Axonal guidance during development of the great cerebral commissures: Descriptive and experimental studies,in vivo,on the role of preformed glial pathways

Do structres exist within the embryonic central nervous system that guide axons across the midline during development of the great cerebral commissures (corpus callosum, anterior commissure)? With the use of serial section and reconstructive computer graphic techniques we have found that during normal ontogeny of the mouse forebrain and before the arrival of the pioneer fibers of the corpus callosum at the midline, a population of primitive glial cells migrates medially (through the fused walls of the dorsal septum) from the ependymal zones of each hemisphere. At the midline, and well rostral to the lamina terminalis, these cells unite to form a bridgelike structure of “sling” suspended below the longitudinal cerebral fissure. The first callosal axons grow along the surface of this cellular bridge as they travel toward the contralateral side of the brain. The “sling” disappears neonatally. The fibers of the anterior commissure grow within the lamina terminalis along a different type of preformed glial structure. Movement of these axons occurs through an aligned system of glial processes separated by wide extracellular spaces. Do these transient glial tissues actually provide guidance cues to the commissural axons? Analyses of three situations in which the glial “sling” is genetically or surgically impaired or nonexistent indicate that this structure does, indeed, play an essential role in the development of the corpus callosum. We have analyzed (1) the embryonic stages of a congenitally acallosal mouse mutant (strain BALB/cCF), (2) several pouch stages of a primitive acallosal marsupial, Didelphys virginiana (opossum), and (3) animals in which the “sling” had been lesioned surgically through the uterine wall in the normal embryo (strain C57BL/6J). In the acallosal mouse mutant fusion of the septal midline is delayed by about 72 hours and the “sling” does not form. Although the would-be callosal axons approach the midline on schedule, they do not cross. Instead, the callosal fibers whirl into a pair of large neuromas adjacent to the longitudinal fissure. Similarly, in the opossum, fusion of the medial septal walls and formation of the glial “sling” are also lacking. However, in this species, instead of traveling dorsally, the “callosal” axons turn ventrally and pass contralaterally by way of the anterior commissure pathway. Surgical disunion of the glial “sling” also resulted in acallosal individuals. The callosal pathology in these affected animals mimicked exactly that of the genetically lesioned mutant. Our observations suggest that many different types of oriented glial tissues exist within the embryonic neural anlage. We propose that such tissues have the ability to influence the directionality of axonal movements and, thereby, play a crucial role in establishing orderly fiber projections within the developing central nervous system.     

Fields of origin and pathways of the interhemispheric commissures in the temporal lobe of macaques

The interhemispheric connections of the cortical areas of the temporal lobe and some neighboring regions were investigated in monkeys (Macaca mulatta and Macaca fascicularis) by anterograde autoradiographic tracing, following injection of radioactively labeled amino acids. The results revealed that the interhemispheric projections of the temporal lobe course through three interhemispheric commissures on their way to the opposite hemisphere. The anterior commissure receives fibers from virtually the entire temporal lobe, including the temporal pole, superior and inferior temporal gyri, and parahippocampal gyrus. Moreover, area 13 of the orbitofrontal cortex, the frontal and temporal subdivisions of the prepiriform cortex, and the cortical and deep nuclei of the amygdala also contribute fibers to the anterior commissure. The heaviest projections arise in the rostral third of the temporal isocortex. These projections become progressively lighter from more caudal regions. By contrast, the corpus callosum receives fibers from the caudal two-thirds of the temporal lobe, including the temporal pole, superior and inferior temporal gyri, and parahippocampal gyrus. The heaviest projections arise in the caudal third of the temporal lobe and cross primarily in the caudal third of the corpus callosum, including the splenium. Progressively lighter projections arise more rostrally. Fibers from proisocortical and isocortical areas of the posterior parahippocampal gyrus cross in the ventralmost part of the splenium (inferior forceps), whereas cortical areas lateral to the occipitotemporal sulcus give rise to fibers that cross in the caudal part of the body of the corpus callosum and dorsal splenium. The dorsal hippocampal commissure receives fibers exclusively from the parahippocampal gyrus. The fibers of the corpus callosum, hippocampal commissure, and, to a lesser extent, the anterior commissure are intimately associated with the ventricular system as they course through the white matter of the temporal lobe. The fields of origin of the anterior commissure and corpus callosum overlap extensively over the caudal two-thirds of the temporal lobe. The posterior parahippocampal gyrus is unique in that it gives rise to fibers that cross in all three commissures.     

