Zones Of Lobules Research Articles

Zonated quantification of immunohistochemistry in normal and steatotic livers.

Immunohistochemical stains (IHC) reveal differences between liver lobule zones in health and disease, including nonalcoholic fatty liver disease (NAFLD). However, such differences are difficult to accurately quantify. In NAFLD, the presence of lipid vacuoles from macrovesicular steatosis further hampers interpretation by pathologists. To resolve this, we applied a zonal image analysis method to measure the distribution of hypoxia markers in the liver lobule of steatotic livers.The hypoxia marker pimonidazole was assessed with IHC in the livers of male C57BL/6J mice on standard diet or choline-deficient L-amino acid-defined high-fat diet mimicking NAFLD. Another hypoxia marker, carbonic anhydrase IX, was evaluated by IHC in human liver tissue. Liver lobules were reconstructed in whole slide images, and staining positivity was quantified in different zones in hundreds of liver lobules. This method was able to quantify the physiological oxygen gradient along hepatic sinusoids in normal livers and panlobular spread of the hypoxia in NAFLD and to overcome the pronounced impact of macrovesicular steatosis on IHC. In a proof-of-concept study with an assessment of the parenchyma between centrilobular veins in human liver biopsies, carbonic anhydrase IX could be quantified correctly as well.The method of zonated quantification of IHC objectively quantifies the difference in zonal distribution of hypoxia markers (used as an example) between normal and NAFLD livers both in whole liver as well as in liver biopsy specimens. It constitutes a tool for liver pathologists to support visual interpretation and estimate the impact of steatosis on IHC results.

A spatiotemporally resolved single-cell atlas of the Plasmodium liver stage.

Malaria infection involves an obligatory, yet clinically silent liver stage1,2. Hepatocytes operate in repeating units termed lobules, exhibiting heterogeneous gene expression patterns along the lobule axis3, but the effects of hepatocyte zonation on parasite development at the molecular level remain unknown. Here we combine single-cell RNA sequencing4 and single-molecule transcript imaging5 to characterize the host and parasite temporal expression programmes in a zonally controlled manner for the rodent malaria parasite Plasmodium berghei ANKA. We identify differences in parasite gene expression in distinct zones, including potentially co-adaptive programmes related to iron and fatty acid metabolism. We find that parasites develop more rapidly in the pericentral lobule zones and identify a subpopulation of periportally biased hepatocytes that harbour abortive infections, reduced levels of Plasmodium transcripts and parasitophorous vacuole breakdown. These 'abortive hepatocytes', which appear predominantly with high parasite inoculum, upregulate immune recruitment and key signalling programmes. Our study provides a resource for understanding the liver stage of Plasmodium infection at high spatial resolution and highlights the heterogeneous behaviour of both the parasite and the host hepatocyte.

Clinicopathological analysis of 16 cases of pyrrolizidine alkaloids-associated hepatic sinusoidal obstruction syndrome

Objective: To observe the histopathological manifestations of liver biopsy in patients with hepatic sinusoidal obstruction syndrome (HSOS) induced by pyrrolizidine alkaloid (PA). Methods: Patients diagnosed with PA-HSOS from 2012 to 2017 were selected, and the general conditions, liver function indexes, medication history, liver biopsy time, histopathological slides of liver biopsy, and follow-up data of clinical prognosis after 6 months of onset were collected. Clinical staging with clinical data was used to observe the histopathological manifestations of patients at different clinical stages. Wilcoxon rank-sum test, unpaired t-test and univariate linear regression analysis were used for data analysis. Results: A total of 16 cases were collected. Alanine transaminase and aspartate transaminase was 59.25 U/L and 25.50 U/L, 108 U/L and 45 U/L, respectively, after 6 months of onset and follow-up, and the differences were statistically significant. Moreover, total bile acids and albumin was 35 μmol/L and 36.15 μmol/L, and 32.45 g/L and 31 g/L, respectively, and the differences were not statistically significant. PA-HSOS pathological development process was divided into early, middle and late stages. In the early stage, the central lobular sinusoidal endothelium integrity was impaired and the entry of erythrocytes had interspersed thin reticular fibers and perisinusoidal space. In the middle stage (hemorrhagic zone), erythrocytes, reticular fibers and collagen fibers were lysed, densely collapsed and deposited. The cavity of the bloodstream was hyperemic and dilated, and the cavity was covered with sinus endothelial cells. The hepatic plate regenerated around the hemorrhagic zone and some of the hepatic sinuses were decompensated. In the late stage, deposited collagen in the hemorrhagic zone had formed a large fibrous scar, and most of the dilated cavity in the bloodstream was covered with vascular endothelium. The marginal zone hepatic cells were regenerated in two rows and gradually inserted into the fibrous septum. Different hepatic lobular lesions obtained from the same patients liver biopsy tissues were changed at different stages. Hepatic lobule injury proportion with severe internal bleeding in liver biopsy tissue had no relation with the prognosis of patients. Conclusion: In the early stage of PA-HSOS, erythrocytes in the central zone of lobules enter the perisinusoidal space through the damaged sinus endothelium, which is manifested as hepatic plate hemorrhagic necrosis. In the middle and late stage, liver plate regeneration and vascular remodeling occurred, so most of the patients' clinical course was self-limited. Pathological staging and liver biopsy time have an apparent correlation, but the prognosis of patients cannot be judged based on the extent of hemorrhage and injury of biopsy samples.

