Types Of Nonpyramidal Cells Research Articles

Layer I neocortical ectopia: Cellular organization and local cortical circuitry

Focal cortical dysplasia (FCD) are associated with neurological disorders and cognitive impairments in humans. Molecular layer ectopia, clusters of misplaced cells in layer I of the neocortex, have been identified in patients with developmental dyslexia and psychomotor retardation. Mouse models of this developmental disorder display behavioral impairments and increased seizure susceptibility. Although there is a correlation between cortical malformations and neurological dysfunction, little is known about the morphological and physiological properties of cells within cortical malformations. In the present study we used electrophysiological and immunocytochemical analyses to examine the distribution of neuronal and non-neuronal cell types within and surrounding layer I neocortical ectopia in NXSMD/EiJ mice. We show that cells within ectopia have membrane properties of both pyramidal and a variety of non-pyramidal cell types, including fast-spiking cells. Immunocytochemical analysis for different interneuronal subtypes demonstrates that ectopia contain nonpyramidal cells immunoreactive for calbindin-D28K (CALB), parvalbumin (PARV), and calretinin (CR). Ectopia also contains astrocytes, positive for glial fibrillary acidic protein (GFAP) and oligodendrocyte precursor cells positive for NG2 proteoglycan (NG2). Lastly, we provide electrophysiological and morphological evidence to demonstrate that cells within ectopia receive input from cells within layers I, upper and deeper II/III, and V and provide outputs to cells within deep layer II/III and layer V, but not layers I and upper II/III. These results indicate that ectopia contain cells of different lineages with diverse morphological and physiological properties, and appear to cause disruptions in local cortical circuitry.

Localization of the GoLoco motif carrier regulator of G‐protein signalling 12 and 14 proteins in monkey and rat brain

Regulator of G-protein signalling (RGS)12 and -14 proteins possess the RGS domain, Ras-binding domains and the GoLoco motif. Emerging evidence suggests that these proteins are involved in several cellular functions in addition to stimulation of GTPase activity of G-protein alpha subunits. However, our understanding of the role of the two proteins in brain function remains marginal. Here, we have studied the expression pattern of RGS12 and RGS14 proteins in brain at regional, cellular and subcellular levels. Both proteins were expressed throughout the brain regions, including cortex, hippocampus, striatum, thalamus and substantia nigra. The most intense immunostaining for RGS12 was seen in cortex and that of RGS14 was found in striatum. In cortex, RGS12 and RGS14 proteins were associated with pyramidal and nonpyramidal cell types. Apical dendrites of pyramidal cells were also labelled. RGS12 was found in both nuclear and cytoplasmic compartments. In contrast to RGS12 protein, RGS14 was localized in astrocytes in addition to neurons. Pyramidal cells in the CA1 area showed labelling for both RGS proteins. The presence of RGS12 was predominantly nuclear in the striatum of rat brain; however, the labelling of this protein was non-nuclear in adult monkey brain. To our surprise, in 1-month-old monkey brain the immunostaining pattern of the same protein was changed to nuclear. Non-nuclear staining for RGS12 was also evident in thalamus of adult monkey brain; however, in 1-month-old monkey brain, it was seen into two different populations, one with nuclear and the other with cytoplasmic staining. Both RGS12 and RGS14 were exclusively localized at postsynaptic sites of excitatory synapses. Our results demonstrate a highly dynamic expression pattern of RGS12 and RGS14 proteins in the central nervous system, and support the view that these proteins may participate not only in G-protein receptor signalling pathways but also in other cellular activities.

