Biomedical subjects

S R Sesack

Publications and source records attributed to S R Sesack.

At least 37 records · Page 2Linked to original sources

PubMed1997

Ultrastructural immunocytochemical localization of the dopamine D2 receptor within GABAergic neurons of the rat striatum.

Classical antipsychotics, which block dopamine (DA) D2 receptors, showing intrastriatal variation in their effectiveness in modulating GABAergic function. To determine the cellular basis for such differences, we examined the electron microscopic immunocytochemical labeling of D2 receptors and GABA in the dorsolateral caudate-putamen (CPn) and the nucleus accumbens (Acb) shell. In both regions, peroxidase reaction product and gold-silver deposits representing D2 receptor immunoreactivity (D2-IR) and GABA immunoreactivity (GABA-IR), respectively, were detected in dendrites and perikarya having characteristics of either spiny projection neurons or aspiny interneurons. Some perikarya in both regions are dually labeled with D2-IR and GABA-IR. Neurons axon terminals in each region also contained one or both markers. However, there were notable regional differences in the immunolabeling patterns. In the CPn, D2-IR was more commonly seen in dendrites/spines than in axon terminals, and proportionally more dendrites were dually labeled than in the Acb. In the Acb shell, D2-IR was detected with similar frequency in terminals and dendrites/spines, but more terminals co-localized D2-IR and GABA-IR in this region compared with the CPn. These results provide the first ultrastructural evidence for direct D2-mediated effects of DA on striatal GABAergic neurons. They further suggest that modulation of GABAergic neurons by DA acting at D2 receptors may be relatively more postsynaptic in the CPn, but more presynaptic in the Acb shell.

Animals↗

PubMed1997

Axosomatic input to subpopulations of cortically projecting pyramidal neurons in primate prefrontal cortex.

Pyramidal cells, the major class of cortical excitatory neurons, can be divided into different subpopulations based upon the target region of their principal axon projection. The activity of pyramidal neurons is regulated in part through inhibitory synaptic inputs to the soma from local circuit neurons. However, little is known about how the density of these axosomatic inputs differs among subpopulations of pyramidal neurons in the prefrontal cortex of primates. In this study, retrograde transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) was used to identify pyramidal neurons in monkey prefrontal cortex (areas 9 and 46), which were labeled via either associational (ipsilateral hemisphere) or callosal (contralateral hemisphere) principal axon projections. Ultrastructural analysis revealed that the relative number of terminals apposed to the somatic membrane did not differ between associational and callosal neurons. However, neurons in the supragranular layers were apposed by a significantly greater number of axon terminals than were neurons in the infragranular layers. These findings suggest that the laminar environment of a neuron may play a more important role than principal axon projection in determining the amount of axosomatic inhibitory input it receives.

Analysis of Variance↗

PubMed1996

Ultrastructural immunocytochemical localization of neurotensin and the dopamine D2 receptor in the rat nucleus accumbens.

The neuroleptic-like effects of neurotensin (NT) are thought to be due to interactions with dopamine (DA) acting primarily at D2 receptors within the nucleus accumbens septi (Acb). Using electron microscopic dual labeling immunocytochemistry, we sought to demonstrate cellular substrates for functional interactions involving NT and DA D2 receptors in the adult rat Acb. Peroxidase reaction product representing D2 receptor-like immunoreactivity (D2-LI) was seen along membranes of Golgi lamellae and multivesicular bodies of perikarya containing immunogold labeling representing NT-LI. Dually labeled somata usually contained highly indented nuclei, a characteristic of aspiny neurons. Dendrites also occasionally colocalized the two immunomarkers. Other somata, dendrites, and all axon terminals were singly labeled with either NT-LI or D2-LI. In distinct sets of terminals, NT-LI was commonly associated with large, dense-cored vesicles, whereas D2-LI was found along the plasmalemma and over nearby small clear vesicles. Each type of terminal comprised approximately 20% of synaptic input to NT-immunoreactive dendrites. Similar proportions of terminals containing NT-LI or D2-LI contacted unlabeled (approximately 55%) or NT-labeled (approximately 35%) dendrites and, occasionally, were observed converging onto common dendrites. Terminals containing NT-LI or D2-LI also were often closely apposed. These findings provide the first ultrastructural evidence that: (1) NT and D2 receptors are colocalized in aspiny neurons and dendrites, (2) NT may produce a direct postsynaptic effect on neurons receiving input from terminals which are presynaptically modulated by DA via D2 receptors, and (3) NT and DA acting at D2 receptors may interact through presynaptic modulation of common axon terminals.