Deletion of the cannabinoid CB1 receptor impacts on the ultrastructure of the cerebellar parallel fiber-Purkinje cell synapses.

The cannabinoid CB1 receptor localizes to the glutamatergic parallel fiber (PF) terminals of the cerebellar granule cells and participates in synaptic plasticity, motor control and learning that are impaired in CB1 receptor knockout (CB 1 -KO) mice. However, whether ultrastructural changes at the PF-Purkinje cell (PC) synapses occur in CB 1 -KO remains unknown. We studied this in the vermis of the spinocerebellar lobule V and the vestibulocerebellar lobule X of CB 1 -KO and wild-type (CB 1 -WT) mice by electron microscopy. Lobule V, but not lobule X, of CB 1 -KO had significantly less and longer synapses than in CB 1 -WT. PF terminals were significantly larger in both lobules of CB 1 -KO with no changes in PC dendritic spines. The PF terminals in lobule V of CB 1 -KO contained less synaptic vesicles and lower vesicle density; by contrast, vesicle density in lobule X of CB 1 -KO remained unchangeable relative to CB 1 -WT. There were as many vesicles in lobule V of CB 1 -KO as in CB 1 -WT, but their distribution decreased drastically at 300 nm of the active zone. In lobule X of CB 1 -KO, less vesicles were found within 150 nm from the presynaptic membrane; however, no vesicles were at 450-600 nm of the active zone. A significant higher amount of synaptic vesicles close to the active zone in lobule V and X of CB 1 -KO was observed. In conclusion, the absence of CB1 receptors strikingly and distinctively impacts on the ultrastructural architecture of the PF-PC synapses located in cerebellar lobules that differ in vulnerability to damage and motor functions.

Spatial sorting enables comprehensive characterization of liver zonation.

The mammalian liver is composed of repeating hexagonal units termed lobules. Spatially resolved single-cell transcriptomics revealed that about half of hepatocyte genes are differentially expressed across the lobule, yet technical limitations impeded reconstructing similar global spatial maps of other hepatocyte features. Here, we show how zonated surface markers can be used to sort hepatocytes from defined lobule zones with high spatial resolution. We apply transcriptomics, miRNA array measurements and mass spectrometry proteomics to reconstruct spatial atlases of multiple zonated features. We demonstrate that protein zonation largely overlaps with mRNA zonation, with the periportal HNF4α as an exception. We identify zonation of miRNAs such as miR-122, and inverse zonation of miRNAs and their hepatocyte target genes, highlighting potential regulation of protein levels through zonated mRNA degradation. Among the targets we find the pericentral Wnt receptors Fzd7 and Fzd8 and the periportal Wnt inhibitors Tcf7l1 and Ctnnbip1. Our approach facilitates reconstructing spatial atlases of multiple cellular features in the liver and other structured tissues.

Spatial heterogeneity in the mammalian liver.

