinoceros (Diceros bicornis).

The dorsal lingual surface of a black rhinoceros (Diceros bicornis) was examined by scanning electron microscopy. The tongue was about 30 cm in length. There were about 60 vallate papillae on both sides. Filiform, fungiform and vallate papillae were found. The filiform papillae were distributed over the entire dorsal surface of the tongue. The papillae had a hair-like shape. The fungiform papillae were round in shape, and more densely distributed on the lingual apex. No foliate papillae were seen on the dorsal surface. The vallate papillae were located on both sides of the posterior end of the lingual body. Each papilla was surrounded by a groove.

Morphology of the dorsal lingual papillae in the bush dog (Speothos venaticus).

The dorsal lingual surface of a bush dog (Speothos venaticus) was examined by scanning electron microscopy (SEM). The tongue was about 7 cm in length. Filiform, fungiform and vallate papillae were found. The filiform papillae were distributed over the entire dorsal surface of the tongue. Each filiform papillae on the apical surface of the tongue had several conical processes, in the midportion were larger than those on the apex in size. In the region of the vallate papillae, the filiform papillae had not the conical processes and more larger than those on the midportion of the tongue. The fungiform papillae were present rounded bodies and more densely distributed on the tip of the lingual apex. There were 5 vallate papillae on both sides. The vallate papillae were located on both sides of the posterior end of the lingual body. Each papilla was surrounded by a groove and a crescent pad. In the dorsal surfaces of the papillae, small conical papillae were observed.

SEM study on the dorsal lingual surface of Microtus montebelli.

The dorsal lingual surface of Microtus montebelli was examined by scanning electron microscopy (SEM). The filiform papillae were distributed over the entire dorsal surface of the tongue. The filiform papillae notably differed in morphology by their location on the tongue and could be classified into 4 types: 1) simple conical papillae, 2) true filiform papillae, 3) giant conical papillae, and 4) saw-like papillae. The fugiform papillae were present rounded bodies scattered over the anterior dorsal surface of the tongue. A vallate papilla was located at the middle end of the lingual body. The surface of the radix zone was smooth and without lingual papillae.

SEM study on the dorsal lingual surface of the flying squirrel, Petaurista leucogenys.

The dorsal lingual surface of the flying squirrel was examined by scanning electron microscopy (SEM). Filiform (FI), fungiform (FU), foliate (FO) and vallate papillae (VA) were observed. The FI were distributed over the entire dorsal surface of the tongue. In the region of the VA, the FI appeared as giant cones. The FU were present as rounded bodies scattered over the dorsal surface of the tongue. They were relatively scarce and appear to be concentrated around the edges of the tongue. The FO were observed on the posterolateral regions of the tongue. The flying squirrel showed the triangular arrangement of the three VA, with the apex of the triangle directed posteriorly.

Electron microscopic study of the parathyroid gland of Rattus rattus.

The ultrastructure of the parathyroid glands of Rattus rattus was investigated. In the chief cells, the nucleus showed deep indentations, and lysosomal inclusions and rod-shaped crystalloids were present. Their ultrastructure is described, and the role of the lysosomal inclusions is discussed.

Targeted delivery and improved therapeutic potential of catalase by chemical modification: combination with superoxide dismutase derivatives.

Four types of bovine liver catalase (CAT) derivatives, succinylated (Suc-CAT), galactosylated (Gal-CAT), mannosylated (Man-CAT), and polyethylene glycol conjugate (PEG-CAT), were synthesized and their pharmacokinetics and therapeutic potential in a hepatic ischemia/reperfusion injury model were studied in mice. About 90% of the CAT enzymatic activity was retained after chemical modification. Biodistribution studies showed that 111indium (111In)-Gal-CAT accumulated selectively in the liver parenchymal cells as 111In-CAT, whereas an increased amount of 111In-Suc-CAT and 111In-Man-CAT was delivered to liver nonparenchymal cells. 111In-PEG-CAT exhibited prolonged retention in plasma. Pharmacokinetic analysis revealed that the hepatic uptake clearances of 111In-Suc-CAT, 111In-Gal-CAT, and 111In-Man-CAT were much greater than that of 111In-CAT, whereas that of 111In-PEG-CAT was very small. In the ischemia/reperfusion injury model, in which hepatic injury was induced by occlusion of the portal vein for 30 min followed by 1 h reperfusion, the elevation of plasma glutamic pyruvic transaminase and glutamic oxaloacetic transaminase levels was slightly inhibited by treatment with native CAT or Gal-CAT. PEG-CAT was less potent. In contrast, Suc-CAT and Man-CAT effectively suppressed the increase in plasma glutamic pyruvic transaminase and glutamic oxaloacetic transaminase. Coinjection of mannosylated superoxide dismutase marginally improved the inhibitory effects of CAT derivatives. These results demonstrate that targeted CAT delivery to liver nonparenchymal cells via chemical modification is a promising approach to prevent hepatic injuries caused by reactive oxygen species. The potential usefulness of combining of CAT and superoxide dismutase derivatives is also demonstrated.