Animals↗

PubMed1996

Hippocampal afferents to the rat prefrontal cortex: synaptic targets and relation to dopamine terminals.

Afferents to the prefrontal cortex (PFC) from the hippocampal formation and from midbrain dopamine (DA) neurons have been implicated in the cognitive and adaptive functions of this cortical region. In the present study, we investigated the ultrastructure and synaptic targets of hippocampal terminals, as well as their relation to DA terminals within the PFC of adult rats. Hippocampal afferents were labeled either by anterograde transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) from the ventral hippocampal formation or by anterograde degeneration following fimbria lesion. Hippocampal terminals in the PFC, identified by either method, formed primarily asymmetric axospinous synapses, with a small percentage forming asymmetric axodendritic synapses. Dopamine terminals in the PFC were identified by peroxidase immunocytochemistry for either tyrosine hydroxylase or DA and formed primarily symmetric synapses onto dendritic spines and small caliber dendritic shafts. Spines that received symmetric synaptic contact from DA terminals invariably also received an asymmetric synapse from an unlabeled terminal, forming a triadic complex. Hippocampal and DA terminals in the PFC were not often observed in the same area of the neuropil, and no examples of convergence of hippocampal and DA terminals onto common postsynaptic targets were observed. Further analysis revealed that spines receiving synaptic contact from hippocampal terminals did not receive additional synaptic contact from any other source. However, when localized to the same area of the neuropil, hippocampal and DA terminals were often in direct apposition to one another, without forming axo-axonic synapses. These results suggest that 1) hippocampal terminals primarily form excitatory synapses onto spiny pyramidal neurons, 2) hippocampal afferents are unlikely to be synaptically modulated by DA or non-DA terminals at the level of the dendritic spine, and 3) appositions between hippocampal and DA terminals may facilitate presynaptic interactions between these afferents to the PFC.

Afferent Pathways↗

PubMed1996

Dopaminergic transmission in the rat retina: evidence for volume transmission.

The study was designed to determine whether dopaminergic neurotransmission in the retina can operate via volume transmission. In double immunolabelling experiments, a mismatch as well as a match was demonstrated in the rat retina between tyrosine hydroxylase (TH) and dopamine (DA) immunoreactive (ir) terminals and cell bodies and dopamine D2 receptor-like ir cell bodies and processes. The match regions were located in the inner nuclear and plexiform layers (D2 ir cell bodies plus processes). The mismatch regions were located in the ganglion cell layer, the outer plexiform layer, and the outer segment of the photoreceptor layer, where very few TH ir terminals can be found in relation to the D2 like ir processes. In similar experiments analyzing D1 receptor like ir processes versus TH ir nerve terminals, mainly a mismatch in their distribution could be demonstrated, with the D1 like ir processes present in the outer plexiform layer and the outer segment where a mismatch in D2 like receptors also exists. The demonstration of a mismatch between the localization of the TH terminal plexus and the dopamine D2 and D1 receptor subtypes in the outer plexiform layer, the outer segment and the ganglion cell layer (only D2 immunoreactivity (IR)) suggests that dopamine, mainly from the inner plexiform layer, may reach the D2 and D1 mismatch receptors via diffusion in the extracellular space. After injecting dopamine into the corpus vitreum, dopamine diffuses through the retina, and strong catecholamine (CA) fluorescence appears in the entire inner plexiform layer and the entire outer plexiform layer, representing the match and mismatch DA receptor areas, respectively. The DA is probably bound to D1 and D2 receptors in both plexiform layers, since the DA receptor antagonist chlorpromazine fully blocks the appearance of the DA fluorescence, while only a partial blockade is found after haloperidol treatment which mainly blocks D2 receptors. These results indicate that the amacrine and/or interplexiform DA cells, with sparse branches in the outer plexiform layer, can operate via volume transmission in the rat retina to influence the outer plexiform layer and the outer segment, as well as other layers of the rat retina such as the ganglion cell layer.

Animals↗

PubMed1995

Axon terminals immunolabeled for dopamine or tyrosine hydroxylase synapse on GABA-immunoreactive dendrites in rat and monkey cortex.

