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Border effects on brightness: a review of findings, models and issues.

This paper presents a summary of experimental findings, theoretical models and unresolved issues regarding border effects on brightness, of which the Cornsweet illusion (Cornsweet, 1970 Visual Perception. Academic Press: New York) is the best-known example. It is argued that no current theoretical model completely accounts for the wide variety of effects described. Contrast sensitivity function (CSF) models can explain many low-contrast, but not high-contrast, border effects. Lightness integration models based on Land and McCann's retinex theory (Land and McCann, 1971. J. Opt. Soc. Am. 61, pp. 1-11) have the advantage over CSF models in that they predict transitivity of border effects where they are found to occur. However, they fail to predict the appearance of a variety of Cornsweet-like figures, have never been tested with relatively high contrast versions of those figures, and have only been implemented by qualitative demonstration. It is argued that edge-detector models are potentially the most promising theoretical candidates but, as with lightness-integration models, they have invariably relied on qualitative demonstrations and have only dealt with low-contrast border effects. A computational edge-detector model which predicts the appearance of both high and low contrast Cornsweet figures is proposed and its advantages over other models, as well as its current limitations, are discussed. The final section discusses the neural locus for border effects in brightness.

Animals↗

Rhombomere rotation reveals that multiple mechanisms contribute to the segmental pattern of hindbrain neural crest migration.

Hindbrain neural crest cells adjacent to rhombomeres 2 (r2), r4 and r6 migrate in a segmental pattern, toward the first, second and third branchial arches, respectively. Although all rhombomeres generate neural crest cells, those arising from r3 and r5 deviate rostrally and caudally (J. Sechrist, G. Serbedzija, T. Scherson, S. Fraser and M. Bronner-Fraser (1993) Development 118, 691-703). We have altered the rostrocaudal positions of the cranial neural tube, adjacent ectoderm/mesoderm or presumptive otic vesicle to examine tissue influences on this segmental migratory pattern. After neural tube rotation, labeled neural crest cells follow pathways generally appropriate for their new position after grafting. For example, when r3 and r4 were transposed, labeled r3 cells migrated laterally to the second branchial arch whereas labeled r4 cells primarily deviated caudally toward the second arch, with some cells moving rostrally toward the first. In contrast to r4 neural crest cells, transposed r3 cells leave the neural tube surface in a polarized manner, near the r3/4 border. Surprisingly, some labeled neural crest cells moved directionally toward small ectopic otic vesicles that often formed in the ectoderm adjacent to grafted r4. Similarly, they moved toward grafted or displaced otic vesicles. In contrast, surgical manipulation of the mesoderm adjacent to r3 and r4 had no apparent effects. Our results offer evidence that neural crest cells migrate directionally toward the otic vesicle, either by selective attraction or pathway-derived cues.

Animals↗

Development of the marginal zone in the rhombenecephalon of Xenopus laevis.

The marginal zone is the most superficial component of the developing central nervous system. In this study we describe the pattern of formation of the marginal zone in Xenopus laevis as seen with the light microscope and correlate its appearance with developing axon bundles seen with the electron microscope. The marginal zone was first identified at stage 26 on the ventrolateral aspects of the neural tube in restrictive foci throughout the medulla. By stage 28 this presumptive fiber area had expanded to occupy a longitudinal zone along the ventrolateral aspect of the rhombencephalon. The marginal zone continued expanding rostrocaudally, mediolaterally, and dorsoventrally coming to occupy a substantial area along the periphery of the central nervous system by the time of hatching (stage 36). Electron microscopic (EM) observations allowed us to identify axons and growth cones on the lateral borders of the neural tube at stage 22. Foci of marginal zones identified in the caudal medulla at stage 24 consisted of several fascicles grouped together on the ventrolateral surface. These fascicles, separated and surrounded by ependymal processes contained axons and growth cones. The pattern of marginal zone development reflected the addition of axons within fascicles and also the addition of new fascicles of axons.

Animals↗

Excess lunatic fringe causes cranial neural crest over-proliferation.

