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PubMed2002

Projection to the inferior colliculus from the basal nucleus of the amygdala.

This report describes a projection from the amygdala, a forebrain center mediating emotional expression, to the inferior colliculus (IC), the midbrain integration center of the ascending auditory system. In the IC of mustached bats (Pteronotus parnellii) and pallid bats (Antrozous pallidus), we placed deposits of retrograde tracers at physiologically defined sites and then searched for retrogradely labeled somata in the forebrain. Labeling was most sensitive in experiments using cholera toxin B-subunit as tracer. We consistently observed retrograde labeling in a single amygdalar subdivision, the magnocellular subdivision of the basal nucleus (Bmg). The Bmg is distinctive across mammals, containing the largest cells in the amygdala and the most intense acetylcholinesterase staining. Labeled amygdalar cells occurred ipsilateral and contralateral to IC deposits, but ipsilateral labeling was greater, averaging 72%. Amygdalar labeling was observed after tracer deposits throughout the IC, including its central nucleus (ICC). In comparison, labeling in the auditory cortex (layer V) was heavily ipsilateral (averaging 92%). Cortical labeling depended on the location of IC deposits: dorsomedial deposits resulted in the most labeled cells, whereas ventrolateral deposits labeled few or no cortical cells. Cortical labeling occurred after several deposits in the ICC. Across experiments, the average number of labeled cells in the amygdala was similar to that in the auditory cortex, indicating that the amygdalocollicular projection is significant. The results demonstrate a direct, widespread projection from the basal amygdala to the IC. They also suggest the presence of a rapid thalamoamygdalocollicular feedback circuit that may impose emotional content onto processing of sensory stimuli at a relatively low level of an ascending sensory pathway.

Acoustic Stimulation↗
PubMed2006

Diversity of laminar connections linking periarcuate and lateral intraparietal areas depends on cortical structure.

Lateral prefrontal and intraparietal cortices have strong connectional and functional associations but it is unclear how their common visuomotor, perceptual and working memory functions arise. The hierarchical scheme of cortical processing assumes that prefrontal cortex issues 'feedback' projections to parietal cortex. However, the architectonic heterogeneity of these cortices raises the question of whether distinct areas have laminar-specific interconnections underlying their complex functional relationship. Using quantitative procedures, we showed that laminar-specific connections between distinct prefrontal (areas 46 and 8) and lateral intraparietal (LIPv, LIPd and 7a) areas in Macaca mulatta, studied with neural tracers, varied systematically according to rules determined by the laminar architecture of the linked areas. We found that axons from areas 46 and rostral 8 terminated heavily in layers I-III of all intraparietal areas, as did caudal area 8 to area LIPv, suggesting 'feedback' communication. However, contrary to previous assumptions, axons from caudal area 8 terminated mostly in layers IV-V of LIPd and 7a, suggesting 'feedforward' communication. These laminar patterns of connections were highly correlated with consistent differences in neuronal density between linked areas. When neuronal density in a prefrontal origin was lower than in the intraparietal destination, most terminations were found in layer I with a concomitant decrease in layer IV. The opposite occurred when the prefrontal origin had a higher neuronal density than the target. These findings indicate that the neuronal density of linked areas can reliably predict their laminar connections and may form the basis of understanding the functional complexity of prefrontal-intraparietal interactions in cognition.

Analysis of Variance↗
PubMed1996

Specificity in the axonal connections of layer VI neurons in tree shrew striate cortex: evidence for distinct granular and supragranular systems.

