Ultrastructure of cells from the liver-endocrine pancreas system in hibernating animals

In hibernated ground squirrel (Citellus erythrogenys) during various periods of hibernation morphological changes in organs participating in insurance of energoplastic homeostasis (liver, endocrine part of the pancreas) have been followed. At the beginning of hibernation certain signs of functional strain in the liver are observed-shortening and swelling of the endoplasmic reticulum (ER) canaliculi, decreases of glycogen resources and activation of the lysosomal apparatus, as well as certain features demonstrating its decreased activity--shortening cisterns in the Golgi complex, poorly manifested granular ER, tightening of mitochondrial matrix. At a deep hibernation, the signs of strain disappear, ultrastructure of hepatocytes corresponds to a new lower level of functioning in comparison to that in the active state and at the beginning of hibernation. During these periods the structure of endocrine cells in the pancreas demonstrates certain changes in the character of functioning of the cells. Ultrastructural transformations in the liver and endocrine part of the pancreas in the ground squirrel are mutually correlated.     

State of the epiphysis during winter hibernation]

It was found that the pineal gland of active ground squirrels (Citellus erythrogenys) differed from this gland in rats by a more pronounced differentiation into cortical and medullary zones, by larger pineocytes with polymorphous nuclei and numerous pseudokariosomes. Hybernation was accompanied by reduced concentration of serotonin in the pineal gland, by the disappearance of differences between the cortical and the medullary zones and by a number of morphological signs of functional depression. The appearing changes were similar to those occurring in denervation of the pineal gland.     

Parenchymal-stromal relations in the myocardium of heterothermal animals in conditions of seasonal biorhythms

The myocardium of 20 Citellus erythrogenys Br. was studied by quantitative morphology in different seasons. Morphometry and stereological methods were used to measure the cardiomyocyte diameter, volume and surface densities of the main tissue structures. The morphological criteria were derived for the seasonal features of the relations in muscular and connective tissues of the hibernating rodent myocardium. Myocardial atrophy (seasonal regression of the heart weight and the cardiomyocyte diameter) was noted during hibernation. It was accompanied by adaptive tissue reorganization. The parenchymal-stromal relations in the myocardium of both active and hibernating animals were found to be stable. In these conditions there occurred significant changes in the architectonics of the myocardium.     

Seasonal changes in the endocrine cells of the gastric epithelium of Rana temporaria

By means of the light and electron microscopy methods histotopography, quantity and ultramicroscopic organization of endocrine cells (EC) have been studied in gastric epithelium of the frog (Rana temporaria) during various periods of its life: hibernation, awakening, active life. At all the periods of life 8 types of EC are revealed. Depending on the season, certain changes both in their amount and ultrastructural organization are observed. Decreasing amount of EC is noted during hibernation and their increase in spring during awakening and in summer. Quantity of enterochromaffin cells is not subjected to seasonal fluctuations. This is explained by the fact that in hibernation these cells are also at their active functional state, since they produce serotonin necessary for maintaining deep hypothermia, specific for hibernating animals.     

Endocrine cells in the large intestine epithelium in various vertebrate species

Amount of argyrophile and argentoffin endocrine cells per 1 mm2 of the large intestine mucous membrane has been counted in the turtle, pigeon, white rat and frog at various periods of their life (winter hibernation, awakening, active life). As the animal's organization becomes more complicated, the amount of the endocrine cells in the epithelium increases both in the initial and in the terminal parts of the large intestine, their amount in the terminal (rectum) part being less than in the initial one. The greatest amount of the endocrine cells is found in simmer frogs at the expense of increasing content of the elements that do not belong to the argentaffin group. The latter do not significantly change during various seasons.     

