Effect of measured chemical sympathectomy on the dynamics of the chromatin functional indices of rat sympathetic neurocytes during their differentiation

Differences have been found between age-related shifts of DNA template activity of sympathetic neuron chromatin of normal and guanethidine-desympathized rats. The mean level of DNA template activity in mature neurons was the higher the heavier the desympathization of rat. In the ganglia of guanethidine-treated rats a subpopulation of neurocytes was revealed with an unusually high nucleolar-to-nucleoplasmic label ratio, never observed in control rats. The maturational disturbances in the course of alterations in histones visualized by ammoniacal silver, as well as shifts in the percentage of specifically stained nuclei are proportional to the level of desympathization. These changes in histones can be interpreted as a preliminary acceleration and aging of guanethidine-treated rat ganglia which are proportional to the number of destructed cells.     

Effect of different fixation methods and partial chemical sympathetic denervation on cell size and 3H-leucine incorporation by rat sympathetic neurocytes

Dimensions of sympathetic nerve cells and of their nuclei were studied using various methods of fixation and embedding media, as well as autoradiographic indices of the protein synthesized system of neurocytes in normal rats, and in rats partially sympathectomized at an early age. On the ground of different changes in the average size of neurocytes seen under various methods of fixation, and of decreased labeling intensity of large neurocytes after the injection of the labeled protein biosynthesis precursor there is some reason to believe that large neurocytes may appear in the late reproductive age mainly due to the nerve cell cytoplasm swelling as a consequence of exhaustion of their growth resources.     

Reverse sphingomyelin-synthase in rat liver chromatin

The chromatin phospholipid fraction is enriched in sphingomyelin content which changes during cell maturation and proliferation. Recently, we have demonstrated that the sphingomyelin variations can be due to chromatin neutral sphingomyelinase and sphingomyelin-synthase activities which differ in pH and K(m) optima from those present in nuclear membranes. The sphingomyelin can be used also as a source of phosphorylcholine for phosphatidylcholine synthesis by reverse sphingomyelin-synthase. In the present work we have studied the possible existence of reverse sphingomyelin-synthase activity in nuclear membrane and chromatin. A very low activity was detected in the homogenate, cytosol and nuclear membrane (0.93+/-0.14, 2.61+/-0.33 and 0.87+/-0.13 pmol/mg protein/min, respectively), whereas the activity present in chromatin was 37.09+/-2.05 pmol/mg protein/min. The reverse sphingomyelin-synthase decreases the intranuclear diacylglycerol pool and increases the intranuclear ceramide pool, whereas sphingomyelin-synthase has an opposite effect. The possible correlation between these enzymes is discussed.     

Intracellular relationships of the oestrogen receptor in the rat uterus and hypothalamus during the oestrous cycle.

Simultaneous measurements were made of the specific oestrogen receptor in the nuclear and cytosol fractions prepared from the uterus and hypothalamus of 50--81-day-old female rats undergoing a 4-day oestrous cycle. In the uterus, the content of nuclear receptor fluctuated in concert with known cyclic changes in the secretion of oestrogen, being maximal at pro-oestrus. Over the period of 50--81 days, the nuclear content at all phases increased with age, again corresponding to known age-related increases in ovarian secretion of oestrogen. This age-related increase in nuclear content, averaged from the values of the different phases in each age group, was related to equivalent increases in uterine wet weight, an increase of 1 pmol of receptor being accompanied by an increase of 80--90 mg. The concentration of cytosol receptor was maintained constant, with respect to wet weight, throughout the cycle and with age, irrespective of changes in nuclear content. In the uterus of normal mature females, translocation of receptor into the nucleus did not lead to depletion of cytosol receptor, suggesting a process of continuous replenishment/synthesis. In the hypothalamus, the nuclear content of oestrogen receptor was also maximal at pro-oestrus. In contrast with the uterus, the content of hypothalamic cytosol receptor was minimal at this phase and reflects depletion of the cytosol receptor, possibly as a result of translocation. The extent of translocation was low compared with that in the uterus and did not alter with age during the age-period studied. This low nuclear binding of the receptor in vivo is discussed in relation to the presence of a cytosol factor, present in limiting amounts, which in vitro mediates the binding of cytosol receptor to oligo(dT)-cellulose. The difference in the physiological response of the uterus and of the hypothalamus to oestrogens may be related to the extent of nuclear binding of receptor.