Separate cortical and hippocampal cell populations target the rat nucleus reuniens and mammillary bodies.

Nucleus reuniens receives dense projections from both the hippocampus and the frontal cortices. Reflecting these connections, this nucleus is thought to enable executive functions, including those involving spatial learning. The mammillary bodies, which also support spatial learning, again receive dense hippocampal inputs, as well as lighter projections from medial frontal areas. The present study, therefore, compared the sources of these inputs to nucleus reuniens and the mammillary bodies. Retrograde tracer injections in rats showed how these two diencephalic sites receive projections from separate cell populations, often from adjacent layers in the same cortical areas. In the subiculum, which projects strongly to both sites, the mammillary body inputs originate from a homogenous pyramidal cell population in more superficial levels, while the cells that target nucleus reuniens most often originate from cells positioned at a deeper level. In these deeper levels, a more morphologically diverse set of subiculum cells contributes to the thalamic projection, especially at septal levels. While both diencephalic sites also receive medial frontal inputs, those to nucleus reuniens are especially dense. The densest inputs to the mammillary bodies appear to arise from the dorsal peduncular cortex, where the cells are mostly separate from deeper neurons that project to nucleus reuniens. Again, in those other cortical regions that innervate both nucleus reuniens and the mammillary bodies, there was no evidence of collateral projections. The findings support the notion that these diencephalic nuclei represent components of distinct, but complementary, systems that support different aspects of cognition.

Visual landmark information gains control of the head direction signal at the lateral mammillary nuclei.

The neural representation of directional heading is conveyed by head direction (HD) cells located in an ascending circuit that includes projections from the lateral mammillary nuclei (LMN) to the anterodorsal thalamus (ADN) to the postsubiculum (PoS). The PoS provides return projections to LMN and ADN and is responsible for the landmark control of HD cells in ADN. However, the functional role of the PoS projection to LMN has not been tested. The present study recorded HD cells from LMN after bilateral PoS lesions to determine whether the PoS provides landmark control to LMN HD cells. After the lesion and implantation of electrodes, HD cell activity was recorded while rats navigated within a cylindrical arena containing a single visual landmark or while they navigated between familiar and novel arenas of a dual-chamber apparatus. PoS lesions disrupted the landmark control of HD cells and also disrupted the stability of the preferred firing direction of the cells in darkness. Furthermore, PoS lesions impaired the stable HD cell representation maintained by path integration mechanisms when the rat walked between familiar and novel arenas. These results suggest that visual information first gains control of the HD cell signal in the LMN, presumably via the direct PoS → LMN projection. This visual landmark information then controls HD cells throughout the HD cell circuit.

Mammillothalamic tract lesions disrupt dead reckoning in the rat.

Debate surrounds the role of the limbic system structures' contribution to spatial orientation. The results from previous studies have supported a role for the mammillary bodies and their projections to the anterior thalamus in rapid encoding of relationships among environmental cues; however, this work is based on behavioral tasks in which environmental and self-movement cues could not be dissociated. The present study examines the effects of mammillothalamic tract lesions on spatial orientation in the food hoarding paradigm and the water maze. Although the food hoarding paradigm dissociates the use of environmental and self-movement cues, both sources of information are available to guide performance in the water maze. Mammillothalamic tract lesions selectively impaired performance on both tasks. These impairments are interpreted as providing further evidence for the role of limbic system structures in processing self-movement cues.

Collateral Projections Innervate the Mammillary Bodies and Retrosplenial Cortex: A New Category of Hippocampal Cells.