Hepatocytes operate in highly structured repeating anatomical units termed liver lobules. Blood flow along the lobule radial axis creates gradients of oxygen, nutrients and hormones, which, together with morphogenetic fields, give rise to a highly variable microenvironment. In line with this spatial variability, key liver functions are expressed non-uniformly across the lobules, a phenomenon termed zonation. Technologies based on single-cell transcriptomics have constructed a global spatial map of hepatocyte gene expression in mice revealing that ~50% of hepatocyte genes are expressed in a zonated manner. This broad spatial heterogeneity suggests that hepatocytes in different lobule zones might have not only different gene expression profiles but also distinct epigenetic features, regenerative capacities, susceptibilities to damage and other functional aspects. Here, we present genomic approaches for studying liver zonation, describe the principles of liver zonation and discuss the intrinsic and extrinsic factors that dictate zonation patterns. We also explore the challenges and solutions forobtaining zonation maps of liver non-parenchymal cells. These approaches facilitate globalcharacterization of liver function with high spatial resolution along physiological and pathological timescales.

Prenatal morphogenesis of compartments of the parenchyma of the lymph nodes of domestic cattle (Bos taurus)

The article analyzes the regularities of the formation and development of the lymphoid lobules of the parenchyma of the somatic (Limphonodi cervicales superficiales) and visceral (L. jejunales) lymph nodes in domestic cattle in the fetal period of ontogenesis. We used routine histological techniques and author's modification of the impregnation of total median sections with silver nitrate. Visualization of various zones of lymphoid lobules was carried out taking into account the specific for different cell zones architectonics of reticular fibers. It has been established that signs of morphological heterogeneity of parenchyma of lymph nodes are first found in three month-old fetuses, which is associated with the concentration of lymphoid tissue along the marginal sinus. Separation of lymphoid lobules and their structural and functional differentiation are first detected in five month fetuses. In the lymphoid lobules of five month-old fetuses all structural and functional cell zones are observable, among which the regions of clonal proliferation of T and B lymphocytes are the least developed, and regions of the transit corridors for lymphocytes migrating medullary and the cords (zone of potential accumulation of plasmocytes and antibody formation) are the most developed. Structural and functional transformations of compartments in the prenatal period of ontogenesis are accompanied by a predominantly moderate increase of the relative volume of specialized T- and B-dependent zones of lobules, against a background of a gradual decrease of the volume of transit corridors for lymphocytes migrating and zone of potential accumulation of plasmocytes and antibody formation. Due to the small volume and relatively low rates of development of the lymphocytes clonal proliferation zones, the quantitative ratios of the cellular zones in lymphoid lobules of the lymph nodes of domestic cattle in prenatal ontogenesis remain relatively stable, while maintaining the maximum indices of the development of transit corridors for lymphocytes migration and medullary cords. Among the zones of lymphocytes clonal proliferation throughout the fetal period, T-dependent zones predominate, the relative volume of which is 5.0–7.5 times greater than the volume of B-dependent zones. Lymphoid lobules in the lymph nodes of the domestic cattle fetuses of all age groups are arranged along the marginal sinus in one row and have a polar structure due to the formation of lymph nodes at one pole of the lobules in the interfollicular zone. In the visceral lymph nodes (L. jejunum) of 8–9 month-old fetuses, individual lymph nodes can form in paracortical strands, on the border with the interfollicular zone.

Organization of cerebral projections to identified cerebellar zones in the posterior cerebellum of the rat.

The cerebrocerebellar connection makes use of two of the largest fiber tracts in the mammalian brain, i.e., the cerebral and medial cerebellar peduncles. Neuroanatomical approaches aimed to elucidate the organization of this important connection have been hindered by its multisynaptic nature, the complex organization of its components, and the dependency of conventional tracers on precisely placed injections. To overcome these problems, we used rabies virus (RV) as a retrograde transneuronal tracer. RV was injected simultaneously with cholera toxin β subunit (CTb) into selected areas of the cerebellar cortex of 18 male Wistar rats. A survival time of 48-50 h resulted in first- and second-order labeling of RV in combination with first-order labeling of CTb. The distribution of CTb-labeled neurons in the inferior olive established the zonal identity of the injection site. In this way, it was possible to examine the cortical distribution of neurons from which disynaptic cerebrocerebellar projections to specific cerebellar loci originate. The results show that this distribution covaries with the identity of the injected cerebellar lobule. More subtle changes were present when different zones of the same lobule were injected. The C1 zone of lobule VIII receives a more prominent projection from the somatosensory cortex compared with the C2/D zones. The laterally positioned D zones receive information from more rostral regions of the cerebral cortex. The vermis of lobule VII receives a prominent input from the retrosplenial and orbitofrontal cortices. Different injection sites also result in differences in laterality of the connections.

