Morphologic changes in the pineal gland of rats exposed to continuous darkness

We examined the pineal structure of rats exposed to constant darkness (DD) at light microscopic level. Two groups of rats were exposed to 12:12 light/dark cycle (LD) or DD from their prenatal ontogenesis and then for 3 months after birth. The gland structure of DD rats was observed to have an active appearance. Some of the observed pinealocytes with light nuclei from DD rats were determined to contain double nucleoli. Nuclear area and perimeter of both dark and light types were greater in rats kept in DD than in LD. Rats exposed to DD had more cells with light nuclei and lesser cells with dark ones than rats kept in LD. No significant differences in nuclear characteristics of intermediate type were found between rats kept in LD and those kept in DD. The activity of mammalian pineal can be altered by light conditions to which the animal is exposed.     

Elimination of cholesterol and bile acids in bile collected by total fistula in the rat. II. Influence of the pituitary gland]

This paper shows the role of the hypophysis gland on the bile elimination of essential tauroconjugated acids and cholesterol. After transauricular hypophysectomies were performed on rats which were kept under total fistula treatment, we employed an original technique of bile recuperation in rats without contention and observed, over a period of 10 or more days, the choleresis and the composition of eliminated bile.     

Effect of iodide-deficiency on rat mammary gland.

When rats are kept iodide-deficient, atrophy and necrosis takes place in the mammary gland and areas of dysplasia and atypia are seen. Administration of estradiol to iodide-deficient rats stimulates cell division in the gland and leads to the formation of alveoli. Continued stimulation by estradiol produces changes in the newly-formed alveolar cells. Their nucleoli are altered and show a separation of components. Ribosomes and lipid droplets increase and the cells synthesize large vacuoles containing protein. The secretion of great quantities of this material into areas of the tissue where regressive changes have occurred undoubtedly contributes to the formation of cysts within the gland. The present findings indicate that iodide-deficiency alters the structure and function of mammary gland alveolar cells and makes them highly sensitive to stimulation by estradiol.     

Change in the metabolism of 7,12-dimethylbenz(a)anthracene under the influence of vitamin A]

The in vitro metabolism of 7,12-dimethylbenz(a)anthracene in its incubation with the liver microsomes, the liver and mammary gland homogenates of rats, kept on vitamin A-enriched diet, was studied. Vitamin A inhibited the formation of lipophilic metabolites and increased the generation of water-soluble metabolites. The amount of lipophilic metabolites extracted from the microsomes and the liver and mammary gland homogenates were decreased by a factor of 2.2 and 5, respectively. The amount of unmetabolized DMBA in the liver microsomes was the same in control and experimental animals.     

The light-dark regimen and cancer development

The role of the modulation of the pineal gland function in development of cancer is discussed in the review. An inhibition of the pineal function with pinealectomy or with the exposure to the constant light regimen stimulates mammary carcinogenesis, whereas the light deprivation inhibits the carcinogenesis. Epidemiological observations on increased risk of breast cancer in night shift workers, flight attendants, radio and telegraph operators and on decreased risk in blind women are in accordance with the results of experiments in rodents. Treatment with pineal indole hormone melatonin inhibits carcinogenesis in pinealectomized rats or animals kept at the standard light/dark regimen (LD) or at the constant illumination (LL) regimen.     

Sex differences in secretion pattern of neonatal rat Harderian gland under various environmental lighting conditions

1. Secretion pattern of Harderian gland of neonatal rats maintained under (a) diurnal lighting conditions; (b) continuous light or (c) continuous darkness was studied at light microscopy level. 2. All animals were placed in especially designed cages at 13:00 hr on day 1 and studied on day 7 at 13:00 and 23:00 hr, respectively. 3. Acini with intraluminal secretion were counted in glands from each animal and the results were separately grouped for male and female animals. 4. A diurnal rhythm in secretion pattern of rat Harderian gland in neonatal period was demonstrated. 5. A statistically significant difference was observed in the gland secretion pattern between males and females at both, 13:00 and 23:00 hr when the animals were kept under diurnal lighting conditions. 6. Under continuous light or continuous darkness, the diurnal rhythm in secretion pattern was lost and no significant differences were seen when data from males were compared to those from female neonates. 7. Results are discussed in terms of the possible function of Harderian gland as element of an extraretinal photoreceptor system involved in the regulation of pineal function in neonatal rats.     

