Maturation of rat visual cortex: IV. The generation, migration, morphogenesis, and connectivity of atypically oriented pyramidal neurons

The generation, migration, and morphogenesis of atypically oriented pyramidal neurons in the rat visual cortex were examined. In the mature cortex, these neurons were distributed through layers II-VI. Moreover, the atypically oriented pyramidal neurons in a particular layer tended to be oriented in a specific way; atypically oriented pyramidal neurons in layer II, layers III-VIa, and layer VIb were obliquely, radially, and obliquely oriented, respectively. Ultrastructurally, the somata of atypically oriented pyramidal neurons contained large euchromatic ovoid nuclei and cytoplasm that was replete with rough endoplasmic reticulum and Golgi apparatus. These somata formed only symmetric axosomatic synapses. Many atypically oriented pyramidal neurons projected axons into the white matter as demonstrated by a Golgi method and by a retrograde tract-tracing technique; however, some of these pyramidal neurons in layers III-V had axons that ascended to layer I. By using a technique which combined retrograde tract tracing with [3H]thymidine autoradiography, it was determined that most atypically oriented pyramidal neurons in layers V and VIa, layer IV, and layer II were generated on gestational days (GD) 15-17, GD 17-19, and GD 20-21, respectively. Atypically oreinted pyramidal neurons were identified during the period from postnatal day 0 (day of birth) to day 30. On day 0, obliquely oriented pyramidal neurons were distributed in the deep cortical plate, i.e., immature layer VI. On day 3, the youngest atypically oriented pyramidal neurons were radially oriented and were located in layer IV. Some obliquely oriented pyramidal neurons were present in layer II on day 6, but the greatest number and the most severely canted pyramidal neurons in layer II were evident on day 9. The orientations of the cell body and the apical dendrite did not change appreciably after migration was complete, except for those in layers V and VI with obliquely oriented cell bodies and radially oriented apical dendrites. The second and third postnatal weeks were marked by substantial morphological differentiation of all pyramidal neurons as noted by the lengthening and branching of dendrites and by the appearance of dendritic spines. By the fourth postnatal week, atypically oriented pyramidal neurons achieved their mature morphology. The generation, migration, and morphogenesis of atypically oriented pyramidal neurons proceed by an inside-to-outside sequence. This development is similar and concurrent with that of typically oriented pyramidal neurons.     

Visual cortex development in the ferret. I. Genesis and migration of visual cortical neurons

The production of ferret visual cortical neurons was studied using 3H-thymidine autoradiography. The genesis of cortical neurons begins on or slightly before embryonic day 20 (E20) of the 41 d gestational period, continues postnatally until 2 weeks after birth (P14), and follows an inside-out radial gradient with neurons for the deeper cortical layers being generated before those for the superficial layers. Layer I neurons are generated both early (E20-E30) and late (P1-P14) in the period of cortical neurogenesis and, thus, provide at least a partial exception to the inside-out gradient of cortical neurogenesis. Tangential gradients of cortical neurogenesis extend across areas 17 and 18 in both the anterior-to-posterior and lateral-to-medial directions. Neither of these gradients bears a meaningful relationship to the cortical representation of the visual field. Most infragranular and granular layer neurons are generated prenatally, while most supragranular layer neurons are produced postnatally. Neurons destined for a given layer are produced over a period of several days, and the neurons generated on any given day contribute to the formation of 2 or more cortical layers. In general, prenatally generated neurons complete their migration in 1 week or less, while most postnatally generated neurons require approximately 2 weeks to complete their migration.     

