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PubMed2004

Three-dimensional structural views of branch migration and resolution in DNA homologous recombination.

The processing of the Holliday junction by various proteins is a major event in DNA homologous recombination and is crucial to the maintenance of genome stability and biological diversity. The proteins RuvA, RuvB and RuvC play central roles in the late stage of recombination in prokaryotes. Recent atomic views of these proteins, including protein-protein and protein-junction DNA complexes, provide new insights into branch migration mechanisms: RuvA is likely to be responsible for base-pair rearrangements, whereas RuvB, classified as a member of the AAA(+) family, functions as a pump to pull DNA duplex arms without segmental unwinding. The mechanism of junction resolution by RuvC in the RuvABC resolvasome remains to be elucidated.

Bacterial Proteins↗

PubMed1996

Role of RuvA in branch migration reactions catalyzed by the RuvA and RuvB proteins of Escherichia coli.

The RuvA and RuvB proteins of Escherichia coli promote ATP-dependent branch migration of Holliday junctions during homologous genetic recombination and DNA repair. In this process, RuvA acts as a specificity factor that targets RuvB, a hexameric ring motor protein, to the junction. Because elevated concentrations of RuvB can promote branch migration in the absence of RuvA, it has been suggested that RuvA acts as a molecular matchmaker. In the studies presented here, we compared the requirements for RuvAB- and RuvB-mediated branch migration reactions and found that reactions catalyzed by RuvB alone were highly sensitive to inhibition by NaCl, temperature, ADP, and ATPgammaS. Our observations indicate that the two reactions occur by distinct mechanisms and support the notion that RuvAB-mediated branch migration is physiologically more relevant than that catalyzed by RuvB. We also show that ongoing RuvAB-mediated branch migration reactions were blocked by the addition of polyclonal antibodies raised against RuvA. The role of RuvA is therefore unlikely to be restricted to RuvB targeting; instead, it is required continually during branch migration. Competition with excess synthetic Holliday junctions, sufficient to sequester released RuvA, failed to cause an immediate block and leads us to suggest that RuvAB promote branch migration by a processive mechanism.

Adenosine Diphosphate↗

PubMed1995

Synapses on axons of sympathetic preganglionic neurons in rat and rabbit thoracic spinal cord.

Axosomatic and axodendritic synapses occur on sympathetic preganglionic neurons, but it is not yet known whether their axons receive synaptic input, which could be particularly effective at regulating sympathetic outflow. Here, we examined retrogradely labelled sympathetic preganglionic axons to see if they received synapses. Cholera toxin B subunit (CTB) or CTB conjugated to horseradish peroxidase (CTB-HRP) was used to label neurons projecting to the rat or rabbit superior cervical ganglion, the rat adrenal medulla, or the rabbit stellate ganglion. At the light microscopic level, small groups of CTB-immunoreactive axons travelled through the ventral horn near its lateral boundary, with occasional axons taking a more medial course. The axons passed through the ventrolateral funiculus to exit at the ventral roots. In parasagittal section, a few axons branched within the ventral horn, sending processes rostrally and caudally for short distances before they turned ventrally to exit the spinal cord. At the ultrastructural level, CTB-immunoreactive rat and rabbit sympathetic preganglionic axons were almost exclusively unmyelinated. In contrast, labelling with CTB-HRP revealed both myelinated and unmyelinated axons in the ventral horn, the ventrolateral white matter, and the ventral roots. CTB-HRP also allowed the detection of the initial segment of a sympathetic preganglionic axon. Synapses, with vesicles clustered presynaptically and membrane specializations postsynaptically, were found on some unmyelinated CTB-immunoreactive axons. Occasional axons received several synapses. Synapses were most common on CTB-containing axons just ventral to the intermediolateral cell column. One synapse was found on an axon within 2 microns of its origin from a proximal dendrite. Rare synapses were found several hundred micrometers ventral to the intermediolateral cell column. One branching axon had synapses just below the branch point on both the main axon and the axonal branch. These findings indicate an extensive synaptic input to the axons of at least some sympathetic preganglionic neurons. These axoaxonic synapses could have a profound effect on sympathetic activity.

Animals↗

PubMed1992

Neurite regeneration from single primary-auditory neurons in vitro.

