Distribution of 3-hydroxybutyrate dehydrogenase in primary lobules of rat liver.

In an attempt to establish the functional organization of the hepatic parenchymal unit, we used histo- and microchemical procedures to assess metabolic liver cell heterogeneity at the level of the primary lobule. Because of the close interrelation of glucogenesis and ketone body formation, and in view of the distinct regional differences of the in vivo activity of glucose-6-phosphatase (G6Pase), these techniques were used on livers from male rats to investigate the distribution of the ketogenic enzyme, 3-hydroxybutyrate dehydrogenase (3-HBDH), during the post-resorptive phase. A close reciprocity was found between the general increase in the activity of 3-HBDH and the decrease of the in vivo activity of G6Pase along the sinusoidal axis, and also with regard to enzyme gradients along sinusoids of different origin. The activity of the ketogenic enzyme was higher throughout septal than portal sinusoids, whereas the opposite applied to the glucogenic enzyme. Histo- and microchemical data support the concept of a lobular parenchymal unit composed of "primary lobules," and show also that hepatocyte function varies with cell location along the sinusoidal axis and with the origin of the sinusoids.     

Heterogeneity of kinetic parameters of enzymes in situ in rat liver lobules

In the present review, metabolic compartmentation in liver lobules is discussed as being dynamic and more complex than thus far assumed on the basis of numbers of mRNA or protein molecules or the capacity (zero-order activity) of enzymes. Isoenzyme distribution patterns and local kinetic parameters of enzymes may vary over the different zones of liver lobules. As a consequence, metabolic fluxes in vivo at physiological substrate concentrations may be completely different from those that are assumed on the basis of the number of molecules or the capacity of enzymes present in zones of liver lobules. For a more correct estimation of the levels of metabolic processes in the different compartments of liver tissue, local kinetic parameters and substrate concentrations have to be determined to calculate local metabolic fluxes. direct measurements of metabolic fluxes in vivo with the use of noninvasive techniques is a promising alternative and the techniques will become increasingly important in future metabolic research.This paper was presented at the symposium Metabolic Zonation of the Liver: New Answers to Old Questions held in honour of Prof. Dr. D. Sasse's 60th birthday, 26 August 1994, in Basel     

Heterogeneous expression of mRNA in rat liver lobules as detected by differential display

Although liver hepatocytes appear to be uniform histologically, they are considerably heterogeneous with respect to their individual physiological capacities. In order to find still unknown genes that are heterogeneously expressed and with the aim of evaluating the usefulness of the differential display technique for this purpose, we performed differential displays with mRNA isolated from hepatocytes from the periportal and pericentral zone of the rat liver. In this way we identified at least two mRNAs exclusively expressed in the pericentral fraction. Sequence analysis revealed that the corresponding genes encode proteins with proline-glutamate dipeptide repeats similar to ones previously identified in rat pheochromocytoma and brain. In situ hybridization confirmed the heterogeneous distribution of the mRNA. Only one to two cell lines surrounding the terminal hepatic venules were positive, strongly resembling the heterogeneous expression of the enzyme glutamine synthetase. Our work demonstrates that the differential display method is a useful tool for the identification of genes that are differentially expressed in individual parenchymal cells. In fact, our results prove that differential display technology can be used for the identification of cellular markers for distinct subpopulations of cells in a given tissue.     

An oscillatory neural network model that demonstrates the benefits of multisensory learning

Since the world consists of objects that stimulate multiple senses, it is advantageous for a vertebrate to integrate all the sensory information available. However, the precise mechanisms governing the temporal dynamics of multisensory processing are not well understood. We develop a computational modeling approach to investigate these mechanisms. We present an oscillatory neural network model for multisensory learning based on sparse spatio-temporal encoding. Recently published results in cognitive science show that multisensory integration produces greater and more efficient learning. We apply our computational model to qualitatively replicate these results. We vary learning protocols and system dynamics, and measure the rate at which our model learns to distinguish superposed presentations of multisensory objects. We show that the use of multiple channels accelerates learning and recall by up to 80%. When a sensory channel becomes disabled, the performance degradation is less than that experienced during the presentation of non-congruent stimuli. This research furthers our understanding of fundamental brain processes, paving the way for multiple advances including the building of machines with more human-like capabilities.     