Lunatic fringe is a vertebrate homologue of Drosophila fringe, which plays an important role in modulating Notch signaling. This study examines the distribution of chick lunatic fringe at sites of neural crest formation and explores its possible function by ectopic expression. Shortly after neural tube closure, lunatic fringe is expressed in most of the neural tube, with the exception of the dorsal midline containing presumptive neural crest. Thus, there is a fringe/non-fringe border at the site of neural crest production. Expression of excess lunatic fringe in the cranial neural tube and neural crest by retrovirally mediated gene transfer resulted in a significant increase ( approximately 60%) in the percentage of cranial neural crest cells 1 day after infection. This effect was mediated by an increase in cell division as assayed by BrdU incorporation. Infected embryos had an up-regulation of Delta-1 in the dorsal neural tube and redistribution of Notch-1 to the lumen of the neural tube, confirming that excess fringe modulates Notch signaling. These findings point to a novel role for lunatic fringe in regulating cell division and/or production of neural crest cells by the neural tube.

Animals↗

The spatial and temporal dynamics of Sax1 (CHox3) homeobox gene expression in the chick's spinal cord.

Sax1 (previously CHox3) is a chicken homeobox gene belonging to the same homeobox gene family as the Drosophila NK1 and the honeybee HHO genes. Sax1 transcripts are present from stage 2 H&H until at least 5 days of embryonic development. However, specific localization of Sax1 transcripts could not be detected by in situ hybridization prior to stage 8-, when Sax1 transcripts are specifically localized in the neural plate, posterior to the hindbrain. From stages 8- to 15 H&H, Sax1 continues to be expressed only in the spinal part of the neural plate. The anterior border of Sax1 expression was found to be always in the transverse plane separating the youngest somite from the yet unsegmented mesodermal plate and to regress with similar dynamics to that of the segregation of the somites from the mesodermal plate. The posterior border of Sax1 expression coincides with the posterior end of the neural plate. In order to study a possible regulation of Sax1 expression by its neighboring tissues, several embryonic manipulation experiments were performed. These manipulations included: removal of somites, mesodermal plate or notochord and transplantation of a young ectopic notochord in the vicinity of the neural plate or transplantation of neural plate sections into the extraembryonic area. The results of these experiments revealed that the induction of the neural plate by the mesoderm has already occurred in full primitive streak embryos, after which Sax1 is autonomously regulated within the spinal part of the neural plate.

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Six3, a medaka homologue of the Drosophila homeobox gene sine oculis is expressed in the anterior embryonic shield and the developing eye.