Times of generation of glutamic acid decarboxylase immunoreactive neurons in mouse somatosensory cortex

The birth dates of neurons showing glutamic acid decarboxylase (GAD) immunoreactivity have been determined in mouse somatosensory cortex. Pregnant C57Bl mice received pulse injections of (3H)thymidine from E10 through E17 (E0 being the day of mating). The distributions of thymidine-labeled, GAD-positive and nonimmunoreactive (non-GAD) cells as a function of depth under the pial surface were recorded in adult animals. The maximum rate of generation of GAD-positive neurons occurred at E14, whereas the generation of non-GAD neurons reached its maximum rate at E13. Except for those in layer I, GAD-positive neurons followed an inside-out sequence of positioning. GAD-positive neurons born at E12 and E13 were located in layers VI-IV. GAD-positive neurons born at E14 were found throughout the cortical thickness, with a maximum in layer IV. The GAD-positive neurons labeled after pulses at E15 or E16 or E17 were limited to the superficial strata, forming a band that became narrower as it moved toward the pial surface with increase in age of pulse labeling. GAD-positive neurons in layer I were generated at a constant rate during the whole embryonic period analyzed. Non-GAD neurons also followed an inside-out spatiotemporal gradient. Two partially overlapping phases were distinguished in non-GAD neurogenesis. During the first phase (from E12 to E14) neurons populating adult layers VI and V originated, while neurons located in layers IV through I were generated during the second phase (from E13 to E17). Since GAD-immunoreactive neurons form a heterogeneous population, we envisage further studies in order to test whether differences exist in birth dates among the classes.     

Ontogeny of cholinergic neurons in the mouse forebrain

The development of cholinergic neurons in the mouse forebrain was studied by immunocytochemistry with a monoclonal antibody to choline acetyltransferase (ChAT), the rate-limiting enzyme for acetylcholine synthesis. Since this antibody stained dividing cells in ventricular germinal zones as well as differentiating neurons, likely routes of migration could be inferred on the basis of the location of immunoreactive (IR) cells at different gestational ages. Germinal zones for cholinergic cells were observed in all ventricular zones of the forebrain with the ventral zones generating the earliest cells by gestational day 13.5 (GD13.5). On GD14, ChAT IR cells were visible in the germinal zones of the eye, olfactory ventricle, anterior horn, and dorsolateral aspect of the lateral ventricle, lateral ganglionic eminence, ventro- and dorsolateral third ventricle, and in the pineal anlage (epiphysis). ChAT IR neurons continued to develop in these and additional germinal zones on GD15, including the medial, dorsal, and dorsomedial walls of the lateral ventricle, and the medial and dorsal ganglionic eminence. On GD16, ChAT IR neurons were located in the prelimbic, pyriform, and parietal cortices and the lamina terminalis, and a cluster of IR cells was observed in the ventricular zone of the caudatopallial angle. On GD17-18, neurons in the anterior olfactory nucleus, olfactory tubercle, horizontal and vertical nucleus of the diagonal band, and medial septal nucleus stained more darkly and were multipolar, whereas immature bipolar neurons appeared to continue their migration into the hippocampus and along major fiber tracts, such as the corpus callosum, external capsule, fornix and anterior commissure. This study provides a comprehensive view of the zones of origin, probable routes of migration, and final destination of cholinergic neurons in the mouse forebrain.     

Comparison of the distributions of ipsilaterally and contralaterally projecting corticocortical neurons in cat visual cortex using two fluorescent tracers