Neurons of the VIIIth cranial nerve in lower vertebrates precisely reconnect with their targets after sustaining injury. It is not known, however, whether the regenerating neurites are guided entirely by external cues or may also be directed by intrinsic mechanisms. To address this issue, single adult primary-auditory neurons were dissected from goldfish and placed in an in vitro environment, devoid of the normal complement of satellite cells, neighboring neurons, and synaptic targets, to observe their patterns of growth. Because acutely isolated neurons showed little neurite outgrowth, neurite regeneration was enhanced by focally crushing the VIIIth cranial nerve 2 to 24 h prior to removal for tissue culture. Neurons that regenerated under identical culture conditions showed growth patterns that could be categorized into three separate groups based on both their morphology and growth patterns. They either 1) remained unbranched (54%), 2) bifurcated or trifurcated into major branches directly from the myelinated stump (V-shaped) (19%), or 3) bifurcated from a regenerated process (Y-shaped), sometimes with a third, smaller branch (27%). Unbranched and V-shaped neurites grew at a constant elongation rate, while Y-shaped neurites grew variably, with alternating retractions and elongations. Neurite elongation was completed in a uniform time period of approximately 15 days despite the differences in elongation rate, maximum length, and latency to growth onset. The neurite branching morphology and manner of growth revealed in this study indicated that adult regenerating neurons can reproduce some elements of the final branching patterns in the absence of extrinsic cues, a capability which may ultimately contribute to the fidelity of reconnection seen in vivo.

Animals↗

PubMed2001

The three-dimensional architecture of retinal blood vessels in KK mice, with special reference to the smooth muscle cells and pericytes.

In the present study, the three-dimensional architecture of retinal vasculature was studied in KK mice, by combined use of resin injection, chemical treatment and scanning electron microscopy (SEM). In particular, Mercox/methylmethacrylate resin-injected eye tissues were subjected in sequence to NaClO immersion, ultrasonication and HCl treatment. The present technological innovation made possible SEM visualization of deeper retinal vasculature. In KK mice, the tunica media of stem arterioles and of first and second order branches consisted of a single layer of spindle-shaped smooth muscle cells provided with spine-like cytoplasmic processes. In addition, there occurred small triangular smooth muscle cells provided with slender cytoplasmic processes. The processes, giving off tiny secondary processes, overlapped with each other, thus forming assemblages around branching sites. Such a structure was particularly prominent for those branching sites where parent arterioles gave rise to their branches in a side arm-like pattern. The third (and occasionally fourth) order branches were surrounded by atypical smooth muscle cells, with considerable dimension of endothelial surface remaining uncovered. Capillary pericytes consisted of fusiform cell bodies and slender cytoplasmic processes. Smooth muscle cells of retinal venules differed from those of arterioles. They were stellate in shape, exhibiting several cytoplasmic processes.

Animals↗

PubMed1993

Spike conduction properties of T-shaped C neurons in the rabbit nodose ganglion.

The electrical activity of C-type neurons was recorded intracellularly in the rabbit nodose ganglion maintained in vitro. The initial segment of their axon is spirally wound close to the cell body and a primary branching point divides it into a central process (CP) projecting to the nucleus of solitary tract in the medulla oblongata and a peripheral process (PP) which conveys sensory inputs from the viscera. Stimulation of the CP induced either somatic ("S") spikes or low-amplitude axonal ("A") spikes ("A1" or "A2"). In some cases abrupt changes in the latency of "S" or "A" spikes (jumps) were observed by gradually increasing the stimulus intensity. They are discussed in relation to a secondary branching on the central axon located inside or near the ganglion. Collision experiments showed that antidromic "A" spikes are blocked at the primary bifurcation of the axon (T-shaped neuron). Stimulation of the PP induced either "S" spikes or high amplitude "A" spikes ("A3" or "A4"). Orthodromic spikes could be blocked either before or after the primary bifurcation. When blocking occurs after the bifurcation on the stem axon, the spike can invade the central axon without invading the soma. The study of the refractory periods of the two processes and the application of high frequency stimulation showed that the PP allows higher frequencies than the soma and the CP, and thus that branching and the CP act as low-pass filters. These data support the view that the primary branching point and the CP of these T-shaped cells represent a strategic area to modulate visceral afferent messages.