Dendritic Branch Typing and Spine Expression Patterns in Cortical Nonpyramidal Cells

To understand the dendritic differentiation in various types of cortical nonpyramidal cells, we analyzed quantitatively their dendritic branching and spine expression. The dendritic internode and interspine interval obeyed exponential distributions with type-specific decay constants. The initial branching pattern, internode interval and spine density at the light microscopic level divided nonpyramidal cells into three dendritic types, correlated with axonal, neurochemical and firing types. The initial branching pattern determined the overall vertical spread of dendrites. Basket cell subtypes with different firing and chemical expression patterns were distinct in the vertical and horizontal spatial spread, providing diverse input territories. Internode densities of dendritic spines, as well as those of axonal synaptic boutons, did not correlate with the tortuosities and intervals, suggesting a tendency to distribute synapses homogeneously over the arbor. Dendritic spines identified at the electron microscopic level were different in length and shape among subtypes. Although the density was lower than that of pyramidal cells, spines themselves were also composed of several morphological types such as mushroom and multihead ones, which were expressed differentially among subtypes. Correlation of dendritic branching characteristics with differences in spine structure suggests distinct ways to receive specific inputs among the subtypes.

A quantitative analysis of parvalbumin neurons in rabbit auditory neocortex

Parvalbumin (PV) is a calcium-binding protein present in GABAergic cells in the cerebral cortex and in thalamic relay neurons. In the present study, parvalbumin immunocytochemistry (PVi) and stereological methods were used to obtain estimates of cortical volume, total neuron number, laminar density, and the percentage of PV-immunoreactive neurons in auditory neocortex. PVi clearly delineated the primary auditory cortex (AI), which was characterized by two PV+ bands: dense terminal-like labeling within lamina III/IV and PV+ somata in lamina VIa. Stereological analysis of Nissl-stained sections revealed that the total number of neurons in rabbit AI was 1.48 x 10(6) with a mean neuronal density of 55 x 10(3)/mm3. Based on a mean cortical thickness of 1.92 mm, there are approximately 106,000 neurons in a 1 mm2 column of auditory cortex. PVi yields an extraordinary Golgi-like staining of nonpyramidal cells in all cortical layers. PV+ nonpyramidal cells constitute approximately 7.0% of the neurons in AI. There were significant differences in the morphology and density of PV+ neurons across layers. Although only 5% of cells in lamina I were PV+, three nonpyramidal cell types were present. Lamina II had the highest numerical density within AI but the lowest percentage of PV+ neurons (3.3%). Lamina II, however, contained the greatest diversity of PV+ nonpyramidal cell types, which included small multipolar cells, bipolar cells, and, less frequently, large cells of the bitufted, bipolar, and stellate varieties. Lamina IV had one of the highest numerical densities (67.6 x 10(3) neurons/mm3) and contributed nearly 27% of the total neuron number in AI. The numerical density of PV+ nonpyramidal cells was also greatest within lamina IV (7.1 x 10(3)/mm3) where they formed 10.4% of the neuronal population. PV+ nonpyramidal cells in lamina IV and lamina III were predominantly large basket-type cells with bitufted dendritic domains and tangentially oriented local axonal plexuses. The terminal-like label within lamina III/IV derived in part from the basket-cell axons, which formed pericellular arrays around unstained somata. Cell-sparse lamina V contained the largest PV+ nonpyramidal cells in AI. These cells, which formed 11% of the neuron population in lamina V, were notable for their tangentially oriented dendritic fields and local axonal arbors. PVi partitioned lamina VI into VIa and VIb. Large multipolar nonpyramidal cells were distributed throughout lamina VI and made up approximately 6% of the total population. Lamina VIa contained a band of lightly labeled PV+ pyramidal neurons that formed 15% of the neuronal population.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of cholinergic agonists on two non-pyramidal cell types in rat hippocampal slices

In the hippocampus, pyramidal cells (PCs) are not the only cell type sensitive to cholinergic stimulation. Two non-pyramidal cell types from animals as young as 8 days demonstrated clear, direct responses to application of cholinergic agonists. These cholinergic actions are excitatory, mostly blocked by muscarinic antagonists, and persist under conditions which block synaptic transmission (TTX, low Ca 2+/high Mg 2+). Cholinergic agonists may affect different conductances in interneurons than in PCs, sometimes resulting in rapid depolarization. Demonstration of direct excitatory cholinergic effects on inhibitory interneurons supports the view that cholinergically-evoked hyperpolarizations in PCs are due to local circuit interactions.

