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PubMed1984

Physiological Studies on Pea Tendrils : XIV. Effects of Mechanical Perturbation, Light, and 2-Deoxy-d-Glucose on Callose Deposition and Tendril Coiling.

When excised tendrils of pea (Pisum sativum L. cv Alaska) are mechanically perturbed there is an immediate and transient increase in callose deposition in the sieve cells. Mechanical perturbation (MP) results in a coiling response in light-grown tendrils and in dark-adapted tendrils, provided, in the latter case, that they receive adequate illumination within a limited period of time after MP. In nonperturbed tendrils the number of callose deposits decreases to some minimum with increasing time in the dark, and their ability to coil in the dark in response to MP diminishes with time in the dark. The transient increase of callose deposition due to MP, however, occurs whether or not tendrils are dark adapted, and whether they receive light or are retained in the dark after MP. This indicates that if callose is directly involved in tendril coiling, then it exerts its effect on the sensory perception of the mechanical stimulus. In the present investigation, there is never tendril coiling without the transient increase in callose, and the time after MP at which the peak of callose deposition occurs precedes the time of the peak amount of coiling.An inhibitor of callose formation, 2-deoxy-d-glucose (DDG), is equally effective at inhibiting tendril coiling and MP-induced callose deposition, indicating, within the limitations of the specificity of DDG, that callose deposition may be required in order for the coiling response to occur. Alternatively, DDG may prevent the availability of some other factor necessary for tendril coiling.

Journal Article↗

PubMed1995

Anomalous patterns of callosal connections develop in visual cortex of monocularly enucleated hamsters.

In this study we analyzed the effect of neonatal monocular enucleation on the pattern of callosal connections in striate cortex of the golden hamster. Callosal connections were revealed in the hemisphere ipsilateral to the remaining eye following multiple injections of either the enzyme horseradish peroxidase or the fluorescent tracer Fluoro-Gold into the contralateral hemisphere. The most salient anomaly induced by the removal of one eye at birth is the appearance of a dense band of callosal connections that runs anteroposteriorly in medial portions of striate cortex. No obvious changes in the laminar distribution of callosal connections were observed. Comparison of our present results with those obtained by Olavarria et al (1987) in monocularly enucleated rats reveals that neonatal enucleation induces remarkably similar anomalies in the callosal patterns of rats and hamsters. This similarity suggests that the role the eyes play in the development of the visual callosal pathway is similar among rodent species. Moreover, the finding of an anomalous callosal band in striate cortex one-eyed hamsters supports the notion that disruption of visual input does not arrest callosal development, but rather leads to the development of entirely new features in the callosal pattern.

Animals↗

PubMed1997

Callosal neurons in the cingulate cortical plate and subplate of human fetuses.

Given the scarcity of data on the development of the cerebral cortex and its connections in man, four brains of human fetuses at 25, 26, 30, and 32 weeks postovulation were used to investigate the following: 1) the radial distribution of callosal neurons in the cingulate cortex at the immediate postmigratory period; 2) the existence of callosally projecting neurons in the cortical subplate; and 3) the dendritic morphology of developing callosal neurons. The carbocyanine dye (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate) (DiI) was used as a fluorescent postmortem tracer for the identification and morphological description of callosal neurons, 4-6 months after the insertion of DiI crystals at the callosal midplane. Sixty-one completely labeled neurons were selected for microscopical analysis, drawn by use of a camera lucida and photographed. The main findings were the following: 1) the human cingulate cortex at 25-32 weeks postovulation contains callosally projecting neurons both in the cortical plate and in the subplate; 2) callosal cells in the plate are mostly spiny pyramids with somata distributed uniformly throughout the depth of the plate, irrespective of rostrocaudal position. They have well-differentiated basal dendrites and apical dendrites that consistently ramify within layer 1; 3) subplate callosal cells are smooth neurons of diverse dendritic morphology, distributed widely throughout the subplate depth. They were classified into four cell types according to the dendritic morphology: radially oriented, horizontally oriented, multipolars, and inverted pyramids. These findings extend to the human brain some of the evidence obtained in animals concerning the development of the cerebral cortex, especially those that are relevant to the formation of a transitory circuitry in the subplate.

