Expression of Nerve Growth Factor,p75, and trk in the Somatosensory and Motor Cortices of Mature Rats: Evidence for Local Trophic Support Circuits

Neurotrophins such as nerve growth factor (NGF) are critical for the maintenance of CNS neurons. We determined the expression of NGF and the neurotrophin receptors p75 and trk in the somatosensory and motor cortices of mature rats with immuno-histochemical techniques. Sections of mature rat cortex were processed immunohisto-chemically with primary antibodies directed against NGF, p75, or trk. The distribution of immunoreactive elements was examined, and stereological techniques were used to determine the density and size of immunoreactive cell bodies. Some sections processed for trk immunoreactivity were examined with an electron microscope.

From the size and morphology of the labeled cells, it appeared that only neurons in the gray matter were NGF-positive. NGF was detected in one-third of the neurons in layers II-III, V, and VI of both somatosensory cortex and motor cortex; however, fewer than 1 in 12 of the layer IV neurons was NGF-positive. With the notable exception of layer V, few cell bodies (2–10% of the total population) were p75– or trk-immunoreactive. Layer Vb was replete with receptor-positive cell bodies; more than one-third of the layer Vb neurons were p75– or trk-positive. All labeled cells appeared to be pyramidal neurons. The distribution of p75 labeling with the two anti-p75 antibodies was indistinguishable. In addition, the neuropil in the supragranular laminae was p75– or trk-positive. Electron microscopy showed that trk immunoreactivity was also expressed by dendrites. Only rarely were immunoreactive axons detected.

In summary, NGF is expressed by cortical neurons throughout cortex, and neurotrophin receptors are widely produced by postsynaptic targets. Thus, NGF appears to participate in an intracortical autoregulatory system. The strong expression of neurotrophin receptors by pyramidal neurons in layer Vb (the origin of brainstem and spinal cord projections) suggests that the neurotrophins are especially critical for the regulation of corticofugal projection systems.     

Comparative electron microscopic study of the visual cortex neurons of the cat in postnatal ontogeny

The maturation of layers II-VI of neurons and perineuronal neuropil of the cat visual cortex (field 17) was studied from postnatal day 1 to day 21. The differentiation of large, small (associate) pyramid and stellate neurons was described. During the first postnatal week, the somata of layers II-VI of neurons undergo significant changes, the perikaryal cytoplasm increases in volume. Cell bodies of large pyramidal neurons mature by day 15. During the second postnatal week and almost till day 15, the rough endoplasmic reticulum of small pyramidal and stellate neurons undergoes proliferation; dendritic processes are branching. In stellate neurons the amount of cytoplasmic organelles increases dramatically only after the second postnatal week, and this is presumably induced by the opening of eyes on day 12. The second postnatal week is the period of greatest growth of dendritic, axonal and glial processes in perineural neuropil of layers V-VI. In the perineuronal neuropil of large pyramidal neurons (layers V-VI) there appear symmetric synapses with pyramidal cells, dendritic processes and dendritic spines. This occurs just at the time when kittens first open the eyes. From this time and during postnatal days 15-21, asymmetric synapses appear in the perineuronal neuropil of large pyramidal neurons. In the perineuronal neuropil of small pyramidal and stellate neurons. (layers II-IV), synapses reveal the mature appearance by day 15. After the opening of the eyes and up to postnatal day 21, dendritic growth and spine production occur in the perineuronal neuropil of small pyramidal and stellate neurons.     

