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PubMed1990

Development of the retinotectal projection in zebrafish embryos under TTX-induced neural-impulse blockade.

The influence of neural activity on the morphology of retinal-axon-terminal arbors and the precision of the developing retinotectal projection in zebrafish embryos was explored. Terminal-arbor morphology and their distribution in the tectum was determined with anatomical fiber-tracing methods using the fluorescent dyes dil and diO. To allow development under activity-deprived conditions, TTX was injected into the eyes of 30-38-hr-old zebrafish embryos at concentrations that effectively blocked neural activity both in retinal ganglion cells and throughout the CNS. Much like axons with normal neural-activity patterns, activity-deprived axons from dorsal and ventral and from temporal and nasal regions in the retina terminated over retinotopically appropriate and nonoverlapping regions of the tectum. Even after ablation of 1 hemiretina at the time of axonal outgrowth, activity-deprived axons from the remaining hemiretina grew directed toward and arborized selectively within their retinotopically appropriate tectal half in the same way as would nondeprived axons. Besides being retinotopic, the area over which small populations of activity-deprived axons from neighboring ganglion cells arborize is as small as that of active axons. The size of terminal arbors of retinal ganglion cell axons was unaffected by blockade of neural activity. The mean terminal-arbor size was 27 x 18 microns for the TTX-injected and 31 x 22 microns for the control embryos. The tectal coverage of TTX-blocked and control axons was equally small, with values of 1.4% and 1.6%, respectively. These data show that a precisely organized retinotopic map in developing zebrafish forms independent of neural-impulse activity.

Animals↗

PubMed1989

Inaccuracies in initial growth and arborization of chick retinotectal axons followed by course corrections and axon remodeling to develop topographic order.

The retinotectal projection is organized in a precise retinotopic manner. We find, though, that during development the growth and arborization of temporal retinal axons within the optic tectum of chick embryos is initially imprecise. Axonal targeting errors occur along the rostral-caudal and medial-lateral tectal axes, and arbors are formed at topographically inappropriate positions. Subsequent course corrections along both tectal axes and large-scale axonal remodeling lead to the retinotopic ordering of terminal arborizations characteristic of the mature projection. The trajectories and branching patterns of temporal retinal axons labeled with Dil or DiO were determined in whole mounts of retina and tectum from chicks ranging in age from embryonic day 9 to posthatching. Within the retina, labeled retinofugal axons travel in a compact bundle but do not maintain strict neighbor relations, as they course to the optic fissure. The axons enter the contralateral tectum at its rostral edge and grow caudally. Many extend well past their appropriate terminal zone within rostral tectum; a proportion of these later reverse their direction of growth. Many axons grow onto the tectum at incorrect positions along the medial-lateral tectal axis. Some correct this error in a directed manner by altering their trajectory or extending collateral branches at right angles. About 80% of the positional changes of this type are made in the direction appropriate to correct axon position, and thus are likely a response to tectal positional cues. After maturation of retinotopic order, about half of the axons that project to a mature terminal zone have made abrupt course corrections along one or both tectal axes, indicating that initially mistargeted axons can establish appropriately positioned arbors and survive. The development of temporal axons within the tectum is characterized by 3 phases: elongation, branch and arbor formation, and remodeling. After considerable rostrocaudal elongation, an axon typically develops numerous side branches and arbors, many at inappropriate locations. Most arbors are formed by side branches that develop as interstitial collaterals; few axons grow directly to their appropriate terminal zone and arborize. Aberrant arbors, and axons and axon segments that fail to form arbors in the appropriate terminal zone, are rapidly eliminated over about a 2 d period. Axon degeneration appears to play a role in this remodeling process.

Animals↗

PubMed1989

Nitrosylated high density lipoprotein is recognized by a scavenger receptor in rat liver.

