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Genesis of interneurons in the dorsal lateral geniculate nucleus of the cat.

Most neurons in the A-laminae of the cat's dorsal lateral geniculate nucleus (LGN) are born between embryonic days 22 and 32. Whereas approximately 78% of these cells are destined to become geniculocortical relay cells, the remaining 22% of LGN neurons do not appear to establish connections with visual cortex, and therefore can be considered interneurons. In the present study we have combined the 3H-thymidine method for labeling dividing neurons with the retrograde horseradish peroxidase (HRP) method for identifying LGN relay cells in order to study specifically the genesis of interneurons in the cat's LGN. Developing LGN interneurons in 12 kittens were labeled with 3H-thymidine by injecting the radioactive label into the allantoic cavity of their pregnant mothers on different embryonic days. Approximately 8-22 weeks after birth LGN relay cells in the A-laminae were labeled retrogradely by injecting large volumes of HRP into visual cortex areas 17 and 18. LGN cells that could not be labeled retrogradely with HRP were considered to be interneurons. Our results show that interneurons are born on each of the embryonic days studied, E24-E30. This period represents approximately the middle two-thirds of the entire period of LGN neurogenesis. Although the birth rate for interneurons is not uniform, there is no indication from our data that interneurons and relay cells in the cat's LGN are born at different times during LGN neurogenesis.

Animals↗

Distribution and morphology of synapses on nonspiking local interneurones in the thoracic nervous system of the locust.

The structure and distribution of synapses on nonspiking local interneurones in the metathoracic ganglion of the locust was revealed by electron microscopy following intracellular injection of horseradish peroxidase (HRP). Before staining, each interneurone was characterized physiologically as nonspiking and its output effects on motor neurones innervating muscles in a hindleg were investigated. Three nonspiking interneurones of different morphologies, each typical of a previously described population, were selected for detailed study. The first has a dorsal soma and ipsilateral neuropilar branches, the second a ventral soma and ipsilateral branches, and the third a ventral soma and contralateral branches. The somata have few trophospongial invaginations, and most of their volume is occupied by the nucleus. The initial parts of the primary neurites are either wrapped in glia or isolated in tracts from the neuropile and thus do not participate in synaptic interactions. Some of the larger secondary neurites are also wrapped in glia, but others both make and receive synaptic contacts. Output synapses have an array of some 500-1,600 round, agranular vesicles (diameter 47.0 +/- 5.7 nm; mean +/- S.D., n = 97) associated with a bar-shaped presynaptic density up to 0.3 micron long. Two postsynaptic processes, whose diameter can vary greatly, are usually associated with each presynaptic density. Processes making input synapses onto nonspiking local interneurones typically contain round, agranular vesicles and often make several contacts within a few microns. Serial reconstructions from one of the interneurones revealed input and output synapses intermingled on the larger processes with outputs dominating by a factor of 3:1, whereas on some of the thinner processes only input synapses are present. In the other two interneurones, however, both input and output synapses are present on the fine branches. No feature of the structure or distribution of synapses observed here on the nonspiking local interneurones distinguishes them from spiking neurones in the same ganglia.

Animals↗

Embryonic development of a population of spiking local interneurones in the locust (Schistocerca gregaria).

We examined the embryonic development of an identified group of thoracic spiking local interneurones in the locust. These interneurones are primary integrators of mechanosensory information from the legs and make inhibitory output connections with motor neurones, nonspiking local interneurones, and intersegmental interneurones. Using intracellular dye injection, we traced the origins of these interneurones and described their morphological development. All of the interneurones in this population are produced by neuroblast NB4-1. The interneurones are produced during the latter stages of the neuroblast lineage and could not be identified before 55% development. Their growth could be divided into three distinct phases: first, a period of initial outgrowth (55-70%) during which the basic skeleton of major neurites is formed; second, a shorter period of rapid growth (70-80%) during which the basic skeleton is elaborated by the addition of many side branches; and third, a period of maturation (80-95%) during which the branches formed during earlier growth appear to be pruned. The possible implications of these results for neural development and synaptogenesis are discussed.

Animals↗

Physiology and morphology of spiking local interneurons in the terminal abdominal ganglion of the crayfish.