Whisker-related circuitry in the trigeminal nucleus principalis: Ultrastructure

Trigeminal (V) nucleus principalis (PrV) is the requisite brainstem nucleus in the whisker-to-barrel cortex model system that is widely used to reveal mechanisms of map formation and information processing. Yet, little is known of the actual PrV circuitry. In the ventral “barrelette” portion of the adult mouse PrV, relationships between V primary afferent terminals, thalamic-projecting PrV neurons, and gamma-aminobutyric acid (GABA)-ergic terminals were analyzed in the electron microscope. Primary afferents, thalamic-projecting cells, and GABAergic terminals were labeled, respectively, by Neurobiotin injections in the V ganglion, horseradish peroxidase injections in the thalamus, and postembedding immunogold histochemistry. Primary afferent terminals (Neurobiotin- and glutamate-immunoreactive) display asymmetric and multiple synapses predominantly upon the distal dendrites and spines of PrV cells that project to the thalamus. Primary afferents also synapse upon GABAergic terminals. GABAergic terminals display symmetric synapses onto primary afferent terminals, the somata and dendrites (distal, mostly) of thalamic-projecting neurons, and GABAergic dendrites. Thus, primary afferent inputs through the PrV are subject to pre- and postsynaptic GABAergic influences. As such, circuitry exists in PrV “barrelettes” for primary afferents to directly activate thalamic-projecting and inhibitory local circuit cells. The latter are synaptically associated with themselves, the primary afferents, and with the thalamic-projecting neurons. Thus, whisker-related primary afferent inputs through PrV projection neurons are pre- and postsynaptically modulated by local circuits.     

SynArfGEF is a guanine nucleotide exchange factor for Arf6 and localizes preferentially at post-synaptic specializations of inhibitory synapses

SynArfGEF, also known as BRAG3 or IQSEC3, is a member of the brefeldin A-resistant Arf-GEF/IQSEC family and was originally identified by screening for mRNA species associated with the post-synaptic density fraction. In this study, we demonstrate that synArfGEF activates Arf6, using Arf pull down and transferrin incorporation assays. Immunohistochemical analysis reveals that synArfGEF is present in somata and dendrites as puncta in close association with inhibitory synapses, whereas immunoelectron microscopic analysis reveals that synArfGEF localizes preferentially at post-synaptic specializations of symmetric synapses. Using yeast two-hybrid and pull down assays, we show that synArfGEF is able to bind utrophin/dystrophin and S-SCAM/MAGI-2 scaffolding proteins that localize at inhibitory synapses. Double immunostaining reveals that synArfGEF co-localizes with dystrophin and S-SCAM in cultured hippocampal neurons and cerebellar cortex, respectively. Both β-dystroglycan and S-SCAM were immunoprecipitated from brain lysates using anti-synArfGEF IgG. Taken together, these findings suggest that synArfGEF functions as a novel regulator of Arf6 at inhibitory synapses and associates with the dystrophin-associated glycoprotein complex and S-SCAM.     

Recovery of Cholinergic Phenotype in the Injured Rat Neostriatum: Roles for Endogenous and Exogenous Nerve Growth Factor

Polyclonal antibodies against recombinant human nerve growth factor (rhNGF) potently inhibited PC12 neurite outgrowth, blocked high-affinity 125I-rhNGF binding but not its receptor, and cross-reacted with rat, mouse, and human nerve growth factor (NGF) but not with brain-derived neurotrophic factor, neurotrophin-3, ciliary neurotrophic factor, insulin-like growth factor, epidermal growth factor, or activin A. Immunocytochemistry revealed many NGF-positive neurons in the rat neostriatum. The NGF-positive neurons disappeared by 3 days after mechanical injury to the neostriatum and were replaced by intensely NGF- and glial fibrillary acidic protein-positive astrocytes. Enzyme-linked immunosorbent assay measurements revealed that the NGF content of the injured striatum was elevated by eightfold 3 days postinjury and by twofold 2 weeks later. The high-affinity choline uptake (HACU) into cholinergic nerve terminals was decreased by 23% at 2 and 4 weeks postinjury, yet choline acetyltransferase (ChAT) activity in these neurons was unchanged at 2 weeks and decreased by 14% at 4 weeks. Daily infusion of 1 microgram of rhNGF into the injury area did not alter the loss of HACU. However, this treatment elevated ChAT activity by 23-29% above intact neostriatal levels and by 53-65% relative to HACU at both survival times. Thus, lesion-induced increases in NGF levels within astrocytes are associated with maintenance of striatal ChAT activity at normal levels following cholinergic injury, even with decreases in HACU. Pharmacologic doses of rhNGF can further augment ChAT activity in damaged cholinergic neurons, showing the usefulness of exogenous NGF even when endogenous NGF is elevated in response to injury.     