Extending the bandwidth of long‐term plasticity at the cerebellar input stage

Extending the bandwidth of long‐term plasticity at the cerebellar input stage

Angiogenesis in chronic hepatitis C is associated with inflammatory activity grade and fibrosis stage

Data regarding the assessment of angiogenesis in liver tissue in chronic hepatitis C (CHC) are rare. The study was performed to explain the association between the histopathological features and the number of new blood vessels in lobules and portal tracts in CHC. The second aim of the study was to define the localization of sprouting and pattern of formation of new vessels by estimating CD 34 antigen expression in the liver. The study involved 74 patients with CHC, infected with viral genotype 1b before antiviral therapy. The number of new-formatted blood vessels was positively associated with fibrosis stage and inflammatory activity grade in the liver biopsy from CHC patients. The relationship was evident in the portal tract, fibrous septa and periportal zones of lobules. The results suggest that inflammatory hepatocyte injury may promote neo-angiogenesis.

Metabolic heterogeneity of glycogen in hepatocytes of patients with liver cirrhosis: the glycogen of the liver lobule zones in cirrhosis

The concentrations of total glycogen (TG) and its labile (LF) and stable (SF) fractions were determined in hepatocytes of portal and central zones of the normal human liver and in the liver of patients with cirrhosis of viral and alcohol aetiologies. Using PAS reaction, TG, LF and SF were revealed in histological sections of the material obtained by the liver punch biopsies. The concentrations of TG and its fractions were measured by televisional cytophotometry. In liver cirrhosis, the concentrations of TG, LF and SF in both zones of the hepatic lobule have been found to be much higher than in the normal liver. It has been shown that the ratio of the hepatocyte TG concentrations in the portal zone to the central zone both in the normal liver and in viral cirrhosis exceeds 1.0, amounting to 1.264 +/- 0.021 and 1.030 +/- 0.009, respectively. The glycogen fraction composition in the cells of both the liver lobule zones in viral cirrhosis does not differ significantly from the norm. On the contrary, in the liver of patients with alcoholic cirrhosis, the ratio of the TG concentrations in the portal zone to the central zone is reduced to 0.815 +/- 0.016 and is accompanied by qualitative changes of the glycogen composition.

Rostrocaudal branching within the climbing fibre projection to forelimb-receiving areas of the cerebellar cortical C1 zone.

The inferior olive climbing fibre projection to two somatotopically corresponding regions of the paravermal C1 zone in lobule V of the anterior lobe and the rostral folia of the paramedian lobule (PML) in the posterior lobe were investigated in cats by using a combined electrophysiological and retrograde double-labelling technique. In each experiment (n = 7), a small injection of rhodamine-tagged beads was made into the forelimb-receiving part of the C1 zone in one region of the cortex, and an injection of fluorescein-tagged beads was made into the other region. The two regions were found to receive olivary input from cells located in a partially overlapping territory within the rostromedial dorsal accessory olive (DAO). At progressively more rostral levels of DAO, the territory providing climbing fibres to the anterior lobe was centred more laterally than the territory projecting to the rostral PML. Where overlaps occurred, cells that provided climbing fibres to one or the other region were intermingled with a smaller population of double-labelled cells that had axons that branched to supply climbing fibres to both regions of the zone. The double-labelled cells represented on average 26% of the smaller total population of labelled cells within the area of overlap and 7% of the total population of neurones projecting to both regions of the zone. Overall, the findings suggested that the C1 zone within spatially separate but somatotopically corresponding regions of the paravermal cerebellar cortex receives at least partially independent olivary inputs, consistent with the presence of distinct microzones at different rostrocaudal levels of the zone.