Using the retrograde fluorescent tracers Fast Blue and Diamidino Yellow we have studied the callosal and ipsilateral corticocortical connections between the cat's area 17/18 border region and the posteromedial lateral suprasylvian visual area (PMLS), as well as the callosal connections of each of these regions with its contralateral homologue. The main goal was to determine whether single cortical neurons project with branching axons to more than one cortical target. In addition, the double-labeling technique enabled us to examine, within a single section of cortical tissue, the relative distributions of neurons with different cortical targets. Most corticocortical neurons labeled in the area 17/18 border region and in area PMLS projected to only one of the cortical injection sites tested. When two callosal neuron types were labeled in the same area, no double-labeled neurons were found. When ipsilateral corticocortical and callosal neurons were labeled in combination, a few double-labeled neurons were found in both cortical regions examined. The most common type of double-labeled neuron was located in area PMLS and projected bilaterally to the area 17/18 border region. Our findings regarding the laminar distributions of ipsi- and contralaterally projecting neurons are in agreement with previous studies. In addition, we have found that, for callosal neurons within the upper layers of areas 17 and 18, neurons projecting to the contralateral area 17/18 border are located in the lower half of layer II/III and in upper layer IV, whereas neurons projecting to contralateral area PMLS are restricted to the lower portion of layer II/III. In addition, for callosal neurons within the deep layers of area PMLS, neurons projecting to contralateral area PMLS are located throughout layers V and VI, whereas neurons projecting to the contralateral area 17/18 border are restricted to layer VI. There are numerous other possible targets for axon collaterals not examined in this paper. However, the scarcity of neurons with multiple projections demonstrated in this study reflects the high degree of specificity of cortical connectivity. This anatomical organization may be the basis for a precise channeling of differential information at the single neuron level.     

Distribution of visual callosal projection neurons in hamsters subjected to transection of the optic radiations on the day of birth

The optic radiations of hamsters were transected on the day of birth and visual callosal projections in these animals were traced using retrograde transport of either horseradish peroxidase (HRP) or the fluorescent tracers True blue (TB) or Diamidino yellow (DY) when the animals reached maturity (greater than 45 days of age). In the hemisphere ipsilateral to the neonatal lesion, the distribution of callosal cells was markedly altered. These neurons were almost completely restricted to a continuous band in lower lamina V and the upper portion of layer VI. Anterograde HRP transport to the deafferented hemisphere also revealed an abnormal distribution of callosal terminals. The band of labelling that is located along the 17-18a border in the normals was much broader than is normally the case. In the hemisphere contralateral to the lesion, the distributions of callosal cells and terminals were essentially normal. Labelled neurons were located in the infragranular layers (primarily lower layer V and the upper part of lamina VI) throughout area 17 and also in layers II-IV in the 17-18a border region. Anterograde labelling was visible in layers V and VI throughout the mediolateral extent of the dorsal posterior neocortex and supragranular labelling was restricted to the lateral portion of area 17 and medial 18a. These results suggest that the normal thalamic projection to the visual cortex is necessary for the establishment of the strip of supragranular callosal projection neurons which is normally located in the 17-18a border region, but not for the establishment (or maintenance) of callosal projections by large numbers of neurons in the infragranular laminae. They show further that neonatal transection of the optic radiations results in reduction in the correspondence between the distributions of callosal cells and terminals in the deafferented hemisphere.     

Effect of prenatal exposure to alcohol on the distribution and time of origin of corticospinal neurons in the rat

The distribution and the time of origin of corticospinal neurons were examined in rats prenatally exposed to ethanol and in control rats. The distribution of corticospinal neurons was determined by tracing the retrograde transport of horseradish peroxidase (HRP) from an injection site in the cervical spinal cord. In control rats, HRP-positive neurons were distributed in layer Vb throughout motor area 4, rostral motor area 6/8, dorsal somatosensory area 3, caudal somatosensory area 2, and various "association" regions including parietal areas 14, 39, and 40, occipital areas 18a and 18b, cingulate areas 24a and 24b, and prefrontal area 32. In ethanol-exposed rats, the distribution of retrogradely labeled neurons was similar to control animals with three notable exceptions: (1) HRP-positive neurons were evident throughout the rostrocaudal extent of area 6/8; (2) occasionally ectopic labeled neurons were identified in the supragranular layers, layers Va and Vc, and superficial layer VI; and (3) the density of HRP-labeled neurons and the ratio of labeled neurons to the total number of neurons in areas 4, 6/8, 3 and 2 were significantly greater (20-48%) in ethanol-exposed rats than in controls. There was, however, no intergroup difference in the area of the cell bodies of HRP-positive neurons. Taken together, these findings indicate that ethanol exposure resulted in an increased number of corticospinal neurons. The time of origin of corticospinal neurons was determined by using a technique that combined tritiated thymidine autoradiography and retrograde transport of HRP. In control animals, HRP-positive neurons were double labeled by an injection of tritiated thymidine on gestational day (GD) 15, 16, or 17. In ethanol-exposed rats, corticospinal neurons were generated on GD 16, 17, and 18, the late-generated ones being distributed in caudal area 6/8. These intergroup differences represent a persisting ethanol-induced alteration of cortical structure that may underlie motor dysfunction and mental retardation in fetal alcohol-affected offspring. Moreover, the increase in the number and the delay in the time of origin of corticospinal neurons suggest that the normal process of paring down exuberant corticospinal projections may be affected by prenatal exposure to ethanol.     