Caudatonigral projections under normal conditions and after an MPTP-induced lesion of the cat nigrostriatal system: Quantitative ultrastructural analysis

Ultrastructural and morphometric studies of caudatonigral synapses located on the nigrothalamic neurons were carried out on intact and N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated adult cats. Three types of synapses with different ultrastructural features were found. Morphometric analysis showed that 11.3% of analyzed junctions were caudatonigral synapses; 5.9% and 5.4% of them were located on the somata of nigrothalamic neurons and on their dendrites respectively. Among axo-somatic synapses, the caudatonigral ones amounted to 11.9%: 7.6% were type-I synapses and 4.3% belonged to type-III synapses. Both types had symmetrical contacts and could be considered inhibitory. Caudatonigral axodendritic synapses amounted to 10.6%: 3.2% were of type I; 4.2%, of type III; and 3.2%, of type II synapses with asymmetric contacts. The labelled type-II synapses were found exclusively on the nigrothalamic dendrites. The ultrastructural changes and the statistically significant decrease in the size of caudate axon terminals following the MPTP treatment took place only in the type I exosomatic synapses. It is suggested that the development of motor disorders in the cat after experimentally induced striatal dopamine insufficiency is due to the decrease in the efficacy of caudatonigral influences, which causes disinhibition of GABA-ergic inhibitory nigrothalamic neurons, and to enhanced influences of the latter on the cells of the motor thalamic nuclei.Neirofiziologiya/Neurophysiology, Vol. 26, No. 2, pp. 150–156, March–April, 1994.     

A quantitative profile of the synapses on the stellate cell body and axon in the cochlear nucleus of the chinchilla

Summary. One of the most numerous neurons in the cochlear nucleus is the type I stellate cell. Previous attempts to understand the structural basis for its signal coding assumed that integration of synaptic potentials arising from axodendritic synapses should account for the generation of its response properties. However, the present study documents the importance of excitatory and inhibitory types of synapses on the soma and axon. Retrograde transport of cholera toxin B subunit, injected in the inferior colliculus of chinchillas, was used to label exclusively type I stellate cells in the anteroventral cochlear nucleus. The relative distribution of terminal types by vesicle morphology was pleomorphic < large spherical < flattened < smaller spherical. The somatic perimeter covered by endings ranged from almost none to nearly half. More flattened-vesicle terminals contacted somata in the high-frequency than in the low-frequency region. Eight of twenty axons received endings that contained large spherical vesicles and made asymmetric junctions; half of these extensively apposed the initial segment, forming a collar of presumed excitatory input. Thus, type I stellate cells are a heterogeneous group. Inhibitory synapses probably compose the majority of terminals. Some cells receive mostly inhibitory synapses near the presumed site of the spike generator, but others also have a prominent excitatory input. These findings call for a new look at the mechanisms for signal coding in stellate cells in the auditory system in particular and raise issues concerning the stochastic nature of information processing in sensory systems in general.     

Ultrastructural characteristics of layer III pyramidal neurons in the cat primary auditory cortex (Al)

Electron microscope studies were made of retrogradely horseradish peroxidase-labeled pyramidal neurons forming transcallosal projections in layer III of the cat primary auditory cortex (Al). These showed a significant proportion of the somatic membrane to be covered with processes of astroglia, while synapses occupy 20% of the synaptic surface on average. Between 4 and 10 axosomatic synapses were identified on the profiles of callosal cell somata. All these were formed by axonal terminals containing small, flattened synaptic vesicles and had symmetrical contacts. Average length of these synaptic contacts equaled 1.6 µm. Numerous anterogradely horseradish peroxidase-labeled axonal terminals of callosal fibers were found in cortical area Al in amongst retrogradely HP-labeled neurons. The ultrastructural pattern of these is described.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 22, No. 4, pp. 520–526, July–August, 1990.     

Intrinsic organization of snake dorsomedial cortex: an electron microscopic and golgi study.