Effects of long-term treatment with ethanol on the ultrastructure of the golden hamster parathyroid gland.

The ultrastructure of the parathyroid gland in golden hamsters after long-term treatment with ethanol was studied. Male hamsters of experimental groups were given ethanol at the concentration of 7% for 3 and 5 months with food and water freely available. In the ethanol-treated hamsters, the Golgi complexes associated with many prosecretory granules were well developed and many secretory granules were located near the plasma membrane as compared with those of the control animals. Exocytotic events were observed in 5-month-treated animals. These findings suggest that the secretory activity of the parathyroid gland is stimulated after long-term treatment with ethanol.

Macrophages in the hamster parathyroid gland: immunohistochemical and ultrastructural investigations.

The distribution and morphology of the parathyroid macrophages in golden hamsters from neonatal to senile periods were investigated using the monoclonal antibody to ED2 and electron microscopy. The results showed that definite ED2-positive cells were hardly detectable in the parathyroid gland of 1-day-old hamsters. A few ED2-positive cells could be identified in the parathyroid gland of 10-day-old hamsters. The ED2-positive cells were more densely and became conspicuous in 1-, 3-, and 12-month-old hamsters. The number of the cells seems to be increased with aging. Ultrastructurally, we did not find any macrophages in parathyroid glands of 1-day-old hamsters. In the 10-day-old hamster parathyroid gland, a few macrophages existed only in the interstitial tissues. In the parathyroid gland of 1-, 3-, and 12-month-old hamsters, many macrophages were found in the perivascular regions, some cells located among the parenchymal chief cells with no obvious vascular association. These cells showed some physical contacts with chief cells. These results suggest that the parathyroid macrophages exhibit dramatical changes in their distribution and morphology from neonatal to senile periods.

Morphology of the dorsal lingual papillae in the blackbuck, Antilope cervicapra.

The dorsal lingual surface of a blackbuck (Antilope cervicapra) was examined by scanning electron microscopy (SEM). The tongue was about 125 mm in length. There were about 30 vallate papillae on both sides. Filiform, conical, fungiform and vallate papillae were found. The filiform papillae were distributed over the entire dorsal surface of the tongue, excepted for the lingual torus where conical papillae were present. The fugiform papillae were present rounded bodies, and more densely distributed on the tip and ventral surface of ligual apex. No foliate papillae were seen on the dorsal surface. The vallate papillae were located on both sides of the midline in the caudal part. Each papilla was surrounded by a groove. These findings indicate that the tongue of the blackbuck is similar to that of the formosan and japanese serow.

Ultrastructure of the parathyroid gland of the young golden hamster after short-term treatment with ethanol.

We studied the ultrastructure of the parathyroid gland of young golden hamsters after short-term treatment with ethanol (1.5 g/kg bw or 6.0 g/kg bw). We did not find any ultrastructural changes of the parathyroid gland after administration of 1.5 g/kg ethanol. In the hamsters, 3 hours after administration of 6.0 g/kg ethanol, the mean serum calcium concentration was significantly low as compared to that of the control animals. In the parathyroid gland 1 hour after administration of 6.0 g/kg ethanol, the Golgi complexes associated with a few prosecretory granules and the volume density occupied by the Golgi complexes decreased compared with that of the control animals. In the parathyroid glands 3 hours after administration of 6.0 g/kg ethanol, the Golgi complexes decreased as compared with those of the control animals, while the large vacuolar bodies increased. These findings suggest that the cellular activity of the parathyroid gland is suppressed after short-term treatment with ethanol. Intracellular lumen was found in the parathyroid chief cells 3 hours after administration of 6.0 g/kg ethanol, and the significance of this structure is discussed.

Fine structure of the parathyroid and ultimobranchial glands of the snake, Elaphe quadrivirgata.