Immunocytochemical expression of monocarboxylate transporters in the human visual cortex at midgestation

Lactate and the other monocarboxylates are a major energy source for the developing brain. We investigated the immunocytochemical expression of two monocarboxylate transporters, MCT1 and MCT2, in the human visual cortex between 13 and 26 post-ovulatory weeks. We used immunoperoxidase and immunofluorescence techniques to determine whether these transporters co-localized with markers for blood vessels (CD34), neurons (microtubule-associated protein 2 [MAP2], SMI 311), radial glia (vimentin), or astrocytes (glial fibrillary acidic protein [GFAP], S100beta protein). MCT1 immunoreactivity was visible in blood vessel walls as early as the 13th week of gestation mainly in the cortical plate and subplate. At this stage, less than 10% of vessels in the ventricular layer expressed MCT1, whereas all blood vessels walls showed this immunoreactivity at the 26th gestational week. Starting at the 19th week of gestation, sparse MCT1 positive cell bodies were detected, some of them co-localized with MAP2 immunoreactivity. MCT2 immunoreactivity was noted in astrocytic cell bodies from week 19 and spread subsequently to the astrocyte end-feet in contact with blood vessels. MCTs immunoreactivities were most marked in the subplate and deep cortical plate, where the most differentiated neurons were located. Our findings suggest that monocarboxylate trafficking between vessels (MCT1), astrocytes (MCT2) and some postmitotic neurons (MCT1) could develop gradually toward 20 gestational weeks (g.w.). These data suggest that lactate or other monocarboxylates could represent a significant energy source for the human visual cortex at this early stage.     

Axonal regrowth of layer II-III visual-projecting cortical neurons in rats fails beyond eye opening

Fetal neurons (embryonic age E16) of occipital origin grafted in the visual cortex of albino rats at increasing postnatal stages (P0, P7, P15, P30, P60, P120) can be activated by photic stimulation. Inputs originate from five major areas of the brain ipsilateral to the graft, namely, the claustrum, the periallocortex/proisocortex, the isocortex, the visual thalamus, and some unspecific subthalamic and hypothalamic nuclei. All inputs decrease in number with the age at which grafting was performed. Isocortical afferents exhibit furthermore a progressive laminar shaping. In neonates, layer II-III and layer V-VI neurons contribute equally to the graft input. In adults, grafts receive prominent input (approximately 70-80%) from layer VI neurons whereas layer II-III neurons account for less than 10%. Proportions of layer IV (approximately 2-4%) and layer V (approximately 15-20%) neurons innervating the graft remain stable, irrespective of the age of the recipient. The adult pattern of connectivity between the host brain and the graft establishes in frontal and temporal areas 1 week earlier than in occipital areas. It is nearly completed in postnatal day 15 (P15) grafted recipients. Supragranular neurons would be thus unable to innervate and to make stable synapses at the graft level beyond P15, i.e., when eyes open. Some infragranular neurons (supposedly remnants of the earliest generated cortical cell population) still have this capacity in adults.     