Identifying players in the functional network around pRB

Comment on: Tschöp K, et al. Genes Dev 2011; 25:814-30.     

Portal vein insulin response of glucose intolerant Yucatan miniature swine to common secretogogues

A group of glucose intolerant miniature swine exhibiting an impaired portal vein insulin response to an IVGTT were examined with respect to their portal vein insulin response to the secretogogues: isoproterenol, arginine and leucine. Equivalent insulin responses to isoproterenol and leucine were noted on the part of the glucose intolerant animals when compared to control subjects. An impaired portal vein insulin response was evident during an infusion of 0.5 g/kg arginine and again when a pulse injection of .25 g/kg glucose was administered in the presence of isoproterenol (.05 microgram/kg . min). The close agreement of these results with those reported for human diabetics suggests that a similar pancreatic defect, most probably associated with the glucoreceptor, is present in this group of glucose intolerant miniature swine.     

A fully coupled porous media and channels flow approach for simulation of blood and bile flow through the liver lobules

Two dimensional, steady state, and incompressible blood and bile flows through the liver lobules are numerically simulated. Two different geometric models A and B are proposed to study the effects of lobule structure on the fluid flow behaviour. In Model A, the lobule tissue is represented as a hexagonal shape porous medium with a set of flow channels at its vertices accounting for the hepatic artery, portal and central veins along with bile ductules. Model B is a channelized porous medium constructed by adding a set of flow channels, representing the bile canaliculies and lobule sinusoids, to Model A. The bile and blood flow through the lobule is simulated by the finite element approach, based on the Darcy/Brinkman equations in the lobule tissue and the Navier-Stokes (or Stokes) equations in the flow channels. In Model B, a transmission factor on the boundaries of the bile canaliculies is introduced to connect the bile and blood flows. First, a single regular lobule is utilized to exhibit the fluid flow pattern through the liver lobule represented by proposed geometric models. Then, the model is extended to a group of liver lobules to demonstrate the flow through a liver slice represented by irregular lobules. Numerical results indicate that the Darcy and Brinkman equations provide nearly the same solutions for Model A and similar solutions with a little difference for Model B. It is shown that the existence of sinusoids and bile canaliculies inside the liver lobules has noticeable effects on its fluid flow pattern, in terms of pressure and velocity fields.     

Sex-specific regionality of liver metabolism during starvation; with special reference to the heterogeneity of the lobular periphery

Staining procedures for glucose-6-phosphatase and 3-hydroxybutyrate dehydrogenase activity and for glycogen were used to investigate adaptive changes in the regionality of hepatic gluconeogenesis and ketogenesis in fasting male and female rats. A reciprocal distribution of gluconeogenic and ketogenic capacities was found in both sexes, but male and female animals were different with respect to: a) the time necessary for full induction of glucose-6-phosphatase activity (24 h in females, 48 h in males); b) the overall activity of 3-hydroxybutyrate dehydrogenase; and c) glycogen content. The activity of the latter enzyme and the glycogen content did increase with time of starvation, but at all times, were higher in males, than in females. Results, thus, indicate that the extent to which ketone bodies replace glucose as major fuel for the brain is larger in males than in females. This may explain the delayed induction of glucose-6-phosphatase activity and the higher glycogen content in the male during starvation. Distributions of enzyme activities and of glycogen, furthermore, revealed the heterogeneity of the lobular periphery, i.e. functional differences among sinusoids dependent upon whether they originate from the portal tract or the vascular septum, and thus confirm the lobular concept proposed by Matsumoto et al. (1979).     

Sex-specific regionality of liver metabolism during starvation; with special reference to the heterogeneity of the lobular periphery