Pyramidal neurons in layer VI of striate cortex are the source of descending projections to the lateral geniculate nucleus (LGN) as well as a major source of axon terminals in the layers of striate cortex that receive LGN projections. This study examines how the connections of layer VI neurons are arranged with respect to the functionally distinct classes of neurons that compose their cortical and subcortical targets. By placing injections of biocytin into layer VI of tree shrew striate cortex, we identified two sublayers that differ in their intracortical and thalamic connections. Neurons in the upper part of layer VI, layer VIa, terminate in cortical layer IV, whereas those in the lower part of layer VI, layer VIb, terminate throughout the supragranular layers, layers I-III. The selectivity of layer VI subdivisions for the granular and supragranular layers is also evident in their descending projections. Neurons in layer VIa terminate preferentially in the LGN layers that supply layer IV (LGN layers 1, 2, 4, and 5), whereas neurons in layer VIb terminate in the LGN layers that supply layers I-III (LGN layers 3 and 6) and in the pulvinar nucleus. Additional subclasses of layer VIa neurons were identified based on the restriction of their terminal fields to narrow subtiers within layer IV. By influencing the activity of distinct populations of cortical neurons and the thalamic neurons that supply them, layer VI neurons could exert a powerful influence on the flow of activity in functionally distinct cortical circuits.

Animals↗
PubMed2005

Downregulation of NANOG induces differentiation of human embryonic stem cells to extraembryonic lineages.

The homeobox transcription factor Nanog has been proposed to play a crucial role in the maintenance of the undifferentiated state of murine embryonic stem cells. A human counterpart, NANOG, has been identified, but its function and localization have not hitherto been described. We have used a combination of RNA interference and quantitative real-time polymerase chain reaction to study NANOG in human embryonic stem and embryonic carcinoma cells. Transfection of NANOG-specific small interfering RNAs reduced levels of NANOG transcript and protein and induced activation of the extraembryonic endoderm-associated genes GATA4, GATA6, LAMININ B1, and AFP as well as upregulation of trophectoderm-associated genes CDX2, GATA2, hCG-alpha, and hCG-beta. Immunostaining of preimplantation human embryos showed that NANOG was expressed in the inner cell mass of expanded blastocysts but not in earlier-stage embryos, consistent with a role in the maintenance of pluripotency. Taken together, our findings suggest that NANOG acts as a gatekeeper of pluripotency in human embryonic stem and carcinoma cells by preventing their differentiation to extraembryonic endoderm and trophectoderm lineages.

Blastocyst↗
PubMed2003

Lung growth and development.

The organogenesis of lung involves several complex mechanisms, including interactions between cells originating from two germ layers--endoderm and mesoderm. Regulation of lung branching morphogenesis with reference to its architecture, growth pattern, differentiation, interactions between epithelium and mesenchyme and / or endothelium, as well as genes regulating these processes have been addressed by the pulmonary biologists through careful molecular biology and genetic experimental approaches. The mammalian lung develops by outpouching from the foregut endoderm as two lung buds into the surrounding splanchnic mesenchyme. Several different regions of the foregut are specified to develop into different thoracic and visceral organs. The lung-buds further elongate and branch, and the foregut longitudinally gets separated into esophagus and trachea. In rodents (mice and rats), this occurs around embryonic day 11, where the right lung bud develops into four different lobes and left lung develops as a single lobe. In humans, these processes occur by 3-4 weeks of embryonic development, where the right lung is a trilobar lung and the left lung is a bilobar lung. Several generations of dichotomous branching occur during embryonic development, followed by secularization and alveolarization pre- and post-natally, which transform a fluid-filled lung into an air-breathing lung able to sustain the newborn. During these different developmental stages from embryonic to newborn stage, the lung architecture undergoes profound changes, which are marked by a series of programmed events regulated by master genes (e.g., homeobox genes), nuclear transcription factors, hormones, growth factors and other factors. These programmed events can be altered by undesirable exposure to overdoses of hormones/vitamins/growth factors, synthetic drugs, environmental toxins, radiation and other agents. In the recent years molecular techniques have opened avenues to study specific functions of genes or their products (proteins) in vivo or in vitro at a cellular or an organelle level, some of these include targeted disruption, knock-in / knock-out genes, in vitro mutagenesis, use of sense and anti-sense oligonucleotides. Some of these aspects with reference to regulation of normal lung development and growth and a specific example of pulmonary hypoplasia as an abnormal lung formation are discussed in this review.

Animals↗
PubMed1985

Identification of a cell surface glycoprotein family of olfactory receptor neurons with a monoclonal antibody.