To understand the hippocampus, it is necessary to understand the subiculum. Unlike other hippocampal subfields, the subiculum projects to almost all distal hippocampal targets, highlighting its critical importance for external networks. The present studies, in male rats and mice, reveal a new category of dorsal subiculum neurons that innervate both the mammillary bodies (MBs) and the retrosplenial cortex (RSP). These bifurcating neurons comprise almost half of the hippocampal cells that project to RSP. The termination of these numerous collateral projections was visualized within the medial mammillary nucleus and the granular RSP (area 29). These collateral projections included subiculum efferents that cross to the contralateral MBs. Within the granular RSP, the collateral projections form a particularly dense plexus in deep Layer II and Layer III. This retrosplenial termination site colocalized with markers for VGluT2 and neurotensin. While efferents from the hippocampal CA fields standardly collateralize, subiculum projections often have only one target site. Consequently, the many collateral projections involving the RSP and the MBs present a relatively unusual pattern for the subiculum, which presumably relates to how both targets have complementary roles in spatial processing. Furthermore, along with the anterior thalamic nuclei, the MBs and RSP are key members of a memory circuit, which is usually described as both starting and finishing in the hippocampus. The present findings reveal how the hippocampus simultaneously engages different parts of this circuit, so forcing an important revision of this network.

A neuromodulatory role for oxytocin within the supramammillary nucleus.

Oxytocin functions as both a neurohypophysial hormone and central neuromodulatory peptide, and has been implicated in reproductive behaviours, anxiety and reward, as well as facilitation of the neuroendocrine milk-ejection reflex. A potential substrate for oxytocin is the supramammillary nucleus (SuM), a structure that contains oxytocin binding sites and serves as an important relay within the limbic system. Hence, this study investigated the neuromodulatory role of oxytocin within the SuM. Firstly, the effect of oxytocin on neuronal firing within the SuM was studied, using in vitro brain slices from virgin female rats. Oxytocin (10(-6)M) excited approximately 50% of SuM neurones, and similar results were obtained with the selective oxytocin agonist, Thr(4) Gly(7) oxytocin (TGOT) (10(-6) and 10(-7)M). The remaining neurones were unaffected. The TGOT response was blocked by application of the oxytocin antagonist, [d(CH(2))51,Tyr(Me)(2),Thr(4),Orn(8),Tyr-NH29]-vasotocin. Repeat doses of TGOT caused diminution of the response, indicative of desensitisation. In the second series of experiments, immunocytochemical techniques were used to study the oxytocinergic innervation of the SuM. The supramammillary decussation was found to contain numerous oxytocinergic fibres, and some could be seen coursing ventrally to enter the SuM. Whereas, some were clearly "en passant" fibres innervating the neurohypophysis, others followed a more convoluted and branching course, and appeared to terminate within the nucleus. Finally, in vivo microinfusion studies investigated whether oxytocin injected into the SuM facilitated the milk-ejection reflex, a well known action of central oxytocin. Oxytocin microinfusion in the region of the SuM caused a pronounced facilitation of the reflex, contrasting with the much smaller effects of microinfusions made rostral or caudal to the nucleus. Collectively, these results strongly support a neuromodulatory role for oxytocin within the SuM. This could have important implications for understanding the diverse neuroendocrine and behavioural functions of central oxytocin, including its role in reward.

Laminar Localization and Projection-Specific Properties of Presubicular Neurons Targeting the Lateral Mammillary Nucleus, Thalamus, or Medial Entorhinal Cortex.

The presubiculum (PrS) is part of an interconnected network of distributed brain regions where individual neurons signal the animals heading direction. PrS sends axons to medial entorhinal cortex (MEC), it is reciprocally connected with anterior thalamic nuclei (ATNs), and it sends feedback projections to the lateral mammillary nucleus (LMN), involved in generating the head direction signal. The intrinsic properties of projecting neurons will influence the pathway-specific transmission of activity. Here, we used projection-specific labeling of presubicular neurons to identify MEC-, LMN-, and ATN-projecting neurons in mice. MEC-projecting neurons located in superficial layers II/III were mostly regular spiking pyramidal neurons, and we also identified a Martinotti-type GABAergic neuron. The cell bodies of LMN-projecting neurons were located in a well-delimited area in the middle portion of the PrS, which corresponds to layer IV. The physiology of LMN projecting, pyramidal neurons stood out with a tendency to fire in bursts of action potentials (APs) with rapid onset. These properties may be uniquely adapted to reliably transmit visual landmark information with short latency to upstream LMN. Neurons projecting to ATN were located in layers V/VI, and they were mostly regular spiking pyramidal neurons. Unsupervised cluster analysis of intrinsic properties suggested distinct physiological features for the different categories of projection neurons, with some similarities between MEC- and ATN-projecting neurons. Projection-specific subpopulations may serve separate functions in the PrS and may be engaged differently in transmitting head direction related information.

