Quantitative histoenzymologic characteristics of the submaxillary salivary glands of white rats during an ovarian cycle]

Ovarian cycle in albino rats was applied to ascertain the problem of the relationship between the salivary and endocrine glands, and also of the extent of participation of individual components of the salivary glands with different functional orientation in the endocrine regulation of individual components of the salivary glands. The content of protein, mucopolysaccharides, DNA, and RNA, the activity of NAD- and NADP-diaphorase, alkaline phosphatase, malate and isocitrate dehydrogenase, alpha-leucine-aminopeptidase was studied. Cytospectrophotometric analysis showed that synchronous changes in the activity of the enzymes under study occurred in all the portions of the salivary glands, depending on the ovarian cycle phases. Of the four successive phases of the cycle the greatest activity of the enzymes and of the protein and mucopolysaccharide content was noted during the proestrus and metaestrus. Different metabolic processes were observed in the salivary ducts in comparison with other parts of the gland; this was apparently connected with peculiarities of the secretion and hormone production.     

Ultrastructural changes of sebaceous glands in castrated and testosterone-treated male rats

Summary The effect of testosterone on the sebaceous gland was studied in the male rat. Biopsies of dorsal skin from intact rats, from rats four weeks after castration, and from castrated rats treated with testosterone propionate for three weeks at a dose of 250 g/kg/day (s.c.) were examined by electron microscopy. In the treated animals intermediate sacrifices were performed on days 4,7,14,21. Stereology was used for a morphometric analysis of the smooth endoplasmic reticulum (SER). The presence of a vesicular endoplasmic reticulum throughout the cytoplasm of differentiating cells was observed in the sebaceous glands of intact rats. Following castration there was a shrinkage of these cells and a striking decrease in the volume density of the endoplasmic reticulum vesicles. The administration of testosterone to gonadectomized rats resulted in an increase in vesicle content above the normal level from the first week as revealed by stereological analysis. This study confirms the trophic effect of the androgen on the sebaceous gland at a subcellular level. Furthermore, it is shown that stereology is a useful method for detecting early hormone-induced changes and could be valuable for studying the effects of anti-androgens on this gland.     

Effects of estrus cycle on thermoregulatory responses during exercise in rats

In female rats, rectal temperature (Tre), tail vasomotor response, oxygen uptake (VO2), and carbon dioxide production (VCO2) were measured in proestrus and estrus stages during treadmill running at two different speeds at an ambient temperature (Ta) of 24 degrees C. Experiments were performed at 2.00-6.00 a.m., when the difference in Tre was greatest between the two stages; Tre at rest in the estrus stage was 0.54 degrees C higher than in the proestrus stage. In a mild warm environment, threshold Tre for a rise in tail skin temperature (Ttail) was also higher in the estrus stage than in the proestrus stage. In contrast, no difference was seen in the threshold Tre and steady state Tre at the end of exercise between proestrus and estrus stages. These values were higher at the higher work intensity. VO2 was also similar between the two stages, except in the second 5 min after the beginning of exercise, when VO2 was greater and Tre rose more steeply in the proestrus stage. These data indicate that deep body temperature during exercise is regulated at a certain level depending on the work intensity and is not influenced by the estrus cycle.     

Orienting behavior of female white rats in estrus cycle and its dependence on handling

Characteristics of orienting behavior of intact and handled female white rats were studied in the "open field" test. Experimental group of animals were formed according to stages of estrous cycle: (1) diestrus, (2) proestrus, (3) estrus, and (4) metestrus. The stage of the cycle was determined by vaginal smears. Over the period of 10 days rats were handled daily for 5 minutes. No behavioral changes over the course of estrous cycle were found in intact rats. Handling revealed some behavioral features related to the levels of steroid hormones in reproductive cycle. Most prominent changes in orienting behavior were observed at transition from estrous to metestrous. At the stage of estrous motor activity was maximal and grooming was minimal. The maximal contrast in the structure of orienting behavior was observed between estrous and diestrous stages.     

Ultrastructural mechanisms of myocardial atrophy in white rats during starvation

By means of complex morphological, morphometrical and stereological analyses the myocardium of Wistar rats was studied in full alimentary starvation during 6 days. The availability of two mechanisms of myocardial atrophy in adaptation to full starvation was revealed, these are the diminution of dimensions of parenchymatous elements and the decrease of muscle cell number. By ultrastructural investigation of cardiomyocytes the signs of structural protein synthesis decline were revealed, this is a simple myocardial atrophy. In quantitative analysis of cardiomyocytes the decrease of their number without changes in proportions of cells with different nuclei number was observed, that indicated systematic character of muscle cell elimination out of myocardium by apoptosis mechanism involvement, this is a numerical myocardial atrophy. Stereological analysis of myocardial atrophy development in conditions of full starvation determined the main regulated indices, these are the absolute total myofibril mass and mitochondrial mass and surface area.     