Neurons of the lateral entorhinal cortex of the rhesus monkey: A golgi, histochemical, and immunocytochemical characterization

This study identifies the neuronal types of the rhesus monkey lateral entorhinal cortex (LEC) and discusses the importance of these data in the context of the connectional patterns of the LEC and the possible role of these cells in neurodegenerative diseases. These neuronal types were characterized with the aid of Golgi impregnation techniques. These characterizations were based upon their spine densities, dendritic arrays, and, where possible, axonal arborizations. The cells could be segregated into only spinous and sparsely spinous types. The most numerous spinous types were pyramidal neurons. Other spinous types included multipolar, vertical bipolar and bitufted, and vertical tripolar neurons. The sparsely spinous neuronal types consisted of multipolar, horizontal bipolar and bitufted, and neurogliaform cells. These cells were further classified with the aid of histochemical stains and immunocytochemical markers. Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry stained multipolar, bipolar, and bitufted neurons. Stain for cytochrome oxidase (CO) was found in pyramidal and nonpyramidal cell types. Immunocytochemical techniques revealed several nonpyramidal neurons that contain somatostatin (Som) or substance P (SP). This study complements previous analyses of the neuronal components described in the LEC and adds further information about the distribution of selected neurochemicals within this cortex.

Demonstration of muscarinic acetylcholine receptor-like immunoreactivity in the rat forebrain and upper brainstem.

The distribution of muscarinic acetylcholine receptor protein (mAChR) in the rat forebrain and upper brainstem was described by using a monoclonal antibody (M35) raised against mAChR purified from bovine forebrain homogenates. A method is investigated for light microscopic (LM) and electronmicroscopic (EM) immunocytochemical visualization of reactivity to mAChR-proteins. Putative cholinoceptive neurons including their dendrites were found immunoreactive in the cortical mantle, hippocampus, basal ganglia, amygdala, thalamus and several midbrain regions. In the neocortex, immunoprecipitate with M35 was mainly present in layer 5 pyramidal cells, some layer 3 pyramidal neurons and layer 2 stellate cells, all including their characteristic dendritic profiles of both basal and apical dendrites. In the hippocampus, a variety of pyramidal, granular and non-pyramidal celltypes were stained in various hippocampal cell layers, in the dentate hilus and in stratum oriens of cornu ammonis. Moreover, positively reacting cells occurred in central and lateral amygdala, all parts of the basal ganglia and ventral pallidum. The thalamus was very richly provided with labeled neurons in several nuclei but notably numerous in the ventrolateral, anteroventral and geniculate nuclei. In cortex and hippocampus also some staining of astrocytes occurred. Electron microscopic study of the intracellular distribution of M35 immunoreactivity in all cases showed dense precipitates in the soma cytoplasm in close association with the golgi apparatus, but conspicuous absence near the endoplasmic reticulum. Immunoprecipitate can be followed within the dendritic tree along the microtubular transport system, up to proximal and distal postsynaptic membrane positions, apposing non labeled presynaptic endings. Muscarinic receptor subtype recognition by M35 will be discussed by comparing M35 distribution with cholinergic innervation patterns, muscarinic receptor ligand binding studies and localization of muscarinic receptor subtype mRNAs.