Dopamine afferents to the cortex regulate the excitability of pyramidal neurons via a direct synaptic input. However, it has not been established whether dopamine also modulates pyramidal cell activity indirectly through synapses on gamma-aminobutyric acid (GABA) interneurons, and whether such inputs differ across cortical regions and species. We sought to address these issues by an immunocytochemical electron microscopic approach that combined peroxidase staining for dopamine or tyrosine hydroxylase (TH) with a pre-embedding gold-silver marker for GABA. In the deep layers of the rat prefrontal cortex and in the superficial layers of the monkey prefrontal and primary motor cortices, terminal varicosities immunoreactive for dopamine or TH formed primarily thin, symmetric synapses on distal dendrites. Both GABA-immunoreactive dendrites as well as unlabeled spines and dendrites were contacted by dopamine- or TH-immunoreactive terminals. Synaptic specializations were detected at some, but not all of these contacts. The relative frequency of these appositional and synaptic contacts did not appear to differ between the rat and monkey prefrontal cortex, or between the monkey prefrontal and motor cortices. Across regions and species, labeled and unlabeled targets of dopamine- or TH-positive terminals received additional synaptic input from unlabeled, and occasionally GABA-immunoreactive terminals. Close appositions between dopamine- or TH-immunoreactive and GABA-positive terminals were observed only rarely. These findings indicate that dopamine afferents provide direct synaptic inputs to GABA local circuit neurons in a consistent fashion across cortical regions and species. Thus, dopamine's cellular actions involve direct as well as modulatory effects on both GABA interneurons and pyramidal projection neurons.

Animals↗

PubMed1995

Ultrastructural associations between dopamine terminals and local circuit neurons in the monkey prefrontal cortex: a study of calretinin-immunoreactive cells.

Dopamine terminals in the monkey prefrontal cortex (PFC) synaptically target the distal dendrites of both pyramidal cells and GABA interneurons. We sought to determine whether the latter input includes the innervation of interneurons that utilize calretinin (CalR) as a calcium-binding protein. Sections through prefrontal area 9 of cynomolgus monkeys were processed by immunoperoxidase for tyrosine hydroxylase (TH) to label dopamine varicosities and by pre-embedding immunogold for CalR. Electron microscopic examination of layers 1-3 revealed numerous TH-immunoreactive (TH-ir) terminals, but few were located in the vicinity of CalR-ir dendrites. Although close appositions were sometimes detected between these labeled processes, no synaptic inputs from TH-ir terminals to CalR-ir dendrites were observed. However, in adjacent sections from the same animals, TH-ir terminals were observed to synapse on GABA-ir dendrites. These findings suggest that dopamine afferents to the monkey PFC target the subclasses of GABA interneurons that do not contain CalR.

Animals↗

PubMed1995

Ultrastructural relationships between terminals immunoreactive for enkephalin, GABA, or both transmitters in the rat ventral tegmental area.

The ventral tegmental area (VTA) receives extensive afferent input from neurons containing the opioid peptide enkephalin (Enk) and/or GABA. We examined the ultrastructural basis for known functional interactions between these inhibitory neuromodulators using a combined immunoperoxidase and immunogold-silver technique. As visualized with either marker in single sections, Enk-immunolabeled terminals contained numerous small clear vesicles and one or more intensely immunoreactive dense-cored vesicles. Enk-labeled terminals formed either symmetric or asymmetric synapses on small or large unlabeled dendrites. The immunoreactive dense-cored vesicles were usually detected away from these sites of synaptic contact. Terminals singly immunoreactive for GABA, or dually labeled for Enk and GABA, showed similar morphological features but formed primarily symmetric axo-dendritic synapses. In many instances, GABA- and/or Enk-immunolabeled terminals were in direct apposition to each other and formed synapses on immediately adjacent parts of a common dendrite. Close appositions were also noted between GABA- and Enk-immunoreactive axons and varicosities that did not form synapses with either common or divergent dendrites in single sections. Immunoreactive dense-cored vesicles were frequently detected at the apposed plasmalemmal surfaces between these axon terminals. The findings suggest that Enk and GABA are released from the same or convergent terminals and co-regulate the activity of common target neurons within the rat VTA. The results are also consistent with potential presynaptic interactions between these transmitters.

Animals↗

PubMed1995

Cellular and subcellular localization of syntaxin-like immunoreactivity in the rat striatum and cortex.