Anterior commissure of the wallaby (Macropus eugenii): Adult morphology and development

In metatheria, all neo- and paleo-cortical commissural connections are made by the anterior commissure. We have examined the adult morphology of this commissure and its development in a diprotodontid metatherian, the wallaby (Macropus eugenii), at both the light and electron microscope level. The total number of axons in the adult anterior commissure was 21.7 million, of which 55–62% were myelinated. The dorsal two thirds of the commissure, containing neocortical commissural axons, showed a higher percentage of larger, myelinated axons than the ventral one third, which contains paleocortical commissural axons. The commissure also showed a topographical gradient, with cells in the dorsal cortex projecting through the dorsal region of the commissure, the fasciculus aberrans. In the rostrocaudal axis, axons from the frontal cortex tended to pass more anteriorly through the commissure and those from the occipital more posteriorly, but there was extensive overlap of projections from different areas. The gestation length of this wallaby is 28.3 days, and all commissural development occurs postnatally. The anterior commissure first appeared at P (postnatal day) 14, at which time commissural fibres were apparently derived from the external capsule exclusively. Commissural fibres passing through the internal capsule, and joining the anterior commissure via the fasciculus aberrans, were first noted at P18. By that age there were 94,000 to 161,000 axons. Peak axon counts of 50 to 63 million occurred between P100 and P150. The number of growth cones in a single midline section peaked at approximately P114 (480,000) and dropped to 0 by P170. The distribution of growth cones was analysed during the early stages of anterior commissure development (P18, P30, P82). At P18, growth cones were concentrated in the dorsal parts of the commissural bundle, suggesting a ventrodorsal sequence of addition of axons. There was no apparent preferential association of growth cones with the periphery of the commissure or glial structures at any of the three ages examined. The results show that axonal overproduction and regression in cortical commissural connections are features of development in diprotodontid metatheria, as in eutheria. © 1996 Wiley-Liss, Inc.     

Efferent Projections to the Host Brain from Intrastriatal Striatal Mouse-to-rat Grafts: Time Course and Tissue-type Specificity as Revealed by a Mouse Specific Neuronal Marker

The developmental time-course and growth characteristics of efferent graft-to-host projections were studied from mouse fetal striatal grafts (E13 - 14) implanted as a cell suspension into the ibotenate-lesioned striatum of immunosuppressed adult rats. A cell surface monoclonal antibody specific for mouse neurons (M6) was used to identify the donor cells and their projections into the host brain. At 3 - 5 days after implantation, sparse fascicles of M6-positive graft-derived fibres extended for approximately 0.3 - 0.4 mm across the graft - host border into the surrounding host striatum. From the beginning they were selectively orientated in one direction, i.e. caudally along the myelinated fibre bundles of the internal capsule. At 8 days, the graft-derived fibres were more numerous and more densely labelled. They ran in dense fascicles inside the myelinated bundles of the host internal capsule and reached the rostral host globus pallidus, a distance of approximately 1.2 mm from the caudal tip of the graft. Two weeks after grafting, the M6-positive fibre fascicles were clearly seen to branch within the globus pallidus to form terminal-like networks. From this time onwards, the immunoreactivity of the outgrowing fibre fascicles gradually diminished, although small but dense terminal-like networks could be found in the host globus pallidus in most, but not all, of the rats at longer survival times (3 - 15 weeks). This is consistent with previous work showing that outgrowing axons lose their M6 immunoreactivity as they mature and become myelinated. Control grafts of fetal neocortical and fetal cerebellar tissue were used to assess the tissue-type specificity of the efferent fibre growth. The neocortical implants projected densely up to about 3 mm into the host brain, along the internal capsule and the corpus callosum and into the overlying cortex. By contrast, although the cerebellar grafts survived well, they showed very little efferent fibre growth. Double immunostaining for DARPP-32 and M6 revealed that all M6-positive fibre fascicles extending from the striatal (but not neocortical) grafts also showed DARPP-32 positivity, and thus that it was the DARPP-32-positive regions of the striatal grafts that projected to the host brain. It is concluded that graft-to-host projections, running along and inside host myelinated bundles, are formed from intrastriatal striatal grafts within 1 - 2 weeks of implantation. Grafts of neocortical tissue grew well along the same trajectory, whereas neurons of a type not normally projecting along the internal capsule, i.e. cerebellum, failed to extend axons over any significant distance along this trajectory.     