Seasonal changes in the cardiomyocytes of heterothermal animals

In 20 Citellus erythrogenys the volumetric and surface densities of myofibrils, mitochondria, sarcoplasmic reticulum, T-system and lipid drops were estimated in cardiomyocytes by means of stereological methods before, during and after hibernation. The volumetric and surface-volumetric ratios for the main intracellular compartments were calculated. Certain morphological criteria were found for the seasonal peculiarities in the structural organization of cardiomyocytes in the hibernating rodents. During hibernation, myocardial atrophy (seasonal regression of the heart weight and decreasing diameter of cardiomyocytes) takes place. It is accompanied with an adaptive rearrangement of the cardiomyocyte ultrastructure. The latter facilitates to myocardial contractility at low body temperatures at the expense of activation of the cellular transport-energic interaction system.     

Seasonal dynamics of the gastric APUD cell count in a hibernator

Light microscopy was applied to study variation in the composition and number of apudocytes in the gastric epithelium of red-cheeked sousliks over time. During hibernation, the content of serotonin-producing EC cells was elevated. This is in agreement with serotonin involvement in the maintenance of the hibernation state. The number of non-EC cells taking part in digestion, which does not occur during hibernation, was lowered. Spring awakening was accompanied with an increase in the number of non-EC cells and the lack of significant changes in cells. In souslics, which were active in winter, the content of non-cells was high, since the animals continued feeding on, and the content of EC-cells was elevated similarly to that in hibernating animals, which could be explained by genetic dependence. The number of apudocytes in the gastric mucosa was discovered to correlate in the main with the features of their function at different seasons.     

Seasonal changes in phosphorylation of myosin regulatory light chains and C-protein in myocardium of hibernating ground squirrel Citellus undulatus

A comparative study concerning the extent of phosphorylation of myosin regulatory light chains and C-protein from the left ventricle of hibernating ground squirrel Citellus undulatus during the periods of hibernation and activity was carried out. During hibernation, regulatory light chains of ground squirrel were found to be completely dephosphorylated. In active animals, the share of phosphorylated light chains averages 40-45% of their total amount. The extent of phosphorylation of the cardiac C-protein during hibernation is about two times higher than that in the active state. Seasonal differences in phosphorylation of the two proteins of ground squirrel myocardium are discussed in the context of adaptation to hibernation.     

Morphofunctional analysis of the chief (peptic) glandulocytes in the stomach of hibernating rodents

Ultrastructure of the chief gastric gland cells has been studied in the ground squirrel (Cytellus erythrogenys Brandt) at various seasons of the year. When the ground squirrel is in the active state, the cells studied do not differ from those in other animals. During winter hibernation, their activity is inhibited. During first days of awakening, the activity of the chief cells increases; the protein-synthesizing apparatus, the Golgi complex become activated, secreating granules accumulate. At the same time, the greatest number of the destroing cellular elements and discharge of the residual bodies out of the activizing cells are observed. Restoration of the normal structure in the chief cells is completed by the end of the second week. No signs demonstrating transformation of additional cells into the chief ones are revealed. Non-differentiated cells in the ground squirrel fundal glands are widely distributed along the whole length of the glandular cervix and occur even in the upper parts of its body.     

Antioxidant defenses in the ground squirrel Citellus citellus. 2. The effect of hibernation

In spring and autumn, the ground squirrel Citellus citellus is awake and active but in winter it usually hibernates. Reawakening from hibernation involves intense metabolic activity in the interscapular brown adipose tissue (IBAT). The IBAT of hibernation animals showed significant increases in the activities of superoxide dismutase (both copper-zinc and manganese-containing enzymes), glutathione peroxidase, and in the amount of ascorbate present. Glutathione peroxidase also increased in the liver, as did ascorbate in the plasma. These changes were not merely a consequence of exposure to low environmental temperatures. It is proposed that antioxidant defenses are increased in the IBAT of ground squirrels at the onset of hibernation in order to protect the tissue from reactive oxygen species generated as a result of the intense metabolic activity sustained by this tissue during reawakening.     