Cerebral Cortex↗

PubMed2006

Development of callosal topography in visual cortex of normal and enucleated rats.

In normal rats callosal projections in striate cortex connect retinotopically corresponding, nonmirror-symmetric cortical loci, whereas in rats bilaterally enucleated at birth, callosal fibers connect topographically mismatched, mirror-symmetric loci. Moreover, retina input specifies the topography of callosal projections by postnatal day (P)6. To investigate whether retinal input guides development of callosal maps by promoting either the corrective pruning of exuberant axon branches or the specific ingrowth and elaboration of axon branches at topographically correct places, we studied the topography of emerging callosal connections at and immediately after P6. After restricted intracortical injections of anterogradely and retrogradely transported tracers we observed that the normal, nonmirror-symmetric callosal map, as well as the anomalous, mirror-symmetric map observed in neonatally enucleated animals, are present by P6-7, just as collateral branches of simple architecture emerge from their parental axons and grow into superficial cortical layers. Our results therefore do not support the idea that retinal input guides callosal map formation by primarily promoting the large-scale elimination of long, nontopographic branches and arbors. Instead, they suggest that retinal input specifies the sites on the parental axons from which interstitial branches will grow to invade middle and upper cortical layers, thereby ensuring that the location of invading interstitial branches is accurately related to the topographical location of the soma that gives rise to the parental axon. Moreover, our results from enucleated rats suggest that the cues that determine the mirror-symmetric callosal map exert only a weak control on the topography of fiber ingrowth.

Animals↗

PubMed1993

Synapses of extrinsic and intrinsic origin made by callosal projection neurons in mouse visual cortex.

Neurons in areas 17/18a and 17/18b of mouse cerebral cortex were labeled by the retrograde transport of horseradish peroxidase (HRP) transported from severed callosal axons in the contralateral hemisphere. Terminals of the local axon collaterals of labeled neurons (intrinsic terminals) were identified in the border regions of area 17 with areas 18a and 18b, and their distribution and synaptic connectivity were determined. Also examined were the synaptic connections of extrinsic callosal axon terminals labeled by lesion-induced degeneration consequent to the severing of callosal fibers. A postlesion survival time of 3 days was chosen because by this time the extrinsic terminals were all degenerating, whereas the intrinsic terminals were labeled by horseradish peroxidase. Both intrinsic and extrinsic callosal axon terminals occurred in all layers of the cortex where, with rare exception, they formed asymmetrical synapses. Layers II and III contained the highest concentrations of intrinsic and extrinsic callosal axon terminals. Analyses of serial thin sections through layers II and III in both areas 17/18a and 17/18b yielded similar results: 97% of the intrinsic (1,412 total sample) and of the extrinsic (414 total sample) callosal axon terminals synapsed onto dendritic spines, likely those of pyramidal neurons; the remainder synapsed onto dendritic shafts of both spiny and nonspiny neurons. Thus, the synaptic output patterns of intrinsic vs. extrinsic callosal axon terminals are strikingly similar. Moreover, the high proportion of axospinous synapses formed by both types of terminal (97%) contrasts with the proportion of asymmetrical axospinous synapses that occurs in the surrounding neuropil where about 64% of the asymmetrical synapses are onto spines. This result is in accord with previous quantitative studies of the synaptic connectivities of callosal projection neurons in mouse somatosensory cortex, and lends additional weight to the hypothesis that axonal pathways are highly selective for the types of elements with which they synapse.

Animals↗

PubMed1995

Two rules for callosal connectivity in striate cortex of the rat.