A monoclonal antibody (Mab) has been developed which recognizes a family of cell surface glycoproteins found in high levels of rat olfactory receptor neurons. This Mab, designated 2B8, was produced by the fusion of X63-Ag8.653 myeloma cells and spleen cells of a mouse immunized with PC12 rat pheochromocytoma cells. Immunofluorescence analyses of cryostat sections of neonatal olfactory epithelium show prominent 2B8 binding to receptor neurons. Within the olfactory bulb only the glomerular and olfactory nerve layers show 2B8 binding. All other neural structures in the main olfactory bulb have background levels of reactivity. Analyses of 2B8 binding to particulate protein preparations from several central and peripheral nervous system components demonstrated highest 2B8 antigen specific activity in olfactory bulb and epithelium and detectable levels in dorsal root ganglia (DRG), whole cerebrum, cerebellum, and brainstem. However, 2B8 antigen could not be detected in non-olfactory structures by immunofluorescence. Some non-neural tissues also had the ability to bind 2B8 Mab in the particulate protein radioimmunoassay. In order to compare the 2B8-reactive molecules found in each tissue, Mab was applied to polyacrylamide gels of unlabeled membrane proteins. A family of molecules with diverse molecular weights was found. Some were unique to individual tissues whereas others were shared among tissues. Olfactory bulb and epithelium had a unique band with Mr = 215,000 and another band with Mr = 142,000. The 142,000-dalton band was also found with PC12 cells. PC12 cells also had several bands of lesser molecular weight, including 51,000 and 43,000. Testes membranes had immunoreactive bands only at Mr = 46,000 and 43,000. Bone marrow, perinatal liver, and DRG each expressed a single 2B8-reactive band with Mr = approximately 114,000. Salivary gland had four reactive bands, two common to it and only PC12 cells, the 114,000-dalton band which is similar to that found in adult rat bone marrow and DRG, and a unique band at Mr = 152,000. 2B8 immunoprecipitates of olfactory bulb and epithelium were analyzed for glycosyl groups by lectin reactivity. Wheat germ agglutinin and Ricinus communus agglutinin I bound the 2B8 antigens using two distinct assay methods. This suggests that the 2B8 antigens recognized in the olfactory system are glycoproteins having sialic acid and D-galactosyl components.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1997

Immunodetection of amelogenin-like proteins in the ganoine of experimentally regenerating scales of Calamoichthys calabaricus, a primitive actinopterygian fish.

BACKGROUND: The account of the present study is to test our previous hypothesis that ganoine, a highly mineralized layer found at the scale surface of primitive actinopterygian fish, could be homologous with the enamel covering the crown of vertebrate teeth. METHODS: Immunocytochemical techniques have been carried out on regenerating scales of a primitive polypterid, Calamoichthys calabaricus, with three antibodies to mammalian amelogenins. RESULTS: The present study provides the first evidence that ganoine contains molecules which cross-react with mammalian amelogenin proteins. CONCLUSIONS: This result is consistent with our previous findings that ganoine and enamel can be considered as homologous tissues. Moreover, the presence in ganoine of a primitive actinopterygian of amelogenin-like proteins, which share epitopes with amelogenins of mammalian enamel, indicates that the gene(s) coding for these proteins appeared earlier than previously suggested and supports the hypothesis that amelogenins show a highly conserved structure through vertebrate evolution.

Amelogenin↗
PubMed1989

Calmodulin immunocytochemistry in rat incisor enamel organ through its life cycle.

The enamel organ of the growing rat incisor was perfusion-fixed with a mixture of formaldehyde and glutaraldehyde and processed for indirect immunogold labeling of calmodulin on post-embedded ultrathin sections. Throughout the zones of presecretion, secretion, and maturation of enamel, specific protein A-immunogold labeling was localized on polyribosomes and those attached to endoplasmic reticulum, mitochondria, nuclear chromatin, phagolysosomes, and cytoplasm adjacent to the plasma membrane, and tonofilaments associated with desmosomes of ameloblasts and cells of outer layer of enamel organ. Golgi membranes, condensing vacuoles, secretion granules, primary lysosomes, and micropinocytotic coated vesicles were hardly labeled. In the presecretion zone, the basal lamina of the preameloblasts and the matrix vesicles and collagen fibrils of the predentin matrix were not immunoreactive. Tomes' process of secretory ameloblast and adjacent enamel crystals were labeled. In addition to the above immunoreactive structures, some phagolysosomes, ferritin granules, and the cytoplasm of the ruffled border zone of maturation ameloblast contained immunogold particles. In control sections incubated with either protein A-gold complex alone, or antiserum preabsorbed with an excess of calmodulin and protein A-gold complex, only a few gold particles were observed to be randomly associated with the tissues. These results indicate that calmodulin is present in the cells of the enamel organ through all stages of amelogenesis. Its wide distribution is consistent with its involvement in various cytoplasmic functions.

