Posteromedial Cortical Nucleus Research Articles

Amygdala Corticofugal Input Shapes Mitral Cell Responses in the Accessory Olfactory Bulb.

Interconnections between the olfactory bulb and the amygdala are a major pathway for triggering strong behavioral responses to a variety of odorants. However, while this broad mapping has been established, the patterns of amygdala feedback connectivity and the influence on olfactory circuitry remain unknown. Here, using a combination of neuronal tracing approaches, we dissect the connectivity of a cortical amygdala [posteromedial cortical nucleus (PmCo)] feedback circuit innervating the mouse accessory olfactory bulb. Optogenetic activation of PmCo feedback mainly results in feedforward mitral cell (MC) inhibition through direct excitation of GABAergic granule cells. In addition, LED-driven activity of corticofugal afferents increases the gain of MC responses to olfactory nerve stimulation. Thus, through corticofugal pathways, the PmCo likely regulates primary olfactory and social odor processing.

The vomeronasal cortex – afferent and efferent projections of the posteromedial cortical nucleus of the amygdala in mice

Most mammals possess a vomeronasal system that detects predominantly chemical signals of biological relevance. Vomeronasal information is relayed to the accessory olfactory bulb (AOB), whose unique cortical target is the posteromedial cortical nucleus of the amygdala. This cortical structure should therefore be considered the primary vomeronasal cortex. In the present work, we describe the afferent and efferent connections of the posteromedial cortical nucleus of the amygdala in female mice, using anterograde (biotinylated dextranamines) and retrograde (Fluorogold) tracers, and zinc selenite as a tracer specific for zinc-enriched (putative glutamatergic) projections. The results show that the posteromedial cortical nucleus of the amygdala is strongly interconnected not only with the rest of the vomeronasal system (AOB and its target structures in the amygdala), but also with the olfactory system (piriform cortex, olfactory-recipient nuclei of the amygdala and entorhinal cortex). Therefore, the posteromedial cortical nucleus of the amygdala probably integrates olfactory and vomeronasal information. In addition, the posteromedial cortical nucleus of the amygdala shows moderate interconnections with the associative (basomedial) amygdala and with the ventral hippocampus, which may be involved in emotional and spatial learning (respectively) induced by chemical signals. Finally, the posteromedial cortical nucleus of the amygdala gives rise to zinc-enriched projections to the ventrolateral septum and the ventromedial striatum (including the medial islands of Calleja). This pattern of intracortical connections (with the olfactory cortex and hippocampus, mainly) and cortico-striatal excitatory projections (with the olfactory tubercle and septum) is consistent with its proposed nature as the primary vomeronasal cortex.

Orexinergic innervation of the extended amygdala and basal ganglia in the rat

The orexinergic system interacts with several functional states of emotions, stress, hunger, wakefulness and behavioral arousal through four pathways originating in the lateral hypothalamus (LH). Hundreds of orexinergic efferents have been described by tracing studies and direct immunohistochemistry of orexin in the forebrain, olfactory regions, hippocampus, amygdala, septum, basal ganglia, thalamus, hypothalamus, brain stem and spinal cord. Most of these tracing studies investigated the whole orexinergic projection to all regions of the intracranial part of the CNS. To identify the orexinergic efferents at the subnuclear level of resolution, we focussed on the orexinergic target in the amygdala, which is substantially involved in the LH output and contributes mostly to the functional outcome of the orexinergic system and the basal ganglia. Immunohistochemical identification of axonal orexin A and orexin B in male adult rats has been performed on serial sections. In the extended amygdala many new orexinergic targets were found in the anterior amygdaloid area (dense), anterior cortical nucleus (moderate), amygdalostriatal transition region (moderate), basolateral regions (moderate), basomedial nucleus (moderate), several bed nucleus of the stria terminals regions (few to dense), central amygdaloid subdivisions (dense), posteromedial cortical nucleus (moderate) and medial amygdaloid subnuclei (dense). Furthermore, the entopeduncular nucleus has been newly identified as another target for orexinergic fibers with a high density. These results suggest that subdivisions and subnuclei of the extended amygdala are specific targets of the orexinergic system.

Oxytocin injected into the ventral subiculum or the posteromedial cortical nucleus of the amygdala induces penile erection and increases extracellular dopamine levels in the nucleus accumbens of male rats