Action Potentials↗

PubMed2002

Attention-deficit/hyperactivity disorder (ADHD) behaviour explained by dysfunctioning reinforcement and extinction processes.

Inattentiveness, overactivity and impulsiveness are presently regarded as the main clinical symptoms of attention-deficit/hyperactivity disorder (ADHD). Inattention is, however, a characteristic of most psychiatric disorders. It is argued that the ADHD Inattentive subtype may have heterogeneous origins and be qualitatively different from the ADHD Hyperactive/Impulsive subtype. At the neurobiological level, ADHD symptoms may to a large extent be caused by a dysfunctioning dopamine system: A dysfunctioning meso-limbo-cortical dopamine branch will produce altered reinforcement and extinction processes, on a behavioural level giving rise to deficient sustained attention, hyperactivity, motor and cognitive impulsiveness. A dysfunctioning nigro-striatal dopamine branch will cause 'extrapyramidal' symptoms. Our model disentangles the behaviours usually explained by 'executive functions' into cognitive impulsiveness, motor impulsiveness and deficient motor control. The various dopaminergic branches may not be equally dysfunctional in all individuals with ADHD. Etiologically, dopamine dysfunctioning will probably mainly be genetically determined while sometimes be induced by environmental factors like drugs of abuse or pollutants, which may explain geographical differences in prevalence rates.

Animals↗

PubMed1989

Laminin and a basement membrane extract have different effects on axonal and dendritic outgrowth from embryonic rat sympathetic neurons in vitro.

We have characterized the effects of laminin and a basement membrane extract (BME) on the morphology of embryonic rat sympathetic neurons maintained in tissue culture in the absence of nonneuronal cells. Neurons were grown on polylysine-coated coverslips in the presence or absence of laminin or BME in serum-free medium. Axons were distinguished from dendrites using intracellular dye injections, immunocytochemistry, and [3H]uridine autoradiography. In short-term (less than or equal to 24 hr) culture, laminin had a potent neurite-promoting effect, causing increases in the number of processes, total neuritic length, and neuritic branching. In long-term (3-35 days) cultures chronically exposed to laminin, most (greater than 75%) neurons maintained supernumerary axons but failed to form dendrites. In contrast, most neurons (greater than 70%) grown in long-term culture on polylysine in the absence of laminin were unipolar, extending a single axon. BME caused sympathetic neurons to extend multiple (range, 1-15) dendrites. Morphometric measurements made after 1 month of exposure to BME indicated that the amount of dendritic growth that occurred in vitro was similar to that normally occurring during a comparable period in situ. BME did not cause changes in the number of axons per neuron or in the uptake of neurotransmitter. Preliminary characterization of the dendrite-promoting activity of BME suggests that it resides in extracellular matrix (ECM) molecules and not in low-molecular weight contaminants. These observations indicate that (1) axonal and dendritic growth may be differentially regulated by various constituents of the ECM, and (2) such process-specific interactions can significantly affect the morphological development of sympathetic neurons.

Animals↗

PubMed2026

Breast Cancer Recurrence Status Assessment in 5 Years Using Multimodal Integrated Learning: A Feasibility Study.

Despite advances in breast cancer detection and treatment, recurrence after curative therapy continues to impact long-term survival and quality of life. Therefore, early identification of high-risk patients is crucial to guide personalized treatment and follow-up strategies. Although genomic assays provide valuable prognostic insights, their high cost and limited accessibility hinder widespread adoption in clinical practice. Recent machine learning or deep learning approaches leveraging clinical, imaging, or multimodal data have shown promise but do not reflect real-world clinical scenarios. This study proposes a deep learning-based multimodal framework for predicting 5-year breast cancer recurrence using routinely collected clinical data. The framework consists of three main components. First, we adopted automated tumor segmentation with MedSAM to extract the tumor region from ultrasound images. The radiomics features are extracted from those tumor regions. Second, report features are extracted using a Med-Contrastive Pre-trained Transformers (MedCPT)-based approach incorporating predefined, clinically informed queries. Third, a multimodal integration model jointly processes image, radiomics, clinical features, and report features through modality-specific branches. The image branch employs the Ultrasound Foundation Model (USFM) as the backbone, while structured tabular data is processed using the FT-Transformer architecture. The features of all branches are fused using a mixture-of-experts (MoE)-based classifier, and the entire model is trained using a progressive fusion training strategy. Experimental results confirm the feasibility of using ultrasound images with tumor mask integration for recurrence prediction and demonstrate the additive value of integrating multiple data modalities through the proposed multimodal integration model. The final model for recurrence prediction achieved an AUC of 0.7540, accuracy of 74.61%, sensitivity of 70.41%, and specificity of 76.44%. This feasibility study's findings underscore the potential of the proposed multimodal deep learning framework to provide accessible, accurate, and generalizable recurrence risk prediction using routinely available clinical data, potentially supporting more informed treatment decisions and personalized post-treatment monitoring in real-world clinical practice.