Syntaxin is a synapse-specific protein previously localized to the plasma membrane of axon terminals. Biochemical and molecular biological studies indicate a prominent role for syntaxin 1A and 1B in synaptic vesicle docking and/or fusion, suggesting that these proteins are localized to active zone regions of most terminal varicosities in the central nervous system. We sought to test this hypothesis by examining the cellular and subcellular immunocytochemical localization of syntaxin 1 proteins in the striatum and frontal cortex of rats. Using either a polyclonal anti-syntaxin antibody, or a monoclonal antibody directed against the identical protein, HPC-1, immunoperoxidase reaction product was localized to preterminal axons and terminal varicosities that made almost exclusively Type I (asymmetric) synapses on dendritic spines or distal shafts. Immunoreactive terminals forming Type II (symmetric) synapses were observed rarely and only in tissue that was pretreated by rapid freeze-thaw to enhance antibody penetration. From a semi-quantitative analysis, it was estimated that at least 48-62% of all vesicle-filled varicosities and 67-69% of all terminals forming Type I synapses were immunoreactive for syntaxin or HPC-1, respectively. Using a pre-embedding immunogold-silver technique to provide a non-diffusible marker for subcellular localization, gold-silver particles for syntaxin or HPC-1 were localized to the cytoplasmic surface of non-synaptic portions of the plasma membrane of preterminal axons and terminal varicosities. Enrichment of presynaptic active zone regions was not observed with immunogold-silver staining. These findings suggest that syntaxin is primarily contained in a subpopulation of terminals that are associated with excitatory amino acid transmitters, but appears not to be ubiquitously expressed in all terminal classes. The results further indicate that syntaxin is localized to non-synaptic regions of axon and terminal membranes, but may not be enriched in presynaptic active zones. The apparent inconsistency between the subcellular localization of syntaxin and its proposed role in vesicle exocytosis is discussed in terms of possible technical limitations and alternative functions for syntaxin.

Animals↗

PubMed1994

Dynorphin-immunoreactive terminals in the rat nucleus accumbens: cellular sites for modulation of target neurons and interactions with catecholamine afferents.

Dynorphin facilitates conditioned place aversion and reduces locomotor activity through mechanisms potentially involving direct activation of target neurons or release of catecholamines from afferents in the nucleus accumbens. We examined the ultrastructural substrates underlying these actions by combining immunoperoxidase labeling for dynorphin 1-8 and immunogold silver labeling for the catecholamine synthesizing enzyme, tyrosine hydroxylase (TH). The two markers were simultaneously visualized in single coronal sections through the rat nucleus accumbens. By light microscopy, dynorphin immunoreactivity was seen as patches of immunoreactive varicosities throughout all rostrocaudal levels of the nucleus accumbens. The dynorphin-immunoreactive terminals identified by electron microscopy ranged from 0.2 to 1.5 microns in cross-sectional diameter, contained numerous small (30-40 nm) clear vesicles, as well as one or more large (80-100 nm) dense core vesicles. From the dynorphin-immunoreactive terminals quantitatively examined in single sections, 74% (173/370) showed symmetric synaptic junctions mainly with large unlabeled dendrites. Of the dynorphin-immunoreactive terminals forming identifiable synapses, approximately 30% contacted more than one dendritic target. In addition, single dendrites frequently received convergent input from more than one dynorphin-labeled terminal. Irrespective of their dendritic associations, dynorphin-immunoreactive terminals also frequently showed close appositions with other axons and terminals; these included unlabeled (41%), TH-labeled (10%) or dynorphin-labeled axons (14%). In contrast to dynorphin-immunoreactive terminals, TH-labeled terminals formed primarily symmetric synapses with small dendrites and spines or lacked recognizable specializations in the plane of section analyzed. In some cases, single dendrites were postsynaptic to both dynorphin and TH-immunoreactive terminals. We conclude that dynorphin-immunoreactive terminals potently modulate, and most likely inhibit, target neurons in both subregions of the rat nucleus accumbens. This modulatory action could attenuate or potentiate incoming catecholamine signals on more distal dendrites of the accumbens neurons. The findings also suggest potential sites for presynaptic modulatory interactions involving dynorphin and catecholamine or other transmitters in apposed terminals.

Animals↗

PubMed1994

Analysis of synaptic inputs and targets of physiologically characterized neurons in rat frontal cortex: combined in vivo intracellular recording and immunolabeling.