Zonal organization of cortico-nuclear and nucleo-cortical projections of the paramedian lobule of the cat cerebellum. 1. the C1 zone.

The cortico-nuclear (C-N) and nucleo-cortical (N-C) projections of the C1 cortical zone in pars anterior (pa) and pars copularis (pc) of the paramedian lobule (PML) in the posterior lobe of the cat cerebellum were investigated with a combined electrophysiological and neuroanatomical technique. In each experiment the mediolateral boundaries of the zone were located on the cortical surface by recording field potentials mediated via climbing fibres and evoked in the zone by activity elicited in spino-olivocerebellar paths through percutaneous stimulation of fore- and hindlimbs; a small (15-30 nl) injection of WGA-HRP was then made into the zone. The distributions in the deep cerebellar nuclei were determined (with light microscopy) both for terminal labelling due to anterograde axonal transport by Purkinje cells and for cell bodies labelled due to retrograde transport in N-C axons. The extent to which injection sites were confined to the C1 zone was assessed both by comparing injection site and zone widths and by determining the distributions of retrogradely labelled neurones within the contralateral inferior olive. The C-N projection from the part of the zone in PML pa (a forelimb part) terminates almost exclusively (perhaps exclusively) in nucleus interpositus anterior (NIA), primarily in caudal and dorsal parts, where it overlaps heavily with the C-N projections from the lobule V parts (also forelimb parts) of the C1 and C3 zones as previously defined. The C-N projection from the part of the zone in PML pc (a hindlimb part) also terminates virtually exclusively in NIA but primarily in almost all parts of the medial third of the nucleus. There is, nevertheless, sufficient overlap between the PML pa and PML pc projections that approximately one third of the termination territory of each projection overlaps that of the other. The PML pc part of the zone is almost entirely lacking in a N-C projection, as previously found for the lobule V part of the C1 zone (and C3 zone). However, the PML pa part of the zone receives N-C projections that arise, in descending order of size, from nucleus interpositus posterior (NIP), from NIA, from the NIA/nucleus lateralis (NL) fusion area and (perhaps) NL. The projection from NIP is similar in size to that provided by the nucleus to the C2 zone in lobule V of the anterior lobe. The findings are discussed, with particular emphasis on their implications for the hypothesis that the cerebellum is divisible into a number of olivo-cortico-nuclear complexes or compartments.

Numerical aspects of pontine, lateral reticular, and inferior olivary projections to two paravermal cortical zones of the cat cerebellum

Two different olivo-cortico-nuclear zones in the cat cerebellum have been compared quantitatively as regards the numbers of cells projecting to them from within several sources of mossy fibres (MFs), namely the basal pontine nuclei (BPN), nucleus reticularis tegmenti pontis (NRTP), and the ipsilateral lateral reticular nucleus (LRN). The zones chosen were the C3 zone in lobule V of the anterior lobe and the C1 zone in pars copularis of the paramedian lobule (PMLpc), localised by recording climbing fibre-mediated potentials evoked on their surface as a result of volleys set up in their spino-olivocerebellar paths. The zones were injected with fluorescent-labelled latex microspheres and cell bodies, retrogradely labelled in the MF source nuclei and in the contralateral inferior olive, were counted and mapped. Evidence was obtained that tracer efficiency was very high in both the MF projections and the olivo-cerebellar projection and that each olivocerebellar axon may provide only one climbing fibre to the upper part of a lobule V folium but an average of nearly two to the same part of a PML folium. When the numbers of labelled cells in each MF source nucleus were expressed as a percentage of the total number of labelled pontine cells, the biggest source for lobule V was the contralateral BPN, followed by LRN, contralateral NRTP, ipsilateral BPN, and ipsilateral NRTP. For PMLpc, the order was similar except that ipsilateral BPNs exceeded contralateral NRTPs, but the dominance of contralateral BPN as a source was much greater. Cell totals were converted into projection densities (i.e., numbers of cells labelled per square millimetre of cortical sheet involved in the injection site); densities for PMLpc were found to be almost three times greater than those for lobule V for contralateral BPN but the two densities were not significantly different for ipsilateral BPN. The three other MF sources projected at higher densities to lobule V than to PML. These findings indicate that two cortical zones, both of which receive climbing fibres from the rostral part of the dorsal accessory olive and project to nucleus interpositus anterior, nevertheless differ markedly in regard to both the relative and the absolute sizes of the projections they receive from several of their most important sources of MFs.