Breast cancer recurrence↗

PubMed1987

Cholinergic neurons in the rabbit retina: dendritic branching and ultrastructural connectivity.

The structure and synaptic connectivity of rabbit retinal cholinergic neurons have been studied with an immunocytochemical technique for the localization of choline acetyltransferase (ChAT). Cholinergic processes ramified in two narrow strata within the inner plexiform layer; viewed in the plane of the retina, each immunoreactive stratum took the form of a polygonal meshwork with openings of about 20 microns. There was frequently an extensive matching of the patterns of the two strata, such that openings in one meshwork lay directly over openings of similar size in the other meshwork. It is hypothesized that the pattern of branching of these cholinergic processes is a reflection of the branching pattern of ganglion cell dendrites in the cholinergic strata. The proximal cholinergic stratum was examined ultrastructurally, and had the following characteristics: (1) ChAT-immunoreactive processes made large numbers of synaptic contacts with ganglion cell dendrites; often there were many such ganglion cell dendrites running past each other at various angles within the plane of the cholinergic stratum; (2) cholinergic processes were never observed presynaptic to any other type of inner plexiform process; (3) the principal input onto cholinergic processes was provided by bipolar cell axon terminals; (4) ganglion cell dendrites within the cholinergic stratum also received direct bipolar input; and (5) unidentified (i.e. non-cholinergic and probably non-GABAergic) amacrine cell processes were often found that were presynaptic to these same ganglion cell dendrites, and that also formed reciprocal contacts with bipolar axon terminals within these synaptic complexes.

Animals↗

PubMed1994

Plasma membrane fluidity during regeneration and atrophy of the rat liver following portal branch ligation.

Dynamic changes in liver plasma membrane fluidity caused by regeneration and atrophy were assessed in rats following portal branch ligation (PBL). The portal branch, which perfuses 70% of the liver, was ligated with 5-0 prolene, and liver plasma membranes were isolated by ultracentrifugation. The membrane fluorescence polarization was measured as an index of membrane fluidity using 1,6-diphenyl-1,3,5-hexatriene (DPH) as the probe dye. In nonligated lobes, a significant decrease in fluorescence polarization was observed 12 and 24 h after PBL (0.171 +/- 0.004, p < 0.01 and 0.165 +/- 0.005, p < 0.001, respectively) as compared to the controls (0.181 +/- 0.002). The fluorescence polarization values then gradually returned to near control levels. In contrast, in the ligated lobes, the fluorescence polarization had increased by 12 hours after PBL (0.196 +/- 0.002, p < 0.01), and remained significantly elevated (p < 0.01) for up to 1 week after PBL, gradually returning to control levels within 3 weeks. The membrane composition was also evaluated by analyzing the cholesterol/phospholipid (C/P) ratio. A significant increase in the C/P ratio was detected in the ligated lobes 12 h and 3 days after PBL, but there was no significant difference in fluorescence polarization values between nonligated lobes and controls. These results suggest that alterations in membrane fluidity play an important role in the regenerative and atrophic processes of the liver following portal branch ligation.

Animals↗

PubMed2003

Type III intermediate filament proteins interact with four-way junction DNA and facilitate its cleavage by the junction-resolving enzyme T7 endonuclease I.