In the rat, callosal cells occupy lateral as well as medial portions of striate cortex. In the region of the border between areas 17 and 18, which contains a representation of the vertical meridian of the visual field, cells projecting through the corpus callosum are concentrated throughout the depth of the cortex. In contrast, in medial portion of striate cortex, where peripheral portions of the visual field are represented, callosal cells are preferentially found in infragranular layers. These differences in topography and laminar distribution suggest that these callosal regions, referred to as medial and lateral callosal regions in the present study, subserve different functions. We explored this possibility by analyzing the patterns of callosal linkages in these two callosal regions. We charted the location of retrogradely labeled cells within striate cortex of one hemisphere after placing restricted injections of one or more fluorescent tracers into selected sites in the contralateral striate cortex. We found the medial and lateral callosal regions have distinctly different topographic organizations. Injections into medial striate cortex of one hemisphere produced labeled cells predominantly in mirror-symmetric loci in medial portions of contralateral striate cortex. The arrangement of these connections suggests that they mediate direct interactions between cortical regions representing visual fields located symmetrically on opposite sides of the vertical meridian of the visual field. In contrast, the mapping in the lateral callosal region is reversed: injections into the 17/18a border produced labeled fields located medial to the contralateral 17/18a border, while injections slightly medial to the 17/18a border produced labeled fields located at the contralateral 17/18a border.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1980

Callosal projections from the two body midlines.

1. Horseradish peroxidase (HRP) was injected within the proximal limb and trunk representation zones of the first somatosensory area (SI) of 16 cats. The tangential and laminar distributions of retrogradely labelled neurones (callosal neurones) of the contralateral homotopic cortex were studied. This cortex was explored with microelectrodes on the day after HRP delivery to relate the distribution of callosal neurones to the electrophysiological map of the trunk. 2. Callosal neurones were found in the contralateral SI area mainly in layer III, but also many in layer VI, especially following large HRP injections, and very few in the other layers. Callosal neurones of layer III were mostly pyramidal, those of layer VI pyramidal and non-pyramidal. Many neurones were intensely stained by HRP, and cell details, such as fine dendritic branchings, spines, and axon collaterals, could be seen. 3. Callosal neurones are grouped within two regions located, respectively, in the rostral and caudal parts of the exteroceptive trunk map. The rostral region overlaps the representation of the dorsal midline (cytoarchitectonic field 3b) and the second one that of the ventral body midline (cytoarchitectonic field 2). The cortex intermediate between these two fields contains rare callosal cells and receives afferences from the lateral trunk surface. Few or no callosal cells were found within the proximal limb zones. Neurones recorded from the two midline zones have bilateral receptive fields straddling either the back or the ventral surface of the trunk. 4. It is concluded that the interhemispheric fusion between the two hemibody representations in areas SI is brought about by the mutual callosal links which the two midline zones entertain with their contralateral homologues.

Animals↗

PubMed2005

Two callose synthases, GSL1 and GSL5, play an essential and redundant role in plant and pollen development and in fertility.

Callose, a beta-1,3-glucan that is widespread in plants, is synthesized by callose synthase. Arabidopsis thaliana contains a family of 12 putative callose synthase genes (GSL1-12). The role of callose and of the individual genes in plant development is still largely uncertain. We have now used TILLING and T-DNA insertion mutants (gsl1-1, gsl5-2 and gsl5-3) to study the role of two closely related and linked genes, GSL1 and GSL5, in sporophytic development and in reproduction. Both genes are expressed in all parts of the plant. Sporophytic development was nearly normal in gsl1-1 homozygotes and only moderately defective in homozygotes for either of the two gsl5 alleles. On the other hand, plants that were gsl1-1/+ gsl5/gsl5 were severely defective, with smaller leaves, shorter roots and bolts and smaller flowers. Plants were fertile when the sporophytes had either two wild-type GSL1 alleles, or one GSL5 allele in a gsl1-1 background, but gsl1-1/+ gsl5/gsl5 plants produced an extremely reduced number of viable seeds. A chromosome with mutations in both GSL1 and GSL5 rendered pollen infertile, although such a chromosome could be transmitted via the egg. As a result, it was not possible to obtain plants that were homozygous for mutations in both the GSL genes. Pollen grain development was severely affected in double mutant plants. Many pollen grains were collapsed and inviable in the gsl1-1/gsl1-1 gsl5/+ and gsl1-1/+ gsl5/gsl5 plants. In addition, gsl1-1/+ gsl5/gsl5 plants produced abnormally large pollen with unusual pore structures, and had problems with tetrad dissociation. In this particular genotype, while the callose wall formed around the pollen mother cells, no callose wall separated the resulting tetrads. We conclude that GSL1 and GSL5 play important, but at least partially redundant roles in both sporophytic development and in the development of pollen. They are responsible for the formation of the callose wall that separates the microspores of the tetrad, and also play a gametophytic role later in pollen grain maturation. Other GSL genes may control callose formation at different steps during pollen development.