Ultrastructural immunocytochemical identification of transmitters in afferent terminals and targets of individual physiologically characterized neurons is essential for understanding the complex circuitry within the mammalian neocortex. For this type of analysis, we examined the utility of combining in vivo intracellular recording and biocytin injections with silver intensified 1 nm immunogold labeling of GABA and the catecholamine synthesizing enzyme, tyrosine hydroxylase (TH). These transmitters are found to local neurons and afferents known to prominently modulate the activity of pyramidal neurons in the neocortex. Individual neurons were physiologically characterized and filled with biocytin in the frontal cortex of anesthetized rats. The brains were then preserved by vascular perfusion with aldehydes. Single vibratome sections through the recording site were reacted (1) for immunoperoxidase detection of biocytin and (2) for immunogold labeling of GABA or TH. Dually labeled sections were processed for light microscopy or embedded in plastic for electron microscopy. The dense peroxidase product for biocytin was detected in pyramidal neurons. These were located in superficial as well as deep cortical laminae, and were readily distinguished from immunogold silver labeling. GABA labeled terminals formed symmetric synapses with larger biocytin filled dendrites, whereas the TH labeled terminals contacted distal dendrites and spines. Peroxidase labeling for biocytin also was seen in a few axon terminals forming synapses with unlabeled and with GABA immunoreactive dendrites. These results suggest that single pyramidal neurons of the rat frontal cortex receive dual input from both GABA and catecholamine terminals. Additionally, this study demonstrates the usefulness of silver enhancement of 1 nm colloidal gold prior to plastic embedding for electron microscopic detection of neurotransmitters within afferents and targets of neurons physiologically characterized in vivo.

Animals↗

PubMed1994

Ultrastructural localization of D2 receptor-like immunoreactivity in midbrain dopamine neurons and their striatal targets.

Potential cellular substrates for functions ascribed to the dopamine D2 receptor were examined in rat brain using immunoperoxidase for localization of a D2 receptor peptide and immunogold staining for the catecholamine biosynthetic enzyme tyrosine hydroxylase (TH). Specificity of the rat polyclonal antiserum, raised against a 15 amino acid fragment from the third intracellular loop of the D2 receptor, was shown by immunoblot analysis and by selective labeling of cultured Chinese hamster ovary cells permanently transfected with the cDNA for the D2 receptor. Although the light microscopic distribution of immunolabeling for the D2 peptide was diffuse, it was selectively localized to regions containing dopamine cells (substantia nigra and ventral tegmental area) or their forebrain projections (dorsal and ventral striatum, nucleus accumbens, and olfactory tubercles). Electron microscopic examination of the medial substantia nigra and ventral tegmental area revealed readily detectable peroxidase immunoreactivity for the D2 peptide, primarily associated with the smooth endoplasmic reticulum and plasmalemmal surfaces of dendrites. Many D2 peptide-immunoreactive dendrites also contained immunogold labeling for TH, although some dendrites were singly labeled for either marker. In the medial and dorsolateral striatum, immunoperoxidase product for the D2 peptide was localized most extensively in dendrites, with the greatest intensity of immunolabeling seen in spines. A number of striatal dendrites exhibiting D2 peptide labeling were contacted by axon terminals immunoreactive for TH. Additionally, D2 peptide immunoreactivity was distributed to some synaptic vesicles and portions of the plasmalemmal surface in unmyelinated axons and in axon terminals. Most D2 peptide-immunoreactive terminals either lacked detectable membrane specializations, or formed thin, symmetric synapses in single sections. A few D2 peptide-labeled terminals formed asymmetric junctions on dendritic spines. In dually labeled sections, most D2 peptide-immunoreactive terminals lacked detectable immunolabeling for TH. However, in fortunate planes of section, peroxidase product for D2 peptide immunoreactivity was occasionally seen in pre-terminal portions of axons whose terminal varicosities contained immunogold labeling for TH. These ultrastructural results are consistent with the localization of a dopamine D2 receptor-like protein that is strategically positioned to subserve (1) autoreceptor functions at the level of dendrites in the midbrain and presynaptic axon terminals in the striatum, as well as (2) postsynaptic actions on striatal spiny dendrites and other nondopamine terminals.

Animals↗

PubMed1993

Cellular substrates for interactions between dynorphin terminals and dopamine dendrites in rat ventral tegmental area and substantia nigra.