The cellular populations present in dorsomedial cortex in the snakes Constrictor constrictor, Natrix sipendon and Thamnophis sirtalis are described at the light microscopic level using Nissl and Golgi preparations as well as at the ultrastructural level. This area plays a central role in cortical organization in snakes by participating in major commissural and association projections. Systematic analyses of Golgi preparations indicate that five populations of neurons are present in dorsomedial area and have a preferential laminar distribution. Layer 1 stellate cells have somata positioned in the center of the outermost cortical layer, layer 1. Their dendrites are confined to this layer. Double pyramidal cells have their somata loosely packed in layer 2. Their dendrites bear a moderate population of spines, ascending through layer 1 to the pial surface and descending partially through layer 3. Some double pyramidal cells have somata displaced downwards into the upper third of layer 3. These neurons closely resemble the layer 2 double pyramidal cells. Layer 3 stellate cells have somata positioned in the middle third of layer 3. Their dendrites extend in all directions throughout layer 3 and through layer 2 into layer 1. Finally, horizontal cells have their somata positioned deep in layer 3, near the ventricle, and dendrites aligned concentric with the ventricle. Comparison of the organization of the known afferents to dorsomedial area with the distribution of the five cell types suggests that the laminations of both afferent fibres and dorsomedial neurons places specific neuronal populations in synaptic contact with specific sets of afferents.     

Electron microscopy examination of galanin-like peptide (GALP)-containing neurons in the rat hypothalamus

Galanin-like peptide (GALP) is a novel peptide which is isolated from the porcine hypothalamus. GALP-containing neurons are present in the arcuate nucleus (ARC), being particularly densely concentrated in medial posterior regions. To observe the ultrastructure and synaptic relationships of GALP-containing neurons in the ARC, light and immunoelectron microscopy techniques were used. At the light microscope level, GALP-containing neurons were observed distributed rostrocaudally throughout the ARC, with the majority present in the posterior, periventricular zones. At the electron microscope level, many immunopositive dense-cored vesicles were evident in the perikarya, dendrites and axon terminals of the GALP-containing neurons. Furthermore, these neurons received synapses from immunonegative axon terminals that were symmetric in the case of synapses made on perikarya, and both asymmetric and symmetric for synapses made on dendrites. Axon terminals of GALP-containing neurons often made synapses on immunonegative dendrites. Such synapses were all symmetric. Synapses were also found between axon terminals and perikarya as well as dendrites of GALP-containing neurons. These findings suggest that the physiological role of the GALP-containing neurons in the ARC is based on complex synaptic relationships between GALP-containing neurons and either GALP-immunopositive or -immunonegative neurons.     

Structural plasticity of synapses in hippocampal slices after oxygen-glucose deprivation

We analyzed structural rearrangements of synaptic contacts in the stratum radiatum of the CA1 area of cultured rat hippocampal slices under conditions of the development of potentiation of synaptic transmission induced by short-term (10 min) oxygen-glucose deprivation (OGD). Studies were carried out using electron microscopy and 3D reconstruction of cellular compartments. Within the 1st h after OGD, we observed increases in the volume of pre-synaptic terminals and post-synaptic spines and also in the area of postsynaptic densities (PSDs) in both asymmetric excitatory and symmetric inhibitory synapses, especially in the case were the PSD was perforated. We also observed significant activation of glial cells (increases in their volume and area of contacts of their processes with the components of synapses). Therefore, OGD results in activationassociated structural rearrangements of both excitatory and inhibitory synapses of the hippocampal CA1 area. Such rearrangements are accompanied by a clearly pronounced reaction of the glia, which correlates with an important role of the latter in modulation of the functioning of neurons.     