Peptide TSKY Decelerates Outcome of the Ground Squirrel from Hibernation

Effect of peptide TSKY on the heart rate was studied in the ground squirrel Citellus undulatus at outcome from hibernation. At provocation of awakening in the first half of the hibernation bout (the 3rd–4th hibernation days), an intraperitoneal peptide injection at a dose of 1 mg/kg produced a deceleration of the HR increase and elongated the time of transfer of the animal to the normothermia state. This effect was not observed in the second half of the hibernation bout (the 5th–7th hibernation days). The study of the peptide cardiotropic activity on an isolated frog heart did not reveal any statistically significant TSKY activity.     

The evaluation of the sizes of erythrocytes of the hibernating ground squirrel Citellus undulatus Pallas

A topographical image of individual erythrocytes of the ground squirrel Citellus undulatus Pallas, in unfixed unstained smears was first obtained by scanning probe microscopy for two states of the animal: hibernation and the active state. The scannig of single discocytes, i.e. erythrocytes having a typical discal form, was fulfilled. For the active male, the diameter of the discocyte was found to be approximately 6500 nm. For the hibernating female, the diameter is approximately 6000 nm. According to the data of light microscopy, the discocyte diameters are: 6610 +/- 100 nm for the active state of animal and 6430 +/- 160 nm for the hibernating state.     

Enterochromaffin and enterochromaffin-like cells of the stomach during aging in rats

In the gastric mucosal membrane of rats--mature, first old and late old age (9.5-20, 26.5-29 and 32.5-37 months) ultrastructure of serotonin-producing EC-cells and histamine-producing ECL-cells has been studied. By means of stereomorphometric methods volumes of nuclei, nucleoli, cytoplasm and its organelles, nuclear cytoplasmic index, amount of endocrine granules have been determined. Increasing volumes of EC- and ECL-cells and their organelles in the old age demonstrate certain development of adaptive changes, directed to maintenance of monoamines secretion at a necessary level. This is reached at the expense of increasing energetic and plastic expenditure. In the late old age intensification of adaptation, further growth of organelles volumes, responsible for energetic and plastic metabolism are kept for EC-cells. Increasing content of active granules in cytoplasm demonstrates an enhanced production of serotonin by every separate cells. In the same age in ECL-cells the adaptive processes decrease. Volumes of the cells and their all organelles sharply drops, the number of the active granules decreases; this should result in decreasing histamine production in the gastric mucous membrane.     

Further studies on opioids and hibernation: delta opioid receptor ligand selectively induced hibernation in summer-active ground squirrels

To examine the possible involvement of multiple opioid receptors in animal hibernation, we infused opioids selective for mu, kappa, and delta opioid receptors into summer-active ground squirrels (Citellus tridecemlineatus). The effects of those opioid treatments on the hibernation induced by HIT (Hibernation Induction Trigger) were also examined. Mu opioids morphine (1.50 mg/kg/day) and morphiceptin (0.82 mg/kg/day) and kappa opioid peptide dynorphin A (0.82 mg/kg/day) did not induce hibernation. On the contrary, morphine, morphiceptin and dynorphin A antagonized HIT-induced hibernation in summer-active ground squirrels. Infusion of delta opioid DADLE (D-Ala2-D-Leu5 enkephalin; 1.50 mg/kg/day), however, induced summer hibernation in a manner comparable to that induced by HIT. It is concluded therefore that delta opioid receptor and its ligand may be intimately involved in animal hibernation. In view of the fact that HIT was obtained from winter hibernating animals and might therefore be responsible for natural hibernation, our results also suggest that naturally occurring mu and kappa opioids may play an important role in the arousal state of hibernation.     

The hypothalamo-hypophysial system in the ground squirrels, Citellus erythrogenys Brandt and Citellus undulatus Pallas. I. Microanatomy and cytomorphology of the gomori-positive neurosecretory system with special reference to its state during hibernation.