Effect of hydrocortisone on amine content in the thymus

Intraperitoneal injection of 2.5% hydrocortisone suspension (0.5 ml) changes the content of biogenic amines in different lobule zones and thymocytes of high animals. The content of norepinephrine and serotonin increases in luminescent cells of the subcapsular and premedullary zones and decreases in mast cells.

Quantitative analysis of glycogen content in hepatocytes of portal and central lobule zones of normal human liver and in patients with chronic hepatitis of different etiology

Glycogen content was determined in hepatocytes of different lobule zones of the normal human liver (23 patients without any liver pathology) and the liver of patients with chronic viral B hepatitis (30 patients) and chronic alcohol hepatitis (28 patients). All the patients were males and aged between 17–50 years. Quantitative analysis of the glycogen content in hepatocytes of portal and central lobule zones was carried out in sections of the human liver (material of punctional biopsies) stained with PAS-reaction. The measurements were carried out using an image analyser ‘Magiscan’ which allows combined cytophotometric analysis of a substance in cells and determination of the cell localization in tissue. The results showed significant differences of the glycogen content in different lobule zones in the normal liver and in the liver in chronic viral and alcohol hepatitis. Ratios of glycogen content in hepatocytes of the portal and the central zones of liver lobule were 1.128 ± 0.004 and 1.061 ± 0.003 in normal human liver, and liver of patients with chronic viral hepatitis respectively, i.e. the glycogen content in hepatocytes of the portal lobule zone was much higher than in the central lobule zone in the normal liver and in the liver of patients with chronic viral B hepatitis. The ratio in patients with chronic alcohol hepatitis was less than 1.0 (0.930 ± .0.003), i.e. a significantly higher glycogen content was found in hepatocytes of the central liver lobule zone. Possible mechanisms of this phenomenon are discussed. Thus, the pattern of the glycogen content in hepatocytes of different lobule zones can be used as an indicator of etiology of chronic hepatitis.

Correspondence between climbing fibre input and motor output in eyeblink-related areas in cat cerebellar cortex.

The purpose of the present work was to identify sites in the cerebellar cortex which are likely to control eyeblink. This work was motivated by findings suggesting that the cerebellum is involved in the learning and/or performance of the classically conditioned eyeblink response. The identification was based on climbing fibre input to the cortex and on the effects of electrical stimulation of the cerebellar cortex in cats decerebrated rostral to the red nucleus. The cerebellar surface was searched for areas receiving short latency climbing fibre input on periorbital electrical stimulation. Four such areas were found in the c1 and c3 zones of lobules VI and VII in the anterior lobe of the cerebellum and in the c3 zone in the paramedian lobule. Electrical stimulation of the cerebellar cortex with trains (150-400 Hz) of at least 10 ms duration evoked two types of EMG response in the orbicularis oculi muscle. An early response, time-locked to the onset of the stimulation, was unrelated to climbing fibre input and a delayed response, time-locked to the termination of the stimulation, could only be evoked from areas which received short latency climbing fibre input from the eye, that is, the c1 and c3 zones. The delayed responses had long latencies (up to 50 ms) after the termination of the stimulus train and could be delayed further by prolonging the stimulation. Both types of response were abolished by injections of small amounts of lignocaine into the brachium conjunctivum. A number of characteristics of the delayed responses are described. They could be inhibited by a further shock to the same area of the cerebellar cortex. Their latency could be increased by increasing the stimulation frequency. The period between stimulation and appearance of the response often showed a decrease in spontaneous EMG activity. There was a close topographical correspondence between input and output. Delayed responses could be evoked from all four of the areas in the c1 and c3 zones which have climbing fibre input from the periorbital area. They could not be evoked from other areas. In contrast, early responses were only evoked from areas without such climbing fibre input. It is proposed that the delayed responses were generated by activation of Purkinje cell axons leading to hyperpolarization and a subsequent rebound depolarization and activation of cells in the interpositus nucleus. The cortical areas are therefore probably involved in the control of the orbicularis oculi muscle.(ABSTRACT TRUNCATED AT 400 WORDS)