The conserved transcription factor Pax6 is essential for eye development in Drosophila and mammals (Hill, R.E., Favor, J., Hogan, B.L.M., Ton, C.C.T., Saunders, G.F., Hanson, I.M., Prosser, J., Jordan, T., Hastie, N.D., van Heyningen, V., 1991. Mouse small eye results from mutations in a paired-like homeobox containing gene. Nature 354, 522-525; Ton, C., Hirvonen, H., Miwa, H., Weil, M., Monaghan, P., Jordan, T., van Heyningen, V., Hastie, N., Meijers-Heijboer, H., Drechsler, M., Royer-Pokora, B., Collins, F., Swaroop, A., Strong, L.C., Saunders, G.F., 1991. Positional cloning and characterization of a paired box- and homeobox-containing gene from the aniridia region. Cell 6, 1059-1074; Matsuo, T., Osumi-Yamashita, N., Noji, S., Ohuchi, H., Koyama, E., Myokai, F., Matsuo, N., Toniguchi, S., Dari, H., Jseki, S., Ninomiya, Y., Fujiwara, M., Watanabe, T., Eto, K., 1993. A mutation at the Pax-6 gene in rat small eye is associated with impaired migration of midbrain crest cells. Nature genet. 3, 299-304; Quiring, R., Walldorf, U., Kloter, U., Gehring, W.J., 1994. Homology of the eyeless gene of Drosophila to the small eye gene in mice and aniridia in humans. Science 265, 785-789). These findings led to the hypothesis that additional genes involved in invertebrate and vertebrate eye development are structurally and functionally conserved (Halder, G., Callaerts, P., Gehring, W.J., 1995. New perspectives on eye evolution. Curr. Opin. Gen. Dev. 5, 602-609; Quiring, R., Walldorf, U., Kloter, U., Gehring, W.J., 1994. Homology of the eyeless gene of Drosophila to the small eye gene in mice and aniridia in humans. Science 265, 785-789). Candidates for such conserved genes are the Drosophila homeobox gene sine oculis (Cheyette, B.N.R., Green, P.J., Martin, K., Garren, H., Hartenstein, V., Zipursky, S.L., 1994. The Drosophila sine oculis locus encodes a homeodomain-containing protein required for the development of the entire visual system. Neuron l2, 977-996) and its murine homologue Six3 (Oliver, G., Mailhos, A., Wehr, R., Copeland, N.G., Jenkins, N.A., Gruss, P., 1995. Six3, a murine homologue of the sine oculis gene, demarcates the most anterior border of the developing neural plate and is expressed during eye development. Development 121, 4045-4055). sine oculis (so) is essential for the development of the larval and adult visual system (Cheyette, B.N.R., Green, P.J., Martin, K., Garren, H., Hartenstein, V., Zipursky, S.L., 1994. The Drosophila sine oculis locus encodes a homeodomain-containing protein required for the development of the entire visual system. Neuron l2, 977-996). Six3 is expressed in the anterior neural plate and optic vesicles, lens, olfactory placodes and ventral forebrain (Oliver, G., Mailhos, A., Wehr, R., Copeland, N.G., Jenkins, N.A., Gruss, P., 1995. Six3, a murine homologue of the sine oculis gene, demarcates the most anterior border of the developing neural plate and is expressed during eye development. Development 121, 4045-4055). Overexpression of mouse Six3 gene in medaka fish embryos (Orvzias latipes) results in the formation of an ectopic lens, indicating that Six3 activity can trigger the genetic pathway leading to lens formation (Oliver, G., Loosli, F., Koster, R., Wittbrodt, J., Gruss, P., 1996. Ectopic lens induction in fish in response to the murine homeobox gene Six3. Mech. Dev. 60, 233-239). We isolated the medaka Six3 homologue and analyzed its expression pattern in the medaka embryo. It is expressed initially in the anterior embryonic shield and later in the developing eye and prosencephalon. The early localized expression of Six3 suggests a role in the regionalization of the rostral head.

Amino Acid Sequence↗

Scanning and transmission electron microscope studies of interkinetic nuclear migration in the cerebral vesicles of the rat.

A scanning electron microscope (SEM) was used to examine the morphology and surface texture of neuroepithelial cells during interkinetic nuclear migration in the cerebral vesicles of the rat at 12, 13 and 14 days of gestation. Serial sections of embryonic material of the same age were also prepared for the transmission electron microscope (TEM). Particular attention was paid to the SEM and TEM appearance of mitotic neuroepithelial cells which occur exclusively along the ventricular border of the neural epithelium. Three distinctly-shaped classes of mitotic cells were recognised in scanning micrographs. (1) Pyriform cells. This type of mitotic cell was characterised by the presence of very long, fine processes radiating from the tip and shoulders of a short external (basal) process. These fine processes were termed "intramitotic filopodia." Microvilli were found on the surface of most pyriform cells. (2) Conical cells. These lacked an external process but there were large numbers of intramitotic filopodia at the basal pole of the cell body, and the perikaryal surface was rich in microvilli. (3) Globular cells. It was possible to subdivide this class of cell into large and small sizes, but usually a few short intramitotic filopdia were present at the basal pole. The perikaryal surfaces of the globular population were raised in coarse lumps and bubble-like protrusions. By pooling TEM and SEM information we were able to deduce that pyriform cells probably possess a prophase or prometaphase chromosome morphology, while conical cells exhibit a chromosome morphology somewhere between prometaphase and early anaphase. Large globular mitotic cells were found to be between metaphase and late anaphase and small globular cells were identified as early telophase cells. On the basis of these findings we have proposed that as a bipolar neuroepithelial cell rounds up for mitosis it passes first through a pyriform stage during which the external process is retracted or broken down, and then through a conical stage when the cell consolidates its position on the ventricular surface. Finally, the cell enters a large globular stage before dividing into two small globular telophase cells. It is not known what part, if any, the intramitotic filopodia play in this process of rounding up.