Not only nonspiking local interneurons but also spiking local interneurons in the terminal abdominal ganglion of the crayfish (Procambarus clarkii Girard) contribute to form the motor output of the uropod. In this study, 14 spiking local interneurons are identified by their gross morphology and physiological properties including the sensory inputs from the uropod and premotor output onto the uropod motor neurons on both sides. Morphologically, they are divided into three groups based on the position of their somata: anterior, medial, and posterior. The main branches of interneurons in each group are usually extended on the side contralateral to the soma, or they have profuse bilateral branches. Physiologically, all of them receive excitatory inputs from the afferents innervating the exopodite on the side ipsilateral their main branches. They are usually silent, and spike only in response to sensory stimulation. The majority of them (11 out of 14 neurons) have output effects on the uropod motor neurons that elicit reciprocally closing pattern. They increased the activity of the closer motor neurons and decreased the activity of the opener motor neurons. Only one interneuron inhibits the closer motor neurons. The remaining two interneurons coactivate both the antagonistic motor neurons. Since the reciprocal closing pattern of the uropod motor neurons is elicited in response to sensory stimulation of the uropod, spiking local interneurons may initiate this local reflex circuit. They detect the initiation of the stimulus and convey signals from the sensory to the motor side.

Animals↗

Immunocytochemically defined interneuron populations in the hippocampus of mouse strains used in transgenic technology.

Transgenic mice are overtaking the role of model animals in neuroscience. They are used in developmental, anatomical, and physiological as well as experimental neurology. However, most results on the organization of the nervous system derive from the rat. The rat hippocampus and its neuronal elements have been thoroughly investigated, revealing remarkable functional and morphological diversity and specificity among hippocampal interneurons. Our aim was to examine the properties of distinct hippocampal interneuron populations, i.e., those immunoreactive for calcium-binding proteins (parvalbumin, calbindin, and calretinin), neuropeptides (cholecystokinin, neuropeptide Y, somatostatin, vasoactive intestinal polypeptide), and certain receptors (metabotropic glutamate receptor 1alpha, cannabinoid receptor type 1) in four strains of mice widely used in transgenic technology, and to compare their properties to those in the rat. Our data indicate that the distribution as well as the dendritic and axonal arborization of mouse interneurons immunoreactive for the different markers was identical in the examined mouse strains, and in most respects are similar to the features found in the rat. The postsynaptic targets of neurons terminating in the perisomatic (parvalbumin), proximal (calbindin), and distal (somatostatin) dendritic region, as well as on other interneurons (calretinin), also matched those found in the rat. However, a few significant differences could also be observed between the two species in addition to the already described immunoreactivity of mossy cells for calretinin: the absence of spiny calretinin-immunoreactive interneurons in the CA3 region, sparse contacts between calretinin-immunoreactive interneurons, and the axon staining for somatostatin and neuropil labeling for cholecystokinin. We can conclude that the morphofunctional classification of interneurons established in the rat is largely valid for mouse strains used in transgenic procedures.

Animals↗

Mu opioid receptors are in discrete hippocampal interneuron subpopulations.

In the rat hippocampal formation, application of mu opioid receptor (MOR) agonists disinhibits principal cells, promoting excitation-dependent processes such as epileptogenesis and long-term potentiation. However, the precise location of MORs in particular inhibitory circuits, has not been determined, and the roles of MORs in endogenous functioning are unclear. To address these issues, the distribution of MOR-like immunoreactivity (-li) was examined in several populations of inhibitory hippocampal neurons in the CA1 region using light and electron microscopy. We found that MOR-li was present in many parvalbumin-containing basket cells, but absent from cholecystokinin-labeled basket cells. MOR-li was also commonly in interneurons containing somatostatin-li or neuropeptide Y-li that resembled the "oriens-lacunosum-moleculare" (O-LM) interneurons innervating pyramidal cell distal dendrites. Finally, MOR-li was in some vasoactive intestinal peptide- or calretinin-containing profiles resembling interneurons that primarily innervate other interneurons. These findings indicate that MOR-containing neurons form a neurochemically and functionally heterogeneous subset of hippocampal GABAergic neurons. MORs are most frequently on interneurons that are specialized to inhibit pyramidal cells, and are on a limited number of interneurons that target other interneurons. Moreover, the distribution of MORs to different neuronal types in several laminae, some relatively far from endogenous opioids, suggests normal functional roles that are different from the actions seen with exogenous agonists such as morphine.