Effect of weightlessness and artificial gravitation on thyroid gland morphology]

The investigation of the thyroid gland was carried out in Wistar rats, SPF colony 4.5--13 h and 25 days after a 18.5 days flight on board the space biosatellite "Cosmos-936". In animals subjected to weightlessness, moderate symptoms of the thyroid hypofunction were observed, statistically significant decrease in number and volume of the nuclei in calcitonin-secreting cells (C-cells) was especially pronounced during 4.5--9 h after landing. Similar but less pronounced changes were observed in C-cells of the rats subjected to artificial conditions of space flight, besides weightlessness. The similarity of the changes in the animals of both groups made it possible to connect the increasing amount of C-cells and the morphological symptoms of their functional inhibition with the effect of weightlessness and hypokinesia. During the space flight, the animals were kept under the conditions of artificial gravitation on board the biosatellite and therefore morphological peculiarities specific for the earth conditions were preserved in C-cells and the thyroid gland. Thus, it was concluded that artificial gravitation prevented the development of the thyroid changes which appeared under the influence of weightlessness.     

Effect of vitamin E on follicular cell proliferation and expression of apoptosis-associated factors in rats with 6-N-propyl-2-thiouracil-induced goitrogenesis

We have investigated immunohistochemically the effect of dl-alpha-tocopherol (vitamin E) on thyroid gland with 6-n-propyl-2-thiouracil (PTU)-induced hypothyroidism in rats. The animals were divided into four groups. Rats in group I were designated as control, rats in group II were treated with injections of PTU (10 mg/kg) for 15 days, rats in group III were treated with injections of PTU+vitamin E (10 mg/100 g) for 15 days. Rats in group IV were treated with injections PTU for 15 days and kept for 15 next days after cessation of PTU treatment. At the end of experiment, the animals were killed by decapitation, blood samples were obtained, thyroid tissues were collected and processed for quantitative evaluation of immunohistochemical PCNA (marker of cell proliferation), Bax (pro-apoptotic marker) and Bcl-2 (anti-apoptotic marker) staining. There was an increase in the number of PCNA-immunopositive cells in follicular epithelial cells of group II rats compared with other groups (p<0.05). After vitamin E treatment, the number of PCNA-immunopositive cells decreased (p<0.05) while the number of Bax-immunopositive cells increased (p<0.05). The number of Bcl-2-positive follicular epithelial cells of group IV rats was higher than in those of other groups (p<0.05). The results of this study indicate that hypothyroidism induces cell proliferation in the thyroid gland and vitamin E may promote involution of the gland.     

大豆黄酮促进妊娠大鼠乳腺发育和泌乳的实验研究

研究大豆黄酮（Ｄａ）对妊娠大鼠乳腺发育和泌乳的作用及其与神经内分泌的关系，结果表明；妊娠期口服大豆黄酮，能显著提高泌乳前期大鼠乳腺重量、ＤＮＡ、ＲＮＡ含量和ＲＮＡ／ＤＮＡ值，同时极显著地提高大鼠的泌乳量、血清ＧＨ和ＰＲＬ含量及乳腺胞浆雌二醇受体的数目和亲和力；相反．妊娠后期口服多巴胺激动剂溴隐亭，能显著降低泌乳前期乳腺重量、ＤＮＡ和ＲＮＡ含量及泌乳量，并显著降低血清ＧＨ和ＰＲＬ含量及乳腺雌二醇受体的数目和亲和力；但连续两周口服Ｄａ后再口服澳隐亭一周，溴隐亭的上述作用被阻断。结果提示：Ｄａ促进大鼠乳腺发育和泌乳的作用与其抑制或站抗多巴胺对垂体ＰＲＬ、ＧＨ分泌的抑制作用有关。     

Long-term effects of cage-cleaning frequency and bedding type on laboratory rat health, welfare, and handleability: a cross-laboratory study

Cage-cleaning is necessary for a hygienic environment, but since rats communicate using scent, they might suffer if their cages are cleaned too frequently. Male rats (Sprague-Dawley and Wistar) were kept for five months across four animal units. Their cages were cleaned twice-weekly, weekly, or every two weeks, and contained either aspen woodchips or absorbent paper bedding. Aggression, injuries and general health, weight gain, chromodacryorrhoea (a stress-related Harderian gland secretion), handleability, and lung pathology were monitored, as was in-cage ammonia. Cleaning frequency had no clear impact on rat welfare, although frequent cleaning decreased ammonia concentrations and handleability, and non-aggressive skirmishing was highest in weekly cleaned rats. Surprisingly, bedding type did not affect ammonia, but all ammonia readings were unexpectedly low. However, rats kept on aspen had greater sneezing rates and lung pathology than those on paper bedding, but also had higher body weights. The results raise concerns about aspen bedding, which is relatively inert compared with other wood beddings, but nevertheless more harmful than paper. Animal unit significantly affected eight of the 11 variables tested, having interactive effects on five of them. The study also demonstrates the interactive effects of different animal units, casting doubt on the feasibility of standardization. We explored multiple variables of interest, so all findings require confirmation through further work. Nevertheless, cage-cleaning rates seem to affect socially housed male rats little, while bedding type has important effects on rat health.     

Studies on submaxillary gland immunoreactive glucagon.