Unmyelinated axons show selective rostrocaudal pathology in the corpus callosum after traumatic brain injury

Axonal injury is consistently observed after traumatic brain injury (TBI). Prior research has extensively characterized the post-TBI response in myelinated axons. Despite evidence that unmyelinated axons comprise a numerical majority of cerebral axons, pathologic changes in unmyelinated axons after TBI have not been systematically studied. To identify morphologic correlates of functional impairment of unmyelinated fibers after TBI, we assessed ultrastructural changes in corpus callosum axons. Adult rats received moderate fluid percussion TBI, which produced diffuse injury with no contusion. Cross-sectional areas of 13,797 unmyelinated and 3,278 intact myelinated axons were stereologically measured at survival intervals from 3 hours to 15 days after injury. The mean caliber of unmyelinated axons was significantly reduced at 3 to 7 days and recovered by 15 days, but the time course of this shrinkage varied among the genu, mid callosum, and splenium. Relatively large unmyelinated axons seemed to be particularly vulnerable. Injury-induced decreases in unmyelinated fiber density were also observed, but they were more variable than caliber reductions. By contrast, no significant morphometric changes were observed in myelinated axons. The finding of a preferential vulnerability in unmyelinated axons has implications for current concepts of axonal responses after TBI and for development of specifically targeted therapies.     

Cellular and molecular tunnels surrounding the forebrain commissures of human fetuses

Glial cells and extracellular matrix (ECM) molecules surround developing fiber tracts and are implicated in axonal pathfinding. These and other molecules are produced by these strategically located glial cells and have been shown to influence axonal growth across the midline in rodents. We searched for similar cellular and molecular structures surrounding the telencephalic commissures of fetal human brains. Paraffin-embedded brain sections were immunostained for glial fibrillary acidic protein (GFAP) and vimentin (VN) to identify glial cells; for microtubule-associated protein-2 (MAP-2) and neuronal nuclear protein (NeuN) to document neurons; for neurofilament (NF) to identify axons; and for chondroitin sulfate (CS), tenascin (TN), and fibronectin (FN) to show the ECM. As in rodents, three cellular clusters surrounding the corpus callosum were identified by their expression of GFAP and VN (but not MAP-2 or NeuN) from 13 to at least 18 weeks postovulation (wpo): the glial wedge, the glia of the indusium griseum, and the midline sling. CS and TN (but not FN) were expressed pericellularly in these cell groups. The anterior commissure was surrounded by a GFAP+/VN+ glial tunnel from 12 wpo, with TN expression seen between the GFAP+ cell bodies. The fimbria showed GFAP+/VN+ cells at its lateral and medial borders from 12 wpo, with pericellular expression of CS. The fornix showed GFAP+ cells somewhat later (16 wpo). Because these structures are similar to those described for rodents, we concluded that the axon guiding mechanisms postulated for commissural formation in nonhuman mammals may also be operant in the developing human brain.     

Increased Axon Number in the Anterior Commissure of Mice Lacking a Corpus Callosum

Relatively few behavioral deficits are apparent in subjects with hereditary absence of the corpus callosum (CC). The anterior commissure (AC) has been suggested to provide an extracallosal route for the transfer of interhemispheric information in subjects with this congenital defect. Anterior commissure size, axon number, axon diameter, and neuronal distribution were compared between normal mice and those with complete CC absence. No difference in midsagittal AC area was found between normals and acallosals, nor were differences found in the numbers or diameters of myelinated axons. However, axon counts indicated an 17% increase or about 70,000 more unmyelinated axons in the AC of acallosal mice, and the mean diameter of unmyelinated axons was slightly less than in normal mice (0.24 vs 0.26 μm). This decrease in axon diameter enabled more axons to pass through the AC without increasing its midsagittal area. The topographical distribution of neurons sending axons through the AC, assessed with lipophilic dyes, was qualitatively similar for almost all the known regions of origin of the anterior commissure in normal and acallosal mice. There was a pronounced deficit of AC cells in the anterior piriform cortex of BALB/c mice, but this occurred whether or not the mouse suffered absent CC. Although the increase in AC axon number is far smaller than the number of CC axons that fail to reach the opposite hemisphere, the higher number of axons present in the AC of acallosal mice may contribute to the functional compensation for the loss of the CC.     