Angular path integration by moving "hill of activity": a spiking neuron model without recurrent excitation of the head-direction system.

During spatial navigation, the head orientation of an animal is encoded internally by neural persistent activity in the head-direction (HD) system. In computational models, such a bell-shaped "hill of activity" is commonly assumed to be generated by recurrent excitation in a continuous attractor network. Recent experimental evidence, however, indicates that HD signal in rodents originates in a reciprocal loop between the lateral mammillary nucleus (LMN) and the dorsal tegmental nucleus (DTN), which is characterized by a paucity of local excitatory axonal collaterals. Moreover, when the animal turns its head to a new direction, the heading information is updated by a time integration of angular head velocity (AHV) signals; the underlying mechanism remains unresolved. To investigate these issues, we built and investigated an LMN-DTN network model that consists of three populations of noisy and spiking neurons coupled by biophysically realistic synapses. We found that a combination of uniform external excitation and recurrent cross-inhibition can give rise to direction-selective persistent activity. The model reproduces the experimentally observed three types of HD tuning curves differentially modulated by AHV and anticipatory firing activity in LMN HD cells. Time integration is assessed by using constant or sinusoidal angular velocity stimuli, as well as naturalistic AHV inputs (from rodent recordings). Furthermore, the internal representation of head direction is shown to be calibrated or reset by strong external cues. We identify microcircuit properties that determine the ability of our model network to subserve time integration function.

The anatomical and computational basis of the rat head-direction cell signal.

As a rat navigates through space, neurons called head-direction (HD) cells provide a signal of the rat's momentary directional heading. Although partly guided by landmarks, the cells also show a remarkable ability to track directional heading based on angular head movement. Theoretical models suggest that the HD cells are linked together to form an attractor network, and that cells which signal angular velocity update the directional setting of the attractor. Recently, cell types similar to those required theoretically have been discovered in the lateral mammillary and dorsal tegmental nuclei. Lesion and anatomical data suggest these nuclei might constitute the postulated attractor-path integration mechanism, and that they provide the HD cell signal to cortical areas where it has been observed.

The effect of descending theta rhythmic input from the septohippocampal system on firing in the supramammillary nucleus.

The supramammillary nucleus (SUM) is part of an ascending pathway conveying behavior-dependent drive to the septal generator of limbic theta rhythm. The SUM is, however, reciprocally connected to the septohippocampal system and there is strong evidence that both septum and SUM are capable of generating theta rhythmic activity. The present study examined the possible role of a descending rhythmic input to the SUM using simultaneously recorded hippocampal EEG and SUM neuronal activity in anesthetized rats. Fourier based phase analysis was performed on recordings in which fast theta rhythmic activity was elicited by tail pinch and in which a slower theta rhythm persisted after cessation of the sensory stimulus. It was found that the firing of a subpopulation of SUM neurons followed the hippocampal theta waves with a constant time delay, rather than a constant phase, suggesting that during deceleration associated with a shift from sensory-elicited theta to spontaneous theta rhythm they followed a descending rhythmic input, most likely from the medial septum. Neurons of a second group, which fired at the hippocampal theta peaks, did not show such relationship demonstrating heterogeneity in the population of rhythmic SUM neurons and their possible roles in theta generation. Combined with previous studies focusing on the role of the ascending theta drive from the SUM, these results demonstrate dynamic bidirectional coupling between subcortical theta generators. Thus, during certain states, rhythmically firing SUM neurons lead the septal theta oscillator, in others the direction may reverse and SUM follows a theta drive of septal origin.