Animals↗

Hyperpolarizing synaptic potentials evoked in CA1 pyramidal cells by glutamate stimulation of interneurons from the oriens/alveus border of rat hippocampal slices. II. Sensitivity to GABA antagonists.

The receptor type mediating the inhibitory postsynaptic potentials (glut-IPSPs), recorded in CA1 pyramidal cells, as a result of glutamate stimulation of interneurons in stratum oriens near the alveus (O/A) was assessed and compared to the type mediating recurrent IPSPs evoked by recurrent activation of interneurons through glutamate stimulation of pyramidal cells in stratum pyramidale (PYR). In response to repetitive electrical stimulation, the peak amplitude of both the O/A glut-IPSP and the PYR glut-IPSP was attenuated (n = 5) in parallel to the reduction in amplitude of the early and late components of the electrically evoked response (stimulus-evoked disinhibition). This suggested the involvement of GABAergic receptors and attested that the interneurons activated during glut-IPSPs were also involved in the circuitry of the electrically evoked IPSPs. The local application of the selective GABAA antagonist bicuculline (100-200 microM) to the slice resulted in a significant reduction in the amplitude of both the O/A (by 76.5%; n = 9) and PYR (by 86.2%; n = 5) glut-IPSPs, in parallel to a decrease of the electrically evoked early IPSP, but not of the late IPSP. The presence of the GABAB antagonist 2-hydroxy-saclofen (1 mM) was able to significantly reduce the amplitude of the O/A glut-IPSPs (by 27.5%; n = 7) and of the electrically evoked late IPSP, but not the PYR glut-IPSP (n = 3). Although the application of phaclofen (20 mM) to the slice reduced the amplitude of the O/A glut-IPSPs (n = 3), the reduction was not statistically significant. These results suggest that recurrent IPSPs elicited from activation of interneurons by stimulation of pyramidal cells are mediated solely via GABAA receptors. Inhibitory postsynaptic potentials elicited from stimulation of interneurons in O/A were also mediated mostly by GABAA receptors, but in addition, displayed a minor component mediated by GABAB receptors. Therefore, since a large proportion of interneurons in O/A are recurrently excited by pyramidal cells (Lacaille J-C et al., 1987, J Neurosci 7: 1979-1993), and since recurrent IPSPs appeared mediated by GABAA receptors, a subpopulation of interneurons activated from O/A might exist that do not receive recurrent excitation but can inhibit pyramidal cells via GABAB receptors.

Animals↗

Unexpected divergence among identified interneurons in different abdominal segments of the crayfish Procambarus clarkii.

The command elements that initiate and coordinate the abdominal movements in crayfish show little similarity between the various abdominal segments. Our criteria for similarity among interneurons were based on both cell morphology and electrophysiology. By contrast, previously published evidence shows much greater intersegmental similarity in the skeletal, muscular, motoneuronal, and sensory components of the abdominal system in crayfish, structures that are controlled by or send information to the command elements. Therefore, unlike the command elements, these structures have retained nearly identical form and function in the various segments. We also found in different ganglia examples of interneurons involved with abdominal positioning behavior that have similar morphology but different function and vice versa. Such interneurons could represent divergent pairs of serial homologues. It is unknown why so many of the abdominal positioning interneurons have become different. The various ganglia may perform subtly different functions, requiring differences in the positioning interneurons but not in the motor neurons or muscles. Alternatively, some of the abdominal positioning interneurons underlie more than one behavior; consequently, selection acting on these multiple functions may have changed these interneurons through evolution.

Abdomen↗

Excitatory responses to serotonin (5-HT) in neurons of the rat piriform cortex: evidence for mediation by 5-HT1C receptors in pyramidal cells and 5-HT2 receptors in interneurons.