Dynorphin and other kappa opioid agonists are thought to elicit aversive actions and changes in motor activity through direct or indirect modulation of dopamine neurons in ventral tegmental area (VTA) and substantia nigra (SN), respectively. We comparatively examined the immunoperoxidase localization of anti-dynorphin A antiserum in sections through the VTA and SN of adult rat brain to assess whether there were common or differential distributions of this opioid peptide relative to the dopamine neurons. We also more directly examined the relationship between dynorphin terminals and dopamine neurons in VTA and SN by combining immunoperoxidase labeling of rabbit dynorphin antiserum and immunogold-silver detection of mouse antibodies against tyrosine hydroxylase (TH) in single sections through the VTA and SN. Light microscopy showed dynorphin-like immunoreactivity (DY-LI) in varicose processes. These were relatively sparse in VTA and were unevenly distributed in the SN, with little labeling in the pars compacta (pcSN) and the highest density of DY-LI in the medial and lateral pars reticulata (prSN). Electron microscopy established that the regional differences were attributed to differences in density (number/unit area) of immunoreactive profiles. The profiles containing DY-LI were designated as axon terminals based on having diameters greater than 0.1 micron, few microtubules and many synaptic vesicles. In both the VTA and SN, the dynorphin-labeled terminals contained primarily small (35-40 nm) clear vesicles. These vesicles were rimmed with peroxidase immunoreactivity and were often seen clustered above axodendritic synapses. These synaptic specializations were usually symmetric; however a few asymmetric densities also were formed by immunoreactive terminals in both VTA and SN. Additionally, most of the dynorphin-labeled terminals contained 1-2, but occasionally 7 or more intensely peroxidase positive dense core vesicles (DCVs). Approximately 60% of the DCVs were located near axolemmal surfaces. The axolemmal surfaces contacted by immunoreactive DCVs were more often apposed to dendrites in the VTA; while in the SN other axon terminals were the most commonly apposed neuronal profiles. In both regions, a substantial proportion of the plasmalemmal surface in contact with the labeled DCVs was apposed to astrocytic processes.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1992

Prefrontal cortical efferents in the rat synapse on unlabeled neuronal targets of catecholamine terminals in the nucleus accumbens septi and on dopamine neurons in the ventral tegmental area.

Physiological and pharmacological studies indicate that descending projections from the prefrontal cortex modulate dopaminergic transmission in the nucleus accumbens septi and ventral tegmental area. We investigated the ultrastructural bases for these interactions in rat by examining the synaptic associations between prefrontal cortical terminals labeled with anterograde markers (lesion-induced degeneration or transport of Phaseolus vulgaris leucoagglutinin; PHA-L) and neuronal processes containing immunoreactivity for the catecholamine synthesizing enzyme, tyrosine hydroxylase. Prefrontal cortical terminals in the nucleus accumbens and ventral tegmental area contained clear, round vesicles and formed primarily asymmetric synapses on spines or small dendrites. In the ventral tegmental area, these terminals also formed asymmetric synapses on large dendrites and a few symmetric axodendritic synapses. In the nucleus accumbens septi, degenerating prefrontal cortical terminals synapsed on spiny dendrites which received convergent input from terminals containing peroxidase immunoreactivity for tyrosine hydroxylase, or from unlabeled terminals. In single sections, some tyrosine hydroxylase-labeled terminals formed thin and punctate symmetric synapses with dendritic shafts, or the heads and necks of spines. Close appositions, but not axo-axonic synapses, were frequently observed between degenerating prefrontal cortical afferents and tyrosine hydroxylase-labeled or unlabeled terminals. In the ventral tegmental area, prefrontal cortical terminals labeled with immunoperoxidase for PHA-L were in synaptic contact with dendrites containing immunogold reaction product for tyrosine hydroxylase, or with unlabeled dendrites. These results suggest that: (1) catecholaminergic (mainly dopaminergic) and prefrontal cortical terminals in the nucleus accumbens septi dually synapse on common spiny neurons; and (2) dopaminergic neurons in the ventral tegmental area receive monosynaptic input from prefrontal cortical afferents. This study provides the first ultrastructural basis for multiple sites of cellular interaction between prefrontal cortical efferents and mesolimbic dopaminergic neurons.

Animals↗

PubMed1992

Cellular basis for interactions between catecholaminergic afferents and neurons containing Leu-enkephalin-like immunoreactivity in rat caudate-putamen nuclei.