Interhemispheric pathways of the hippocampal formation, presubiculum, and entorhinal and posterior parahippocampal cortices in the rhesus monkey: the structure and organization of the hippocampal commissures

The interhemispheric pathways originating in the hippocampal formation, presubiculum, and entorhinal and posterior parahippocampal cortices and coursing through the fornix system were investigated by autoradiographic tracing in 29 rhesus monkeys (Macaca mulatta). The results revealed that crossing fibers are segregated into three contiguous systems. A ventral hippocampal commissure lies at the transition between the body and anterior columns of the fornix in the vicinity of the subfornical organ and the interventricular foramina of Monro; it is formed by axons arising in the most anterior (uncal and genual) subdivisions of the hippocampal formation. A dorsal hippocampal commissure lies inferior to the posterior end of the body of the corpus callosum; it is formed by axons arising in the presubiculum and entorhinal cortex of the anterior parahippocampal gyrus and the proisocortical and neocortical subdivisions of the posterior parahippocampal gyrus but not in the hippocampal formation. A hippocampal decussation lies between the ventral hippocampal commissure and dorsal hippocampal commissure; it is formed by axons arising in the body of the hippocampal formation. In contrast to the fibers of the ventral hippocampal commissure and dorsal hippocampal commissure, which terminate in contralateral cortical areas, these decussating fibers terminate in the contralateral septum. Thus, the ventral hippocampal commissure and dorsal hippocampal commissure of the rhesus monkey appear to be homologous to similarly designated structures in other mammals. To the extent that these observations also apply to the interhemispheric fibers of the human hippocampal formation and parahippocampal areas, their possible preservation must be considered when interpreting the effect of callosal transection on seizures and the results of "split-brain" studies, since callosal transection may fail to sever the hippocampal commissures in their entirety.     

Locomotor damage in rats after x-irradiation in utero.

Alterations in gait were found in rats after whole-body irradiation with 125 r on day 14, 15, and 16 of gestation. No effects on locomotion were detected after irradiation on day 17 with 125 r or after irradiation on day 14 with 50 r. A technique was set up for quantitative evaluation of locomotion based on a modification of other methods. Walking patterns of irradiated rats were recorded, when they were adults, by requiring them to walk up a 10° incline through a corridor after their feet had been dipped in ink. Rats irradiated on gestational day 14 had an in-phase, hopping gait with the sine of the angle between the hind feet and the direction of progression over 0.9. Rats irradiated on gestational days 15 and 16 had an alternating, waddling gait with wider stance and broader angle than control rats. Histologic examination of serial sections of the brains of these rats showed that the 14-day rats lacked all telencephalic commissures except for a few fibers which crossed in some rats. There was a progressive improvement in the condition of the anterior and ventral hippocampal commissures up to day 17, but the corpus callosum and dorsal hippocampal commissure were lacking or markedly reduced in all day 17 rats. No animals showed damage to the mesencephalic posterior commissure. Since rats which used the in-phase mode of locomotion were never observed to use alternating gait, the possible causal relationship of the commissural damage to the altered locomotor patterns was considered. In view of the restricted period of damage found for the anterior and ventral hippocampal commissures and the restriction of altered locomotion to damage in the same period, primary involvement of the corpus callosum and dorsal hippocampal commissure could be excluded, but a possible role for the other telencephalic commissures remained.     