In order to assess the presence of specific recognition sites for high density lipoprotein (HDL) in vivo, HDL was nitrosylated with tetranitromethane and the decay and liver uptake were compared with that of native HDL. The association of intravenously injected nitrosylated HDL (TNM-HDL) with liver was greatly increased as compared to native HDL. Using a cold cell isolation method, it became evident that the liver endothelial cells were responsible for the increased uptake of the modified HDL. The involvement of the endothelial cells in the uptake of TNM-HDL from the circulation could also be demonstrated morphologically by using the fluorescent dye dioctadecyl-tetramethyl-indocarbocyanine perchlorate (Dil) to label HDL. In vitro competition studies with isolated liver endothelial cells indicated that unlabeled modified HDL and acetylated LDL displaced iodine-labeled TNM-HDL, while no competition was seen with LDL and a slight displacement was seen with unlabeled native HDL. Nonlipoprotein competitors of the scavenger receptor such as fucoidin and polyinosinic acid blocked the interaction of TNM-HDL with the liver endothelial cells. Also the degradation of TNM-HDL was blocked by low concentrations of chloroquine. It can be concluded that a scavenger receptor on liver endothelial cells is involved in the clearance of tetranitromethane-modified HDL, which excludes the possibility of using TNM-HDL in vivo to assess the non-receptor-dependent uptake of HDL. The use of nitrosylated HDL in vitro as a low affinity control is limited to cell types that do not possess scavenger receptors, because cell types with scavenger receptors will recognize and internalize TNM-HDL by a high affinity scavenger pathway.

Animals↗

PubMed1989

[Development of a small-lumen vascular prosthesis coated with autologous endothelial cells].

A new compliant prosthesis with a monolayer of autologous endothelial cells (ENC) has been developed. It consists of a porous polyurethane-siloxane-copolymer reinforced by a polyester network to prevent excessive dilatation. On the inner surface an ENC monolayer is established before implantation by a cell culture procedure. The prosthesis displays compliance (13.2 +/- 3.0 x 10(-4) mmHg-1) comparable to native arteries. It is non-kinkable (minimal radius of curvature less than 5 mm). Burst resistance, remaining deformation, cut out force and tensile strength are superior to standard values. ENC-coverage in excess of 95% of the inner surface was produced in vitro using a lining procedure. The monolayer of confluent cells was demonstrated to consist of endothelial cells by their characteristic cobblestone morphology, the expression of factor VIII related antigen and the specific uptake of Dil-Ac-LDL. The unstimulated prostacyclin production was similar both in native veins as well as in lined prostheses. Antithrombogenicity of the endothelial cell lining was demonstrated in 24 h animal implants.

Animals↗

PubMed1988

Retinotopic organization of the developing retinotectal projection in the zebrafish embryo.

Developing retinal axons in the zebrafish embryo were stained with HRP or with the fluorescent dyes dil and diO to study the formation of the retinotectal projection. Retinal axons leave the eye at 34-36 hr postfertilization (PF), invade the tectum at 46-48 hr PF, and innervate the tectal neuropil at 70-72 hr PF. Dorsal and ventral axons occupy separate aspects of the optic nerve and tract and pass into their retinotopically appropriate ventral and dorsal hemitectum, respectively. Nasal and temporal axons are segregated in the nerve, mixed in the tract, and are coextensive over the rostral half of tectum until 56 hr PF. They then segregate again, due to the progression of nasal axons into the open caudal tectum. Thus, at 70-72 hr PF, dorsal and ventral as well as temporal and nasal axons occupy their retinotopically appropriate tectal quadrants. After ablation of the temporal retina prior to the time of axonal outgrowth, the nasal axons bypass the vacant rostral tectum to terminate in the caudal tectal half. Temporal axons in the absence of nasal axons remain restricted to their appropriate rostral tectal half, suggesting that nasal and temporal axons possess a preference for their retinotopically appropriate tectal domains. Measurements of individual terminal arbors and the tectal areas in embryos and in adult zebrafish showed that individual arbors are large with respect to the embryonic tectum but are about 14-15 times smaller than in the adult. However, the proportion of tectum covered by embryonic arbors is about 7 times larger than in the adult, suggesting that a higher precision of the adult projection is achieved as a result of a greater enlargement of the tectum than of the arbors.

