Akihiko Maekawa

Disruption of Spermatogenic Cell Adhesion and Male Infertility in Mice Lacking TSLC1/IGSF4, an Immunoglobulin Superfamily Cell Adhesion Molecule

ABSTRACT TSLC1/IGSF4, an immunoglobulin superfamily molecule, is predominantly expressed in the brain, lungs, and testes and plays important roles in epithelial cell adhesion, cancer invasion, and synapse formation. We generated Tslc1/Igsf4-deficient mice by disrupting exon 1 of the gene and found that Tslc1−/− mice were born with the expected Mendelian ratio but that Tslc1−/− male mice were infertile. In 11-week-old adult Tslc1−/− mice, the weight of a testis was 88% that in Tslc1+/+ mice, and the number of sperm in the semen was approximately 0.01% that in Tslc1+/+ mice. Histological analysis revealed that the round spermatids and the pachytene spermatocytes failed to attach to the Sertoli cells in the seminiferous tubules and sloughed off into the lumen with apoptosis in the Tslc1−/− mice. On the other hand, the spermatogonia and the interstitial cells, including Leydig cells, were essentially unaffected. In the Tslc1+/+ mice, TSLC1/IGSF4 expression was observed in the spermatogenic cells from the intermediate spermatogonia to the early pachytene spermatocytes and from spermatids at step 7 or later. These findings suggest that TSLC1/IGSF4 expression is indispensable for the adhesion of spermatocytes and spermatids to Sertoli cells and for their normal differentiation into mature spermatozoa.

Long-term toxicity/carcinogenicity of musk xylol in B6C3F1 mice

The long-term toxicity/carcinogenicity of musk xylol, a synthetic nitro musk, was examined in B6C3F1 mice of both sexes. Musk xylol was administered at dietary levels of 0 (control), 0.075 or 0.15% for 80 wk. The overall tumour incidences in all treated groups of both sexes were significantly higher than those in the corresponding controls. Combined malignant and benign liver cell tumours were clearly increased in both sexes, and in males a positive significant trend was also noted for the occurrence of hepatocellular carcinomas. In males the incidence of Harderian gland tumours was also significantly greater in treated groups than in controls. Some other neoplasms, such as lung tumours in both sexes and Harderian gland tumours and lymphomas in females, occurred in greater numbers in the treated groups, although the differences were not statistically significant in comparison with the controls. In addition, the incidences and total numbers of malignant tumours were significantly increased in treated groups of both sexes, although the increases were not dose dependent. The results demonstrated that musk xylol is carcinogenic in B6C3F1 mice when given at dose levels of 0.075 or 0.15% in the diet for 80 wk.

Tumor-Promoting Effects of Both Iodine Deficiency and Iodine Excess in the Rat Thyroid

Thyroid tumor-promoting effects of iodine deficiency and iodine excess were investigated in a rodent 2-stage model to estimate an optimal iodine intake range that would not effectively promote development of thyroid neoplasia. Six-week-old male F344 rats were given a single subcutaneous injection of 2,800 mg/kg body weight N-bis(2-hydroxypropyl)-nitrosamine (DHPN) or saline vehicle, maintained on Remingtons iodine-deficient diet (21 ± 2 ng/g iodide), and supplemented with various amounts of potassium iodide up to 260 mg/liter in drinking water to generate conditions ranging from severe iodine deficiency to severe iodine excess. In DHPN-treated rats, both conditions significantly increased thyroid follicular tumorigenesis. In DHPN-untreated rats, iodine deficiency produced diffuse thyroid hyperplasia, characterized by small follicles with tall epithelium and reduced colloid, together with a decrease in thyroxine (T4) and an increase in thyroid-stimulating hormone (TSH). On the other hand, iodine excess produced colloid goiter, characterized by large follicles with flat epithelium and abundant colloid admixed with normal or small-sized follicles lined by cpithelium of normal height, together with normal serum T4 and slightly decreased TSH. These effects were directly proportional to the severity of iodine deficiency or extent of iodine excess and suggest that each condition has a different thyroid tumor promotion mechanism. Iodine intakes that showed the least tumor promotion were 2.6 and 9.7 μg/rat/day in this study. Promoting mechanisms and the problem of statistically estimating recommended daily iodine intake range are briefly discussed.