MYELINATION OF THE MOUSE CORPUS CALLOSUM

Sturrock R. R. (1980) Neuropathology and Applied Neurobiology 6, 415–420
Myelination of the mouse corpus callosum
Myelination has been studied in the corpus callosum of the mouse brain between birth and 240 days-of-age. Myelin sheaths were first seen at 11 days. The most rapid phase of myelination occurred between 14 and 45 days when 13–5% of axons were myelinated, but myelination continued at a reduced rate up to 240 days when 28% of axons were myelinated. The mean diameter of unmyelinated axons was more or less constant throughout the study with an overall mean diameter of 0–25 ± 0–01 μn. Similarly myelinated axon diameter showed little variation with age with a mean diameter of 0–46 ± 001 μn. This suggests that in the corpus callosum axons do not increase in size until they begin to myelinate.     

Bilateral field advantage and evoked potential interhemispheric transmission in commissurotomy and callosal agenesis.

The role of the corpus callosum versus other cerebral commissures in the interhemispheric integration of visual information was studied in four individuals with complete agenesis of the corpus callosum, two individuals with partial agenesis, one total commissurotomy patient, and normal individuals. Evoked potential (EP) indices of interhemispheric transmission of visual sensory responses were observed during matching of unilateral and bilateral visual field letters and patterns. Neither the commissurotomy nor any of the acallosal patients had ipsilateral hemisphere visual EPs (P1 and N1), demonstrating that the posterior callosum is necessary for interhemispheric transmission of these components of visual evoked potentials. While the commissurotomy patient could not compare bilaterally presented letters, the anterior commissure of the acallosal patients appeared to be sufficient for interhemispheric comparison of single letters. However, bilateral comparison of more complex visual patterns resulted in considerable difficulty for complete agenesis patients, while comparison of patterns was more nearly normal when anterior callosal fibers were present (partial agenesis).     

Effects of prenatal gamma irradiation on the development of the corpus callosum of Swiss mice

The temporal sequence of events related to the effects of prenatal gamma irradiation on the development of the corpus callosum and cerebral cortex was studied in Swiss mice. Pregnant females on gestational day 16 were exposed to a 60Co source receiving total doses of 2 or 3 Gy. The offspring were analyzed at both prenatal and postnatal days. One day after irradiation, a great number of pyknotic figures was seen along the whole extension of the cerebral wall, especially in the proliferative zones. At perinatal ages, the thickness of the proliferative zones was reduced and the glial sling was never identified. From 5 days after birth onwards, we observed a severe shrinkage of layers II + III and IV. The majority of the irradiated mice were totally acallosal (particularly when the 3 Gy dose was used), but some animals presented callosal remnants. These remnants were identified above the ventral hippocampal commissure, except for two animals in which a larger callosal remnant extended from the columns of the fornix to the dorsal hippocampal commissure. The presence of callosal remnants in animals irradiated with 3 Gy was dependent on the age at which the animals were analyzed since remnants were observed in some animals analyzed at perinatal ages, but never in older animals. Callosal defects can be explained at least by three factors: (1) Death of a great part of callosal neurons located at layer III. (2) Postnatal axonal elimination. (3) Absence of the glial sling. The callosal agenesis in the absence of the glial sling indicates that this structure may play a crucial role in guiding callosal axons. However, the presence of callosal remnants indicates that surviving callosal axons can use structures other than the sling to cross the midplane. Our data indicate that axons of the middle portion of the callosum can cross the midplane using the ventral hippocampal commissure as a guide. Additionally, the dorsal hippocampal commissure may play a role in directing axons of the posterior part of the corpus callosum.     

The anterior commissure in man: functional variation in a multisensory system

The anterior commissure, which has been presumed to play a minor role in interhemispheric communication, was tested for the transfer of visual, auditory, and olfactory information in patients with complete sections of the corpus callosum. Four of five patients tested with presumed intact anterior commissures demonstrated interhemispheric transfer of verbal and pictorial stimuli presented visually to a single hemisphere. Evidence was also obtained for auditory and olfactory transfer, although successful interhemispheric communication in all three modalities was not found for any one patient. The data suggest that the human anterior commissure is capable of mediating multisensory, interhemispheric messages of a complex nature and provide evidence of functional plasticity in a phylogenetically early cerebral structure.     