Ultrastructure and morphometric parameters of glutamatergic synapses on nigrothalamic and unidentified neurons of the reticular zone of theSubstantia nigra in cats

Nigrothalamic neurons were identified into thesubstantia nigra by their retrograde labelling with horseradish peroxidase. Axon terminals that contain glutamate (the excitatory transmitter) were revealed immunocytochemically with an immunogold electron microscopic technique. Ultrastructural parameters (the large and small diameters of axon terminals, area of their profiles, coefficient of form of profiles, large and small diameters of synaptic vesicles) were analyzed in all 240 synapses under study. Synaptic contacts localized on both nigrothalamic and unidentified neurons belonged to three morphologically specific groups. Synapses of the groups I and III, according to classification by Rinvik and Grofova, were characterized by a symmetric type of synaptic contact and contained polymorphic synaptic vesicles. Contacts in group-II synapses were asymmetric, and respective terminals contained round vesicles. Among the studied synapses, 65.8% were classified as group-I contacts, 25.0% belonged to group II, and 9.2% belonged to group III. Glutamate-positive axon terminals formed predominantly group-II synapses; such connections constituted 70% of this group's synapses. Sixty percent of glutamate-positive synapses were localized on the distal dendrites and 23% on the proximal dendrites, while 17% of such synapses were distributed on the somata of nigral neurons. Such a pattern of distribution of glutamate-positive synapses was observed on both nigrothalamic and unidentified nigral neurons. About 7% of glutamate-positive synapses were formed by very large axon terminals containing round synaptic vesicles; yet, the contacts of these terminals were of a symmetric type. Twenty percent of group-I synapses, i.e., synapses considered inhibitory connections, were found to manifest a weak immune reaction to glutamate.Neirofiziologiya/Neurophysiology, Vol. 28, No. 6, pp. 285–295, November–December, 1996.     

Synaptic interactions between ghrelin- and neuropeptide Y-containing neurons in the rat arcuate nucleus

Morphological relationships between neuropeptide Y- (NPY) like and ghrelin-like immunoreactive neurons in the arcuate nucleus (ARC) were examined using light and electron microscopy techniques. At the light microscope level, both neuron types were found distributed in the ARC and could be observed making contact with each other. Using a preembedding double immunostaining technique, some NPY-immunoreactive axon terminals were observed at the electron microscope level to make synapses on ghrelin-immunoreactive cell bodies and dendrites. While the axo-somatic synapses were mostly symmetric in nature, the axo-dendritic synapses were both symmetric and asymmetric. In contrast, ghrelin-like immunoreactive (ghrelin-LI) axon terminals were found to make synapses on NPY-like immunoreactive (NPY-LI) dendrites although no NPY-like immunoreactive perikarya were identified receiving synapses from ghrelin-LI axon terminals. NPY-like axon terminals were also found making synapses on NPY-like neurons. Axo-axonic synapses were also identified between NPY- and ghrelin-like axon terminals. The present study shows that NPY- and ghrelin-LI neurons could influence each other by synaptic transmission through axo-somatic, axo-dendritic and even axo-axonic synapses, and suggests that they participate in a common effort to regulate the food-intake behavior through complex synaptic relationships.     

Synaptic dimorphism in Onychophoran cephalic ganglia

The taxonomic location of the Onychophora has been controversial because of their phenotypic and genotypic characteristics, related to both annelids and arthropods. We analyzed the ultrastructure of the neurons and their synapses in the cephalic ganglion of a poorly known invertebrate, the velvet worm Peripatus sedgwicki, from the mountainous region of El Valle, Mérida, Venezuela. Cephalic ganglia were dissected, fixed and processed for transmission electron microscopy. The animal has a high degree of neurobiological development, as evidenced by the presence of asymmetric (excitatory) and symmetric (inhibitory) synapses, as well as the existence of glial cell processes in a wide neuropile zone. The postsynaptic terminals were seen to contain subsynaptic cisterns formed by membranes of smooth endoplasmic reticulum beneath the postsynaptic density, whereas the presynaptic terminal showed numerous electron transparent synaptic vesicles. From the neurophylogenetic perspectives, the ultrastructural characteristics of the central nervous tissue of the Onychophora show important evolutionary acquirements, such as the presence of both excitatory and inhibitory synapses, indicating functional synaptic transmission, and the appearance of mature glial cells.     