Arabidopsis↗

PubMed1989

Functional consequences of modification of callosal connections by perinatal enucleation in rat visual cortex.

The effects of neonatal monocular enucleation (right eye) on the callosal connections in the rat visual cortex were studied by physiological and morphological methods. Evoked activity was recorded in the left hemisphere, i.e. contralaterally to the enucleated eye. After enucleation, trans-callosally evoked responses were recorded in a widened stripe of the lateral visual cortex. Compared with the controls, the responsive area was expanded laterally and medially, i.e. into the lateral part of the primary visual area and within the secondary visual cortex (lateral part). Within about 0.5 mm of the expansion, the responses did not differ from those recorded in areas with "normal" callosal connections. Morphological evidence is presented suggesting that this expansion of evoked responses with high amplitudes and short latencies corresponds to an extension of callosal connections with a high density of axon terminals in layers two and three. Further medially within the primary visual cortex, callosally evoked responses with low amplitudes and longer latencies were recorded. The main types of unit responses and characteristic interactions between visually and callosally evoked responses are shown and discussed. These results suggest that following neonatal enucleation (1) the callosal connections expand and form functional synapses in the lateral part of the visual cortex, (2) these connections can activate cortical neurons either directly or by mediation of associational connections between the lateral secondary and primary visual cortex areas and (3) callosal connections can interact with visually evoked potentials and unit responses.

Animals↗

PubMed1995

Migrating neurons in the developing cerebral cortex of the mouse send callosal axons.

The presence of migrating callosal neurons during the development of the murine cerebral cortex was studied using biocytin and the lipophilic dye, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate as retrograde tracers. After injections of biocytin in the presumptive somatosensory cortex of newborn mice which were analysed one day later, many anterogradely labelled fibres coursed towards the contralateral hemisphere through the corpus callosum. Retrogradely labelled callosal cells were also observed. Most callosal neurons corresponded to immature pyramidal cells. In addition, a few biocytin-labelled callosal neurons displayed extremely fusiform shapes, vertical orientation and a short, single process emerging from the apical side of the perikaryon. At the electron microscopic level, these cells had features identical to those described for migrating callosal neurons. Twenty-four hours after birth, these migrating neurons were almost exclusively observed in the upper, dense aspect of the cortical plate (presumptive layers II-III) and only very exceptionally in the infragranular layers. No retrogradely labelled cell resembling migrating neurons were noticed after injections on postnatal days 2 or 5. To study migrating callosal neurons at embryonic stages, crystals of the lipophilic dye were injected in the corpus callosum or the contralateral white matter in embryos aged 17, 18 and 19 days, corresponding to the initial development of the corpus callosum in mice. Whereas callosal migrating neurons were not detected at embryonic days 17 and 18, injections of the lipophilic dye on embryonic day 19 revealed the presence of labelled migrating neurons in the infragranular layers. To corroborate further that these cells are migrating neurons, [3H]thymidine was administered on embryonic days 16 and 17, and labelled mice were injected with biocytin on embryonic day 19 or the first postnatal day. Retrogradely labelled callosal neurons resembling migrating neurons were autoradiographically labelled. These results indicate that the specification of certain neuronal types and the emergence of their cell type-specific characteristics occur shortly after postmitotic neurons leave the ventricular zone, before being positioned within the cortical plate.

Animals↗

PubMed2002

Dendritic morphology of callosal and ipsilateral projection neurons in monkey prefrontal cortex.