Animals↗

PubMed1987

Enhanced percutaneous absorption of ionizable water-soluble drugs.

The percutaneous absorption of diltiazem hydrochloride (DIL) and disodium cromoglycate (DSCG) [representative, respectively, of cationic and anionic water-soluble drugs] was studied in rabbits, using films prepared from water-soluble components. When corresponding fat-soluble counter-ions were added to films prepared using the electrically neutral components polyvinylalcohol and glycerol, the percutaneous absorption of the drugs was enhanced. Furthermore, when non-electrolytes such as BL-9EX or urea were added to the films together with the counter-ions, the bioavailability of the drugs increased. Consequently, it is postulated that ionizable water-soluble drugs are absorbed through skin by forming fat-soluble ion-pairs and additionally that the barrier function against absorption is reduced by non-electrolytes such as BL-9EX or urea. Such non-electrolytes must not hinder the formation of ion-pairs between the drugs and corresponding counter-ions.

Animals↗

PubMedDate not supplied

[Statistical analysis of results in the determination of steroid hormone receptors].

Statistical evaluation of the data of steroid hormone receptors estimation involved the principle of dependence of each experimentally obtained magnitude of total binding (in presence of a single labelled ligand) on the magnitude of unspecific binding (at the similar content of labelled ligand but in addition of unlabelled substance excess). The principle enabled to calculate: individual values of specific binding as a difference between each value for total and unspecific binding independently on amount of parallel probes of total and unspecific bindings, mean value of specific binding using the calculated magnitudes, standard deviation (dispersion) and standard error of mean value. Statistical method allowed to determine differences between magnitudes of specific binding (content of receptors) at saturating concentrations of ligand as well as to estimate standard deviation of KD and Vmax in a plot. The statistical principles were used for estimation of specific binding of 3H-estra dil-17 beta, 3H-dihydrotestosterone and 3H-5 alpha-and-rostan-3 beta, 17 beta-diol in various rat tissues (hypophysis, prostate, uterus) and human tissues (mammary gland tissue tumor, prostate adenoma). The principle developed may be used for estimation of various receptors of biologically active substances, where the specific binding is calculated as a difference between total and unspecific binding.

Animals↗

PubMed1983

Photobleaching recovery studies of T-independent antigen mobility on antibody-bearing liposomes.

A physicochemical model for the antigen-specific B cell membrane was prepared by incorporating palmitate-conjugated MOPC 315, an anti-DNP IgA, into 8-micron diameter liposomes prepared from phosphatidylcholine, cholesterol, and cardiolipin (2:2:1). With this model system, we examined how the purely passive cross-linking of membrane-bound immunoglobulin molecules by antigen affects the lateral mobility of antigen-receptor complexes. At 37 degrees C, liposome-bound tetramethylrhodamine isothiocyanate (TRITC)-labeled anti-DNP antibody diffuses at 1.4 X 10(-8) cm2 sec-1, a rate comparable to that observed for the phospholipid analog dil-C18-(3). Both substances exhibit complete fluorescence recovery after bleaching. The binding of TRITC-conjugates of the antigens DNP-polymerized flagellin (DNP-POL) and DNP-dextran (DNP-DEX) to liposomes bearing nonfluorescent palmitoyl-MOPC 315 was then examined. The diffusion coefficients D observed for bound antigens decrease monotonically with increased antigen dose and epitope density. For low epitope density antigens, the DNP-DEX and DNP-POL complexes are almost completely mobile. At higher epitope densities, a fraction of bound antigen appears immobile on the time scale of the experiment. This fraction is dependent on antigen concentration and epitope density and on the amount of palmitoyl MOPC 315 incorporated. The immobile fraction is 31.5% for DNP4.0-POL at 30 micrograms/ml on liposomes bearing 150,000 immunoglobulin molecules. Under these conditions D for mobile antigen is 5.0 X 10(-10) cm2 sec-1. The observed immobile fractions may represent formation of a two-dimensional gel phase of antigen-immunoglobulin aggregates. The results obtained in this study are compared with those obtained in previous work on antigen-specific mouse B lymphocytes.