Dopaminergic afferents to the dorsal striatum, caudate-putamen nuclei, are known to modulate the levels and synthesis of endogenous opiate peptides (Leu5 and Met5-enkephalins). We examined the dual immunocytochemical localization of antisera raised against Leu5-enkephalin and the catecholamine-synthesizing enzyme, tyrosine hydroxylase (TH), to determine the cellular substrates for these and/or other functional interactions. The antisera were identified by combined immunogold-silver and immunoperoxidase labeling in single coronal sections through the caudate-putamen nuclei of adult rats. These animals were given intraventricular injections of colchicine, and the brains were fixed by acrolein perfusion prior to immunocytochemical labeling. By light microscopy, perikarya and processes containing enkephalin-like immunoreactivity (ELI) were seen in close proximity to varicose processes immunoreactive for TH. Electron microscopy further demonstrated that the ELI was localized to perikarya, dendrites, and axon terminals, whereas the TH was exclusively in axons and terminals. The dendrites containing ELI were postsynaptic to terminals that were either (1) without detectable immunoreactivity, or (2) immunoreactive for TH or enkephalin. Nonsynaptic portions of the dendrites containing ELI were covered with astrocytic processes or were in direct apposition to unlabeled dendrites. Terminals containing ELI were densely immunoreactive and were in direct contact with (1) unlabeled and occasionally enkephalin-labeled proximal dendrites, and (2) TH-labeled and unlabeled terminals. In comparison with the opiate terminals, most catecholaminergic terminals were lightly immunoreactive for TH and usually contacted more distal unlabeled dendrites or spines and, more rarely, dendrites containing ELI. In a few favorable planes of section, the terminals containing ELI and those containing TH (1) converged on common unlabeled dendrites, or (2) formed dual contacts on two different labeled or unlabeled targets. Junctions formed by terminals containing ELI and TH were sometimes characterized by symmetric synaptic densities. However, numerous other dendritic and all axonal appositions were without recognized membrane densities. The findings of the study provide anatomical substrates for multilevel interactions between catecholamines, mostly dopamine, and enkephalin in rat dorsal striatum. These include (1) monosynaptic input from dopaminergic terminals to neurons containing enkephalin, (2) presynaptic modulation of transmitter release through axonal appositions, and (3) dual regulation of common targets through convergent input. In addition, the findings suggest that both enkephalin and dopamine may have similar modulatory roles in synchronizing the activity of dual targets postsynaptic to individual axon terminals. Alterations in any one of these multiple types of interactions could account for noted motor or sensory symptoms in neurological disorders characterized by depletion of dopamine or endogenous opiate peptides, or both.

Animals↗

PubMed1992

Dual ultrastructural localization of enkephalin and tyrosine hydroxylase immunoreactivity in the rat ventral tegmental area: multiple substrates for opiate-dopamine interactions.

Endogenous opiates modulate activity in the mesocorticolimbic dopaminergic system, and this interaction is thought to underlie major aspects of motoric, reward-seeking, and stress-coping behaviors. We sought to determine the ultrastructural substrate for this modulatory action at the level of dopaminergic perikarya in the rat ventral tegmental area (VTA). Using a dual-labeling, immunoperoxidase and immunogold-silver method, we localized antisera directed against leu5-enkephalin (ENK) and the catecholamine-synthesizing enzyme tyrosine hydroxylase (TH) in acrolein-fixed sections through the VTA. ENK-like immunoreactivity (ENK-LI) was visualized within unmyelinated axons and in axon terminals. In terminals, ENK-LI was densely localized to one or more dense-cored vesicles and either densely or lightly detected surrounding small clear vesicles. Immunoreactive dense-cored vesicles were occasionally associated with the presynaptic specialization but were more frequently detected at distant sites along the plasmalemmal surface, often in apposition to astrocytic processes. ENK-immunoreactive terminals formed both symmetric and asymmetric synapses, most frequently on large proximal dendrites. Direct appositions without glial separation were also detected between terminals containing ENK-LI and other ENK-labeled or unlabeled terminals. In contrast to ENK-LI, immunolabeling for TH was primarily detected within perikarya and dendrites in the VTA. Of the ENK-immunoreactive terminals that formed synaptic contacts in single sections, approximately 50-60% were in association with TH-labeled dendrites. The remainder formed synapses on dendrites lacking detectable immunoreactivity for TH. Multiple ENK-immunoreactive terminals occasionally formed convergent synaptic contacts on single TH-labeled or unlabeled dendrites. Furthermore, individual ENK-labeled terminals sometimes formed divergent contacts on two TH-labeled or unlabeled dendrites. When a single ENK-immunoreactive terminal made dual synaptic contacts on TH-labeled dendrites, the latter were usually in close apposition to one another. These findings represent the first ultrastructural demonstration that opioid peptide-containing terminals provide a direct synaptic input to dopaminergic, as well as nondopaminergic, neurons in the VTA. In addition, the morphological evidence suggests that endogenous opioid peptides (1) may be released from nonsynaptic sites, (2) may modulate the release of transmitters from other terminals, and/or (3) may synchronize the activity of multiple neuronal targets in the VTA. These results provide a number of morphological substrates through which opiates may directly or indirectly regulate activity in mesocorticolimbic dopaminergic pathways.