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The onset of dopaminergic innervation during ontogeny decreases melanotrope proliferation in the intermediate lobe of the rat pituitary.

The onset of dopaminergic innervation and its effects on melanotrope proliferation were investigated in the rat pituitary intermediate lobe. Dopamine, and its synthetic rate-limiting enzyme tyrosine hydroxylase, were first detected immunohistochemically on late post-natal day 3 or early postnatal day 4. Axon density was highest at the neural lobe/intermediate lobe border, and decreased toward the pituitary cleft. By postnatal day 10, the adult pattern of tyrosine hydroxylase immunoreactivity was established and remained through post-natal day 14. Neurointermediate lobe dopamine levels, measured by HPLC, correlated well with the increased axon density observed in the immunohistochemical studies. Dopamine could not be measured by our assay (100 fg limit) until post-natal day 3 (439.32 fg/NIL). Dopamine concentration increased to 2.09 +/- 0.425 ng at PN 4, 86.31 +/- 20.42 ng at PN 7, 168.72 +/- 18.37 ng at PN 10. Melanotrope proliferation was determined by [3H]thymidine incorporation before and after innervation. Concomitant with the onset of innervation, the proliferation index dropped from 13.4 +/- 0.01% to 6.5 +/- 0.002% at PN 4, and continued to decrease until a level of 3 +/- 0.003% was established by PN 10. To confirm the inhibitory action of dopaminergic innervation on melanotrope proliferation, rat neonates were injected intracisternally with 150 mg 6-hydroxydopamine to destroy dopaminergic axons within the intermediate lobe. Measurement of dopamine concentrations in neurointermediate lobes of injected animals showed a decrease in dopamine levels as compared to controls. From PN 4 (0.88 +/- 0.165 ng), DA levels gradually increased during development: at PN 5, [DA] = 0.689 +/- 0.104 ng; PN 6 [DA] = 11.60 +/- 2.24 ng; PN 7 [DA] = 20.93 +/- 3.80 ng; and PN 10 [DA] = 27.95 +/- 3.46 ng. Melanotrope proliferation also increased in 6-hydroxydopamine-treated animals. At PN 4, the onset of innervation reduced the pre-innervation proliferation index to 8.75 +/- 0.002%, only a 30% reduction in contrast to the greater than 50% decrease observed in control animals. A stable proliferation level of approximately 7.5% persisted in all subsequent stages with 6-OHDA administration. Our results demonstrated the time of dopamine innervation onset and a characteristic developmental pattern for axons within the rat intermediate lobe. The onset of innervation and increased dopamine concentration suggests increased dopaminergic control of the melanotropes, illustrated specifically by a decrease in their level of proliferation. This is the first presentation of evidence showing that dopaminergic innervation within the intermediate lobe of the rat pituitary regulates melanotrope proliferation.

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Expression of the somatolactin beta gene during zebrafish embryonic development.

Somatolactin (Sl) is a pituitary hormone closely related to prolactin (Prl) and growth hormone that was until now only found in various fish species. We isolated the cDNA coding for zebrafish Slbeta and we identified the gene encoding this hormone. We also obtained a 1kb genomic fragment corresponding to the slbeta upstream promoter region. Furthermore, the slbeta expression pattern was examined during zebrafish embryogenesis using whole-mount in situ hybridization. Slbeta mRNA is first detected in a single cell at the anterior border of the neural plate starting at 23h post fertilization (hpf). Slbeta-expressing cells also express the transcription factor pit1 and are located close to prl-expressing cells. Using combined fluorescent in situ hybridization, we show that slbeta- and prl-expressing cells are clearly distinct at 29 hpf. Starting at 30 hpf, the number of slbeta positive cells increases and their location becomes more clearly distinct from lactotrope cells, in a more posterior position. At later stages (48 hpf), slbeta expression was observed posterior to growth hormone expression, again in a distinct cell type. We show that zebrafish mutants aal, as well as mutants in the pit1 gene, are deficient in slbeta expression. In conclusion, slbeta expression defines a new, additional cell type in zebrafish pituitary that depends on pit1 and aal for its differentiation.