Cellular organization and development of slice cultures from rat visual cortex

Slice cultures from the visual cortex of young rats were prepared using the roller culture technique (Gähwiler 1984). After 10 days in vitro the cortical cultures flattened to 1-3 cell layers, surviving for up to 12 weeks. The cultures were organotypically organized, the typical layered structure of the cortex was preserved. The neuronal composition of slice cultures was studied using intracellular staining, Golgi impregnation and GABA immunohistochemistry. Both pyramidal cells and several types of nonpyramidal cells were identified in the slice cultures. Electrophysiological recordings showed that the electrical properties of cells in culture were similar to those measured in acute slice preparations; for some cells, however, the spontaneous activity was higher. The maintained activity was strongly increased by application of the GABA antagonist bicuculline and decreased by GABA, suggesting that GABAergic inhibition is present in these preparations. We could observe the postnatal maturation of some characteristic morphological features in culture. For example, pyramidal cells in 6 day-old rats in situ have very short basal dendrites with growth-cones, and the dendrites are free of spines. After 2-3 weeks in culture growth-cones were no longer observed. Instead, the cells had developed a large basal dendritic field and the dendrites were covered with spines. Slice cultures therefore may provide a useful tool for physiological, anatomical, pharmacological and developmental studies of cortical neurons in an organotypical environment.

Auditory cortical responses to neonatal deafening: pyramidal neuron spine loss without changes in growth or orientation.

Neonatal rabbits were unilaterally deafened at birth by surgical removal of the stapes, aspiration of the cochlear lymph, and kanamycin injection into the oval window. At 60 days of age, all rabbits were screened with brain stem evoked response tests in order to establish the efficacy of the deafening procedure. The auditory cortex contralateral to the destroyed cochlea was processed according to Golgi-Cox/Nissl procedures. Temporal bone histology revealed nearly complete outer hair cell loss in the damaged cochlea. The dendritic system of lamina III/IV pyramidal neurons contralateral to the deafened ear was digitized from frontal sections using a computer microscope system. Spine counts were also made along the basal dendrites. Spine counts revealed that neonatally deafened rabbits and 38.7% fewer spines along their basal dendrites. No differences between experimental and control rabbits were found in terms of soma cross-sectional area, total number of basal dendrites, total number of dendritic branches and total basal dendritic length. A fan-in projection of the dendritic system revealed no changes in the radial growth of basal dendrites resulting from the early acoustic trauma. In a prior study, spine-free nonpyramidal neurons in the same sections revealed altered dendritic growth and abnormally recurved dendrites. The separate response of pyramidal and nonpyramidal cell types to early cochlear damage is evidence for the different role of epigenetic determinants of dendritic form and orientation in sensory neocortical neurons.

Serotoninergic innervation of the cat cerebral cortex.

Serotoninergic axons in the cat cerebral cortex were demonstrated immunohistochemically with a monoclonal antibody to serotonin (5-HT). Three types of 5-HT axons are distinguished at the light microscopic level by differences in their morphology. Small varicose axons are fine (less than 0.5 micron) and bear fusiform varicosities that are generally less than 1 micron in diameter. These axons extend throughout the width of the cortex and branch frequently, giving rise to widely spreading collaterals. Nonvaricose axons are smooth, show a relatively large and constant caliber (about 1 micron), travel in straight, horizontal trajectories, and branch infrequently. Large varicose axons are distinguished by large round or oval varicosities (1 micron or more in diameter) borne on fine-caliber fibers. These axons often form basket-like arbors around the somata of single neurons. In the simplest basket-like arbors, several large, round varicosities from a small number of axons contact the soma. In complex baskets intertwining collaterals contact the soma and apparently climb along and outline the cell's major dendrites. The patterns revealed by the climbing axons suggest that a variety of nonpyramidal cell types selectively receive dense 5-HT innervation. Serial reconstructions of the 5-HT axons within the cortex show that the large varicose axons arise as infrequent collaterals from the nonvaricose axons. A single nonvaricose parent axon gives rise to several large varicose axon collaterals that may contribute to different basket-like arbors. Conversely, a single basket-like arbor may be formed by large varicose axon collaterals from more than one nonvaricose parent axon. The small varicose axons do not appear to be related within the cortex to either the nonvaricose or large varicose axon types. The results support the hypothesis that the 5-HT projection to the cortex is organized into two subsystems, one of which may exert widespread influence in the cortex via highly divergent branches, while the other, with a more restricted distribution, acts on specific classes of cortical neurons.

Intracortical connections and their physiological correlates in the primary auditory cortex (AI) of the cat.