Animals↗

PubMed1995

Development and regulation of dendritic stratification in retinal ganglion cells by glutamate-mediated afferent activity.

In the mature retina, the dendrites of retinal ganglion cells (RGCs) are segregated into either ON or OFF sublaminae of the inner plexiform layer (IPL), but early in development the dendritic processes of these cells are multistratified, ramifying throughout the IPL. We examined the time course of dendritic stratification in developing beta cells, the largest class of ganglion cells in the cat retina, by retrograde labeling of fixed tissue with Dil. Dendritic stratification begins in the central and peripheral retina by embryonic day 50, about 2 weeks before birth and is not fully completed until 5 months postnatally. A clear central-to-peripheral gradient in the incidence of stratified beta cells first becomes evident shortly after birth. This stratification process was effectively halted by short-term intraocular injections (4-11 d) of the glutamate analog 2-amino-4-phosphonobutyrate (APB), which hyperpolarizes rod bipolar cells and ON cone bipolar cells, thereby preventing the release of glutamate by these interneurons. APB treatment did not alter the somal sizes or the tangential extent of the dendrites of developing beta cells, nor did it cause abnormal loss of these neurons. The organization of the inner nuclear layer, containing the APB-sensitive bipolar cells, was also not compromised by such injections. When APB treatment was discontinued there was a rapid resumption of dendritic stratification resulting in a normal incidence of stratified RGCs. Thus, short-term APB treatment causes a delay rather than a permanent arrest of the stratification process.(ABSTRACT TRUNCATED AT 250 WORDS)

Afferent Pathways↗

PubMed1995

Tenascin knockout mice: barrels, boundary molecules, and glial scars.

In light of a previous report suggesting that the brains of tenascin-deficient animals are grossly normal, we have studied the somatosensory cortical barrel field and injured cerebral cortex in postnatal homozygous tenascin knockout, heterozygote, and normal wild-type mice. Nissl staining, cytochrome oxidase, and Dil axonal tracing of thalamocortical axonal projections to the somatosensory cortex, all reveal the formation of normal barrels in the first postnatal week in homozygous knockout mice that cannot be distinguished from heterozygote or normal wild-type barrels. In addition to confirming the absence of tenascin in knockout animals, and reporting apparently reduced levels of the glycoprotein in barrel boundaries of heterozygote animals using well-characterized antibodies and immunocytochemistry, we also studied the DSD-1-PG proteoglycan, another developmentally regulated molecule known to be associated with transient glial/glycoconjugate boundaries that surround developing barrels; DSD-1-PG was also found to be expressed in barrel boundaries in apparently normal time frames in tenascin knockout mice. Peanut agglutinin (PNA) binding of galactosyl-containing glycoconjugates also revealed barrel boundaries in all three genotypes. We also examined the expression of tenascin-R, a paralog of tenascin-C (referred to here simply as tenascin). As previously reported, tenascin-R is prominently expressed in subcortical white matter, and we found it was not expressed in the barrel boundaries in any of the genotypes. Thus, the absence of tenascin does not result in a compensatory expression of tenascin-R in the barrel boundaries. Finally, we studied wounds of the cerebral cortex in the late postnatal mouse. The astroglial scar formed, for the most part, in the same time course and spatial distribution in the wild-type and tenascin knockout mice. However, there may be some differences in the extent of gliosis between the knockout and the wild type that warrant further study. Roles for boundary molecules like tenascin during brain pattern formation and injury are reconsidered in light of these findings on barrel development and cortical lesions in tenascin-deficient mice.

Animals↗

PubMed1995

Relationships between dendritic fields and functional architecture in striate cortex of normal and visually deprived cats.