Experimental inhibition of peroxisomal beta-oxidation in rats: influence on brain myelination

Oral administration of thioridazine, an inhibitor of peroxisomal beta-oxidation, to normal rats from weaning till day 60 causes a small increase of the very long chain fatty acid C26 in brain lipids. Myelination in the brain is decreased. In the genu of the corpus callosum the ratio of non-myelinated/myelinated axons is increased. In the commissura anterior the myelin sheaths of the axons are significantly thinner in treated than in control animals. Undernourishment caused by the drug is minimal in this experiment. Area and total DNA of glial nuclei are unaltered in both the genu and the commissura anterior of treated rats. The distribution of chromatin (texture), however, shows small differences in the corpus callosum.     

Patterned neuropathologic events occurring in hyh congenital hydrocephalic mutant mice

Hyh mutant mice develop long-lasting hydrocephalus and represent a good model for investigating neuropathologic events associated with hydrocephalus. The study of their brains by use of lectin binding, bromodeoxyuridine labeling, immunochemistry, and scanning electron microscopy revealed that certain events related to hydrocephalus followed a well-defined pattern. A program of neuroepithelium/ependyma denudation was initiated at embryonic day 12 and terminated at the end of the second postnatal week. After the third postnatal week the denuded areas remained permanently devoid of ependyma. In contrast, a selective group of ependymal areas resisted denudation throughout the lifespan. Ependymal denudation triggered neighboring astrocytes to proliferate. These astrocytes expressed particular glial markers and formed a superficial cell layer replacing the lost ependyma. The loss of the neuroepithelium/ependyma layer at specific regions of the ventricular walls and at specific stages of brain development would explain the fact that only certain brain structures had abnormal development. Therefore, commissural axons forming the corpus callosum and the hippocampal commissure displayed abnormalities, whereas those forming the anterior and posterior commissures did not; and the brain cortex was not homogenously affected, with the cingular and frontal cortices being the most altered regions. All of these telencephalic alterations developed at stages when hydrocephalus was not yet patent at the lateral ventricles, indicating that abnormal neural development and hydrocephalus are linked at the etiologic level, rather than the former being a consequence of the latter. All evidence collected on hydrocephalic hyh mutant mice indicates that a primary alteration in the neuroepithelium/ependyma cell lineage triggers both hydrocephalus and abnormalities in telencephalic development.     

Caprine beta-mannosidosis: development of glial and myelin abnormalities in optic nerve and corpus callosum

In caprine beta-mannosidosis, an inherited dysmyelinating disorder, the myelin deficit shows substantial variation throughout the nervous system. In this study morphometric analysis of optic nerve and corpus callosum sections at selected developmental stages was conducted in order to investigate development and persistence of myelin sheaths, the population of axons ensheathed, and the extent of myelin deficits and glial cell abnormalities. The results show that the myelin deficit is severe at very early stages of development and persists to about the same extent into postnatal life. The corpus callosum, much more severely involved than the optic nerve, contains a substantially smaller percentage of myelinated axons when compared to control. In both regions, larger axons are preferentially myelinated. In the corpus callosum before myelination begins, many glial cells appear abnormal, suggesting an early cellular defect. In the postnatal, myelin-deficient corpus callosum, there is a substantial decrease in glial cell density as compared to control, with abnormal appearance of many of the remaining cell profiles. These results define developmental characteristics of the dysmyelination in caprine beta-mannosidosis and document both the early appearance and the persistence of glial cell body and myelin abnormalities.     

Contributions of the corpus callosum and the anterior commissure to visual activation of inferior temporal neurons.

Most neurons in the inferior temporal cortex of the rhesus monkeys have visual receptive fields that extend across the vertical meridian well into both the contralateral and ipsilateral visual half-fields. We examined the role of different portions of the forebrain commissures in providing the ipsilateral input with the following results. (1) Combined section of the splenium and anterior commissure eliminated visual activation from the ipsilateral visual half-field. (2) Section of the splenium, with sparing of the anterior commissure, reduced the incidence of ipsilateral activation by about one-half. (3) Section of the anterior commissure, with sparing of the splenium, did not alter the incidence of ipsilateral activation. (4) Section of the non-splenial portions of the corpus callosum had no effect on the laterality of the receptive fields. Thus, both the splenium and the anterior commissure but not the non-splenial callosum can provide information from the ipsilateral visual field to neurons in inferior temporal cortex. These results are interpreted as suggesting that the converging input from the two visual half-fields onto single inferior temporal neurons provided by the forebrain commissures may mediate interhemispheric transfer of visual habits.     