We studied the functional and anatomical properties of the intrinsic connections in the primary auditory cortex (AI) of the cat by using physiological mapping and retrograde tracing methods. Our results revealed that a focal microinjection of tracer labeled as many as five intracortical patches in AI. The patches contained labeled pyramidal and non-pyramidal cell types, most of which were clustered in the middle layers. A densely distributed anterograde-like reaction product was present in the superficial layers above the labeled cells. The distribution of the patches was anisotropic, with most patches occurring dorsal, ventral, and anterior to the injection site. We examined the correlation between the characteristic frequency (CF) and binaural response properties of the injected and labeled regions. We found local labeling in regions possessing CFs equivalent to or slightly greater than that of the injected area. This appears to be a specific connection since we were able to predict the general location of many of the patches on the basis of the organization of the isofrequency domains. Patches were more numerous dorsoposterior to the injection site when the isofrequency contours ran obliquely (i.e., dorsoposterior to ventroanterior) across AI. The binaural response properties of the injected and labelled regions, however, were unrelated.

Physiological heterogeneity of nonpyramidal cells in rat hippocampal CA1 region.

Functional differentiation of nonpyramidal cells was studied by intracellular recording and staining of cells located in the stratum pyramidale or along the border between the stratum radiatum and the stratum lacunosum-moleculare of slices prepared from rat hippocampal CA1 region. In the stratum pyramidale, nonpyramidal cells (fast-spiking cells, type I cells) exhibited brief-duration action potentials (mean spike-width at one-half amplitude = 0.28 ms, N = 9) and little or no frequency adaptation of spike discharge to depolarizing current pulse. These cells ramified axon collaterals mainly in the stratum pyramidale or in the apical side of the stratum oriens. The HRP-injected nonpyramidal cells located between the stratum radiatum and the stratum lacunosum-moleculare (type II cells) showed different physiological characteristics from fast-spiking cells in the stratum pyramidale. The spike width was longer than that of fast-spiking cells (mean duration measured at one-half amplitude = 0.61 ms, N = 11) and these cells exhibited adaptation of spike discharge in response to depolarizing current pulses. Following hyperpolarizing current pulses, a depolarizing potential was produced in some type II cells. Although most cells of this group sent axon collaterals into the stratum radiatum or into the stratum lacunosum-moleculare, there were also cells whose axon collaterals extended to and ramified in the stratum pyramidale. In contrast to pyramidal cells, spikes of both types of nonpyramidal cells did not broaden during repetitive firing evoked by large depolarizing current pulses. Stimulation of the stratum radiatum caused excitatory and inhibitory postsynaptic potentials in both type I and II cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Two subtypes of non-pyramidal cells in rat hippocampal formation identified by intracellular recording and HRP injection

Intracellular recording and injection of horseradish peroxidase in rat hippocampal slice preparations revealed the existence of two physiologically distinct types of non-pyramidal cells: one a fast spiking type and the other a non-fast spiking type. In the CA1 pyramidal cell layer and the subgranular zone of the dentate gyrus fast spiking cells were found, while in the border area between the stratum lacunosum-moleculare and the stratum radiatum, non-fast spiking cells were found exclusively. These two types of non-pyramidal cells may play different roles in the functions of hippocampal neuronal networks.

Simultaneous recordings from two types of hippocampal nonpyramidal cells during electrically induced paroxysmal discharges

To reveal relationship between hippocampal nonpyramidal cells and seizure activity, single-unit activities of nonpyramidal cells were studied during electrically induced paroxysmal discharges in the hippocampal CA1 region of lightly anesthetized rabbits. Type I nonpyramidal cells ( N = 21) that were located in or near the pyramidal cell layer tended to cease firing during the early part of paroxysmal discharges, but refired during the late part. In contrast, type II nonpyramidal cells ( N = 14) which were found in the inferior subzone of the stratum oriens continued to fire during the entire course of the paroxysmal discharge.