Regulation of neurons in the suprachiasmatic nucleus of Xenopus laevis

In the amphibian Xenopus laevis, suprachiasmatic melanotrope-inhibiting neurons (SMINs) play an important role in the regulation of the background adaptation process. In this study, we investigated the innervation of the SMINs at the light- and electron- microscopical level. Immunocytochemistry in combination with confocal laser scanning microscopy revealed co-existence of neuropeptide Y (NPY) and synaptobrevin in spots in the direct vicinity of the SMINs, suggesting the existence of NPY-containing synapses on these cells. At the ultrastructural level, the SMINs showed a high degree of plasticity, containing more electron-dense vesicles and a larger extent of RER in white- than in black-adapted animals. In black-adapted animals, symmetric synapses (Gray type II) were observed on the soma of the SMINs, suggesting an inhibitory input to these cells. The synaptic profiles contained electron-lucent and electron-dense vesicles, indicating the involvement of both a classical neurotransmitter and a neuropeptide (possibly NPY) in this input. In white-adapted animals, synapses were only found at some distance from the SMIN somata. Our findings indicate a striking plasticity of the innervation of the SMINs in relation to background adaptation and support the hypothesis that the SMINs are innervated by NPY-containing interneurons that inhibit SMIN activity in black-adapted animals.     

Calretinin-containing interneurons innervate both principal cells and interneurons in the CA1 region of the human hippocampus

Hippocampal interneurons consist of functionally diverse cell types, most of them target the dendrites or perisomatic region of pyramidal cells with a few exceptions, like the calretinin-containing cells in the rat: they selectively innervate other interneurons. However, no electron microscopic data are available about the synaptic connections of calretinin-immunoreactive neurons in the human hippocampus. We aimed to provide these data to establish whether interneuron-selective interneurons indeed represent an essential feature of hippocampal circuits across distant species. Two types of calretinin-immunostained terminals were found in the CA1 region: one of them presumably derived from the thalamic reuniens nucleus, and established asymmetric synapses on dendrites and spines. The other type originating from local interneurons formed symmetric synapses on both pyramidal and interneuron dendrites. Distribution of postsynaptic targets showed that 26.8% of the targets were CR-positive interneuron dendrites, and 25.2% proved to be proximal pyramidal dendrites. CR-negative interneuron dendrites were also contacted (12.4%). Small caliber postsynaptic dendrites were not classified (28%). Somata were rarely contacted (7.6%). The present data suggest that calretinin-positive boutons do show a preference for other interneurons, but a considerable proportion of the targets are pyramidal cells. We propose that interneuron-selective inhibitory cells exist in the human Ammon's horn, and boutons innervating pyramidal cells derive from another cell type that might not exist in rodents.     

Glutamatergic components of the retrosplenial granular cortex in the rat

The ultrastructural characteristics, distribution and synaptic relationships of identified, glutamate-enriched thalamocortical axon terminals and cell bodies in the retrosplenial granular cortex of adult rats is described and compared with GABA-containing terminals and cell bodies, using postembedding immunogold immunohistochemistry and transmission electron microscopy in animals with injections of cholera toxin- horseradish peroxidase (CT-HRP) into the anterior thalamic nuclei. Anterogradely labelled terminals, identified by semi-crystalline deposits of HRP reaction product, were approximately 1 μm in diameter, contained round, clear synaptic vesicles, and established asymmetric (Gray type I) synaptic contacts with dendritic spines and small dendrites, some containing HRP reaction product, identifying them as dendrites of corticothalamic projection neurons. The highest densities of immunogold particles following glutamate immunostaining were found over such axon terminals and over similar axon terminals devoid of HRP reaction product. In serial sections immunoreacted for GABA, these axon terminals were unlabelled, whereas other axon terminals, establishing symmetric (Gray type II) synapses were heavily labelled. Cell bodies of putative pyramidal neurons, containing retrograde HRP label, were numerous in layers V–VI; some were also present in layers I–III. Most were overlain by high densities of gold particles in glutamate but not in GABA immunoreacted sections. These findings provide evidence that the terminals of projection neurons make synaptic contact with dendrites and dendritic spines in the ipsilateral retrosplenial granular cortex and that their targets include the dendrites of presumptive glutamatergic corticothalamic projection neurons.     