Ameloblasts↗
PubMedDate not supplied

Cortical structure predicts the pattern of corticocortical connections.

Cortical areas are linked through pathways which originate and terminate in specific layers. The factors underlying which layers are involved in specific connections are not well understood. Here we tested whether cortical structure can predict the pattern as well as the relative distribution of projection neurons and axonal terminals in cortical layers, studied with retrograde and anterograde tracers. We used the prefrontal cortices in the rhesus monkey as a model system because their laminar organization varies systematically, ranging from areas that have only three identifiable layers, to those that have six layers. We rated each prefrontal area based on the number and definition of its cortical layers (level 1, lowest; level 5, highest). The structural model accurately predicted the laminar pattern of connections in approximately 80% of the cases. Thus, projection neurons from a higher-level cortex originated mostly in the upper layers and their axons terminated predominantly in the deep layers (4-6) of a lower-level cortex. Conversely, most projection neurons from a lower-level area originated in the deep layers and their axons terminated predominantly in the upper layers (1-3) of a higher-level area. In addition, the structural model accurately predicted that the proportion of projection neurons or axonal terminals in the upper to the deep layers would vary as a function of the number of levels between the connected cortices. The power of this structural model lies in its potential to predict patterns of connections in the human cortex, where invasive procedures are precluded.

Animals↗
PubMed1986

A transient pyramidal tract projection from the visual cortex in the hamster and its removal by selective collateral elimination.

During the early postnatal development of the neocortex in rats there is an axonal projection from the occipital cortex (which includes the visual cortex) to the spinal cord which is subsequently completely removed through a process of selective collateral elimination. In order to determine whether a similar phenomenon occurs during the development of the hamster cortex, we have injected the retrogradely transported fluorescent dye Fast Blue (FB) into the pyramidal decussation of hamsters at various ages. In adult hamsters such an injection results in a band of labeled neurons confined to layer V and to about the rostral two-thirds of the neocortex; no labeled cells are seen in the occipital cortex. However, a similar FB injection made during the first postnatal week results after a 4-day survival in a continuous band of FB-labeled layer V neurons spread throughout the tangential extent of the neocortex, including the occipital cortex. A similar continuous band of FB labeled layer V neurons is seen throughout the tangential extent of the neocortex including the occipital region in hamsters injected during the first postnatal week but allowed to survive until the fourth week (i.e., after the restriction of the widespread neonatal pattern has occurred). Injections of the anterograde tracer wheat germ agglutinin conjugated to horseradish peroxidase made into the occipital cortex, or for comparison, into more rostral cortical regions in hamsters ranging in age from neonates to adults, reveal that the extension of pyramidal tract axons is staggered along the anterioposterior axis of the cortex such that axons originating from the posterior regions lag behind those arising from more rostral areas. The transient occipital projection appears to reach a maximum around the end of the first postnatal week: a large number of labeled occipital axons is seen in the medullary pyramidal tract, and some of these can be followed through the pyramidal decussation and into the dorsal funiculus of the spinal cord. Injections into the occipital cortex on P16 label only a few fibers in the medullary pyramidal tract, and none is labeled in hamsters injected as adults.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1999

Differential expression of VegT and Antipodean protein isoforms in Xenopus.