Neurogenetic and morphogenetic heterogeneity in the bed nucleus of the stria terminalis

Neurogenesis and morphogenesis in the rat bed nucleus of the stria terminalis (strial bed nucleus) were examined with [3H]thymidine autoradiography. For neurogenesis, the experimental animals were the offspring of pregnant females given an injection of [3H]thymidine on 2 consecutive gestational days. Nine groups of embryos were exposed to [3H]thymidine on E13-E14, E14-E15,... E21-E22, respectively. On P60, the percentage of labeled cells and the proportion of cells originating during 24-hour periods were quantified at six anteroposterior levels in the strial bed nucleus. On the basis of neurogenetic gradients, the strial bed nucleus was divided into anterior and posterior parts. The anterior strial bed nucleus shows a caudal (older) to rostral (younger) neurogenetic gradient. Cells in the vicinity of the anterior commissural decussation are generated mainly between E13 and E16, cells just posterior to the nucleus accumbens mainly between E15 and E17. Within each rostrocaudal level, neurons originate in combined dorsal to ventral and medial to lateral neurogenetic gradients so that the oldest cells are located ventromedially and the youngest cells dorsolaterally. The most caudal level has some small neurons adjacent to the internal capsule that originate between E17 and E20. In the posterior strial bed nucleus, neurons extend ventromedially into the posterior preoptic area. Cells are generated simultaneously along the rostrocaudal plane in a modified lateral (older) to medial (younger) neurogenetic gradient. Ventrolateral neurons originate mainly between E13 and E16, dorsolateral neurons mainly between E15 and E16, and medial neurons mainly between E15 and E17. The youngest neurons are clumped into a medial "core" area just ventral to the fornix. For morphogenesis, pregnant females were given a single injection of [3H]thymidine during gestation, and their embryos were removed either 2 hours later (short survival) or in successive 24-hour periods (sequential survival). The embryonic brains were examined to locate areas of intensely labeled cells in the putative neuroepithelium of the strial bed nucleus, to trace migratory waves of young neurons, and to establish their final settling locations. Two different neuroepithelial sources produce neurons for the strial bed nucleus. The anterior strial bed nucleus is generated by a neuroepithelial zone at the base of the inferior horn of the lateral ventricle from the anterior commissural decussation area forward to the primordium of the nucleus accumbens.(ABSTRACT TRUNCATED AT 400 WORDS)     

Treatment with methylazoxymethanol at different gestational days: two-way shuttle box avoidance and residential maze activity in rat offspring.

Pregnant rats were injected with a single dose of methylazoxymethanol (MAM, 25 mg/kg) on gestational days 14, 15, 16, 17, 18 or 19 which resulted in various degrees of microencephaly. Offspring were tested on a two-way shuttle box avoidance and residential maze activity at 60-90 days of age. Rats treated on gestational day 19 (GD19) were severely impaired in the acquisition of the two-way shuttle box task whereas the other groups did not show any significant difference from controls. Spontaneous activity measured for 23 hr in the residential maze was altered as total, time-course and pattern depending on the time of MAM administration: treatment on GD14 prolonged exploratory behavior, treatment on GD15 and GD16 increased nocturnal activity, treatment on GD16 and GD17 induced changes in locomotion patterns and treatment on GD18 and GD19 decreased total activity. These findings indicate that treatment with MAM results in selective deficits in the acquisition of a shuttle box avoidance and alterations of locomotion patterns in the offspring which are dependent on the time of administration.     

Immunohistochemical localization of a membrane-associated, 4.1-like protein in the rat visual cortex during postnatal development