Expression of phospho‐Ca2+/calmodulin‐dependent protein kinase II in the pre‐Bötzinger complex of rats

The pre‐Bötzinger complex (pre‐BötC) in the ventrolateral medulla oblongata is a presumed kernel of respiratory rhythmogenesis. Ca²⁺‐activated non‐selective cationic current is an essential cellular mechanism for shaping inspiratory drive potentials. Ca²⁺/calmodulin‐dependent protein kinase II (CaMKII), an ideal ‘interpreter’ of diverse Ca²⁺ signals, is highly expressed in neurons in mediating various physiological processes. Yet, less is known about CaMKII activity in the pre‐BötC. Using neurokinin‐1 receptor as a marker of the pre‐BötC, we examined phospho (P)‐CaMKII subcellular distribution, and found that P‐CaMKII was extensively expressed in the region. P‐CaMKII‐ir neurons were usually oval, fusiform, or pyramidal in shape. P‐CaMKII immunoreactivity was distributed within somas and dendrites, and specifically in association with the post‐synaptic density. In dendrites, most synapses (93.1%) examined with P‐CaMKII expression were of asymmetric type, occasionally with symmetric type (6.9%), whereas in somas, 38.1% were of symmetric type. P‐CaMKII asymmetric synaptic identification implicates that CaMKII may sense and monitor Ca²⁺ activity, and phosphorylate post‐synaptic proteins to modulate excitatory synaptic transmission, which may contribute to respiratory modulation and plasticity. In somas, CaMKII acts on both symmetric and asymmetric synapses, mediating excitatory and inhibitory synaptic transmission. P‐CaMKII was also localized to the perisynaptic and extrasynaptic regions in the pre‐BötC.     

Pyramidal Cells Make Specific Connections onto Smooth (GABAergic) Neurons in Mouse Visual Cortex

One of the hallmarks of neocortical circuits is the predominance of recurrent excitation between pyramidal neurons, which is balanced by recurrent inhibition from smooth GABAergic neurons. It has been previously described that in layer 2/3 of primary visual cortex (V1) of cat and monkey, pyramidal cells filled with horseradish peroxidase connect approximately in proportion to the spiny (excitatory, 95% and 81%, respectively) and smooth (GABAergic, 5% and 19%, respectively) dendrites found in the neuropil. By contrast, a recent ultrastructural study of V1 in a single mouse found that smooth neurons formed 51% of the targets of the superficial layer pyramidal cells. This suggests that either the neuropil of this particular mouse V1 had a dramatically different composition to that of V1 in cat and monkey, or that smooth neurons were specifically targeted by the pyramidal cells in that mouse. We tested these hypotheses by examining similar cells filled with biocytin in a sample of five mice. We found that the average composition of the neuropil in V1 of these mice was similar to that described for cat and monkey V1, but that the superficial layer pyramidal cells do form proportionately more synapses with smooth dendrites than the equivalent neurons in cat or monkey. These distributions may underlie the distinct differences in functional architecture of V1 between rodent and higher mammals.     

Electron cytochemical localization of gamma-aminobutyric acid catabolism in rat cerebellar cortex

Summary An electron cytochemical technique is described for the localization of GABA-T, the enzyme which degrades the neurotransmitter GABA, in rat cerebellar cortex. The technique allows ultrastructural demonstration of GABA-T activity by the final deposition of an electron dense formazan precipitate at reaction sites, whilst maintaining adequate ultrastructural preservation for recognition of cellular and subcellular structures. Numerous electron dense precipitates are evident as discrete punctate deposits situated mainly in mitochondria of stellate cells, basket cells and astrocytic glial cells; they are also seen in axonal or dendritic profiles at some synaptic junctions. The technique enables the first cytochemical demonstration of the mitochondrial localization of GABA-T activity in nervous tissue to be presented. It establishes that GABA-T is present in supposed GABA neurones, in pre- or post-synaptic endings, or both, of presumed inhibitory synapses and in glial cells which may be associated with these synapses. From this seemingly ubiquitous distribution, functional aspects of GABA-T in these cells is considered.