Animals↗

Expression pattern of the winged helix factor XFD-11 during Xenopus embryogenesis.

We have identified a cDNA encoding a novel Xenopus winged helix transcription factor termed as XFD-11 (Xenopus fork head domain). The DNA binding domain is most closely related to those of human or murine FREAC-3 (FKHL7/MF-1/FKH-1) proteins. The XFD-11 gene is activated at late blastula/early gastrula and transcription proceeds throughout embryogenesis. Early expression is found in ventral and lateral but not in dorsal mesoderm. At neurula stages, transcripts are found in posterior mesoderm except for the dorsal midline, and the gene is also transcribed at the lateral border of the neural plate and within anterior neuroectoderm. At later stages of development, transcripts are detected within the pronephros, the heart, within neural crest cells surrounding the eye, in the mandibular, hyoid and branchial arches, and within the tail.

Amino Acid Sequence↗

Cloning and embryonic expression of Hrsna, a snail family gene of the ascidian Halocynthia roretzi: implication in the origins of mechanisms for mesoderm specification and body axis formation in chordates.

snail family genes encode a transcription factor with specific zinc finger motifs. In vertebrates, they are expressed in the entire mesoderm in early embryogenesis and later in the paraxial mesoderm and the tail-bud, suggesting roles in specification and morphogenesis of the paraxial mesoderm. In the present study, a snail family gene Hrsna from a member of the chordates, an ascidian (Halocynthia roretzi), was cloned to obtain an insight into the origin of the mechanisms of mesoderm specification and body axis formation as observed in vertebrates. Expression of Hrsna during ascidian embryogenesis was found to be quite similar to that of vertebrate snail genes. First, before gastrulation, Hrsna was initially expressed in most precursors of mesodermal tissues including the notochord where As-T, the ascidian homolog of brachyury, was expressed. Hrsna expression persisted in the paraxial mesoderm, the mesenchyme and muscle, but not in the notochord precursors. Also, just as vertebrate snail family genes are expressed in the border of the neural plate that develops into dorsal neural tube and neural crest cells, so Hrsna expression was detected in the precursors of lateral and dorsal regions of the neural tube. However, Hrsna expression was not detected in the tip of the tail, unlike in vertebrate counterparts. In the light of the present findings, similarity and dissimilarity of mechanisms governing mesoderm specification and body axis formation between ascidians and vertebrates are discussed.

Amino Acid Sequence↗

New approaches to shock and trauma research: learning from multidisciplinary exchange.

BACKGROUND: Our understanding of the complex network of pathophysiology after multiple injury is limited. It is proposed to overcome the limitations of the traditional linear reductionism approach by merging the expertise of biology and medicine with other disciplines such as mathematics, physics and computer science. METHODS: We organized a two-days-workshop, where surgeons and surgical scientists explained the problem from the medical (pathophysiological) perspective to a well selected group of German applied mathematicians and computer scientists. Vice versa they presented and discussed their approaches to complex system analysis. RESULTS AND CONCLUSIONS: Physicians found it difficult to develop questions and concepts that go beyond the classic mechanistic thinking. Well formulated questions are the most important prerequisites for successful application of mathematical tools. The possibilities and borders of Artificial Neural Networks (ANN), Hidden Markow Models (HMM), Agent Based Models (ABM), differential equations for problem solving were discussed. There is no master model for all aspects of pathophysiology, however, application of the models to specific problems is mandatory. CONCLUSIONS: Future breakthroughs can only be expected if we overcome language problems between disciplines. This cross talk was considered by all participants as a most important step.

Communication↗

Functional characteristics of cochlear nucleus in behaving cat examined by acoustic masking of electrical stimuli.