Cholinergic innervation of ferret visual system

The distribution of acetylcholinesterase and choline acetyltransferase in primary visual areas of adult pigmented ferret was determined with cholinesterase histochemistry and choline acetyltransferase immunohistochemistry. In all visual areas the distribution of acetylcholinesterase in the neuropil closely matches that of choline acetyltransferase. In the cerebral cortex acetylcholinesterase and choline acetyltransferase are associated with axons found in every cortical layer and in the white matter. Area 17, identified by Nissl architectonics and cytochrome oxidase histochemistry, is distinguished by having a relatively low density of choline acetyltransferase- and acetylcholinesterase-stained axons in layer IV. Certain cortical non-pyramidal cell types show moderate staining for acetylcholinesterase after relatively long incubations, but no choline acetyltransferase-positive cells are observed in the cortex. In the lateral geniculate nucleus and superior colliculus the levels of choline acetyltransferase and acetylcholinesterase are considerably higher than in cerebral cortex and choline acetyltransferase-stained axons there display prominent varicosities. The distribution of choline acetyltransferase and acetylcholinesterase in the neuropil of lateral geniculate nucleus and superior colliculus of ferret shows marked laminar variation. For instance, in the lateral geniculate nucleus, the levels of acetylcholinesterase and choline acetyltransferase in the “On” sublaminae of laminae A and A1 are higher than the “Off” sublaminae. In the superficial layers of the superior colliculus the levels of choline acetyltransferase and acetylcholinesterase are highest in the stratum zonale and lowest in the stratum opticum; in the intermediate gray layer of the superior colliculus acetylcholinesterase- and choline acetyltransferase-stained fibres are distributed into dense patches. As in cortex, choline acetyltransferase-positive cell bodies are not found in the lateral geniculate nucleus or superior colliculus and acetylcholinesterase-stained cell bodies are visible only after long incubations. Cell bodies staining positively for choline acetyltransferase are found in a satellite of the superior colliculus, the parabigeminal nucleus.

Synaptic connections, axonal and dendritic patterns of neurons immunoreactive for cholecystokinin in the visual cortex of the cat

A subpopulation of γ-aminobutyric acid (GABA) containing neurons was reported to contain cholecystokinin-immunoreactive material in the visual cortex of cat [Somogyi et al., J. Neurosci. (1984) 4, 2590–2603]. In the present study pre-embedding immunocytochemistry was used to identify which of the several types of presumed GABAergic nonpyramidal cells in areas 17 and 18 contain cholecystokinin immunoreactivity. Most of the cholecystokinin-immunoreactive somata were found in layers II-III, they were less frequent in layers I and VI, and relatively rare in layers IV and V. The distribution and density of the axon terminals resembled that of the cell bodies. Two well defined types of cholecystokinin-immunoreactive neuron were distinguished: (1) double bouquet cells in layers II–III with vertically projecting axons, and (2) small basket cells with local axons either restricted to layers II–III, or descending to layer V. Additional cholecystokinin-positive cells showed features of bitufted or multipolar neurons in layers II–VI and horizontal cells in layer I, but these cells could be defined less well due to partial staining. Cholecystokinin-immunoreactive dendrites were found to run horizontally in layer I for several hundred micrometers. Some of the cholecystokininimmunoreactive cells in layer VI had very long dendrites ascending radially up to layer III, as did their axons. A few cholecystokinin-immunoreactive cells appeared to have two axons and this was confirmed by electron microscopy. All cholecystokinin-immunoreactive neurons and terminals were separated from the basal lamina of blood vessels by glial endfeet. Random samples of boutons from each layer as well as identified terminals traced to their origin from local neurons were examined in the electron microscope. All of the boutons established symmetrical (type II) synaptic contacts with dendritic shafts, spines or somata. Quantitative electron microscopy of the postsynaptic targets of double bouquet cells and small basket cells demonstrated clear differences between these two types of neuron; basket cells having a higher proportion of their terminals in synaptic contact with somata. The findings that several distinct types of cortical neurons, as defined by their synaptic connections, contain cholecystokinin-immunoreactive material and that identified axons of all examined neurons form type II synaptic contacts suggests that the majority, if not all cholecystokinin-positive boutons forming type II contacts originate from local cortical cells. The distribution of targets postsynaptic to cholecystokinin-positive neurons is compared to those of cells labelled by other methods.