Subpopulations of cortical pyramidal neurons have been distinguished based on the projection target of their principal axons or by their dendritic morphology. In this study, we sought to test the hypothesis that pyramidal neurons in monkey prefrontal cortex that furnish callosal or ipsilateral projections have distinctive dendritic morphologies. Retrogradely-labeled, Fast Blue-containing callosal and ipsilateral neurons were intracellularly filled with Lucifer Yellow, immunoconverted, and reconstructed. Quantitative measurements of the size and complexity of the dendritic arbor, including total dendritic length, horizontal extent, number of branch points, maximum branch order, and number of segments, as well as spine density, were made. In general, callosal neurons had larger and more complex dendritic arbors for both apical and basilar dendritic trees than did ipsilateral neurons. The greatest difference was in total dendritic length; the apical and basilar trees of callosal neurons were 34 and 25% longer, respectively. In addition, spine density was also significantly greater on the apical and basilar dendrites of callosal neurons. These findings could not be explained by differences in somal size or completeness of dendritic filling between callosal and ipsilateral neurons. Our observations support the hypothesis that callosal and ipsilateral neurons differ on a number of measures of dendritic size and complexity. Furthermore, these findings imply that these two subpopulations of pyramidal cells differ in the number and perhaps types of excitatory inputs that they receive. Finally, differences in the dendritic morphology of callosal and ipsilateral neurons have implications for understanding the functional attributes of these two populations of cells, as well as for the characterization of pyramidal neurons in human disease states.

Amidines↗

PubMedDate not supplied

Development of the visual callosal cell distribution in the rat: mature features are present at birth.

In the present study, the early postnatal distribution and subsequent fate of visual callosal neurons were studied in neonatal rat pups. Previous studies had indicated that the adult pattern of visual callosal neurons was sculpted from an initially uniform distribution in the neonatal cortex. To reexamine this issue, we used a sensitive tracer, latex microspheres conjugated either to rhodamine or fluorescein, that was injected into the occipital cortex of one hemisphere in pups on the day of birth (PND 1), PND 6, or PND 12. Examination of the resulting retrograde labeling of cortical neurons in the opposite hemisphere indicates that features of the mature visual callosal pattern are present as early as PND 1. At all stages of postnatal development, the relative density of callosal projection cells varies consistently across the mediolateral extent of primary visual cortex-it is always highest in the region of the 17/18a border and lowest in the body of area 17. These data strongly suggest that, from the outset, visual cortical neurons in the region of the 17/18a border preferentially make connections with the opposite hemisphere. The results of experiments in which callosal neurons were labeled on the day of birth indicate that only those neurons that have migrated to their final cortical destinations have extended callosal axons into the vicinity of the visual cortex in the opposite hemisphere. The initial pattern of callosal neurons resembles a dense, compact version of the mature one, and the present study suggests that much of the remaining change in the appearance of this pathway may be accounted for by the decrease in the overall density of neurons that is due to expansion of the cortical gray matter during postnatal life. Taken together, these results suggest that the development of the visual callosal pathway in the rat may be more similar to that in the monkey than has been reported previously.

Animals↗

PubMed2001

Plant callose synthase complexes.

Synthesis of callose (beta-1,3-glucan) in plants has been a topic of much debate over the past several decades. Callose synthase could not be purified to homogeneity and most partially purified cellulose synthase preparations yielded beta-1,3-glucan in vitro, leading to the interpretation that cellulose synthase might be able to synthesize callose. While a rapid progress has been made on the genes involved in cellulose synthesis in the past five years, identification of genes for callose synthases has proven difficult because cognate genes had not been identified in other organisms. An Arabidopsis gene encoding a putative cell plate-specific callose synthase catalytic subunit (CalS1) was recently cloned. CalS1 shares high sequence homology with the well-characterized yeast beta-1,3-glucan synthase and transgenic plant cells over-expressing CalS1 display higher callose synthase activity and accumulate more callose. The callose synthase complex exists in at least two distinct forms in different tissues and interacts with phragmoplastin. UDP-glucose transferase, Rop1 and, possibly, annexin. There are 12 CalS isozymes in Arabidopsis, and each may be tissue-specific and/or regulated under different physiological conditions responding to biotic and abiotic stresses.

Gene Expression Regulation, Enzymologic↗

PubMed2005

Callosal connections of dorsal versus ventral premotor areas in the macaque monkey: a multiple retrograde tracing study.