Minocycline reduces microgliosis and improves subcortical white matter function in a model of cerebral vascular disease

Chronic cerebral hypoperfusion is a key mechanism associated with white matter disruption in cerebral vascular disease and dementia. In a mouse model relevant to studying cerebral vascular disease, we have previously shown that cerebral hypoperfusion disrupts axon‐glial integrity and the distribution of key paranodal and internodal proteins in subcortical myelinated axons. This disruption of myelinated axons is accompanied by increased microglia and cognitive decline. The aim of the present study was to investigate whether hypoperfusion impairs the functional integrity of white matter, its relation with axon‐glial integrity and microglial number, and whether by targeting microglia these effects can be improved. We show that in response to increasing durations of hypoperfusion, the conduction velocity of myelinated fibres in the corpus callosum is progressively reduced and that paranodal and internodal axon‐glial integrity is disrupted. The number of microglial cells increases in response to hypoperfusion and correlates with disrupted paranodal and internodal integrity and reduced conduction velocities. Further minocycline, a proposed anti‐inflammatory and microglia inhibitor, restores white matter function related to a reduction in the number of microglia. The study suggests that microglial activation contributes to the structural and functional alterations of myelinated axons induced by cerebral hypoperfusion and that dampening microglia numbers/proliferation should be further investigated as potential therapeutic benefit in cerebral vascular disease.     

Sex and environmental influences on the size and ultrastructure of the rat corpus callosum

A contested report of sex differences in the size of the splenium of the corpus callosum in humans prompted the present examination of the corpus callosum in the rat. We have previously found that sex differences can vary with the rearing environment. Consequently, male and female rats were raised from weaning to 55 days of age in either a complex or an isolated environment. There were no sex differences in the size of the corpus callosum in sagittal cross section in these rats; however, rats of both sexes had a larger posterior third of the corpus callosum if they were raised in the complex environment. Because the corpus callosum continues to grow in size past 55 days of age, we examined socially housed rats at 113 days and again found no sex differences. The splenium was examined with electron microscopy in complex and isolation reared rats at 55 days of age. The ultrastructural analysis revealed differences at were not apparent from gross size measures. Females had more unmyelinated axons regardless of environment, and females from the complex environment had more myelinated axons than comparably housed males. In contrast, males in the complex environment had larger myelinated axons than females. Rats of both sexes from the complex environment had larger and more unmyelinated axons than isolated rats. In addition in myelinated axons, plasticity in the females occurred through changes in axon number and in males, through axon size. Thus sex differences exist in axonal number and size and the environment influences these differences.     

EFFECTS OF UNDERNUTRITION DURING THE SUCKLING PERIOD ON GROWTH OF THE ANTERIOR AND POSTERIOR LIMBS OF THE MOUSE ANTERIOR COMMISSURE

The anterior and posterior limbs of the anterior commissure were examined quantitatively in normal mice, and mice undernourished during the suckling period. There was no change in the mean length of the anterior commissure in either group. The total number of axons, the percentage of myelinated axons, and the mean number of myelin lamellae remained unchanged between each group in both limbs. There was, however, a highly significant fall in the total number of cells in both the anterior ( P <0·001) and posterior ( P <0·001) limbs, mainly in the oligodendrocyte fraction of the population. There was no change in the relative proportion of cells in each limb. The cross-sectional area of both the limbs was significantly less in the undernourished animals, with the posterior limb being affected more than the anterior limb. These differences were found to be due to a significant ( P <0·001) fall in the mean diameter of myelinated axons in the anterior limb and to a significant ( P <0·001) fall in the mean diameter of unmyelinated axons in posterior limb. The mean diameters of the unmyelinated axons in the anterior limb, and of the myelinated axons in the posterior limb remained unchanged. The possible reasons for these changes, and their place in the general pathology of deficits and distortions following early undernutrition are discussed.