Ultrastructural changes in the oviduct of the laying hen during the laying cycle

The ultrastructural changes occurring in the fully functional oviduct of Isa Brown laying hens were studied during various stages of the laying cycle. Hens were killed at different positions of the egg in the oviduct. The oviduct was lined by ciliated and non-ciliated cells (also referred to as granular cells). The granular cells in the infundibulum contributed to secretion during egg formation, whereas ciliated cells showed little evidence of secretion. Ultrastructural changes were recorded in the granular and glandular cells of the distal infundibulum. In the magnum, the surface ultrastructure revealed glandular openings associated with the ciliated and granular cells. Cyclic changes were recorded in the glandular cells of the magnum. With respect to the three observed types of glands, the structure of gland type A and C cells varied at different egg positions in the oviduct, whereas type B cells represented a different type of gland cell containing amorphous secretory granules. The surface epithelium of the isthmus was also lined by mitochondrial cells. Two types of glandular cell (types 1 and 2) were recorded in the isthmus during the laying cycle. Intracisternal granules were found in type 2 cells of the isthmus. A predominance of glycogen particles occurred in the tubular shell gland. The granular cells in the shell gland contain many vacuoles. During egg formation, these vacuoles regressed following the formation of extensive rough endoplasmic reticulum; the reverse also occurred. The disintegrated material found in the vacuoles may have been derived from the disintegrating granules. The Physiology Teaching Unit, University of New England, provided financial support to K. Chousalkar for this study.     

Ultrastructural changes in the oviductal epithelium of Merino ewes during the estrous cycle

Isthmic and ampullary oviductal epithelia sampled from Merino ewes at days -1, 1, 3, and 10 of the estrous cycle (estrus = day 0) were studied by scanning and transmission electron microscopy after fixation by vascular perfusion. Secretory cells, ciliated cells, and lymphocytelike basal cells were observed in both isthmic and ampullary epithelium at all stages of the estrous cycle studied and their ultrastructural features were analyzed. Synthesis of lamellated secretory granules occurred in the ampullary secretory cells during the follicular and early luteal phases, and their contents were released by exocytosis into the oviductal lumen during the luteal phase. Granule release was associated with nucleated apical protrusion of these cells into the oviductal lumen. No such secretory activity was displayed by isthmic secretory cells even though a few cells contained nonlamellated granules. Apocrine release of apical vesicles and accompanying cytoplasmic material from apical protrusions of ciliated cells occurred in the isthmus around estrus but not in the ampulla. This unexpected feature has not previously been reported in any other mammal. Dendritic basal cells were distinguished in the lower part of the epithelium by their heterochromatic nuclei, electron-lucent cytoplasm, and lack of attachment zones. No migration of basal cells was observed, and their ultrastructural features were similar in the ampulla and isthmus and at all stages of the estrous cycle examined. The function of these lymphocytelike cells in the epithelium is uncertain, but the presence of phagocytic bodies and lysosomes in 20% of them may indicate a phagocytic role.     

Ultrastructural changes in guinea pig endometrial cells during the estrous cycle.

In the guinea pig, the estrous cycle is characterized by a constant measurable level of plasma progesterone with two peaks: the first one associated with the peak of plasma estradiol-17 beta occurring at proestrus and the second, during diestrus, more pronounced at the time at which the level of estradiol-17 beta is undetectable. The progesterone receptor content is the highest on day 1 and the lowest on day 10 of the estrous cycle, which lasts 16.3 +/- 1.5 days (n = 37; mean +/- SD). There is a positive correlation between the plasma level of estradiol-17 beta and the progesterone receptors detected immunocytochemically in both endometrial epithelial and stromal cells. The general morphology of the endometrium during proestrus and estrus is consistent with an estrogenic stimulation, i.e., a smooth and regular surface of the endometrium and the presence of numerous microvilli on the cell surface. However, a moderate secretory activity also occurs in proestrus and estrus. During postestrus, the glandular cells display an increase in characteristic secretory features which parallels the rise of progesterone in the plasma.     