The supragenual nucleus: a putative relay station for ascending vestibular signs to head direction cells.

Head direction (HD) cells located in several regions of the brain, including the postsubiculum, retrosplenial cortex, lateral dorsal thalamic nucleus, anterior dorsal thalamic nucleus, and lateral mammillary nucleus, provide a signal of the rat's momentary directional heading. Experimental evidence suggests that vestibular inputs are critical for the maintenance these cells' directional sensitivity. However, it is still unclear how vestibular information is conveyed to the HD cell-related circuitry. In a recent study, the supragenual nucleus (SG) was suggested as a putative relay of vestibular inputs to this circuitry. In the present study, using anterograde and retrograde tract-tracing methods, we first investigated whether the SG is in a position to convey vestibular inputs. Next, we examined the projections of the SG with the Phaseolus vulgaris leucoagglutinin method. Our results indicate that the SG receives direct inputs from the medial vestibular nucleus and projects to elements of the HD cell-related circuitry, providing a massive input to the contralateral dorsal tegmental nucleus and a moderately dense projection to the shell region of the lateral mammillary nucleus. Overall, the present findings serve to clarify how vestibular inputs reach the HD cell-related circuit and point out the SG as an important interface to this end.

Distribution of urocortin 3 neurons innervating the ventral premammillary nucleus in the rat brain.

Urocortin 3 (Ucn 3) is a recently described peptide of the corticotropin-releasing factor family. Neurons expressing Ucn 3 mRNA and peptide are distributed in specific brain areas, including the median preoptic nucleus, the perifornical area (PFx), and the medial nucleus of the amygdala (MEA). Fibers immunoreactive to Ucn 3 are confined to certain brain nuclei, being particularly dense in the ventral premammillary nucleus (PMV). In studies involving electrolytic lesions and analysis of Fos distribution according to behavioral paradigms, the PMV has been potentially implicated in conspecific aggression and sexual behavior. However, the role that Ucn 3 plays in this pathway has not been explored. Therefore, we investigated the origins of the urocortinergic innervation of the PMV of Wistar rat in an attempt to map the brain circuitry and identify likely related functions. We injected the retrograde tracer cholera toxin b subunit into the PMV and found that 88% of the Ucn 3-immunoreactive fibers in the PMV originate in the dorsal MEA, and that few originate in the PFx. As a control, we injected the anterograde tracer biotin dextran amine into both regions. We observed that the PMV is densely innervated by the MEA, and scarcely innervated by the PFx. The MEA is a secondary relay of the vomeronasal system and projects amply to hypothalamic nuclei related to hormonal and behavioral adjustments, including the PMV. Although physiological studies should also be performed, we hypothesize that Ucn 3 participates in such pathways, conveying sensory information to the PMV, which in turn modulates behavioral and neuroendocrine responses.

Astroglial distribution and sexual differences in neural metabolism in mammillary bodies.

The sexual differences in cerebral nuclei are produced by the organizational and the activational function of gonadal hormones. The different performances by male and female rats in memory tasks requiring use of the mammillary bodies (MBs), could be due to structural and functional sexual dimorphic differences. Our work quantifies the number of glial fibrillary acidic protein immunoreactive (GFAP-IR) astrocytes, and neuronal metabolic activity measured by the cytochrome oxidase (CO) histochemistry in the MBs in rats of both sexes. We find that there is no difference in astroglial number in the medial mammillary nucleus (MMN) and in the lateral mammillary nucleus (LMN) of males, females in estrus and diestrus adult rats. However, we do find statistically significant differences between the sexes in the neuronal oxidative metabolism influenced by the estrous cycle. We, therefore, conclude that there are functional and not structural sex differences in the MBs.

Female odors stimulate CART neurons in the ventral premammillary nucleus of male rats.