Carcinogenicity studies of sodium nitrite and sodium nitrate in F-344 rats*

The carcinogenicity of sodium and of sodium nitrate was examined in F-344 rats. Sodium nitrite was administered in the drinking-water for 2 yr at levels of 0.125 or 0.25%. Sodium nitrate was given in the diet at levels 2.5 or 5%. A variety of tumours occurred in all groups including the controls. The only significant difference between treated and control groups in the total number of tumours detected in either of the studies was a significant decrease in tumour incidence in the high-dose females given nitrite compared with controls. There was no positive dose-response relationship either in the incidence or in the induction time of tumours in either of the studies. The only significant result was a reduction in the incidence of mononuclear cell leukaemias in the experimental groups in both studies. It is concluded that sodium nitrite and sodium nitrate did not exert a carcinogenic effect that could be detected under the conditions of this study in which the animals showed a high incidence of spontaneous tumours.

Spontaneous uterine adenocarcinomas in aged rats and their relation to endocrine imbalance.

SummaryIn addition to spontaneous uterine endometrial adenocarcinomas at a high incidence (35.1 %), development of endometrial hyperplasia/adenoma was also frequently detected in rats of the Donryu strain. The total yield of all observed proliferative endometrial lesions was very high (60.6%). The tumors arose commonly in the uterine horn of aged rats. Histologically, most demonstrated glandular structures, consisting of cuboidal or columnar cells with weak eosinophilic or basophilic cytoplasm ↭d large nuclei. In about half of the animals with adenocarcinomas, metastasis to remote organs such as the lung was observed. Histological examination of the ovary and vaginal epithelium revealed ovarian cysts, atrophy of the ovary and cornification of the vaginal epithelium more frequently in rats with endometrial carcinomas than in animals without tumors. These findings indicate that adenocarcinoma development in Donryu rats is associated with endocrine imbalance [increased serum estrogen: progesterone (E2∶P)ratios]. By comparative investigation of strain differences, it was confirmed that irregular estrous cycles began earlier with higher incidence in Donryu rats than in F344 rats, a low-incidence strain. Histological findings of the ovary and vaginal epithelium also suggested relatively increased estrogen levels in Donryu rats compared to F344 rats. Estimated plasma values of gonad steroids showed that the E2∶P ratio in Donryu rats at 12 months of age was about five times that in F344 rats. These results therefore indicate that hormone imbalance, particularly an increased E2∶P ratio, may play an important role in the spontaneous occurrence of endometrial adenocarcinoma in Donryu rats.

Development of invasive adenocarcinoma in a long-standing diverted ileal J-pouch for ulcerative colitis : Report of a case

We report a case of a 50-year-old male with ulcerative colitis who developed well-differentiated adenocarcinoma in the ileal J-pouch, which had been defunctioning for 18 years. The extension of the carcinoma in the pouch suggested that it had recently appeared in the pouch. Monitoring by endoscopic examination and biopsy or pouch excision seems to be an appropriate action if a pouch is out of the fecal stream.

Sequential Observation of Spontaneous Endometrial Adenocarcinoma Development in Donryu Rats

Sequential observation of spontaneous endometrial adenocarcinoma development revealed a clear, hormone-dependent, histogenetic pathway in Donryu rats. The first histological changes of the uterine endometrium appeared in both the lining epithelium and uterine gland of the endometrium at 6 mo of age, along with the beginning of persistent estrus. These changes included areas of tall columnar epithelium and gland formation in the lining epithelium as well as metaplastic change in the uterine gland. At 8 mo of age, endometrial hyperplasias were found, with subsequent increase in both incidence and degree. At 8–10 mo of age, hyperplasias were all within the limit of grade ++. After 12 mo of age, however, severe hyperplasias (grade + + +) began to increase markedly, and adenocarcinomas developed at 15 mo of age. The findings thus suggest that uterine endometrial adenocarcinomas arise from hyperplastic lesions, which should therefore be regarded as preneoplastic, as in the human case. Sequential analysis of plasma gonad steroids also ascertained a link between the appearance of these lesions and an increased estrogen: progesterone ratio, suggesting that estrogen may play an important role in development of both hyperplastic and neoplastic lesions. In Fischer-344 rats used for comparative assessment of strain differences, neither advanced histological changes nor hormonal changes were evident.