BACKGROUND: The lateral premotor cortex plays a crucial role in visually guided limb movements. It is divided into two main regions, the dorsal (PMd) and ventral (PMv) areas, which are in turn subdivided into functionally and anatomically distinct rostral (PMd-r and PMv-r) and caudal (PMd-c and PMv-c) sub-regions. We analyzed the callosal inputs to these premotor subdivisions following 23 injections of retrograde tracers in eight macaque monkeys. In each monkey, 2-4 distinct tracers were injected in different areas allowing direct comparisons of callosal connectivity in the same brain. RESULTS: Based on large injections covering the entire extent of the corresponding PM area, we found that each area is strongly connected with its counterpart in the opposite hemisphere. Callosal connectivity with the other premotor areas, the primary motor cortex, prefrontal cortex and somatosensory cortex varied from one area to another. The most extensive callosal inputs terminate in PMd-r and PMd-c, with PMd-r strongly connected with prefrontal cortex. Callosal inputs to PMv-c are more extensive than those to PMv-r, whose connections are restricted to its counterpart area. Quantitative analysis of labelled cells confirms these general findings, and allows an assessment of the relative strength of callosal inputs. CONCLUSION: PMd-r and PMv-r receive their strongest callosal inputs from their respective counterpart areas, whereas PMd-c and PMv-c receive strong inputs from heterotopic areas as well (namely from PMd-r and PMv-r, respectively). Finally, PMd-r stands out as the lateral premotor area with the strongest inputs from the prefrontal cortex, and only the PMd-c and PMv-c receive weak callosal inputs from M1.

Animals↗

PubMed1976

Callosal transfer of impulses originating from superficial and deep nerves of the cat forelimb.

1. Experiments were performed in 18 chloralose-anaesthetized, curarized cats in order to study the callosal transfer of somatic information originated in exteroceptive and proprioceptive receptors. Several cutaneous and deep nerves of the forelimb were prepared and stimulated with graded intensities, so as to activate selectively afferent fibres pertaining to the different groups of Lloyd's classification. Simultaneous records were taken (and averaged on-line by means of a multichannel analyzer) from the distal end of a cut dorsal rootlet (C7-C8), from the cerebral cortex (SI, SII or area 3a, according to the experiment) and from the somesthetic callosal region (SCR). 2. The low-threshold afferent fibres (Group II) of cutaneous origin were found to have a wide projection to the SCR, with the maximal density in its middle portion. Some of the fastest corticocallosal impulses are relayed monosynaptically at cortical level. Plots of the amplitude of cortical and callosal responses as a function of stimulus strength showed that both central responses have the same threshold and exhibit a parallel, sharply-rising amplitude increase, thus suggesting that the cortico-callosal re-transmission system for afferent impulses of cutaneous origin is very powerful in nature. Impulses elicited in afferent fibres of higher threshold (Group III) do not enhance the cortical and callosal positive waves provoked by Group II afferent volleys. 3. Afferent fibres of deep origin were also found to send a wide projection to the SCR, although less substantial than that of cutaneous fibres. Stimulation of the deep radial nerve elicited mass responses in the whole SCR, provided the strength of stimuli was high enough to engage the Group II fibres. Only in the central portion of the SCR were small potentials recorded in response to pure Group I volleys of DRN. Experiments performed with selective stimulation of pure muscular branches of forelimb deep nerves as well as of articular and mixed (muscular and articular) branches gave evidence making it possible to ascertain the origin of deep afferent fibres projecting to the SCR. Stimulation of the forelimb muscular branches with strength provoking full activation of Group I afferent and additional engagement of those of Group II, did not provoke mass responses in the whole extent of the SCR. In order to obtain callosal potentials upon stimulation of pure muscular nerves, it was necessary to increase the stimulus strength at or above the threshold for Group III fibres. On the contrary, the same callosal foci unresponsive to Group I and II muscular afferent volleys exhibited clear-cut responses to stimulation of the lowest-threshold Group I and/or Group II afferents of articular and mixed nerves. From the results it might be inferred that only proprioceptive information originating from articular receptors and from extrafusal muscular afferents has access to the callosal interhemispheric transfer.

Animals↗

PubMed1990

Identification of the UDP-glucose-binding polypeptide of callose synthase from Beta vulgaris L. by photoaffinity labeling with 5-azido-UDP-glucose.