1. Cats were trained, using an operant procedure, to detect and respond to electrical stimulation delivered in the vicinity of the cochlear nucleus. The electrical stimuli were presented both in silence and in synchrony with repeated noise bursts to determine whether detection thresholds for the electrical stimuli were elevated by the acoustic masking noise. 2. For stimulation sites centered within auditory structures (cochlear nucleus or acoustic nerve root), the acoustic maskers caused a consistent elevation of the electrical detection thresholds. For stimulation sites that were in or bordered on nonacoustic neural structures (e.g., vestibular), the acoustic maskers caused little or no elevation of electrical detection thresholds. 3. The magnitude of the acoustic masking effect was monotonically related to the intensity of the acoustic masker across the range of intensities tested. 4. The magnitude of the masking effect was strongly dependent on the relative timing of the stimulus pulse and the masker noise burst. Maximum masking occurred when the pulse just followed the onset of the neural activity in cochlear nucleus evoked by the masker burst. Less masking occurred when the electrical pulse occurred at the middle or end of the masker burst, and still less when the pulse occurred just prior to the onset (backward masking) or just after the offset (forward masking) of the masker burst. 5. The magnitude of the masking effect also depended on the frequency of the acoustic masker. For tone bursts, masking was maximal for each electrode at a particular frequency and declined monotonically for masker frequencies above or below the optimal frequency. 6. It is concluded that the masking of an electrical stimulus by an acoustic stimulus depends on a direct interaction between the neural responses evoked by the two stimuli, and that similar central, neural interactions may contribute to acoustic masking of acoustic stimuli. It is also concluded that the technique of masking an electrical stimulus by an acoustical stimulus is a precise and useful tool for the study of sensory-neural organization in intact behaving animals.

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Formation of cadherin-expressing brain nuclei in diencephalic alar plate divisions.

During the formation of brain nuclei, the vertebrate neural tube is partitioned into distinct embryonic divisions. In this study, the expression of three members of the cadherin family of adhesion molecules (cadherin-6B, cadherin-7, and R-cadherin) was mapped to study the differentiation of gray matter in the divisions of the diencephalic alar plate of chicken embryos from embryonic day 3 (E3) to E10. At early stages of development (E3-E4), each cadherin is expressed in restricted regions of the diencephalic wall of the neural tube. The borders of some of the expression domains coincide with divisional boundaries. As the mantle layer is formed and increases in thickness from E4 to E8, morphologically discernible aggregates of cells appear that express the three cadherins differentially. These aggregates represent the anlagen of specific diencephalic brain nuclei, e.g., the lateroanterior nucleus, the ventral geniculate nucleus, the nucleus rotundus, the perirotundic area, the principal precommissural nucleus, and the lateral spiriform nucleus. Most of the cadherin-expressing diencephalic nuclei studied in this work apparently derive from a single embryonic division and remain there. The divisional boundaries are replaced gradually by the borders of cadherin-expressing brain nuclei. The current results support the idea that cadherins confer differential adhesiveness to developing structures of gray matter in the diencephalic alar plate. Moreover, they suggest that each cadherin plays a role in the formation of specific brain nuclei within the diencephalic divisions.

Age Factors↗

Formation of cadherin-expressing brain nuclei in diencephalic alar plate divisions.

During the formation of brain nuclei, the vertebrate neural tube is partitioned into distinct embryonic divisions. In this study, the expression of three members of the cadherin family of adhesion molecules (cadherin-6B, cadherin-7, and R-cadherin) was mapped to study the differentiation of gray matter in the division so that diencephalic alar plate of chicken embryos from embryonic day 3 (E3) to E10. At early stages of development (E3-E4), each cadherin is expressed in restricted regions of the diencephalic wall of the neural tube. The borders of some of the expression domains coincide with divisional boundaries. As the mantle layer is formed and increases in thickness from E4 to E8, morphologically discernible aggregates of cells appear that express the three cadherins differentially. These aggregates represent the anlagen of specific diencephalic brain nuclei, e.g., the lateroanterior nucleus, the ventral geniculate nucleus, the nucleus rotundus, the perirotundic area, the principal precommissural nucleus, and the lateral spiriform nucleus. Most of the cadherin-expressing diencephalic nuclei studied in this work apparently derive from a single embryonic division and remain there. The divisional boundaries are replaced gradually by the borders of cadherin-expressing brain nuclei. The current results support the idea that cadherins confer differential adhesiveness to developing structures of gray matter in the diencephalic alar plate. Moreover, they suggest that each cadherin plays a role in the formation of specific brain nuclei within the diencephalic divisions.