Biologically active immunoreactive glucagon is present in submaxillary gland of rat, mouse, guinea pig and human and can be extracted by saline adjusted to pH 2.8 with HCl. Chromatography on Sephadex G-150 indicates its molecular weight to be 29,000. It has similar immunologic characteristics as pancreatic glucagon. It is biologically active and elevates plasma glucose and insulin when injected intraperitoneally into rats. Compared to pancreatic glucagon, the hyperglycemic effect persists much longer. It competes with pancreatic glucagon for binding to specific glucagon receptors of rat liver plasma membranes. It is stable to pH changes, however, urea dissociates it into several smaller molecular weight fragments including that of 3500. It appears to be an aggregate of smaller glucagon molecules and is not responsible for immunoreactive glucagon in totally eviscerated rats. $\text{In vitro}$, the submaxillary gland does not release immunoreactive glucagon in response to arginine or glucose.     

Direct angiotensin II formation by rat submandibular gland kallikrein

Submandibular gland kallikrein [EC 3.4.21.8] of male Sprague-Dawley rats was purified by chromatography on soybean trypsin inhibitor (SBTI)-CH-Sepharose 4B, DEAE-Sephadex A-50, aprotinin-CH-Sepharose 4B and Sephadex G-100 columns and preparative isoelectrofocusing. The molecular weight of the kallikrein was estimated to be 30,000 by Sephadex G-100 gel filtration and 29,000 by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Isoelectric points ranged from pH 3.55 to 4.30. The kinin formed at pH 8 by this kallikrein from bovine low molecular weight (LMW) kininogen showed the same behavior as lysyl-bradykinin on HPLC in a solution of ammonium biphosphate containing acetonitrile. At physiological pH, this kallikrein also generated angiotensin II, a potent vasopressor, from human plasma protein. Rat submandibular gland kallikrein differs from tonin in the isoelectric point, the optimal pH for angiotensin II formation and the type of kinin formed. The tissue kallikrein might play a role in the regulation of local blood flow in view of its ability to form both vasoconstrictive and vasodilatory peptides.     

Effects of Iodine-131 in sheep depending on the content of stable iodine in the diet]

The biological effect of 131I was studied in sheep kept on a diet deficient in stable iodine. An increased capture and accretion of iodine in the thyroid gland and in the whole body were observed. The disturbances in the structure and function of the thyroid gland, liver and haemopoietic organs were more pronounced in the animals kept on the iodine deficient diet.     

Fine structure and function of the digestive cell of Sepia officinalis (Mollusca: Cephalopoda)

The post–embryonic development of the digestive gland has been studied by light and electron microscopy on animals reared in the laboratory. The gland reaches its "adult" structure around the end of the first month after hatching.
The infrastructural changes of the digestive gland have been followed during digestion at various ages (5,10,20,30 days and "adult"); ferritin was used for a tracer to study absorption.
These experiments have produced evidence of endocytosis intake of large proteins and further intracellular digestion inside the digestive cell. The residues of digestion are excreted in a "brown body" while the metabolites are used in new syntheses for the cell itself. These results demonstrate that "ancestral" digestive processes have been kept in at least one cephalopod.     

Central monoamine oxidase and phenolsulfotransferase activities in spontaneously hypertensive rats

The activities of monoamine oxidase and phenolsulfotransferase in the hypothalamus and anterior pituitary gland of spontaneously hypertensive rats and the normotensive control (Wistar Kyoto rat) rats were investigated. The monoamine oxidase activity (determined using dopamine as substrate) in both these tissues was not significantly different between the normo- and hypertensive animals. Hypothalamic phenolsulfotransferase does not sulfate-conjugate dopamine at pH of 6.5 and pituitary phenolsulfotransferase does not sulfate-conjugate dopamine or 3,4-dihydroxyphenylacetic acid at the same pH. Hypothalamic phenolsulfotransferase activity determined using 3,4-dihydroxyphenylacetic acid as substrate was significantly higher in the spontaneously hypertensive than the Wistar Kyoto rats, while pituitary enzyme (determined using phenol as substrate) was the same in both strains of animals. We proposed that in the spontaneously hypertensive rats the higher level of hypothalamic phenolsulfotransferase could (by removing 3,4-dihydroxyphenylacetic acid as sulfated acid) increase the deamination of dopamine by monoamine oxidase. This could in turn result in the presence of high amount of sulfated 3,4-dihydroxyphenylacetic acid in the anterior pituitary gland reported in our earlier study, and be partly responsible for the reduced central dopaminergic activity found in the hypertensive rats.     