Zonal Organization Within the Projection from the Inferior Olive to the Rostral Paramedian Lobule of the Cat Cerebellum

The projection from the inferior olivary nucleus to the forelimb-related regions of the c1, c2 and c3 zones within rostral folia of the paramedian lobule of the cat cerebellum was studied using a combined electrophysiological and neuroanatomical tracing technique. In each experiment, the cerebellar cortical zones were identified by their receipt of spino-olivocerebellar input evoked by percutaneous electrical stimulation of the limbs. A small (15-30 nl) injection of wheatgerm agglutinin-horseradish peroxidase (WGA-HRP) was then made into the centre of one of the zones. The olivary regions projecting to each zone were compared with the regions within the olive that have previously been shown to project to the corresponding cortical zones within lobule V of the anterior lobe. The results show that some (but not all) of the paravermal zones in the anterior lobe are also represented in the paramedian lobule (PML). The most medial (c1) zone in rostral PML receives climbing fibre input from an area in the rostral dorsal accessory olive which overlaps partially with the area that projects to the medial half of the c1 zone in lobule V. A zone equivalent to the lateral half of the c1 zone (the cx zone) in lobule V does not appear to be present within rostral PML. A middle (c2) zone within rostral PML receives olivary input from a region within the rostral medial accessory olive that overlaps partially with the area that projects to the c2 zone in lobule V. The presence of a c3 zone within rostral PML was found to be variable between animals and it could be identified electrophysiologically in only two out of a possible nine cases. In summary, the results demonstrate that, although a number of similarities exist between the olivocerebellar projections to corresponding cortical zones in the two forelimb-receiving regions of the cat paravermal cortex, the differences in olivocerebellar connectivity between the two regions suggest that functional differences between them may also exist.

Immunohistochemical detection of ras and myc oncogene expression in regenerating rat liver

The expression of ras and myc oncogene proteins was examined in formalin-fixed paraffin-embedded sections of male Sprague-Dawley rat liver at various times after liver regeneration was induced by either a necrogenic dose of CCl 4 or a 2/3 partial hepatectomy. Antibody specific for these proteins and immuno-histochemical (IHC) procedures were used to detect oncogene expression in the liver. Increased expression of both oncogene proteins was detected at times comparable to those reported by others using either Northern or Western blot methods. However, the IHC procedure provided additional information since oncogene expression was detected only in hepatocytes, mainly occupying liver lobule zones known to contain proliferating hepatocytes during regeneration. Also, ras and myc stain intensity of individual hepatocytes did not increase during regeneration. Rather, a greater number of hepatocytes were positive at several time points in the regeneration process. This suggested that the increased ras and myc expression detected in Northern and Western blot methods probably represents a greater number of hepatocytes expressing the oncogene proteins rather than increased expression by individual hepatocytes. Overall, this study provides additional information on the cells involved in the expression of the ras and myc oncogenes during regeneration and further substantiates their role in a normal physiological process.

Vagal and somatic representation by the climbing fiber system in lobule V of the cat cerebellum

The organization of the climbing fiber representation of the vagal afferents and the body surface in the vermal and intermediate zones of lobule V was examined in cats anesthetized with α-chloralose. Extracellular single-unit recordings were made from 428 Purkinje cells. Electrical stimulation of the vagus nerve elicited climbing fiber responses in 40% of the cells, most of which had convergent somatic input. Activation of A σ vagal afferent fibers accounted for 65% of the responses, whereas the A β fibers involved 27% and the C fibers included 8% of the responses. The responses driven by vagal nerve stimulation were encountered throughout the lobule, although a significantly increased representation of the vagus was identified for 3 longitudinal 0.5 mm wide sectors (two in the vermis and one in the intermediate region). In the vermis, the fine-grain organization consisted of a mixture of representations of the various parts of the body surface with and without convergent vagal input, although there was little convergence in the medial vermis where many of the responses were elicited by only vagal nerve stimulation. In the intermediate cortex, most of the vagal climbing fiber representation was convergent with forelimb input. These results suggest that vagal input into the cerebellum could have important modulatory effects on the cerebellar somatosensory input.