Summary Staining procedures for glucose-6-phosphatase and 3-hydroxybutyrate dehydrogenase activity and for glycogen were used to investigate adaptive changes in the regionality of hepatic gluconeogenesis and ketogenesis in fasting male and female rats. A reciprocal distribution of gluconeogenic and ketogenic capacities was found in both sexes, but male and female animals were different with respect to: a) the time necessary for full induction of glucose-6-phosphatase activity (24 h in females, 48 h in males); b) the overall activity of 3-hydroxybutyrate dehydrogenase; and c) glycogen content. The activity of the latter enzyme and the glycogen content did increase with time of starvation, but at all times, were higher in males than in females. Results, thus, indicate that the extent to which ketone bodies replace glucose as major fucl for the brain is larger in males than in females. This may explain the delayed induction of glucose-6-phosphatase activity and the higher glycogen content in the male during starvation. Distributions of enzyme activities and of glycogen, furthermore, revealed the heterogeneity of the lobular periphery, i.e. functional differences among sinusoids dependent upon whether they originate from the portal tract or the vascular septum, and thus confirm the lobular concept proposed by Matsumoto et al. (1979).Parts of this study were carried out in the Anatomisches Institut, Universität Freiburg, Federal Republic of GermanySupported by grants from the Deutsche Forschungsgemeinschaft, the Bochringer-Ingelheim Fonds and Grant AM 32654 from the National Institutes of Health     

On the structure of liver ribosomes

The morphology of liver ribosomes and their subparticles, large and small, has been investigated. Analysis of the images has been carried out by successive selection of models and by X-raying them under conditions simulating negative staining. The relation between the main views has been checked by tilting the specimens in an electron microscope through ± 30 °.The small subparticle consists of an elongated body, to one of the ends of which a short “head” is attached. A model has been proposed, whose projections on rotation with respect to the longitudinal axis would satisfy all observable types of images. According to the proposed model, the “head” is tilted with respect to the elongated portion. The length of the subparticle is 230 Å. The dimensions of the elongated portion in the transverse direction are 110 Å × 75 to 80 Å.The large subparticles in lateral view resemble short “rods” 220 to 240 Å long and about 70 to 95 Å wide. At a distance of about 60 Å from the left end of the particles a projection (60 Å in length) is seen, on the inner side of which a depression, or “channel”, filled with the contrasting substance is always observed. Next to this depression a second projection is located, whose height is about 30 Å. The channel is either a cavity in the body of the large subparticle or a part of the RNA without protein. The length of the channel is about 80 Å, the width is about 50 to 60 Å. The left end of the particles is characterized by two sharpened portions; as a result, a cavity that shows an obtuse angle profile makes its appearance. The opposite end of the particles is cut off at an angle of 45 °. In another view, the subparticles appear to be almost rectangular in shape; they are characterized by dimensions of 150 Å × 220 to 240 Å. It is likely that the large projection is displaced sideways with respect to the longitudinal axis of the particles. The asymmetry associated with this displacement gives rise to preferred arrangements of the subparticles on the supporting film. An analysis has been made of the most typical images of monomeric ribosomes, on the basis of which a suggestion is made about mutual orientation of subparticles in a monomer.     

Fractal geometry of mosaic pattern demonstrates liver regeneration is a self-similar process.

Partial hepatectomy causes compensatory, nonneoplastic growth and regeneration in mammalian liver. Compensatory liver growth can be used to examine aspects of patterns of cell division in regenerating tissue. Chimeric animals provide markers of cell lineage which are independent of growth and can be used to follow cell division patterns. Previous experimental evidence suggests that compensatory liver growth is uniform, without focal centers of proliferation. In this study we have extended that observation to include genes important in regeneration and cell cycle control in order to establish that nascent growth centers are not present in regenerating liver. There is a uniform spatial distribution of expression of these genes which is not related to mosaic pattern in the chimeras. While these genes may help regulate hepatocyte proliferation they do not appear to regulate patch pattern in the chimeras. With this information confirming uniform growth it was possible to use fractal analysis to test various hypothesized patterns of regenerative growth in the liver. The results of this analysis indicate that mosaic pattern does not change substantially during the regenerative process. Patch area and perimeter (the area occupied by or perimeter around cells of like lineage) increase during compensatory liver growth in chimeric rats without alteration of the geometric complexity of patch boundaries (boundaries around cells of like lineage). These tissue findings are consistent with previously reported computer models of growth in which repetitive application of simple decisions assuming uniform growth created complex mosaic patterns. They support the notion that an iterating (repeating), self-similar (a pattern in which parts are representative of, but not identical to the whole) cell division program is sufficient for the regeneration of liver tissue following partial hepatectomy. Iterating, self-similar cell division programs are important because they suggest a way in which complex patterns (or morphogenesis) can be efficiently created from a small amount of stored information.