Animals↗

PubMed1990

In the rat medial nucleus accumbens, hippocampal and catecholaminergic terminals converge on spiny neurons and are in apposition to each other.

The nucleus accumbens septi (Acb) represents an interface between limbic and motor systems and a site for modulation of these integrative functions by ascending catecholaminergic, principally dopaminergic, axons. This modulatory regulation is most likely attributed to pre- or postsynaptic associations between limbic telencephalic and brainstem afferents. In the present investigation, we examined the ultrastructure and synaptic associations of hippocampal afferents, as well as their relation to catecholaminergic terminals, in the medial Acb of adult rats. Hippocampal afferents were identified by anterograde transport of wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) injected in the ventral subiculum, and by anterograde degeneration seen 2-3 days following lesion of the fimbria. Specific comparisons between these methods were made (1) to determine whether similar populations of terminals were labeled and (2) to assess the feasibility of combining degeneration with immunoperoxidase labeling for the catecholamine synthesizing enzyme, tyrosine hydroxylase (TH). Hippocampal afferents labeled with HRP were finely myelinated or unmyelinated and gave rise to small terminals (mean diameter 0.58 micron) containing mostly clear, round vesicles. Of the HRP-labeled terminals which made recognizable junctions, 85% (104/122) formed asymmetric synapses with the heads of dendritic spines. The remainder either formed asymmetric axodendritic synapses or symmetric junctions. Degenerating terminals were significantly smaller (mean diameter 0.35 micron) than terminals labeled with HRP. However, these also formed principally asymmetric axospinous synapses (89/102, 87%). Whether identified by HRP transport or anterograde degeneration, the hippocampal afferents comprised approximately 10% of all terminals and 30% of all asymmetric axospinous synapses in the medial Acb. In contrast to hippocampal afferents, TH-labeled terminals formed primarily symmetric contacts with dendritic shafts and the heads and necks of spines. Quantitative analysis of sections containing both anterograde degeneration and TH-immunoreactivity showed that 25% (26/104) of associations formed by degenerating hippocampal terminals involved convergent inputs with TH-labeled terminals on the same postsynaptic structure. These included dual input either to the same spine head or to different parts of the same dendrite. In addition, the plasma membranes of hippocampal and TH-labeled terminals were often directly apposed to each other (10/58, 17% of axo-axonal associations formed by degenerating terminals), without recognizable synaptic specializations.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗

PubMed1989

Topographical organization of the efferent projections of the medial prefrontal cortex in the rat: an anterograde tract-tracing study with Phaseolus vulgaris leucoagglutinin.

The purpose of the present investigation was to examine the topographical organization of efferent projections from the cytoarchitectonic divisions of the mPFC (the medial precentral, dorsal anterior cingulate and prelimbic cortices). We also sought to determine whether the efferents from different regions within the prelimbic division were organized topographically. Anterograde transport of Phaseolus vulgaris leucoagglutinin was used to examine the efferent projections from restricted injection sites within the mPFC. Major targets of the prelimbic area were found to include prefrontal, cingulate, and perirhinal cortical structures, the dorsomedial and ventral striatum, basal forebrain nuclei, basolateral amygdala, lateral hypothalamus, mediodorsal, midline and intralaminar thalamic nuclei, periaqueductal gray region, ventral midbrain tegmentum, laterodorsal tegmental nucleus, and raphe nuclei. Previously unreported projections of the prelimbic region were also observed, including efferents to the anterior olfactory nucleus, the piriform cortex, and the pedunculopontine tegmental-cuneiform region. A topographical organization governed the efferent projections from the prelimbic area, such that the position of terminal fields within target structures was determined by the rostrocaudal, dorsoventral, and mediolateral placement of the injection sites. Efferent projections from the medial precentral and dorsal anterior cingulate divisions (dorsomedial PFC) were organized in a similar topographical fashion and produced a pattern of anterograde labeling different from that seen with prelimbic injection sites. Target structures innervated primarily by the dorsomedial PFC included certain neocortical fields (the motor, somatosensory, and visual cortices), the dorsolateral striatum, superior colliculus, deep mesencephalic nucleus, and the pontine and medullary reticular formation. Previously unreported projections to the paraoculomotor central gray area and the mesencephalic trigeminal nucleus were observed following dorsomedial PFC injections. These results indicate that the efferent projections of the mPFC are topographically organized within and across the cytoarchitectonic divisions of the medial wall cortex. The significance of topographically organized and restricted projections of the rat mPFC is discussed in light of behavioral and physiological studies indicating functional heterogeneity of this region.

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