Neurons accumulating [ 3Hgamma-aminobutyric acid (GABA) in supragranular layers of cat primary auditory cortex (AI)

The classes of neurons accumulating exogenously injected, tritiated gamma-aminobutyric acid ([ 3HGABA) were studied in the supragranular layers in the primary auditory field of the adult cat. The size, laminar locus, and somatodendritic profiles of labeled neurons were studied light microscopically in frozen- or Vibratome-sectioned, 30 μm thick material, and in semithin, 1–2 μm thick, plastic-embedded high-resolution autoradiographic preparations. The chief goals of the study were to determine which types of cells could be identified as accumulating [ 3HGABA in layers I, II and III, and to establish possible relationships between these cells and (i) neurons described in Golgi studies of these layers, and (ii) the neurons found, in parallel investigations of the connections of the primary auditory field, to participate as ipsilateral corticocortical and commissural cells of origin. The principal findings are: that neurons in every layer in the primary auditory field take up tritiated gamma-aminobutyric acid; that their Nissl-counterstained somata have a smaller average area, and a smaller range of areas, than do the unlabeled cells; that more than one type of labeled neuron—as defined by somatic size and shape, height : width ratios, and nuclear membrane morphology—could be identified in each layer; that none of the labeled neurons had a soma with a pyramidal configuration; that the labeled cells are comparable in size, shape, and laminar distribution to some populations of non-pyramidal ipsilateral corticocortical cells of origin in layers II and III, and perhaps to certain classes of commissurally projecting, layer III non-pyramidal neurons; and finally, that only a rather small proportion—perhaps 10% or less, except in layer I—of the supragranular cells appear to accumulate labeled material. With regard to the identity of particular classes of neurons accumulating silver grains above background in the individual layers, in layer I, 2 of the 4 types of neurons characterized in Golgi preparations take up gamma-aminobutyric acid and the remaining 2 types may also, and the relative number of labeled cells appears to be higher than in the other layers; in layer II, 2 of the 9 varieties are labeled, and 4 other types may also be; and in layer III, 2 of the 11 types take up gamma-aminobutyric acid, and 5 other varieties may as well. Three types of non-pyramidal layer II cells that project ipsilaterally from AI to the second auditory cortical field, AII, possibly accumulate gamma-aminobutyric acid; 3 types of commissural non-pyramidal cells of origin linking AI to AI appear to be labeled by gamma-aminobutyric acid. Thus, of the total of 24 types of neurons characterized in previous morphological and connectional studies of layers I–III in AI, 6 are likely candidates for a gamma-aminobutyric acid-modulated inter- or intracortical role, 11 others may be, while 7 others are unlikely to be. The neurons in AI which are positive for gamma-aminobutyric acid are characterized by their somatodendritic heterogeneity, broad laminar distribution, and distinctive patterns of cortical connectivity. Somata accumulating gamma-aminobutyric acid are probably intermingled among ipsilateral corticocortical and commissural cells of origin in AI. These differences imply that more than one cortical pathway or local circuit arising in AI may use gamma-aminobutyric acid as a neurotransmitter, although its precise functional role remains to be demonstrated physiologically in auditory cortex.

The neuronal composition of area 17 of rat visual cortex. III. Numerical considerations.