Olfactory information is known to influence both male and female sexual behavior. Chemosensory compounds known as pheromones activate distinct brain pathways, inducing innate and stereotyped behaviors, as well as hormonal changes. Studies have shown that female odors induce Fos expression in various brain nuclei of conspecific males, including the ventral premammillary nucleus (PMV). Although poorly investigated, previous studies have suggested that the PMV plays a role in aggressive and sexual behavior. In this study, we used Fos protein expression as a marker for neurons responsive to female odors in sexual inexperienced male rats exposed to soiled bedding. We observed that female odors induced intense Fos immunoreactivity throughout the PMV. Most of these neurons also express cocaine- and amphetamine-regulated transcript (CART) immunoreactivity. In addition, we used in situ hybridization and observed that, following exposure to female odors, CART mRNA increased only in the ventral PMV. Our results suggest that female odors stimulate CART production in the PMV of inexperienced males. Considering that the PMV CART neurons also express the leptin receptor, as well as the fact that they project to areas related to reproduction, we hypothesize that PMV CART neurons integrate nutritional and environmental (olfactory) information, being apt to modulate male reproductive behavior.

How do mammillary body inputs contribute to anterior thalamic function?

It has long been assumed that the main function of the mammillary bodies is to provide a relay for indirect hippocampal inputs to the anterior thalamic nuclei. Such models afford the mammillary bodies no independent role in memory and overlook the importance of their other, non-hippocampal, inputs. This review focuses on recent advances that herald a new understanding of the importance of the mammillary bodies, and their inputs from the limbic midbrain, for anterior thalamic function. It has become apparent that the mammillary bodies' contribution to memory is not dependent on afferents from the subicular complex. Rather, the ventral tegmental nucleus of Gudden is a vital source of inputs that support memory processes within the medial mammillary bodies. In parallel, the lateral mammillary bodies, via their connections with the dorsal tegmental nucleus of Gudden, are critical for generating head-direction signals. These two parallel, but distinct, information streams converge on the anterior thalamic nuclei and support different aspects of spatial memory.

Immunohistochemical screening for antibodies in recent onset type 1 narcolepsy and after H1N1 vaccination.

Narcolepsy type 1 patients typically have undetectable hypocretin-1 levels in the cerebrospinal fluid (CSF), as a result of a selective loss of the hypocretin containing neurons in the hypothalamus. An autoimmune attack targeting hypothalamic hypocretin (orexin) neurons is hypothesised. So far, no direct evidence for an autoimmune attack was found. One of the major limitations of previous studies was that none included patients close to disease onset. We screened serum of 21 narcolepsy type 1 patients close to disease onset (median 11 months), including 8 H1N1 vaccinated patients, for antibodies against hypocretin neurons using immunohistochemistry. No autoantibodies against hypocretin neurons could be detected.

Changes in metabolic activity and estrogen receptors in the human medial mamillary nucleus: relation to sex, aging and Alzheimer's disease.

The medial mamillary nucleus (MMN) is situated caudally in the human hypothalamus and is involved in memory processes. In search for putative sites of action in estrogen replacement therapy on memory both in aging and Alzheimer's disease (AD), we aimed at determining whether changes would occur in estrogen receptors (ER) or metabolic activity in the MMN neurons under these conditions in a sex-dependent way. The Golgi apparatus (GA) and cell size, that were previously shown to be good measures of changes in neuronal metabolic activity, were measured in the MMN of 10 young (20-50 years old), 11 elderly (56-76 years old) control men and women and 11 AD patients (54-78 years old). In addition, we investigated whether estrogen receptor alpha or beta (ERalpha or ERbeta) immunoreactivity was altered in the MMN in aging or AD. There were no sex- or AD-related differences in the GA or cell size in the MMN. Both the GA and cell size of the MMN neurons were found to be increased in postmenopausal compared to young control women accompanied by a decrease in the amount of nuclear ERbeta. The percentage of nuclear ERalpha-positive MMN neurons was markedly enhanced in AD patients compared to controls and most prominently in AD men. In AD patients the proportion of nuclear ERalpha-positive neurons was positively correlated to the Braak stages that indicate the progression of the disease. No differences in the proportion of ERbeta-positive neurons were observed between AD and control patients. We propose that estrogens play an inhibitory role with respect to the metabolic activity of human MMN, which is mediated via ERbeta. This inhibitory effect is diminished in postmenopausal women. The role of the enhanced nuclear ERalpha staining in AD, that was also found in other brain areas, remains to be elucidated.