Expression and localization of a membrane-associated protein, an analog of erythrocyte protein 4.1, in the visual cortex were immunohistochemically studied in the rat, ranging in age from newborn to adult. In the adult, dendrites and somas of layer V pyramidal cells were stained by the antiprotein 4.1 antibody. In most of these immunoreactive neurons, the plasma membrane seemed to be preferentially stained. Neurons located in layers II and III of the cortex were only faintly stained, and those in layers IV and VI were not stained. At birth, the immunoreactivity was already present in pyramidal cells located in the upper part of the cortical subplate. Immature neurons located in the cortical plate were not stained by the antibody, suggesting that the 4.1-like protein is expressed only in the neurons that have differentiated or are differentiating. At postnatal days 2-8, immunoreactive neurons were dramatically increased in layers V and VI and intense labeling was seen at the apical dendrites of layer V pyramidal cells. Most of the stained processes of these and other neurons showed a sign of rapid dendritic growth, i.e., growth cones and filopidia. At days 10-17, the basal dendrites of pyramidal cells in layers II and III became detectable, although still slender. At days 20-37, these dendrites in layers II, III, and V became intensely immunoreactive, and dendritic spines were visualized by the antibody. Throughout all the ages, axons of neurons and neuroglia were not stained by the antibody. Also, most of the neurons in layer IV of the cortex were not immunoreactive. These results suggest that the 4.1-like protein is abundantly expressed in growing parts of the dendrites and spines. A hypothesis that this protein may play a role in synaptic plasticity in the developing visual cortex is discussed.     

Overlap of somatic and visual response areas in the Wulst of pigeon.

The somatic and visual response areas of the Wulst were investigated electrophysiologically in pigeons. Somatosensory neurons are distributed in the hyperstriatum accessorium (HA), the hyperstriatum intercalatus superior (HIS) and the hyperstraitum dorsal (HD), mainly in HA. The radial nerve response area is relatively larger and overlaps the sciatic nerve area. Visual neurons are located in HA, especially the more superficial part of HA. In the Wulst, the somatic response area overlaps the visual area and there is somatosensory-visual convergence.     

Treatment with methylazoxymethanol at different gestational days: physical, reflex development and spontaneous activity in the offspring

Pregnant rats were injected with a single dose of methylazoxymethanol (MAM, 25 mg/kg) on gestational day 14, 15, 16, 17, 18 or 19 and offspring were tested for their physical development, reflex development and spontaneous activity. MAM treatment did not affect gestational and litter parameters at any of the time of administration studied. Treatment at gestational day 14 (GD14) had the most severe effect on functional neurodevelopment until weaning: righting reflex at surface, chimney test, horizontal wire test resulted altered. Administration at GD15, 16, 18, 19 did not affect the performance in these tests. Offspring treated at GD17 showed a delayed eye opening and an impaired performance in the horizontal wire test. When tested at 50 days of age on the rotarod, all the treated groups performed worse than controls with the exception of GD19 treated offspring. Administration at GD14 and GD15 resulted in increased spontaneous activity of the offspring at 21 days but not at 60 days of age. Different degrees of microencephaly were observed for all treated groups. The results indicate that alterations of physical and behavioral development induced by MAM treatment are dependent on the time of MAM administration, and specific behavioral tests are able to detect different abnormalities and differentiate among treatment groups. Some alterations observed in MAM rats undergo to adaptive changes during maturation of the CNS.     

Laminar distribution of receptive field properties in the primary visual cortex of the mouse

We studied the receptive field properties of single neurons in the primary visual cortex (area 17) of the mouse and the distribution of receptive field types among the cortical laminae. Three basic receptive field types were found: 1) Cells with oriented receptive fields, many of which could be classified as simple or complex, were found in all layers of the cortex, but occurred with greater frequency in layers II and III and less commonly in Layer IV. 2) Cells with non-oriented receptive fields had ON, OFF, or ON-OFF centers; they were found in all layers but were predominant in layer IV. Two subclasses of non-oriented receptive fields were characterized based on their responses to stationary and moving stimuli. One group of cells with non-oriented receptive fields responded vigorously with sustained firing to stationary flashing stimuli, and also responded well to moving stimuli over a wide range of stimulus velocities. A second group of non-oriented cells, termed motion-selective, responded poorly or not at all to stationary stimuli and responded optimally to moving stimuli over a restricted range of velocities. 3) A distinct group of neurons, termed large field, non-oriented (LFNO) cells, were found almost exclusively in layer V. LFNO cells had receptive fields that were larger than those of the other two major classes at all visual-field locations; they also had higher rates of spontaneous activity and responded to higher stimulus velocities than the other classes. In these respects, LFNO cells resembled the layer V cells of area 17 in the cat and the layer V and VI cells of area 17 in the monkey that project to the superior colliculus. We injected horseradish peroxidase into the superior colliculus, and determined that corticotectal cells in the mouse were also located in layer V, the layer where we recorded LFNO cells. Additional evidence that some LFNO cells project to the superior colliculus was provided by preliminary experiments in which we stimulated the superior colliculus and antidromically activated cortical cells with LFNO receptive fields. Neurons with LFNO receptive fields thus constitute a class that is functionally distinct, with cell bodies that are located in a single layer (V) of area 17 in the mouse.     