Age Factors↗

Localization of organic anion transporting polypeptide 3 (oatp3) in mouse brain parenchymal and capillary endothelial cells.

Organic anion transporting polypeptide 3 (oatp3) transports various CNS-acting endogenous compounds, including thyroid hormones and prostaglandin E2, between extra- and intracellular spaces, suggesting a possible role in CNS function. The purpose of this study was to clarify the expression and localization of oatp3 in the mouse brain. RT-PCR analysis revealed that oatp3 mRNA is expressed in brain capillary-rich fraction, conditionally immortalized brain capillary endothelial cells, choroid plexus, brain and lung, but not in liver or kidney, where oatp1, 2 and 5 mRNAs were detected. Immunohistochemical analysis with anti-oatp3 antibody suggests that oatp3 protein is localized at the brush-border membrane of mouse choroid plexus epithelial cells. Furthermore, intense immunoreactivity was detected in neural cells in the border region between hypothalamus and thalamus, and in the olfactory bulb. Immunoreactivity was also detected in brain capillary endothelial cells in the cerebral cortex. These localizations in the mouse brain suggest that oatp3 plays roles in blood-brain and -cerebrospinal fluid barrier transport of organic anions and signal mediators, and in hormone uptake by neural cells.

Animals↗

Rhombomeric origin and rostrocaudal reassortment of neural crest cells revealed by intravital microscopy.

Neural crest cell migration in the hindbrain is segmental, with prominent streams of migrating cells adjacent to rhombomeres (r) r2, r4 and r6, but not r3 or r5. This migratory pattern cannot be explained by the failure of r3 and r5 to produce neural crest, since focal injections of the lipophilic dye, DiI, into the neural folds clearly demonstrate that all rhombomeres produce neural crest cells. Here, we examine the dynamics of hindbrain neural crest cell emigration and movement by iontophoretically injecting DiI into small numbers of cells. The intensely labeled cells and their progeny were repeatedly imaged using low-light-level epifluorescence microscopy, permitting their movement to be followed in living embryos over time. These intravital images definitively show that neural crest cells move both rostrally and caudally from r3 and r5 to emerge as a part of the streams adjacent to r2, r4, and/or r6. Within the first few hours, cells labeled in r3 move within and/or along the dorsal neural tube surface, either rostrally toward the r2/3 border or caudally toward the r3/4 border. The labeled cells exit the surface of the neural tube near these borders and migrate toward the first or second branchial arches several hours after initial labeling. Focal DiI injections into r5 resulted in neural crest cell contributions to both the second and third branchial arches, again via rostrocaudal movements of the cells before migration into the periphery. These results demonstrate conclusively that all rhombomeres give rise to neural crest cells, and that rostrocaudal rearrangement of the cells contributes to the segmental migration of neural crest cells adjacent to r2, r4, and r6. Furthermore, it appears that there are consistent exit points of neural crest cell emigration; for example, cells arising from r3 emigrate almost exclusively from the rostral or caudal borders of that rhombomere.

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Multiple neural cell types are infected in vitro by border disease virus.

Border disease (BD) of sheep results from a congenitally acquired nonarbotogavirus infection which causes a highly selective central nervous system (CNS) pathological lesion consisting of diffuse decreased myelination without inflammation or neuronal destruction. Thus, a selective disruption of oligodendroglial function appears to occur. In order to investigate the in vitro cell tropism of BD virus, primary cultures derived from fetal and adult ovine CNS and peripheral nervous system were inoculated with BD virus. Infected cell types were determined by dual immunofluorescent labeling for viral and cell type specific antigens. Infection of all the major cell types represented in these cultures, including oligodendrocytes, astrocytes, fibroblasts, dorsal root ganglion neurons and Schwann cells was found. Oligodendrocytes were only infected earlier and appeared to remain infected longer than astrocytes and fibroblasts. Infectious virus was produced by all cultures and continued to be produced even after the disappearance of nearly all immunocytochemically detectable viral antigen within cells. These studies suggest that the selective dysfunction of the oligodendrocyte in BD is not based on a selective viral tropism.

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