The ultrastructure of the parathyroid gland and ultimobranchial gland of the snake, Elaphe quadrivirgata, was studied. The parenchyma of the parathyroid gland was consisted of chief cells arranged in cords. Oxyphil cells and water-clear cells were not recognized. The chief cells contained irregular shaped nucleus, moderately-developed mitochondria, granular endoplasmic reticulum and Golgi complexes. A few secretory granules of 100-300 nm in diameter distributed in the cytoplasm. Small aggregations of glycogen particles and lipid droplets were dispersed widely in the cytoplasm. The ultimobranchial glands consisted of follicles and interfollicular aggregates of C-cells. Follicles were invaginated and composed of C-cells, goblet cells and ciliated cells. C-cells were located in the basal position of the follicle and possessed variety of secretory granules of 100-300 nm in diameter in the basal region of the cell. C-cells presented various secretory cycles. Goblet cells projected microvilli into follicular lumen. At the apical region the goblet cells had large granules of 300-1,200 nm in diameter. Ciliated cells projected cilia intermixed with micovilli from the apical surface of the cell into the follicular lumen.

Effects of beta-adrenoceptor stimulation on contractility, [Ca2+]i, and Ca2+ current in diabetic rat cardiomyocytes.

The mechanism of the diminished inotropic response to beta-adrenoceptor stimulation in diabetic hearts was studied in enzymatically isolated diabetic rat ventricular myocytes in comparison with age-matched controls. The increases in contractions and intracellular Ca2+ concentration ([Ca2+]i) transients produced by isoproterenol were markedly diminished in diabetic myocytes. The inotropic and [Ca2+]i responses to forskolin and dibutyryl cAMP (DBcAMP) were also reduced. No significant difference was found in the stimulating effects of isoproterenol, forskolin, and DBcAMP on the L-type Ca2+ current (ICa) between control and diabetic myocytes. The rise of [Ca2+]i in response to rapid caffeine application, an index of sarcoplasmic reticulum (SR) Ca2+ content, was significantly decreased in diabetic myocytes. Isoproterenol, forskolin, and DBcAMP enhanced this [Ca2+]i response to caffeine in control myocytes more markedly than in diabetic myocytes. The changes in the isoproterenol responses observed in diabetic myocytes were prevented by insulin therapy. We conclude that 1) diabetes causes an impairment of the contractile and [Ca2+]i responses of cardiac myocytes when stimulated at both beta-adrenoceptors and the postreceptor level without affecting the ICa response and 2) altered SR functions of uptake and/or release of Ca2+ may primarily contribute to the diminished beta-adrenergic response.

The parathyroid glands of two species of dolphin--Risso's dolphin, Grampus griseus, and bottlenose dolphin, Tursiops truncatus.

Although there have been many reports regarding the structure of the parathyroid glands of various terrestrial mammals, little is known about the parathyroid glands of marine mammals including Cetacea. The morphology of the parathyroid glands of three Risso's dolphins, Grampus griseus (about 3 m in length and 300 kg in weight), and three bottlenose dolphins, Tursiops truncatus (about 3 m in length and 300 kg in weight), was examined macroscopically and microscopically. The dolphins examined in the present study had two or four parathyroid glands that varied in size and location on the thyroid gland. Each parathyroid gland was encapsulated by fibrous tissue on the dorsal surface of the thyroid gland, and was divided into several lobules by interlobular connective tissue which contained numerous capillaries. The parenchymal cells consisted of pale staining chief cells. Each cell was polygonal and about 15 microm in diameter, and had one round or oval nucleus. Oxyphil cells were not found. Considering their greater body size, the parathyroid glands were rather small. By electron microscopic observation, the parathyroid gland of the bottlenose dolphin had sparse granular endoplasmic reticulum, poorly developed Golgi complexes, and abundant secretory granules in the cytoplasm of the chief cells. These results support a possibility that the activity of the parathyroid gland is suppressed to adapt to a sea habitat.

Effect of immobilization on the ultrastructure of the golden hamster parathyroid gland.

To investigate the morphological changes of the parathyroid gland of the immobilized hamsters, we studied the ultrastructure of the parathyroid gland of golden hamsters kept in special small cage (Ballman cage II). All hamsters of the control group were kept in one ordinary cage. Each hamster of the isolated group was kept in ordinary cage individually. Each hamster of the immobilized group was kept in Ballman cage II individually. All hamsters were kept for 5 days. On the first and fifth day of the experiment, bone mineral content (BMC) and bone mineral density (BMD) of whole body were measured by dual energy X-ray absorptiometry (DXA). In the control and isolated groups, BMD of the fifth day was significantly increased as compared to that of the first day. In the immobilized group BMC and body weight were significantly decreased. There was no significant difference among 3 groups concerning the mean serum calcium level. Volume density of the cell organelles and inclusions was estimated and compared among 3 groups. Volume density of the lysosomes and large vacuolar bodies of the isolated and immobilized groups was significantly higher than that of the control group. Much more lipid droplets were observed in the immobilized group than the control and isolated groups. No particular differences were observed as to the Golgi complex in the isolated and the immobilized groups compared to the control group. These findings suggest that the cellular activity of the parathyroid gland is suppressed with immobilization.