Ascending projections to the mammillary nuclei in the rat: a study using retrograde and anterograde transport of wheat germ agglutinin conjugated to horseradish peroxidase.

Cells of origin of ascending afferents to the mammillary nuclei and the afferents' fields of termination within these nuclei were studied by using retrograde and anterograde transport of wheat germ agglutinin conjugated to horseradish peroxidase in the rat. The pars compacta of the superior central nucleus projects bilaterally to the median region of the medial mammillary nucleus. The ventral tegmental nucleus projects ipsilaterally to the medial mammillary nucleus, except for its median region, in a topographic manner such that the rostrodorsolateral part of the ventral tegmental nucleus projects to the medial quadrant of the medial mammillary nucleus; the rostroventromedial part projects to the dorsal quadrant; the caudodorsolateral part projects to the ventral quadrant; and the caudoventromedial part projects to the lateral quadrant. These projection fields extend throughout the longitudinal axis of the medial mammillary nucleus, except for its most caudal region, to which only the dorsolateral part of the ventral tegmental nucleus projects. This nucleus also projects topographically to the ipsilateral dorsal premammillary nucleus; the rostral part of the ventral tegmental nucleus projects to the dorsal part of the dorsal premammillary nucleus, whereas the caudal part projects to the ventral part. The periaqueductal gray around the dorsal tegmental nucleus projects bilaterally to the supramammillary nucleus. The pars alpha of the pontine periaqueductal gray projects bilaterally to the peripheral part of the lateral mammillary nucleus, whereas the pars ventralis of the dorsal tegmental nucleus projects ipsilaterally to the lateral mammillary nucleus. The results show that the tegmentomammillary projections are organized in a gradient fashion, with the rostral to caudal position of cells of origin within the tegmental nuclei of Gudden being reflected by the medial to lateral position of fields of termination within the mammillary nuclei.

Retrograde double-labeling study of the mammillothalamic and the mammillotegmental projections in the rat.

Collateral axonal branching from the medial or lateral mammillary nuclei to the anterior thalamus, Gudden's tegmental nuclei, the nucleus reticularis tegmenti pontis, and the medial pontine nucleus was studied using the fluorescent retrograde double-labeling method. One day after injection of Fast Blue into the anterior thalamic nuclei or Gudden's tegmental nuclei, Nuclear Yellow was injected into Gudden's tegmental nuclei or the nucleus reticularis tegmenti pontis and the medial pontine nucleus. Following 1 day survival, single- and double-labeled neurons were examined in the mammillary nuclei. The lateral mammillary nucleus contains neurons whose collateral fibers project to both the dorsal tegmental nucleus of Gudden and the ipsilateral or contralateral anterodorsal thalamic nucleus, to both the medial pontine nucleus and the anterodorsal thalamic nucleus, and to both the dorsal tegmental nucleus of Gudden and the medial pontine nucleus. The pars medianus and pars medialis of the medial mammillary nucleus contain neurons whose collateral fibers project to both the anteromedial thalamic nucleus and the ventral tegmental nucleus of Gudden, to both the anteromedial thalamic nucleus and the medial part of the nucleus reticularis tegmenti pontis, and to both the ventral tegmental nucleus of Gudden and the medial part of the nucleus reticularis tegmenti pontis. The dorsal half of the pars posterior of the medial mammillary nucleus contains a few neurons whose collateral fibers project to both the anteromedial thalamic nucleus and the rostral part of the ventral tegmental nucleus of Gudden, and to both the caudal part of the anteroventral thalamic nucleus and the rostral part of the ventral tegmental nucleus of Gudden, while the pars lateralis of the medial mammillary nucleus contains no double-labeled neurons and projects only to the anteroventral thalamic nucleus.