Characteristics of the recovery processes in the resected thyroid of adult rats after sympathectomy

The investigation was carried out on 65 adult white male rats (12-month-old) after normal development or chemical sympathectomy by guanethidine and subtotal thyroidectomy. By means of electronic microscopic, autoradiographic and radioimmunological methods the increased functional activity in thyroid gland of chemically sympathectomized rats was found. This is mainly due to thyrotropin hyperproduction and intensification of proliferative potential of the organ. Partial resection of sympathectomized rats thyroid gland leads to exhaustion of adenohypophysis compensatory potential manifested in disjunction of integrative connections in hypothalamus-hypophysis-thyroid gland system and delay of thyroid status normalization.     

Actions of oestrogens and antioestrogens on rat mammary gland development: relevance to breast cancer prevention

The proliferative actions of a series of antioestrogens on the development of the second thoracic mammary gland of ovariectomized immature Sprague-Dawley rats have been investigated. Evidence is presented that shows trans-tamoxifen, LY 117018 and LY 139481, like oestradiol-17 beta and cis-tamoxifen, promote full mammary gland ductal development and induce a high rate of cell proliferation in the undifferentiated epithelial cells of the terminal end buds, the main growth region for ductal growth. Conversely, ICI 164,384, a new antioestrogen, is without effect on ductal elongation. In vivo exposure of trans-tamoxifen and LY 117018 treated glands in medically castrated animals to the carcinogen DMBA, results in a high rate of mammary tumour development. Indeed, the actions of these so-called antioestrogens are equivalent to those observed in oestradiol-treated rats.     

Androgenic activity in 15-day-old male rats: role of the maternal pineal gland.

Female Sprague-Dawley rats, exposed to a long (18L:6D) or a short (6L:18D) photoperiod from 21 days of age, were mated when they reached 55 days of age. On Day 2 of gestation, dams were pinealectomized or sham-operated. Pre- and postnatal photoperiods were identical, and offspring were killed at 15 days of age. Maternal pinealectomy had no effect when rats were kept on 18L:6D. Rats born to sham-operated mothers and kept on 6L:18D had higher testicular testosterone and androstenedione content than offspring raised on the long photoperiod. This stimulatory effect of the short photoperiod was blocked by maternal pinealectomy and was not dependent on the offspring's own pineal since it was observed in both sham-operated and neonatally (on Day 5 after birth) pinealectomized rats. When sham-operated mothers housed on 18L:6D were treated daily during pregnancy and lactation by s.c. melatonin injection, there was an increase in the testicular testosterone content of offspring. It was concluded that when rats are maintained on a 6L:18D cycle the maternal pineal gland enhances the testicular testosterone and androstenedione content in 15-day-old offspring. This effect is probably mediated by maternally derived melatonin. At 15 days of age, the pineal of the offspring had no influence on testicular function.     

Effect of neonatal sympathectomy on the postnatal differentiation of the submandibular gland of the rat

Summary Right superior cervical sympathectomy was performed in one-day old rats. This operation had a small but definite effect on the postnatal development of the submandibular gland. The gland on the sympathectomized side weighed less and contained smaller amounts of DNA, RNA and protein than the contralateral intact gland. The postnatal development of acinar cells and granular convoluted ductal cells was retarded in the sympathectomized gland. The acinar cells which differentiated after the ganglionectomy were smaller than those in the contralateral intact gland and were filled with secretory granules but devoid of basal basophilia. The rate of cellular proliferation in the sympathectomized gland was, however, similar to that in the intact gland at various ages studied.Supported by Public Health Service Research Grant CA 17038 from the National Cancer Institute to Dr. T. Barka. The authors are indebted to Miss H. van der Noen and Mr. I. Borcsanyi for their assistance and to Dr. T. Barka for his suggestions regarding this paper     

Pineal synaptic ribbons in blinded rats

Summary To study the effects of bilateral ophthalmectomy on the circadian rhythm of the pineal gland, the number of the synaptic ribbons and ribbon fields in the pineal gland of female rats were determined by electron microscopy. In a preliminary experiment, the pineal gland was taken from rats at 15 days, and 1, 2, 3, 4, and 6 months after bilateral ophthalmectomy. It was found that the numbers of both synaptic ribbons and ribbon fields decreased markedly and reached a minimum at 1 month postoperatively, but recovered completely after 4 months. In the main experiment, the number of these intracellular elements was counted in both control and enucleated animals killed at 4 h intervals between 02:00 h and 22:00 h at either 1 month or 6 months after operation. All animals were kept under a lighting regimen of 12 h of illumination (06:00 to 18:00) and 12h of darkness (18:00 to 06:00). After 1 month, a remarkable decrease in the number of both the synaptic ribbons and ribbon fields was again noted but the circadian rhythm still remained. Complete quantitative and qualitative recovery in the circadian rhythm was obtained 6 months later.Supported by a grant-in-aid from the Japanese Ministry of Education, Science and Culture The authors wish to thank Mrs. Kazuko Moriwaki for her technical assistance