We examined relationships between the pattern of geniculocortical innervation and the dendritic fields of cells in layer 4 of in cat primary visual cortex. Experiments were performed on normal animals and on cats in which the geniculocortical projection was altered by monocular deprivation or by the induction of divergent squint during the critical period. Thalamic afferents providing the input from the contralateral eye were anterogradely labeled by injecting the fluorescent tracer Dil into lamina A of the lateral geniculate nucleus. Intracellular staining with Lucifer yellow in slice preparations allowed simultaneous visualization of the morphology of individual cells and the thalamic afferents. Our results demonstrate that spiny stellate cells close to the upper and lower margin of the geniculocortical input have highly asymmetric dendritic fields, and thereby confine their dendrites to the termination zone of these afferents. This effect was specific for the cell class; it was not observed in pyramidal neurons. These dendritic asymmetries perpendicular to the laminar borders of spiny stellate cells were not altered by monocular deprivation or strabismus. In contrast, visual deprivation strongly influenced the dendritic arbors of spiny stellate cells near the borders between adjacent ocular dominance columns. In normal animals, the dendrites of cells near columnar borders remained preferentially within one column. These dendritic asymmetries became much more pronounced in strabismic animals. Monocular deprivation weakened the influence of the columnar borders on dendritic fields. Spiny stellate cells within the columns of the open eye exhibited a slight tendency to confine their dendrites to these columns. Cells in the columns of the deprived eye showed the opposite effect; they extended their dendrites preferentially into the adjacent columns of the open eye. These results demonstrate that the segregation of geniculocortical afferents into ocular dominance columns and its perturbation by manipulation of the visual input plays an important role in defining the morphology of cortical target cells. Thus, activity-dependent structural changes not only occur at the level of the presynaptic terminals, but also at the level of the postsynaptic target cells, and thereby contribute to build up the functional architecture of the cortex.

Animals↗

PubMed1995

Expression and in vitro function of beta 1-integrin laminin receptors in the developing avian ciliary ganglion.

In chick development, ciliary ganglion (CG) neurons go through a period of axon extension from approximately embryonic day (E)4 to E8, followed by a period of synaptogenesis and neuronal cell death. By examining the immunohistochemical localization of laminin, in conjunction with Dil labeling of the ciliary nerve projection, we have determined that the pathway taken by these neurons is rich in laminin expression. Therefore, laminins are good candidate molecules for mediating outgrowth of these neurons in vivo. In vitro, the ability of CG neurons to extend neurites on laminin-1 (EHS laminin, alpha 1 beta 1 gamma 1) is maximal up to E8, then declines dramatically. CG neuron outgrowth on laminin-1 requires the activity of beta 1-class integrins. We have used subunit-specific antibodies to determine which of the five beta 1-containing heterodimers known to be laminin receptors (alpha 1 beta 1, alpha 2 beta 1, alpha 6 beta 1, alpha 7 beta 1) are expressed, and which mediate neurite outgrowth. While we could not detect expression of alpha 2 or alpha 7, we have found that alpha 1, alpha 3 beta 1, and alpha 6 beta 1 are expressed on the surface of ciliary ganglion neuron cell bodies and axons, both in vitro and in vivo. Furthermore, antibodies against alpha 3 and alpha 6, but not alpha 1, interfered with CG neurite outgrowth on laminin-1 in vitro. Taken together, these data suggest that interactions of cell surface alpha 3 beta 1 and alpha 6 beta 1 integrins with laminin-1 are likely to mediate growth of CG neurons during pathfinding in vivo.

Animals↗

PubMedDate not supplied

Skeletal muscle ventricles seeded with autogenous endothelium.