A dopaminergic projection to the rat mammillary nuclei demonstrated by retrograde transport of wheat germ agglutinin-horseradish peroxidase and tyrosine hydroxylase immunohistochemistry.

The presence and distribution of dopaminergic neurons and terminals in the hypothalamus of the rat were studied by tyrosine hydroxylase (TH) immunohistochemistry. Strongly labelled TH-immunoreactive neurons were seen in the dorsomedial hypothalamic nucleus, periventricular region, zona incerta, arcuate nucleus, and supramammillary nucleus. A few TH-positive neurons were also identified in the dorsal and ventral premammillary nucleus, as well as the lateral hypothalamic area. TH-immunoreactive fibres and terminals were unevenly distributed in the mammillary nuclei; small, weakly labelled terminals were scattered in the medial mammillary nucleus, while large, strongly labelled, varicose terminals were densely concentrated in the internal part of the lateral mammillary nucleus. A few dorsoventrally oriented TH-positive axon bundles were also identified in the lateral mammillary nucleus. A dopaminergic projection to the mammillary nuclei from the supramammillary nucleus and lateral hypothalamic area was identified by double labelling with retrograde transport of wheat germ agglutinin-horseradish peroxidase and TH-immunohistochemistry. The lateral mammillary nucleus receives a weak dopaminergic projection from the medial, and stronger projections from the lateral, caudal supramammillary nucleus. The double-labelled neurons in the lateral supramammillary nucleus appear to encapsulate the caudal end of the mammillary nuclei. The medial mammillary nucleus receives a very light dopaminergic projection from the caudal lateral hypothalamic area. These results suggest that the supramammillary nucleus is the principal source of the dopaminergic input to the mammillary nuclei, establishing a local TH-pathway in the mammillary complex. The supramammillary cell groups are able to modulate the limbic system through its dopaminergic input to the mammillary nuclei as well as through its extensive dopaminergic projection to the lateral septal nucleus.

Laminar organization of the pyramidal cell layer of the subiculum in the rat.

The distribution of neurons in the subiculum of the rat that give rise to subcortical connections was studied using retrograde labeling with horseradish peroxidase conjugated to wheat germ agglutinin. Injections were made into the anteroventral thalamic nucleus, the medial mammillary nucleus, the nucleus accumbens, and the lateral septal nucleus. To facilitate the analysis, the hippocampal formation with adjacent cortices were "flattened," which allowed the cutting of sections perpendicular to the full septotemporal axis. Cells projecting to the anteroventral thalamic nucleus (AV cells), the medial mammillary body (MMB cells), and the nucleus accumbens (ACC cells) were observed consistently throughout the entire septotemporal (dorsoventral) and transverse extent of the subiculum (from field CA1 of the hippocampus to the presubiculum). In the transverse plane, the three kinds of projection cells were arranged in a laminar fashion: The AV cells were observed in the deepest portion of the subicular pyramidal cell layer, the ACC cells were observed in the most superficial portion of the layer, and the MMB cells were observed in the middle portion of the layer. Although this laminar arrangement was observed at all septotemporal levels of the subiculum, it was most apparent at the septal level. At more temporal levels, the "laminae" shifted such that the superficially located ACC cells were more prominent in the proximal half of the subiculum, whereas the AV cells were shifted toward the distal half of the subiculum. The average size of somata of the AV cells was 72.3 microm(2), that of the ACC cells was 105.2 microm(2), and that of the MMB cells was 121.8 microm(2). The connectional and cytoarchitectonic data indicate that there is a distinct sublamination of the subicular pyramidal cell layer, suggesting that the subiculum may be analogous to the infragranular layer (layers V and VI) of the isocortex.