The photoaffinity probe 5-azidouridine 5'-[beta-32P]diphosphate glucose (5N3[32P]UDP-Glc) was used to identify a 57-kDa polypeptide as a strong candidate for the UDP-Glc-binding polypeptide of UDP-glucose: (1,3)-beta-glucan (callose) synthase from red beet (Beta vulgaris L.) storage tissue. Unlabeled 5N3UDP-Glc was a competitive inhibitor of callose synthase with a Ki of 310 microM. Callose synthase was purified from plasma membranes by a two-step solubilization with 3-[(3-cholamidopropyl)dimethylammonio]-1-propane-sulfonate, followed by product entrapment, and photoincorporation of radioactivity from 5N3[32P]UDP-Glc was used to identify UDP-Glc-binding polypeptides that copurified with callose synthase activity. Photoinsertion into the 57-kDa band was closely correlated with all catalytic properties examined. Photolabeling of the 57-kDa polypeptide was enriched upon purification of callose synthase by product entrapment, was abolished with increasing levels of unlabeled UDP-Glc, was dependent upon the presence of divalent cations, and the pH dependence of photolabeling correlated with the pH activity profile of callose synthase. In addition, photolabeling of the 57-kDa band did not occur after phospholipase treatment, which destroys enzyme activity. The extent of labeling of this polypeptide thus correlates closely with the activity of callose synthase under a wide variety of conditions. These results imply that the polypeptide at 57 kDa represents the substrate-binding and cation-regulated component of the callose synthase complex of higher plants.

Affinity Labels↗

PubMed1982

The afferent and efferent callosal connections of retinotopically defined areas in cat cortex.

We compared the callosal afferent and efferent connections of different retinotopic loci within a given visual cortical area as well as the connectivity patterns among similar retinotopic loci in different visual areas. Small injections (75 nl) of a mixture of horseradish peroxidase and [3H]leucine were made through a recording pipette at injection sites identified by retinotopic mapping. A small locus of cortex within a callosally connected region had precise reciprocal connections with the homotopic locus in the contralateral hemisphere. This small locus also was callosally connected with a variable number of heterotopic loci. Both reciprocal and nonreciprocal heterotopic callosal connections were found. Homotopic and heterotopic connections appeared to have a high degree of retinotopic fidelity. Precisely homotopic connections were present not only between locations on the vertical meridian representations at the left and right area 17/18 borders but also, for example, between mirror-symmetrical points on the peripheral horizontal meridian representation in the left and right posteromedial lateral suprasylvian areas. In several experiments, we found that both callosal neurons and terminals in the homotopic cortex were grouped into two to three distinct clusters ranging from 600 to 900 micrometers in width. Callosal neurons with homotopic connections were primarily pyramidal cells in lower layer III and upper layer IV. Outside of areas 17 and 18, there was a significant number of pyramidal and fusiform-shaped callosal neurons in layers V and VI. The majority of callosal terminals were located in layers II, III, and IV.

Afferent Pathways↗

PubMed1993

Functional and magnetic resonance imaging correlates of callosal involvement in multiple sclerosis.

To investigate functional and anatomical features of callosal involvement in multiple sclerosis (MS), performances of 90 patients with definite MS and 25 matched normal control subjects were compared on three tasks exploring interhemispheric transfer of auditory, sensory, and motor information: a verbal dichotic listening task, a crossed tactile finger localization task, and an alternate finger tapping task. Each patient also underwent a magnetic resonance imaging (MRI) scan (1) to appreciate the extent of white-matter changes by a semiquantitative evaluation of hemispheric brain MRI hyperintensities and (2) to measure the degree of total and regional callosal atrophy using an automatized method of partition of the midsagittal callosal area. Interhemispheric transfer and/or integration was impaired in patients with MS for all modalities explored and proportional to both degree of callosal atrophy and diffusion of white-matter lesions. Moreover, in good agreement with data obtained from partial commissurotomy studies, performance on each functional task was predominantly associated with atrophy of one part of the callosum, namely left-ear dichotic suppression with the posterior callosal region, alternate finger tapping with the anterior region, and cross-localization with midanterior and posterior regions. Finally, a subgroup of patients without MRI white-matter hyperintensities also showed significant impairment of callosal function and relative atrophy of the callosum. These findings suggest the potential clinical value of callosal involvement in MS and the usefulness of MS as a model of interhemispheric disconnection.

Adolescent↗