Skeletal muscle ventricles (SMVs) are muscular pumping chambers constructed from skeletal muscle. Previously, SMVs were connected to the systemic circulation with vascular conduits and used to assist the heart. In this study, SMVs were constructed from the latissimus dorsi muscle in eight dogs. The SMVs were seeded with autologous endothelial cells, but not connected to the circulation. Endothelial cells were harvested enzymatically from autogenous external jugular vein and grown in tissue culture. After 9 weeks, 6 electrically conditioned SMVs were seeded with endothelial cells by injecting 4-5 ml of culture medium containing 5-8 x 10(6) autogenous endothelial cells into each SMV lumen adjacent to the mandrel. Conditioning was stopped at the time of endothelial seeding. One week after seeding, electrical conditioning was resumed. Two weeks after seeding, the animals were killed and the SMVs excised. Histologic examination confirmed the presence of a confluent monolayer of cells covering 80-100% of the luminal surface in each seeded SMV. The endothelial nature of the cells lining the SMV lumen was established by fluorescent microscopy. Endothelial cells were pre labeled with the cellular marker PKH before seeding; the SMVs were also incubated with the endothelial marker dil-acetylated LDL. Endothelial cells also were identified by staining with fluorescently labeled antibodies to von Willebrand factor. Based upon these data, electrically conditioned SMVs can be seeded successfully with a near-complete, autologous endothelial monolayer. Additionally, this endothelial monolayer can be maintained on the luminal surface of a contracting SMV. In-circulation studies will determine whether endothelial cell seeding of SMVs can decrease or eliminate the incidence of thromboembolism.

Animals↗

PubMed1995

Repeated confocal imaging of individual dendritic spines in the living hippocampal slice: evidence for changes in length and orientation associated with chemically induced LTP.

Using confocal microscopy in conjunction with microdrop application of Dil, we have imaged and measured individual dendritic spines of living hippocampal CA1 pyramidal neurons in acute brain slices, before and approximately 3 hr after induction of long-term potentiation by chemical means. Statistical analysis of changes in the length of individual spines, and comparison with results of Monte Carlo simulations, suggests that two forms of structural change occur in chemically induced long-term potentiation: growth of a subpopulation of small spines, and angular displacement of spines. These changes could provide a structural basis for the expression of long-term potentiation.

Algorithms↗

PubMed1994

A study of the motor neuron pool of the superior rectus muscle in albino rats by retrograde fluorescent double labeling technique.

Recent investigations revealed the localized distribution of the motoneuron subgroups within the mammalian oculomotor nucleus. In this study, we examined the motor neuron pool of the superior rectus muscle (SR) in 12 albino rats by injecting the retrograde fluorescent tracers Fluoro-Gold (FG) into one SR and Dil into the contralateral SR. We also examined the topographic and functional correlation between the neurons controlling the muscles of the upward gaze, i.e., SR and the inferior oblique muscle (IO) in another 5 albino rats by means of the same tracers. Our results demonstrated that: 1) the average total number of the neurons in the motor neuron pool of SR was 322.7 +/- 40.1; 2) topographically, about 94.65% of the motoneurons controlling SR were located in the contralateral side, while the remaining 5.35% lay ipsilaterally; 3) all the IO motoneurons were present on the same side of the muscle they innervate; 4) the motoneurons of SR and IO did not form separate subnuclei, but were intermingling with each other; 5) functionally, about 94.59%, 4.26% and 1.15% of the total motoneurons in the SR subnucleus of either oculomotor nucleus were projecting contralaterally, ipsilaterally and bilaterally, respectively; 6) although the distribution of the perikarya was not homogeneous, the bilaterally projecting (i.e., to both SR) cells were disposed centrally, and their double labeling indicated that their axons innervate the bilateral SR via axonal bifurcation and/or collateral branching; and 7) the motoneurons of SR and IO were functionally segregated from each other as no bilaterally projecting neurons (i.e., to SR and IO) were found.

Animals↗

PubMed1995

Retinal axon divergence in the optic chiasm: uncrossed axons diverge from crossed axons within a midline glial specialization.