Microanatomy of the peptidergic hypothalamo-hypophysial neurosecretory system in the ground squirrels, C. erythrogenys and C. undulatus, is similar to that of other rodents, but differs in details. Bundles of neurosecretory fibres with single neurosecretory cells along their course connect the hypothalamic neurosecretory centers into the one system. The dorso-caudal portions of the NPV reach the level of the columna fornix. The NPO is a special indepedent formation and appears to have some specific functions in the HHNS. The infundibular recess penetrates deeply ihto the hypophysial stem, but does not reach the posterior pituitary. The glandular epithelium of the tuberal portions of the adenohypophysis surrounds the median eminence and a considerable part of the lateral surface of the tuber cinereum. Different functional types of neurosecretory cells reflect certain phases of their secretory cycle. Correlations between their size, structure and the content of NSM are found. A low activity of the HHNS is observed in both C. erythrogenys and C. undulatus during December-January. A higher activity of the HHNS in torpid C. undulatus than in C. erythrogenys is established. This correlates with a difference in deepness of torpor in C. erythrogenys and C. undulatus. An important role of the HHNS in hibernation, i.e. in realization of adaption of species in ontogenesis is suggested.     

Pancreatic hormones in the nonhibernating and hibernating golden mantled ground squirrel

The effects of hibernation on pancreatic insulin, glucagon, somatostatin, and pancreatic polypeptide was investigated in the golden mantled ground squirrel (Citellus lateralis). During hibernation, pancreatic organ weight decreased to 57% of the nonhibernating weight. The content of all four pancreatic hormones during hibernation was significantly reduced. The concentrations of insulin, pancreatic polypeptide and somatostatin, but not of glucagon, were significantly reduced during hibernation. The maintenance of the pancreatic glucagon concentration during hibernation may be related to its role in counter-regulation and carbohydrate homeostasis during fasting.     

季节、环境温度与黄鼠冬眠的关系

观察了达乌尔黄鼠在实验室内冬眠的一般情况。常温黄鼠的体温有着规律性的年周期,与环境温度的年周期变动不完全呈依从关系。出眠初期(4月下旬),动物体温高而稳定。4月至6月常温黄鼠的平均体温(皮温)为35.6℃,波动菹围32—37.5℃。随着体重达到顶峰,体温逐渐降低。8月份部分黄鼠出现低于32℃的低常体温,表明部分黄鼠自8月盛夏开始冬眠。但就整个种群而言,北京地区实验室内黄鼠冬眠季自9月下半月开始。3月底止,共6.5个月。秋季室温下降,动物入眠趋势增长。浅低体温(31.9—15℃)的比数逐渐升高。9月至12月,低体温(低于31.9℃)的百分比从47％增至84.8％,反映了动物从浅冬眠向深冬眠过渡。1月至2月份,低体温占85％以上,深低体温(低于15℃)占绝对优势。标志着动物种群的深眠月份。秋季动物从常温期向冬眠期转化的界线是不清的,而春季从冬眠期向常温相转化的界限却比较明显。     

An ultrastructural stereological analysis of the cardiomyocytes in a hibernating rodent (the red-cheeked suslik Citellus erythrogenys, Brandt)

The quantitative morphological study of cardiomyocytes in heterothermic animals (red-cheeked susliks Citellus erythrogenys Br.) was quantitatively morphologically studied. The features of spatial ultrastructural cardiomyocyte reorganization in hibernating susliks were shown to manifest in pronounced increase of volume and surface densities of sarcoplasmic reticulum and relative volume of lipid droplets.     

Phosphorylation of myosin light chains and C-protein in the myocardium of hibernating ground squirrel Citellus undulatus

A comparative study was made of the extent of phosphorylation of myosin regulatory light chains and C-protein from the left ventricle of the hibernant ground squirrel Citellus undulatus during the periods of hibernation and activity. During hibernation, the light chains were found to be completely dephosphorylated. In active animals, the share of phosphorylated light chains averaged 40–45%. The extent of cardiac C-protein phosphorylation in hibernation was about twice higher than in the active state. Seasonal differences in phosphorylation of the two proteins of ground squirrel myocardium are discussed in the context of adaptation to hibernation.