The isolation from proliferating mouse and human embryo fibroblasts of SDS-stable crosslinkage products of vimentin with DNA fragments containing inverted repeats capable of cruciform formation under superhelical stress and the competitive effect of a synthetic Holliday junction on the binding of cytoplasmic intermediate filament (cIF) proteins to supercoiled DNA prompted a detailed investigation of the proteins' capacity to associate with four-way junction DNA and to influence its processing by junction-resolving endonucleases. Electrophoretic mobility shift analysis of reaction products obtained from vimentin and Holliday junctions under varying ionic conditions revealed efficient complex formation of the filament protein not only with the unstacked, square-planar configuration of the junctions but also with their coaxially stacked X-conformation. Glial fibrillary acidic protein (GFAP) was less efficient and desmin virtually inactive in complex formation. Electron microscopy showed binding of vimentin tetramers or octamers almost exclusively to the branchpoint of the Holliday junctions under physiological ionic conditions. Even at several hundredfold molar excess, sequence-related single- and double-stranded DNAs were unable to chase Holliday junctions from their complexes with vimentin. Vimentin also stimulated bacteriophage T7 endonuclease I in introducing single-strand cuts diametrically across the branchpoint and thus in the resolution of the Holliday junctions. This effect is very likely due to vimentin-induced structural distortion of the branchpoint, as suggested by the results of hydroxyl radical footprinting of Holliday junctions in the absence and the presence of vimentin. Moreover, vimentin, and to a lesser extent GFAP and desmin, interacted with the cruciform structures of inverted repeats inserted into a supercoiled vector plasmid, thereby changing their configuration via branch migration and sensibilizing them to processing by T7 endonuclease I. This refers to both plasmid relaxation caused by unilateral scission and, particularly, linearization via bilateral scission at primary and cIF protein-induced secondary cruciform branchpoints that were identified by T7 endonuclease I footprinting. cIF proteins share these activities with a variety of other architectural proteins interacting with and structurally modulating four-way DNA junctions. In view of the known and hypothetical functions of four-way DNA junctions and associated protein factors in DNA metabolism, cIF proteins as complementary nuclear matrix proteins may play important roles in such nuclear matrix-associated processes as DNA replication, recombination, repair, and transcription, with special emphasis on both the preservation and evolution of the genome.

Bacteriophage T7↗

PubMed1981

Ultrastructure of the crayfish stretch receptor in relation to its function.

The crayfish slow-adapting abdominal stretch receptor was fixed under the relaxed or stretched condition. During this procedure action potentials of the sensory neuron were recorded by a suction electrode. The receptor organ consists of a receptor muscle and a sensory neurons with its dendrites embedded in the connective tissue zone in the receptor muscle. From the cell body of the neuron, several "primary dendrites" arise, branch successively into "dendritic branches", and finally terminate as "dendritic tips," which are cylindrical processes of fairly uniform diameter. In contrast to the primary dendrites and the dendritic branches, the dendritic tips have neither mitochondria nor sheaths and are embedded in the connective tissue zone or apposed to the receptor muscle with a gap of about 15 nm. Microtubules and smooth ER are seen in all parts of the dendrites. When the receptor is stretched and then fixed with 1.6% glutaraldehyde in 0.12 M phosphate buffer (total osmolarity of this solution is isosmotic with the physiological solution), dendritic tips became more parallel to the long axis of the receptor muscle and showed marked deformation consisting of alternate regions of swelling and shrinkage, resulting in a bead-like appearance. When fixed with 1.6% glutaraldehyde in 0.2 M phosphate buffer (total osmolarity of this solution is hyperosmotic), the dendritic tips showed less tendency toward such deformation. These results suggest that the dendritic tip membrane is susceptible to stretch and might be the region where the generator potential is produced.

Animals↗

PubMed2000

Fatty-acid biosynthesis in a branched-chain alpha-keto acid dehydrogenase mutant of Streptomyces avermitilis.