The VegT/Antipodean (Apod) gene is important for germ layer formation in Xenopus. To investigate the role of this gene at the protein level, as opposed to the RNA level, we have generated affinity purified polyclonal antibodies to Apod, and for comparison, to the other early T-box proteins Xbrachyury and Eomesodermin. An anti-VegT/Apod antibody reveals that there are two protein isoforms in Xenopus, one that we refer to as VegT and a smaller molecular weight isoform that we refer to as Apod. These isoforms have different N-terminal domains resulting from developmentally regulated alternative splicing of a primary transcript arising from a single VegT/Apod gene. VegT is maternally expressed. Its translation is blocked during oogenesis but the protein is present from the egg until gastrulation in the presumptive endoderm. There is no evidence for zygotic expression of this isoform. Conversely, the Apod protein isoform is expressed only after the onset of zygotic transcription in the presumptive mesoderm and is inducible by activin. We conclude that the developmental role of VegT/Apod is mediated by two different proteins, with entirely different patterns of expression and response to growth factors.

Activins↗
PubMed1983

Immunolocalization of a novel, cytoskeleton-associated polypeptide of Mr 230,000 daltons (p230).

Antibodies were raised against a cytoskeleton-associated, nonphosphorylated, 230,000-dalton bovine lens polypeptide (designated p230), and rendered monospecific by using a novel immunoaffinity technique. In immunofluorescence and electron microscopy of cultured fibroblasts, as well as of various other cells (endothelial, epithelial, lenticular, monocytes, neuroblastoma cells) and tissues (human kidney and liver), p230 was localized as a distinct subplasmalemmal layer in the peripheral cytoplasm of the cells. It constituted less than 0.3% of the total cellular protein in cultured fibroblasts and was not extractable with Triton X-100. In detergent-extracted cytoskeletal preparations of cultured fibroblasts, p230 remained as an elaborate peripheral network that showed a distribution distinctly different from that of the major cytoskeletal structures, stress fibers, cortical myosin, vinculin, and intermediate filaments (IF). The distribution was not dependent on the presence of intact stress fibers or microtubules, as shown by double-fluorescence microscopy of cells exposed to cytochalasin B or cultured in the presence of monensin and of cold-treated cells. Upon demecolcine-induced reorganization of intermediate filaments, however, the localization of p230 was rapidly altered to a dense plaque underneath the perinuclear aggregate of intermediate filaments. On the other hand, p230 seemed to colocalize with the detergent-resistant cell surface lamina, visualized in fluorescence microscopy with fluorochrome-coupled wheat germ agglutinin-lectin. The results suggest that p230 is part of a cell surface- and cytoskeleton-associated subplasmalemmal structure that may play an important role in cell surface-cytoskeleton interaction in various cells both in vitro and in vivo.

Animals↗
PubMed1985

Expression sequences of cell adhesion molecules.

A reexamination of the expression of cell adhesion molecules (CAMs) during the development of the chicken embryo was carried out using more sensitive immunocytochemical techniques than had been used previously. While the previously determined sequence of CAM expression was confirmed, neural CAM (N-CAM) was also detected on endodermal structures such as the lung epithelium, gut epithelium, and pancreas and on budding structures such as the pancreatic duct and gall bladder. It was also found on ectodermal derivatives of the skin. In most of these sites, N-CAM expression was transient, but in the chicken embryo lung, the epithelium remained positive for N-CAM and liver CAM (L-CAM) into adult life. Thus, at one time or another, both of these primary CAMs can be expressed on derivatives of all three germ layers. At sites of embryonic induction, epithelial cells expressing both L-CAM and N-CAM, or L-CAM only, were apposed to mesenchymal cells expressing N-CAM. Examples included epiblast (NL) and notochord (N); endodermal epithelium (NL) and lung mesenchyme (N); Wolffian duct (NL) and mesonephric mesenchyme (N); apical ectodermal ridge (NL) and limb mesenchyme (N); and feather placode (L) and dermal condensation (N). The cumulative observations indicate that cell surface modulation of the primary CAMs at induction sites can be classified into two modes. In mode I, expression of N-CAM (or both CAMs) in mesenchyme decreases to low amounts at the cell surface, and then N-CAM is reexpressed. In mode II, one or the other CAM disappears from epithelia expressing both CAMs. As a result of the primary processes of development, collectives of cells linked by N-CAM and undergoing modulation mode I are brought into the proximity of collectives of cells linked by L-CAM plus N-CAM or by L-CAM undergoing modulation mode II. Such adjoining cell collectives or CAM couples were found at all sites of embryonic induction examined.