Enzymic changes in rabbit and rat mammary gland during the lactation cycle

1. Activities of glucose 6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate dehydrogenase (EC 1.1.1.44), isocitrate dehydrogenase (EC 1.1.1.42), malate dehydrogenase (EC 1.1.1.37), malate dehydrogenase (decarboxylating) (EC 1.1.1.40), and pyruvate carboxylase (EC 6.4.1.1) were determined in subcellular fractions of mammary gland from rabbits during pregnancy, at different stages of lactation and during weaning. The results were compared with those obtained in similar experiments with rat mammary gland. 2. Three bases of expression of the activity of enzymes in the particle-free supernatant fraction of mammary gland were compared. During lactation, activity expressed per mg. of particle-free supernatant protein (uncorrected for milk protein) correlated well with that expressed per mug. of DNA phosphorus. The disadvantages of expressing activities per g. wet wt. are discussed. 3. The major differences between the two tissues were: (a) neither malate dehydrogenase (decarboxylating) nor a soluble form of pyruvate carboxylase could be detected in rabbit mammary gland at any stage of the lactation cycle; (b) isocitrate dehydrogenase increased in activity during lactation in rabbit mammary gland, but not in that of the rat. 4. Pyruvate carboxylase in the mitochondrial fraction of rabbit mammary gland, and in both the mitochondrial and the soluble fractions of rat mammary gland, did not change in activity during lactation. 5. For each tissue, the NADP-dependent dehydrogenases studied had a high activity at all stages of the lactation cycle compared with the rate of fatty acid synthesis at mid-lactation. The significance of these results is discussed with respect to the supply of NADPH via NADH.     

Ultrastructural changes in cells of the white pulp of the spleen during experimental salmonella infection

The electron microscopic investigation of the immuno-competent cells in various zones of the spleen has revealed certain subcellular bases of the organ's reaction to the experimental salmonellosis. Alterations at early stage of the experiment (3-24 h after infection with Salmonella) are predominantly of destructive character: pyknosis of nuclei, dilated cisterns of the karyotheca, lysis of crests in mitochondria of some cells. On the 3d-14th day hyperplasia and hypertrophy of organelles in the immune-competent cells are revealed; it is connected with increasing processes of immunogenesis. At late stages of the investigation, normalization of the spleen cells ultrastructure and increased functional fibroblastic activity are noted.     

Oscillatory changes in cells of the submandibular glands in mice during the secretory cycle]

By means of 3H-leucine radioautography, ultrastructural and morphometrical analysis, it has been demonstrated that in the cells of the acinar (Ac) and granular (Gr) parts of the submandibular gland (SMG) in mice with a synchronized for 3 h nutrition cycle there are endogenic fluctuations (for about 1 h) of the secretory process. They are demonstrated as alterations in intensity of protein synthesis in the cells, their areas and ultrastructure. In the Ac cells at the moment of feeding and in 50, 110, 150 min after feeding maximal leucine incorporation is observed. The changes in the cell area during the first hour occurs with 10 minutes' overtake of the incorporation intensity, and during the following 2 h an equilibrium is reached between these two parameters, when the maximal contents of the label corresponds to the minimal area of the cells. In the Gr cells one maximum of 3H-leucine incorporation occurs in 60-80 min after feeding. The minimal content of the label takes place in 10 and 120 min after feeding. During the time mentioned inverse ratio between the intensity incorporation of the labelled isotope and the change of the cell volume is observed. This is connected with autoregulatory processes in the cells. It is possible that in the Ac cells together with removal of the secretory granules by means of exocytosis (in 10-20 min after feeding) the secretory material is removed before it is formed as granules by parvivesicular extrusion. In the Gr cells apocrinic and endocrinic secretion takes place. Presence of certain specific formations (secretory lysosomes) explains peculiarities of the secretory process in the SMG.     

Transgene expression in mammary glands of newborn rats

Transgene expression in the mammary glands of newborn rats was studied to establish an early selection system for transgenic animals producing exogenous proteins in their milk during lactation. A fusion gene composed of the bovine alpha S1 casein gene promoter and the human growth hormone gene was microinjected into rat embryos. Transgenic lines that produced human growth hormone in their milk were established and used in this study. Immediately after birth, and without any hormone treatment, human growth hormone was found in the extracts of mammary glands from both male and female rats derived from the line secreting human growth hormone in their milk. The expression of the transgene in mammary glands of newborn rats was also detected by the presence of human growth hormone mRNA. Nontransgenic newborn rats did not express the human growth hormone gene in their mammary glands, while the mRNA for rat alpha casein, an endogenous milk protein, was found in all mammary glands from both transgenic and nontransgenic neonates. These results show that analyzing the expression of transgenes in the mammary glands of neonates is a valuable tool to select the desired transgenic animals and to shorten the selection schedules establishing the transgenic animals. © 1996 Wiley-Liss, Inc.     

Ultrastructural changes in somatosensory cortex of albino rats during space flight

For the first time, mammalian brain has been studied during space flight aboard NASA orbital laboratory Spacelab-2. The main ultrastructural differences in the somatosensory cortex of the brain fixed under microgravity conditions and after landing include an increased number of degenerating presynaptic axon terminals after landing. Apparently, this is due to a sharp increase in afferent impulsation in the cortex during and after landing.