The neuronal population of area 17 of rat visual cortex has been examined by using tissue from brains fixed by perfusion. The tissue was osmicated and embedded in plastic so that the same neurons could be examined by both light and electron microscopy. In these preparations area 17 was 1.49 mm thick and by stereological procedures it was calculated that there are about 120,000 neurons beneath 1 mm2 of cortical surface. If one assumes area 17 in each hemisphere of the rat to occupy between 7.1 and 9.4 mm2 of cortical surface, then in each hemisphere the area contains between 850,000 and 1,128,000 neurons. Of these neurons 85% are pyramidal cells and 15% are nonpyramidal cells. About one-third of the nonpyramidal cells occur in layers I and VIb, both of which contain only this kind of neuron. The remaining two-thirds of the nonpyramidal cells are in layers II-VIa. Within these layers it has been possible to differentiate bipolar cells from other types of nonpyramidal cells and in each of these two nonpyramidal cell groups to recognize both small and large neurons. The greatest concentration of nonpyramidal cells occurs in layer II/III. To confirm the validity of the stereologically derived data direct counts were made of the medium and large pyramidal cells in layer V.

The termination of callosal fibres in the auditory cortex of the rat. A combined Golgi--electron microscope and degeneration study.

When the corpus callosum of the rat is sectioned, the callosal fibres in the cerebral cortex undergo degeneration. In the auditory cortex (area 41) the degenerating axon terminals form asymmetric synapses, and the vast majority of them synapse with dendritic spines. Some other synapse with the shafts of both spiny and smooth dendrites, and a few with the perikarya of non-pyramidal cells. The degenerating axon terminals are contained principally within layer II/III, in which they aggregate in patches. Using a technique in which neurons within the cortex are Golgi-impregnated, then gold-toned and examined in the electron microscope, it has been shown that the dendritic spines of pyramidal neurons with cell bodies in different layers receive the degenerating callosal afferents. The spines arise from the main apical dendritic shafts and their branches, from the dendrites of the apical tufts, and in some cases from the basal dendrites of the pyramidal neurons. The shafts of some pyramidal cell apical dendrites also form asymmetric synapses with callosal afferents. Since we have encountered no spiny non-pyramidal neurons in Golgi preparations of rat auditory cortex, and because other types of non-pyramidal cells have few dendritic spines, it is concluded that practically all of the dendritic spines synapsing with callosal afferents originate from pyramidal neurons.

Structure of the piriform cortex of the opossum. I. Description of neuron types with Golgi methods.

AbstractThe piriform cortex was studied in the adult opossum with rapid Golgi and Golgi‐Cox techniques.Most pyramidal cells in the deep part of layer II and layer III resemble those in other parts of the cerebral cortex by virtue of a single apical dendritic trunk, multiple basal dendrites, a large number of small to medium dendritic spines, and a deeply directed axon. Pyramidal cells in the superficial part of layer II are similar with the exception that “secondary” apical dendrites often emerge directly from the cell body rather than from a single primary trunk.With conservative criteria for categorization, nine different types of nonpyramidal cells were distinguished, four of which have not been previously described. Layer I contains a small number of neurons with both smooth and spiny dendrites including distinctive fusiform cells with large somatic appendages. As in other species, the most common type of nonpyramidal neuron in layer II is the semilunar cell which has only apically directed dendrites. These cells have distinctive large spines confined to their distal dendritic segments. The mid to deep portion of layer III contains multipolar neurons with smooth dendrites that resemble the well‐known large stellate cells in neocortex. In addition, layer III contains three non‐pyramidal neuron types with spiny dendrites: (1) fusiform and multipolar cells with complex, branched dendritic appendages and somatic spines, (2) very large multipolar cells (up to 35 μm mean diameter) with large‐diameter dendrites that give rise to abruptly tapering side branches and filiform spines, and (3) multipolar cells with profusely spiny dendrites. In all three layers, small neurons have been found with spherical cell bodies and “axoniform” dendrites that resemble the so called neurogliaform neurons described in a variety of brain areas. A striking feature of the organization of the piriform cortex is that, with the exception of the neurogliaform neurons, the different types of nonpyramidal cells tend to be segregated in individual layers or sublayers.Physiological implications of the results are discussed. Remarks are also made concerning the potential of the piriform cortex as a model cortical system.