Subcutaneous treatment of p-tert-octylphenol exerts estrogenic activity on the female reproductive tract in normal cycling rats of two different strains

Effects of p-tert-octylphenol (OP), an endocrine disrupting chemical (EDC), on the female reproductive tract of normal cycling Fischer 344 (F344) and Donryu rats were investigated. OP was subcutaneously injected at concentrations of 12.5, 25, 50 or 100 mg/kg for 28 days. The most notable changes were the disappearance of normal cyclicity in the 50 mg/kg or more OP-treated groups of both the strains, and the appearance of persistent estrus (PE) evident on examination of vaginal smears in the 100 mg/kg groups of both the strains, the effects being time- and dose-dependent. In PE rats of both the strains, the uterine morphology deviated from the normal for each estrous stage of the cycling rats, and proliferation in the endometrium was slightly increased. The data for uterine weights, luminal epithelial cell-heights and/or numbers of epithelial cells in the endometrium demonstrated equivocal alteration. In both the strains, the serum 17beta-estradiol (E2) levels were decreased with 50 mg/kg of OP or more. Serum concentrations of the administrated chemical were dose and duration-dependently increased in all the treated groups of both the strains. The results demonstrate that subcutaneous administration of OP at doses of 50 mg/kg or more exerts time- and dose-dependent estrogenic activity on the reproductive tract of normal cycling female Donryu and F344 rats, indicating similar qualitative sensitivity to the effects in both the strains. Vaginal cytology may be the most sensitive endpoint for the detection of estrogenic activity of potential EDCs using adult cycling rats.

Lack of carcinogenicity of tartrazine (FD & C Yellow No. 5) in the F344 rat

The carcinogenicity of tartrazine (C. I. Food Yellow No. 4, FD & C Yellow No. 5), a food, drug and cosmetics colouring, was examined in F344 rats. Tartrazine was dissolved in distilled water at levels of 0, 1 or 2%, and groups of about 50 male and 50 female rats were given one of these solutions ad lib. as their drinking-water for up to 2 yr. No toxic lesions specifically caused by tartrazine were detected in any treated group of either sex. Many tumours developed in all groups including the control group, and the organ distribution of these tumours and their histological characteristics were similar to those of the spontaneous tumours that are known to occur in this strain of rats. Except for mesothelioma in males and endometrial stromal polyp in females, there were no significant increases in the incidences of any tumours over those in the corresponding control group. In males, mesotheliomas were found only in the group given 1% tartrazine and the incidence of this lesion was statistically significant (Fishers exact test) in comparison with the other two groups (P less than 0.02). The incidence of endometrial stromal polyp was also significantly higher among females given the 1% dose than in the controls (P less than 0.05). However, no positive trend was noted in the occurrence of these two tumours using an age-adjusted statistical analysis. Mesothelioma and endometrial stromal polyp are frequently observed spontaneous tumours in this strain of rats, and their incidences in our historical controls are 4.1 and 21.9%, respectively. However in the present study mesothelioma occurred in none of the male control rats and the incidence of endometrial stromal polyp was only 10.6% in the female control group. Moreover, there was no significant difference between the control and treated groups in hyperplastic or pre-neoplastic changes in the mesothelium or endometrium. From these findings, we concluded that the significant increases in the incidences of mesothelioma and endometrial stromal polyp that occurred in the groups given 1% tartrazine were not attributable to tartrazine administration. Thus, it is concluded that tartrazine was not carcinogenic in F344 rats when administered continuously at doses of up to 2% in the drinking-water for up to 2 yr.

Pathological characteristics of mucopolysaccharidosis VI in the rat

The histological and electron microscopical characteristics of the pathology of rats with arylsulphatase B-deficient mucopolysaccharidosis (mucopolysaccharidosis VI; MPS VI) were investigated. In affected animals, intracytoplasmic vacuoles were prominent in chondrocytes, the macrophage system, cardiac valve fibroblasts, cornea, connective tissues, vascular smooth muscle cells and uterine stromal cells. Tissues containing glucosaminoglycans stored in lysosomes were positive to Mowrys colloidal iron and alcian blue stains. By electron microscopy, the lysosomes were seen to be distended by electron lucent or fine fibrillary storage material, and lysosomal storage was also detected in the endothelial cells of the arteries and cornea. In the central and peripheral nervous system abnormalities were restricted to the connective tissue. Lesions in the affected rats resembled those described in human and feline mucopolysaccharidosis VI. These results indicate that MPS VI of the rat may be a useful animal model for human MPS VI (Maroteaux-Lamy syndrome).