As a prerequisite to pharmacological analysis of the excitatory effects of serotonin (5-HT) on piriform pyramidal cells and interneurons, this study first examined the physiological characteristics of these two cell types. Intracellular recordings confirmed that the subpopulation of 5-HT-activated cells located at the border of layers II and III are indeed interneurons. Voltage clamp recordings in pyramidal cells showed that the increase in excitability produced by 5-HT in these cells was the result of voltage- and Ca(2+)-dependent outward currents with the characteristics of IM and IAHP. Pharmacological studies were designed to discriminate 5-HT2 from 5-HT1C responses in interneurons and pyramidal cells of piriform cortex. The 5-HT antagonist spiperone, which has a much higher affinity for 5-HT2 receptors than for 5-HT1C receptors, blocked the excitatory effect of 5-HT at lower concentrations in interneurons (IC50 = 31 nM) than in pyramidal cells (IC50 = 2.1 microM). Similarly, ritanserin, a drug which also has a higher affinity for 5-HT2 than 5-HT1C receptors, blocked the effect of 5-HT at lower concentrations in interneurons (IC50 = 400 nM) than in pyramidal cells (IC50 = 8.1 microM). In contrast, LY 53857, an antagonist with higher affinity for 5-HT1C than for 5-HT2 receptors, blocked the effect of 5-HT at lower concentrations in pyramidal cells (IC50 = 26 nM) than in interneurons (IC50 = 364 nM). The 5-HT1C partial agonist/5-HT2 antagonist mCPP produced agonist-like effects in only 66% of pyramidal cells tested indicating that not all pyramidal cells may express 5-HT1C receptors. In that both spiperone and ritanserin have higher affinity for 5-HT2 receptors than for 5-HT1C receptors and LY 53857 has a higher affinity for 5-HT1C receptors than for 5-HT2 receptors, these data suggest that in piriform cortex excitatory effects of 5-HT are mediated by 5-HT1C receptors in pyramidal cells an by 5-HT2 receptors in interneurons.

Animals↗

Output connections of a wind sensitive interneurone with motor neurones innervating flight steering muscles in the locust.

The output connections of a bilaterally symmetrical pair of wind-sensitive interneurones (called A4I1) were determined in a non-flying locust (Schistocerca gregaria). Direct inputs from sensory neurones of specific prosternal and head hairs initiate spikes in these interneurones in the prothoracic ganglion. The interneurone with its axon in the right connective makes direct, excitatory connections with the two mesothoracic motor neurones innervating the pleuroaxillary (pleuroalar, M85) muscle of the right forewing, but not with the comparable motor neurones of the left forewing. The connections can evoke motor spikes. The interneurones also exert a powerful, but indirect effect on the homologous metathoracic pleuroaxillary motor neurones (muscle 114), and a weaker, indirect effect on subalar motor neurones of the hindwings. No connections or effects were found with other flight motor neurones, or motor neurones innervating hindleg muscles, including common inhibitor 1 which also innervates the pleuroaxillary muscle. One thoracic interneurone with its cell body in the right half of the mesothoracic ganglion and with its axon projecting ipsilaterally to the metathoracic ganglion receives a direct input from the right A4I1 interneurone. These restricted output connections suggest a role for the A4I1 interneurones in flight steering.

Animals↗

Integration in trigeminal premotor interneurones in the cat. 1. Functional characteristics of neurones in the subnucleus-gamma of the oral nucleus of the spinal trigeminal tract.

The profile of integration in a sample of 183 interneurones localized in the subnucleus-gamma of the oral nucleus of the spinal trigeminal tract (NVspo-gamma) has been analyzed. 134 neurones were tested for inputs from primary afferents of the trigeminal, facial and cervical nerves as well as for inputs from the midbrain and from the cervical spinal cord. The remaining 49 neurones were tested for inputs from the primary afferents and for descending convergence from defined sites within the oro-facial primary projections of the cerebral cortex. It was found that the interneurones, mainly recorded in the dorsal and dorsomedial aspect of the NVspo-gamma, receive short latency inputs from the low threshold oral and perioral afferents and longer latency inputs from the high threshold jaw and neck muscle afferents. There was evidence for convergence from the cervical segmental level (29%) and some of the neurones had axon terminals in the superior colliculus. However, the interneurones did not receive a descending tectal input. About 80% of the NVspo-gamma interneurones were activated from the orofacial primary projection fields within cytoarchitectonic areas 3a and 3b of the coronal gyrus. This input was topographically organized and the neurones were activated from the same oral and perioral region of the periphery as the cortical region from which the descending projections themselves originated. Minimum latencies indicated a monosynaptic connection. The convergence profile onto the NVspo-gamma interneurones appeared unique as compared with interneurones located in the intertrigeminal area. Aspects of the possible functional roles of the NVspo-gamma neurones are discussed in relation to ongoing oro-facial ("masticatory") movements. The properties of a selected sample of NVspo-gamma interneurones, which were antidromically activated from the digastric subnucleus of the trigeminal motor nucleus, are reported in a companion paper (Olsson and Westberg 1991).