Fatty-acid biosynthesis by a branched-chain alpha-keto acid dehydrogenase (bkd) mutant of Streptomyces avermitilis was analyzed. This mutant is unable to produce the appropriate precursors of branched-chain fatty acid (BCFA) biosynthesis, but unlike the comparable Bacillus subtilis mutant, was shown not to have an obligate growth requirement for these precursors. The bkd mutant produced only straight-chain fatty acids (SCFAs) with membrane fluidity provided entirely by unsaturated fatty acids (UFAs), the levels of which increased dramatically compared to the wild-type strain. The levels of UFAs increased in both the wild-type and bkd mutant strains as the growth temperature was lowered from 37 degrees C to 24 degrees C, suggesting that a regulatory mechanism exists to alter the proportion of UFAs in response either to a loss of BCFA biosynthesis, or a decreased growth temperature. No evidence of a regulatory mechanism for BCFAs was observed, as the types of these fatty acids, which contribute significantly to membrane fluidity, did not alter when the wild-type S. avermitilis was grown at different temperatures. The principal UFA produced by S. avermitilis was shown to be delta 9-hexadecenoate, the same fatty acid produced by Escherichia coli. This observation, and the inability of S. avermitilis to convert exogenous labeled palmitate to the corresponding UFA, was shown to be consistent with an anaerobic pathway for UFA biosynthesis. Incorporation studies with the S. avermitilis bkd mutant demonstrated that the fatty acid synthase has a remarkably broad substrate specificity and is able to process a wide range of exogenous branched chain carboxylic acids into unusual BCFAs.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗

PubMed2005

Comparison of immunoreactivity to serotonin, FMRFamide and SCPb in the gut and visceral nervous system of larvae, pupae and adults of the yellow fever mosquito Aedes aegypti.

In all life stages, the gut of the mosquito is innervated by a small number (typically 4) of central neurons immunoreactive to serotonin (SI). The serotonergic system appears to pass through metamorphosis largely intact, despite extensive remodeling of the gut. Axons immunoreactive to antibodies raised against molluscan FMRFamide (RF-I) constitute peptidergic innervation that anatomically parallels the serotonergic system. In the larva, two clusters of 3 neurons project to the anterior regions of the gut, whereas in the pupa and adult, typically two large RF-I neurons located next to the esophagus send several processes posteriorly. In adults, these neurons branch throughout the diverticula and anterior stomach. In pupae, but not in larvae or adults, the gut RF-l system coexpresses reactivity to antibodies raised against a member of another peptide family, molluscan small cardioactive peptide b (SCP-I). SCP-I immunoreactivity is localized independently of RF-l immunoreactivity in the ganglia of all stages and in neurons that project along the gut of the adult. We did not find any colocalization of S-I and the peptide markers. Distinct populations of enteroendocrine cells populate different regions of the gut at different life stages. Changes in staining pattern suggest that these cells are replaced at metamorphosis along with the other gut cells during the extensive remodeling of the tract. Distributed in the gut epithelium are subpopulations that express either RF-I or SCP-I; a small fraction of these cells bind antibodies to both peptides. The stomachs of adult females are larger than those of males, and the numbers of SCP-I and RF-I enteroendocrine cells are proportionately greater in females. In all the life stages, the junctions between different regions of the gut are the focus of regulatory input. The larval cardiac valve possesses a ring of cells, the necklace cells, which appear to receive extensive synaptic inputs from both the serotonergic system and the peptidergic system. Another focus of control is the pyloric valve, which is encircled by axon-like processes. The immunoreactive pattern of this region differs across life stages, expressing SCP-I in larvae, S-I in pupae, and both SCP-I and RF-I in adults.

Aedes↗

PubMed1975

Dynamics of liver glycogen: the topochemistry of glycogen synthesis, glycogen content and glycogenolysis under the experimental conditions of glycogen accumulation and depletion.