A long-standing question is how fiber pathways in the mammalian CNS project to both sides of the brain. Static and real-time analyses of dye-labeled retinal axons (Godement et al., 1990, 1994) have demonstrated that at embryonic day 15-17 in the mouse, crossed and uncrossed axons from each eye diverge in a zone 100-200 microns proximal to the midline of the optic chiasm. In this study, we identify cellular specializations in this zone that might serve as cues for retinal axon divergence. Second, using growth cone morphology as an indicator of growth cone destination, we analyzed how crossed and uncrossed retinal growth cones related to these cellular components. Monoclonal antibody RC2, a marker for radial glia in embryonic mouse CNS, revealed a palisade of radial glia straddling the midline. At the midline, a thin raphe of cells that appear morphologically distinct from the radial glia express a free carbohydrate epitope, stage-specific embryonic antigen 1 (SSEA-1). Sections containing Dil-labeled axons and immunolabeled cells indicated that all axons enter the radial glial palisade. Uncrossed axons turn within the palisade, but never beyond the raphe of SSEA-1-positive cells. In addition, ultrastructural analysis indicated that all growth cones contact radial glia, with projections of the growth cone interdigitating with glial fibers. These results demonstrate that retinal axons diverge within a cellular specialization centered around the midline of the developing optic chiasm, consistent with the hypothesis that cues for divergence are located in this zone.

Animals↗

PubMed1995

Soluble factors from the olfactory bulb attract olfactory Schwann cells.

Olfactory Schwann cells (OSCs) extend processes that ensheathe bundles of olfactory axons as they course from the olfactory epithelium to the olfactory bulb (OB). Results of morphological and immunohistochemical studies have led to speculation that OSCs may be involved in guiding the olfactory axons to their target tissue. In this study we have explored this possibility by investigating the relationship between OSCs and the OB. Olfactory Schwann cells labelled with 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate (Dil) were injected into the nasal region of E14 rat embryos and entire embryos were cultured for 24 hr. It was found in some embryos, that the OSCs had migrated toward the presumptive OB. Cocultures of neonatal OB explants on OSC monolayers showed that the OSCs were attracted to the OB and formed a ring-like aggregate around the explant after 48 hr culture. This attraction was absent when a piece of cerebrum was used in place of the OB. When medium conditioned by neonatal OBs was placed in the lower compartment of the chemotaxis chamber, OSCs seeded in the upper compartment migrated through the pores of the nucleopore filter to reach the underside which was in contact with the conditioned medium. After 6 hr of incubation, scanning electron microscopy was performed on the underside of the nucleopore filters. Cell counts of OSCs showed that the cell density was significantly higher when medium conditioned by OBs was used instead of unconditioned medium or medium conditioned by cerebrum. The results of these experiments show that the OSCs migrate toward the OB under the influence of soluble factor(s) secreted by the target tissue.

Animals↗

PubMed1995

Age-dependent specification of the corticocortical connections of cerebral grafts.

To investigate further the factors involved in the development of cerebral cortical circuitry, we examined the organization of corticocortical connections of heterotopic grafts of fetal cortex placed into neonatal rat cortices. Presumptive perirhinal or sensorimotor areas of the cerebral wall were removed as slabs from embryonic day 12 (E12), E13, or E14 rats and transplanted heterotopically into either rostral perirhinal or sensorimotor cortical areas of neonatal rats. Two weeks later, the afferent cortical connections of the grafts were labeled by placing DilC18(-3) (Dil) into each transplant site. Both the E12 and E13 heterotopic transplants of presumptive perirhinal cortex contain mostly precursor cells. When these grafts are placed into sensorimotor cortex, callosal connections are formed primarily with the contralateral sensorimotor (Sml) area, the normal projection of Sml cortex. In contrast, the E14 heterotopic transplants of the perirhinal cortical wall, containing many more postmitotic neurons, developed contralateral connections with both sensorimotor and rostral perirhinal cortices. Thus, when precursor cells are transplanted heterotopically, by using E12/E13 donor tissue, the grafts receive projections that are similar to those of the host cortical area. When older cortical neurons, together with precursors, are transplanted into a heterotopic cortical area, by using E14 donor tissue, their cortical connections exhibit both host and original donor phenotypes. The data are consistent with our previous analysis of thalamocortical connections of grafts (Barbe and Levitt, 1992b) and suggest the existence of a cell-cell recognition system for thalamocortical and corticocortical circuit formation, whose mechanisms of action may be linked to the timing of neurogenesis.

Animals↗