Afferent Pathways↗

Evidence for mutual inhibition of opposite Ia interneurones in the human upper limb.

The H-reflex technique was used to collect indirect evidence for changes in excitability of the interneurones mediating reciprocal Ia inhibition between wrist extensors and flexors. Stimulating the radial nerve results in an inhibition of the flexor carpi radialis (FCR) H-reflex and evidence has previously been presented that the early phase of inhibition is mediated by extensor-coupled Ia interneurones (Ext Ia INs), i.e. by inhibitory interneurones fed by muscle spindle Ia afferents from wrist extensors. Variations in the level of this inhibition were used to assess changes in excitability of Ext Ia INs. Stimulation of group I fibres from flexors was shown to depress the "reference" Ia inhibition, i.e. to inhibit the Ext Ia INs. The central latency of this interneuronal inhibition was compatible with a disynaptic linkage between flexor Ia afferents and Ext Ia INs. Its threshold and time course profile could almost exactly be superimposed on those of reciprocal Ia inhibition from flexors to extensor carpi radialis (ECR) motoneurones (MNs). This suggests that the Ia inhibitions to extensor MNs and extensor Ia INs are collateral effects mediated by the same flexor-coupled Ia interneurones. In two subjects, in whom it was possible to elicit an H-reflex in the ECR, inhibition of flexor-coupled Ia interneurones by activation of extensor Ia interneurones could similarly be demonstrated.

Adult↗

Integration in descending motor pathways controlling the forelimb in the cat. 12. Interneurones which may mediate descending feed-forward inhibition and feed-back inhibition from the forelimb to C3-C4 propriospinal neurones.

Extra- and intracellular recording was made from cells in the C3-C4 segments with the aim of finding interneurones of previously described inhibitory pathways to the C3-C4 propriospinal neurones, which may mediate descending feed-forward inhibition and feed-back inhibition from the forelimb, respectively. The lateral interneurones were found in the lateral part of lamina VII interspersed among the C3-C4 PNs and like them they receive convergent monosynaptic EPSPs and disynaptic IPSPs from the cortico-, rubro-, tecto- and reticulospinal tracts. Disynaptic IPSPs, but only rarely monosynaptic EPSPs, are evoked in them from forelimb nerves. The lateral interneurones do not project to the lateral reticular nucleus (LRN). The medial interneurones were found medially in laminae V and VI in a region where volleys in forelimb nerves evoke extracellular monosynaptic focal potentials (Rosén 1969). There is somatotopic organization of the projection from the forelimb to this region. Many neurones are strongly monosynaptically excited from group I muscle or/and cutaneous forelimb afferents. In addition, late discharges are evoked in many cells from cutaneous afferents and high threshold muscle afferents. Corticospinal volleys evoked monosynaptic excitation in the great majority of these cells and usually also late EPSPs or IPSPs. Typically, rubrospinal and tectospinal volleys evoked neither monosynaptic excitation nor late effects as those elicited from corticospinal fibres. In some of the interneurones, IPSPs were evoked from forelimb nerves. About 20% of the medial "interneurones" have an ascending projection to the caudal brain stem. Threshold mapping for antidromic stimulation revealed termination in the main cuneate nucleus, the external cuneate nucleus and/or the LRN and also a branch projecting to more rostral levels in the brain. A few of the neurones in the medial region are PNs projecting to the forelimb segments. It is postulated that interneurones both of the lateral and medial type are inhibitory and project to the C3-C4 PNs. It is further postulated that the former are intercalated in the descending feed-forward inhibitory pathway to the C3-C4 PNs and the latter in the feed-back inhibitory pathway from the forelimb to these PNs. The role of feed-forward and feed-back inhibition of transmission from the brain to forelimb motoneurones via the C3-C4 PNs is discussed.