Using histochemical techniques the glycogen content and the activities of glycogen synthetase (UDPGGT) and phosphorylase were studied in the livers of 106 golden hamsters under following experimental conditions; a) starvation of 16, 36, 48, 72, and 96 hours: b) alloxan-diabetes. Starvation leads to a depletion of liver glycogen during the first 48 hours, which is finally restricted to zone 3 of the liver acinus. After starvation of 72 and 96 hours a new glycogen accumulation is demonstrable in the microvasculatory periphery of the acinus (zone 3 and 2). The process of glycogen depletion is characterized in the beginning by a high phosphorylase activity in all zones of the acinus, later only in the forefield of glycogen content. The weak activity of glycogen synthetase is mainly restricted to zone 3. All phases of glycogen depletion are to be found in alloxan diabetic animals, too. Out of 45 hamsters 23 showed an extreme depletion of glycogen; typical for this situation is a weak or absent glycogen synthetase activity in zone 3 and a broad field of phosphorylase activity in zones 1 and 2. The short stimulation by insulin leads to a considerable increase of glycogen synthetase activity at the portally oriented border of the glycogen area and to a shift of the moderate phosphorylase activity of zone 1. Thus the histochemical characteristics of glycogen depletion are: a shift of the reduced glycogen content in direction of the microvasculatory periphery of the liver acinus (zone 3), caused by a high phosphorylase activity in the portal forefield, while glycogen synthetase activity is low in the glycogen area. The histochemical characteristics of glycogen accumulation are: after a short phase of glycogen synthesis in all hepatocytes a moderate phosphorylase activity in zone 1 leads to a mobilization of the portal glycogen deposits and to an increasing accumulation of glycogen in the peripheral part of the acinus. At the portally oriented border of the glycogen area a high synthetase activity leads to a broadening of the glycogen area in direction of the portal branches. At the end of this process the "normal" pattern of the liver acinus occurs: all hepatocytes are filled with glycogen, the glycogen enzymes are restricted to the periportal border of zone 1.

Animals↗

PubMed1989

Postnatal development of glial fibrillary acidic protein (GFAP) immunoreactivity in pituicytes and tanycytes of the Mongolian gerbil (Meriones unguiculatus).

The postnatal development (day of birth up to the end of the third month) of neurohypophyseal pituicytes and tanycytes of the median eminence (ME) and the medial basal hypothalamus (MBH) was studied immunohistochemically in the Mongolian gerbil (Meriones unguiculatus) with antibodies directed against glial fibrillary acidic protein (GFAP; the major protein subunit of glial filaments). Weak GFAP-immunoreactivity (IR) was scattered in the neural lobe (NL), the ME and the lining of the ventral 3rd ventricle at the first postnatal days. By the end of the second postnatal week, the intensity of the IR had reached a level comparable to that of adult animals. Generally, in the whole neurohypophysis a cytoarchitectonic pattern, which essentially corresponded to adult conditions, was reached around the beginning of the second month. During the first week postnatum, solely perinuclear stainings, mostly unipolar pituicytes with short processes and isolated fibers were discernible in the NL. In the course of the second and third postnatal week, a growing number of the densely arranged pituicytes appeared in form of bi- and multipolar cells. Thickness and length of pituicyte processes, as well as their degree of branching, increased progressively in the first month. The number of GFAP-positive tanycytes in the ventral 3rd ventricle and in the ME most markedly augmented in the first week postnatum. In the MBH, long tanycyte processes emerged from the ventricular lining to cross the arcuate nucleus in large bows, delimiting groups of neurons. Ependymal and subependymal tanycytes in the ME gave rise to radial processes extending to the external zone.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed2006

Cotranscriptional recognition of human intronic box H/ACA snoRNAs occurs in a splicing-independent manner.

Processing from pre-mRNA introns is a widespread mechanism to generate human box C/D and H/ACA snoRNAs. Recent studies revealed that an optimal position relative to the 3' splice site is important for efficient processing of most box C/D snoRNAs and that assembly of box C/D snoRNPs is stimulated by splicing factors likely bound to the branch point region. Here we have investigated the processing of another major class of human intron-encoded RNAs, the box H/ACA snoRNAs. Analysis of 80 H/ACA RNA genes revealed that human H/ACA RNAs possess no preferential localization close to the 3' or 5' splice site. In vivo processing experiments confirmed that H/ACA intronic snoRNAs are processed in a position-independent manner, indicating that there is no synergy between H/ACA RNA processing and splicing. We also showed that recognition of intronic H/ACA snoRNAs and assembly of pre-snoRNPs is an early event that occurs during transcription elongation parallel with pre-mRNA splice site selection. Finally, we found that efficient processing and correct nucleolar localization of the human U64 H/ACA snoRNA requires RNA polymerase II-mediated synthesis of the U64 precursor. This suggests that polymerase II-associated factors direct the efficient assembly and determine the correct subnuclear trafficking of human H/ACA snoRNPs.

Cell Cycle Proteins↗