Neurons immunoreactive for vasoactive intestinal polypeptide in the rat primary somatosensory cortex: morphology and spatial relationship to barrel-related columns

Vasoactive intestinal polypeptide (VIP) in neocortex affects neuronal excitability as well as cortical blood flow and metabolism. Interneurons immunoreactive for VIP (VIP-IR neurons) are characterized by their predominantly bipolar appearance and the radial orientation of their main dendrites. In order to determine whether the morphology of VIP-IR neurons is related to the functional organization of the cortex into vertical columns, we combined both immunostaining of neurons containing VIP and cytochrome oxidase histochemistry for visualizing barrels, morphological layer IV correlates of functional columns, in the primary somatosensory (barrel) cortex of rats. VIP-IR neurons were localized in supragranular (48%), granular (16%), and infragranular layers (36%) as well as in the white matter. In the granular layer, a clear trend that more neurons were located in interbarrel septa rather than in barrels could be observed, resulting in a neuronal density which was about one-third higher in the septal area. VIP-IR neurons from the different cortical layers were three-dimensionally reconstructed from serial sections by using a computer microscope system. The neurons were mostly bipolar. Striking morphological differences in both axonal and dendritic trees were found between neurons whose cell bodies were located in supragranular, granular, and the upper part of infragranular layers, and those whose cell bodies were located in the area below. The former had dendrites which often reached layer I, where they bifurcated several times, and axonal trees which were particularly oriented vertically, with a tangential extent smaller than the width of barrels. Therefore, these neurons were mostly confined to either a barrel- or septum-related column. By contrast, the dendrites of neurons of the latter group did not reach the granular layer. Furthermore, these neurons had axons with sometimes very long horizontal collaterals, which often spanned two, in one case three, barrel columns. It is proposed that the differential morphology of neurons with different locations as stated above parallels to some extent the divergence of input streaming into the corresponding layer-defined areas. As a possible consequence of this, VIP-IR neurons may be capable of adapting the excitability and metabolism of cortical compartments either in a spatially limited or more extensive way.     

Proportion of glutamate- and aspartate-immunoreactive neurons in the efferent pathways of the rat visual cortex varies according to the target.

Immunohistochemistry, with antisera directed against glutamate (Glu) or aspartate (Asp), was combined with wheat germ agglutinin-horseradish peroxidase (WGA-HRP) histochemistry to examine the distribution, morphology, and proportions of Glu- and Asp-containing neurons that give rise to corticofugal and callosal projections of the rat visual cortex. WGA-HRP injections in the dorsal lateral geniculate nucleus resulted in retrograde labelling of small and medium-sized cells throughout layer VI of the visual cortex. Of these cells, 60% were also Glu-immunoreactive and 61% Asp-positive. WGA-HRP injections in the superior colliculus labelled large and medium-sized neurons in the upper portion of layer V of the visual cortex. Of these cells, 46% were also stained for Glu and 66% for Asp. Injections in the pontine nuclei resulted in retrograde labelling of cells in the deeper part of cortical layer V. Retrogradely labelled cells, which were also immunoreactive for Glu or Asp, were large pyramidal cells. Corticopontine neurons, which were also Glu-positive, accounted for 42% of the total number of WGA-HRP labelled cells, whilst for Asp-positive neurons this percentage was 51%. Finally, after injections in the visual cortex, retrogradely labelled small and medium-sized cells were found throughout layers II-VI in the contralateral visual cortex. Of these neurons, 38% were also labelled for Glu while 49% were also Asp-immunoreactive. The present results demonstrate that substantial proportions of projection neurons in the rat visual cortex are immunoreactive for Glu or Asp, suggesting that these excitatory amino acids are the major transmitters used by the cortical efferent systems examined. Furthermore, the proportions of these immunoreactive neurons in the efferent pathways vary according to the target.     