Afferent Pathways↗

Recurrent inhibition of individual Ia inhibitory interneurones and disinhibition of their target alpha-motoneurones during muscle stretches.

The effects of ramp stretches applied to triceps surae muscle on the discharge patterns of single Ia inhibitory interneurones, monosynaptically invaded from various nerves, were studied in either decerebrate or anesthetized cats. Interneurones which received direct excitatory Ia input from the stretched muscle exhibited augmented activity both during the dynamic and static phase of stretch, which was, however, interrupted by a transient inhibitory influence during the dynamic phase of stretch. The influences on Ia inhibitory interneurones, monosynaptically invaded from hamstring or tibial nerve, were exclusively inhibitory. These stretch-induced inhibitions were better demonstrable in decerebrate than in anesthetized preparations. The timing of the discharge patterns of additionally recorded Renshaw cells during stretch, and the disappearance or reduction of the above described inhibitory effects after administration of DHE, strongly support the idea that these inhibitory actions are caused by Renshaw inhibition. In Ia inhibitory interneurones, monosynaptically activated from the antagonistic peroneal nerve, stretch induced also pronounced inhibitory effects, which were most probably caused by mutual inhibition between Ia inhibitory interneurones. The suppression of agonistic Ia inhibitory interneurone activity below the tonic resting activity corresponded to an enhancement of the monosynaptic reflex amplitude of the antagonistic motoneurone pool. The findings suggest that normal orthodromic activation of Renshaw cells, and consequently the recurrent inhibition of the Ia inhibitory interneurones, is predominantly linked with rapid phasic, rather than slow tonic, motoneuronal firing. The functional role of this mechanism for the performance of rapidly alternating movements and the damping of ballistic agonist contractions is discussed.

Animals↗

Sources of inputs to longitudinal muscle motor neurons and ascending interneurons in the guinea-pig small intestine.

Light- and electron-microscopic studies were used to investigate connections between specific subgroups of neurons in the myenteric plexus of the guinea-pig small intestine. Inputs to two classes of calretinin-immunoreactive (IR) nerve cells, longitudinal muscle motor neurons and ascending interneurons, were examined. Inputs from calbindin-IR primary sensory neurons and from three classes of descending interneurons were studied. Electron-microscopic analysis showed that calbindin-IR axons formed two types of inputs, synapses and close contacts, on calretinin-IR neurons. About 40% of inputs to the longitudinal muscle motor neurons and 70% to ascending interneurons were calbindin-IR. Approximately 50% of longitudinal muscle motor neurons were surrounded by bombesin-IR dense pericellular baskets and 40% by closely apposed varicosities. At the electron-microscope level, the bombesin-IR varicosities were found to form synapses and close contacts with the motor neurons. Dense pericellular baskets with bombesin-IR surrounded 36% of all ascending interneurons, and a further 17% had closely apposed varicosities. Somatostatin- and 5-HT-IR descending interneurons provided no dense pericellular baskets to calretinin-IR nerve cells. Thus, calretinin-IR, longitudinal muscle motor neurons and ascending interneurons receive direct synaptic inputs from intrinsic primary sensory neurons and from non-cholinergic, bombesin-IR, descending interneurons.

Animals↗

Postural interneurons in the abdominal nervous system of lobster. I. Organization, morphologies and motor programs for flexion, extension and inhibition.

Using intracellular recording and dye-filling techniques, a survey of postural interneurons was undertaken by impaling their somata in the 2nd abdominal ganglion of lobster. During the course of study approximately fourty different intersegmental interneurons in this ganglion were sampled. Of these, 8 evoked unique, patterned responses in the postural (superficial) motoneurons; each could be identified morphologically. Five of the 8 interneurons had caudally directed axons; 4 of these projected beyond the 4th abdominal ganglion. The remainder projected rostrally, beyond the 1st abdominal ganglion. The postural interneurons were classified according to the motor program they elicited. Five were flexion producing interneurons (FPIs), one was extension producing (EPI), and two generated only inhibitory motor outputs. All motor responses were bilateral and occurred in several segments, including A2. Two neurons, FPIs 201 and 301, produced the full motor reciprocity that typically is observed when flexion command fibers are stimulated. However, three of the FPIs and the single EPI did not express complete reciprocity in synergistic and antagonistic motoneurons. The results indicate that some interneurons displaying all of the properties of command neurons are located entirely within the abdominal nervous system. The overall organization of posture-evoking interneurons appears to be similar to that found in crayfish, suggesting an even more fundamental homology in the neuronal connectivities of these two species than has been established previously.