Amino Acid Sequence↗
PubMed1981

Morphogenetic behaviour of the rat embryonic ectoderm as a renal homograft.

Halves of transversely or longitudinally cut primary ectoderm of the pre-primitive streak and the early primitive streak rat embryonic shield developed after 15-30 days in renal homografts into benign teratomas composed of various adult tissues, often in perfect organ-specific associations. No clear difference exists in histological composition of grafted halves of the same embryonic ectoderm. The primary ectoderm of the pre-primitive streak rat embryonic shield grafted under the kidney capsule for 2 days displayed an atypical morphogenetic behaviour, characterized by diffuse breaking up of the original epithelial layer into mesenchyme. Some of these cells associated into cystic or tubular epithelial structures. The definitive ectoderm of the head-fold-stage rat embryo grown as renal homograft for 1-3 days gave rise to groups of mesenchymal cells. These migrated from the basal side of the ectoderm in a manner which mimicked either the formation of the embryonic mesoderm or the initial migration of neural crest cells. This latter morphogenetic activity was retained in the entire neural epithelium of the early somite embryo but was only seen in the caudal open portion of the neural groove at the 10- to 12-somite stage. The efficient histogenesis in grafts of dissected primary ectoderm and the atypical morphogenetic behaviour of grafted primary and definitive rat embryonic ectoderm were discussed in the light of current concepts on mosaic and regulative development, interactive events during embryogenesis and positioning and patterning of cells by controlled morphogenetic cell displacement.

Animals↗
PubMed1983

Continuous-flow cultures as in vitro models of the ecology of large intestinal flora.

An anaerobic continuous-flow (CF) culture method has been developed which reproduces a number of bacterial interactions that occur in the large intestine of mice. These were determined in the following ways. (i) Bacterial counts in smears stained with 37 specific fluorescent antisera showed that the numeric balance between 37 strict anaerobes isolated from conventional mice was maintained in CF culture of conventional mouse flora in the same manner as in conventional mice. (ii) Mixed populations of various complexity of bacteria isolated from conventional mice were able to suppress Escherichia coli populations to similar levels in gnotobiotic mice and in CF cultures. (iii) Contents of CF cultures when fed to germfree mice were found to redress the germfree abnormalities studied, namely, cecal size and size of the E. coli population. Furthermore, dense layers of bacterial growth formed on the wall of CF cultures of mouse cecal flora, in a manner analogous to the colonization of mouse large intestinal mucosa. In the absence of such bacterial layers, the culture no longer exhibited these interactions. Because of the complexity and diversity of the interactions studied it is highly probable that at least the major underlying ecological control mechanisms operating in the culture model resemble those of the mouse intestine. We speculate that the somewhat surprising similarity between the ecology of the mouse large intestine and that of a CF culture in a glass vessel is due to the fact that both are dominated by thick layers of complex bacterial flora, the composition of which is controlled by their metabolic activities and by their relative ability to adhere to each other.

Anaerobiosis↗
PubMed2000

Auditory thalamocortical projections in the cat: laminar and areal patterns of input.