Visual activity in macaque area V4 depends on area 17 input

It is known that some direct projections from the lateral geniculate nucleus terminate in area V4 of the macaque monkey. Retinal information can also bypass area 17 and reach V4 through relays in the superior colliculus and pulvinar. This raises the question whether area V4 is visually responsive in the absence of input from area 17. We tested this possibility by recording in area V4 while inactivating a region of area 17 by cooling. This led to a complete abolition of the visual responses of practically all the neurons whose receptive fields were included in the visual field region coded by the inactivated zone in area 17. In contrast, neurons whose receptive fields were outside this region remained visually responsive.     

Cortical projections to the superior colliculus in the macaque monkey: A retrograde study using horseradish peroxidase

The topical and laminar distribution of corticotectal cells, as well as their size and morphology, were studied in the macaque monkey with the horseradish peroxidase (HRP) technique. After HRP injections restricted primarily to the superficial layers of the colliculus, labelled cells were found in visual cortex (areas 17, 18, and 19) and both in the frontal eye field (area 8) and the adjacent part of premotor cortex (area 6). The clustering of labelled cells in visual cortex indicated that each of the anatomically and functionally distinct visual areas has its own set of collicular projections. When intermediate and deeper layers of the colliculus were injected, labelled cells were found also in posterior parietal cortex (area 7) where they were concentrated mainly on the posterior bank of the intraparietal fissure, in inferotemporal cortex (areas 20 and 21), in auditory cortex (area 22), in the somatosensory representation SII (anterior bank of sylvian fissure, area 2), in upper insular cortex (area 14), in motor cortex (area 4), in premotor cortex (area 6), and in prefrontal cortex (area 9). In the motor and premotor cortex, labelled cells formed a continuous band which appeared to stretch across finger-hand-arm-shoulder-neck representation. Similarly, the cluster of labelled cells in area 2 may correspond to the finger-hand representation of SII. The cortical regions not containing labelled cells were the somatosensory representation SI (areas 3, 1 and 2) and the infraorbital cortex. Labelled cells were restricted to layer V of all cortical areas except in the primary visual cortex, where labelled cells were found in both layer V and layer VI. The size spectrum of corticotectal cells ranged from 14.8 μm (average diameter) in area 17 to 27.8 μm in area 6, comprising cells as small as 8 μm and as large as 45 μm. Labelled cells in posterior parietal (area 7), in auditory (area 22), and in motor cortex (area 4) were small and distributed over only a narrow range of sizes. Those in premotor cortex (area 6) were often large and had a wide range in size distribution. The differences in size and morphology of corticotectal neurons suggest that they do not form a uniform class of neurons.     

Elevation of fetal dopamine following exposure to methamphetamine in utero

The effect of methamphetamine on fetal dopaminergic function was examined following treatment of pregnant mice twice daily with 40 mg/kg methamphetamine from either gestational day (GD) 7-13 or from GD 7-15. Dopamine levels were elevated in fetal GD 16 corpus striatum and rostral mesencephalon following both treatment regimens. This increase in fetal dopamine is consistent with our findings that exposure to methamphetamine in utero results in adult dopaminergic neurons which are more responsive in terms of methamphetamine induced release of the neurotransmitter and more sensitive to the neurotoxic effects of the drug.