Abdomen↗

Ischemia changes the coexpression of somatostatin and neuropeptide Y in hippocampal interneurons.

Transient cerebral ischemia causes extensive cell death in hippocampal CA1 pyramidal cells and selective loss of interneurons in the dentate hilus. Many hippocampal interneurons can be classified by their contents of somatostatin (SS) and/or neuropeptide Y (NPY). Following ischemia in the rat, most of the NPY immunoreactivity is permanently lost in hippocampus. Furthermore, SS interneurons in the dentate hilus die, whereas CA1 interneurons survive and their expression of SS mRNA and peptide returns to preischemic levels within 16 days after ischemia. We have addressed the following questions: (1) Does the loss of NPY involve a specific downregulation in surviving CA1 interneurons that pre-ischemically expressed both SS and NPY? (2) Can the subpopulation of dying interneurons in hilus be identified from their preischemic coexpression of SS and NPY? We investigated the coexpression of SS mRNA and NPY peptide using combined in situ hybridization and immunocytochemistry. Cells containing one or both markers were counted in control sections and sections taken 2-16 days after ischemia from the hippocampal formation. In CA1, a decrease in the number of neurons containing NPY alone as well as a decrease in the number of neurons coexpressing NPY and SS was observed, whereas the number of neurons containing SS alone increased 16 days after ischemia. We conclude that neurons coexpressing SS and NPY before ischemia added to the number of neurons containing SS alone after ischemia, because NPY expression was selectively down regulated in the coexpressing population. In hilus, we demonstrated both survival and ischemic cell death of neurons expressing either SS, NPY or both, indicating that hilar interneurons dying from ischemia cannot unequivocally be identified from their preischemic colocalization of SS and NPY.

Animals↗

Segmental and propriospinal projection systems of frog lumbar interneurons.

Spinal interneuronal networks have been implicated in the coordination of reflex behaviors and limb postures in the spinal frog. As a first step in defining these networks, retrograde transport of horseradish peroxidase (HRP) was used to examine the anatomical organization of interneuronal circuitry in the lumbar spinal cord of the frog. Following neuronal degeneration induced by spinal transection and section of the dorsal and ventral roots, HRP was placed at different locations in the spinal cord and the positions of labeled neuronal cell bodies plotted using a Eutectics Neuron Tracing System. We describe four spinal interneuronal systems, three with cell bodies located in the lumbar cord and one with descending projections to the lumbar cord. Interneurons with cell bodies located in the lumbar cord include: (1) Lumbar neurons projecting rostrally. Those projecting to thoracic segments tended to be located in the lateral and ventrolateral gray and in the lower two-thirds of the dorsal horn, with projections that were predominantly uncrossed. Those projecting to the brachial plexus and beyond were located in the dorsal part of the dorsal horn (uncrossed) and in the lateral, ventrolateral, and ventromedial gray (crossed). (2) Lumbar neurons with segmental projections within the lumbar cord. These neurons, which were by far the most numerous, had both uncrossed and crossed projections and were distributed throughout the dorsal, lateral, ventrolateral, and ventromedial gray matter. (3) Lumbar neurons projecting to the sacral cord. This population, which arose mainly from the dorsal horn and lateral or ventrolateral gray, was much smaller than in the other systems. Neuronal density of some of these populations of lumbar interneurons appeared to vary with rostrocaudal level. Finally, a population of neurons with cell bodies in the brachial and thoracic segments that projects to the lumbar cord is described. The most rostral of these neurons were multipolar cells with uncrossed projections, while those with crossed projections were confined almost exclusively to the ventral half of the cord. The distribution of spinal interneurons reported here will provide guidance for future studies of the role of interneuronal networks in the control of movements using the spinal frog as a model system.

Animals↗