Thalamocortical projections were studied in adult cats using biotinylated dextran amines, wheat germ agglutinin conjugated to horseradish peroxidase, and autoradiography with tritiated leucine and/or proline. The input from 7 architectonically defined nuclei to 14 auditory cortical fields was characterized qualitatively and quantitatively. The principal results were that 1) every thalamic nucleus projected to more than 1 field (range, 4-14 fields; mean, 7 fields); 2) only the projection from the ventral division to some primary fields (primary auditory cortex and posterior auditory cortex) had a periodic, clustered distribution, whereas the input from other divisions to nonprimary areas was continuous; 3) layers III-V received >85% of the total axonal profiles; 4) in most experiments, five or more layers were labeled; 5) the projections to nonprimary auditory areas had many laterally oriented axons; 6) the heaviest input to layer I in all experiments was usually in its upper half, suggesting a sublaminar arrangement; 7) the largest axonal trunks (up to 6 microm in diameter) arose from the medial division and ended in layer Ia, where they ran laterally for long distances; 8) there were three projection patterns: type 1 had its peak in layers III-IV with little input to layer I, and it arose from the ventral division and the dorsal superficial, dorsal, and suprageniculate nuclei of the dorsal division; type 2 had heavy labeling in layer I and less in layers III-IV, arising from the dorsal division nuclei primarily, especially the caudal dorsal and deep dorsal nuclei; and type 3 was a trimodal concentration in layers I, III-IV, and VI that originated chiefly in the medial division and had the lowest density of labeling; and 9) the quantitative profiles with the three methods were very similar. The results suggest that the subdivisions of the auditory thalamus have consistent patterns of laminar distribution to different cortical areas, that an average of five or more layers receive significant input in a specific area, that a given thalamic nucleus can influence areas as far as 20 mm apart, that the first information to arrive at the cortex may reach layer I by virtue of the giant axons, and that several laminar patterns of auditory thalamocortical projection exist. The view that the auditory thalamus (and perhaps other thalamic nuclei) serves mainly a relay function underestimates its many modes for influencing the cortex on a laminar basis.

Animals↗
PubMed1989

Lectinohistochemistry of human bladder cancer: loss of lectin binding structures in invasive carcinomas.

With the purpose of studying changes in the expression of glycoconjugate structures in urothelium, nine different lectins (PNA, WGA, VFA, GSA II, STA, UEA I, LCA, DBA and HPA) with specificity for mono- or oligo-saccharides were used on formalin-fixed, paraffin-embedded tissue sections from 47 patients who had undergone surgical resection for bladder tumors and on normal urothelial biopsies from 10 patients. The tumors were graded and a lectinohistochemical method using biotinylated lectins and avidin-biotin-peroxidase complex was used to demonstrate the lectin binding. Positive staining reactions of cells in cytoplasm and on membranes were evaluated in the basal, the intermediate, and the luminal cell layers, respectively. In both normal and atypical urothelium lectin binding predominated in the luminal cell layer and decreased towards the basal cell layer. In normal urothelium all lectins stained greater than 66% of the cells in the luminal cell layer in cytoplasm and between 5 and 100% of the cells on membranes depending on the lectin used. A gradual loss of lectin-binding structures was seen with increasing grade of atypia. The range of this decrease varied considerably from one lectin to another, but it was consistently found that the percentage of cells stained in cytoplasm and on membranes decreased. A significantly lower percentage of cells stained in cytoplasm was found in invasive tumor cell-islands compared to normal urothelium. In invasive tumor cell-islands staining of cells on membranes was completely absent, except for HPA lectin that stained less than 10% of the cells. In conclusion, we demonstrate a dramatic decrease in lectin-binding carbohydrate structures associated with urothelial malignant progression.

Carcinoma, Transitional Cell↗
PubMed2005

Identification of cis-regulatory elements involved in transcriptional regulation of the sea urchin SpFoxB gene.

The SpFoxB gene is transiently expressed first in the mesoderm, then in the endoderm and oral ectoderm during sea urchin gastrulation. Perturbations of a number of proteins involved in endomesoderm specification have been shown to alter the mRNA levels of SpFoxB, but the cis-regulatory elements required for expression of SpFoxB have not been examined. In order to investigate this, we have screened the SpFoxB gene for sequences that can drive its expression. Both positive and negative cis-regulatory elements were found to be present. An enhancer was found that contains four GATA sites and four YY1 sites clustered within 210 base pairs (bp), as well as three lef/tcf binding sites. Electrophoretic mobility shifts indicate that the lef/tcf sites bind a complex of proteins that include beta-catenin in early cleavage, but not during subsequent stages of development. The GATA and YY1 sites bind nuclear proteins prior to SpFoxB transcription, and this binding diminishes coincident with cessation of transcription. Deletion of the GATA/YY1 sites causes a significant decrease in transcription. The DNA binding site of the SpFoxB protein has been determined, and Fox binding sites are found within the 5' UTR of SpFoxB.

Animals↗