Oxytocin (20-100 ng) was found to be able to induce penile erection when injected unilaterally into the ventral subiculum or the posteromedial cortical nucleus of the amygdala of male rats. The pro-erectile effect started mostly 30 min after treatment and was abolished by the prior injection of d(CH(2))(5)Tyr(Me)(2)-Orn(8)-vasotocin (1-2 microg), an oxytocin receptor antagonist, into the ventral subiculum or posteromedial cortical nucleus. Oxytocin-induced penile erection occurred 15 min after the increase in the concentration of extracellular dopamine and its metabolite 3,4-dihydroxyphenylacetic acid in the dialysate obtained from the nucleus accumbens, which was also abolished by d(CH(2))(5)Tyr(Me)(2)-Orn(8)-vasotocin. The pro-erectile effect of oxytocin was also reduced by cis-flupentixol (2 and 5 microg), a dopamine receptor antagonist, injected into the nucleus accumbens, and by (+)MK-801 (5 microg), a noncompetitive N-methyl-d-aspartate receptor antagonist, injected into the ventral tegmental area, but not into the nucleus accumbens. Together with studies showing that glutamatergic efferents from the ventral subiculum/posteromedial cortical nucleus of the amygdala to other areas of the limbic system modulate the activity of mesolimbic dopaminergic neurons, these findings suggest that oxytocin injected into these areas increases glutamatergic neurotransmission in the ventral tegmental area. This, in turn, activates mesolimbic dopaminergic neurons, leading to penile erection. These results provide evidence that the ventral subiculum and the posteromedial cortical nucleus of the amygdala participate in a neural circuit that controls not only the consummatory aspects of sexual behaviour (e.g. penile erection and copulatory performance), but also its motivational/reward aspects, confirming a key role of oxytocin and dopamine in these processes.

A centrifugal pathway to the mouse accessory olfactory bulb from the medial amygdala conveys gender‐specific volatile pheromonal signals

We previously found that female mice exhibited Fos responses in the accessory olfactory bulb (AOB) after exposure to volatile opposite-sex, but not same-sex, urinary odours. This effect was eliminated by lesioning the main olfactory epithelium, raising the possibility that the AOB receives information about gender via centrifugal inputs originating in the main olfactory system instead of from the vomeronasal organ. We asked which main olfactory forebrain targets send axonal projections to the AOB, and whether these input neurons express Fos in response to opposite-sex urinary volatiles. Female mice received bilateral injections of the retrograde tracer cholera toxin B (CTB), into the AOB, and were exposed to either same- or opposite-sex volatile urinary odours 1 week later. We found CTB-labeled cell bodies in several forebrain sites including the bed nucleus of the accessory olfactory tract, the rostral portion of the medial amygdala (MeA) and the posteromedial cortical nucleus of the amygdala. A significant increase in the percentage of CTB/Fos co-labeled cells was seen only in the MeA of female subjects exposed to male but not to female urinary volatiles. In Experiment 2, CTB-injected females were later exposed to volatile odours from male mouse urine, food, or cat urine. Again, a significant increase in the percentage of CTB/Fos co-labeled cells was seen in the MeA of females exposed to male mouse urinary volatiles but not to food or predator odours. Main olfactory-MeA-AOB signaling may motivate approach behaviour to opposite-sex pheromonal signals that ensure successful reproduction.

Species‐specific chemosignals evoke delayed excitation of the vomeronasal amygdala in freely‐moving female rats

Male rat chemosignals attract females and influence their reproductive status. Through the accessory olfactory bulb and its projection target, the posteromedial cortical nucleus of the amygdala (PMCo), species-specific chemosignals detected by the vomeronasal organ (VNO) may reach the hypothalamus. To test this hypothesis in vivo, behavioural activation and neurotransmitter release in the PMCo were simultaneously monitored in freely moving female oestrus rats exposed to either rat or mouse urinary stimuli, or to odorants. Plasma levels of the luteinizing hormone were subsequently monitored. All stimuli induced an immediate behavioural activation, but only species-specific chemosignals led to a delayed behavioural activation. This biphasic behavioural activation was accompanied by a VNO-mediated release of the excitatory amino acids, aspartate and glutamate, in the PMCo. The late behavioural and neurochemical activation was followed by an increase in the levels of circulating luteinizing hormone. In conclusion, these data show that only species-specific chemosignals induce a delayed behavioural activation and excitatory activation of the PMCo, which is dependent on an intact VNO.

Retrograde labelling of mitral/tufted cells in the mouse accessory olfactory bulb following local injections of the lipophillic tracer DiI into the vomeronasal amygdala

It has recently become apparent that there are two classes of vomeronasal receptor neurons that project to functionally separate anterior and posterior sub-regions of the mammalian accessory olfactory bulb. However, anterograde tracing of the projections from these sub-regions, in the mouse, has revealed that the processing pathways are not segregated at the level of the vomeronasal amygdala. Both sub-regions have overlapping projections to the superficial lamina of the medial and posterior medial cortical nuclei of the amygdala. However, differential projections have been found in the opossum, in which only the posterior sub-region projects to the deeper laminae of the medial amygdala. Therefore, there may be species differences in these projections that are important for the control of reproductive behaviour. This study used an alternative approach of retrogradely tracing mitral/tufted cell projections from different nuclei of the vomeronasal amygdala back to the accessory olfactory bulb of mice. Local injections of the lipophillic tracer DiI were made into the antero-dorsal and postero-ventral divisions of the medial amygdala, and into the postero-medial cortical amygdala. In each case, provided the DiI affected the superficial lamina Ia, labelled mitral/tufted cells were found distributed throughout the anterior-posterior extent of the accessory olfactory bulb. These results confirm that mitral/tufted cells of the anterior and posterior sub-regions of the accessory olfactory bulb project to both the medial and postero-medial cortical nuclei of the amygdala. There was no evidence for differential projections from the anterior and posterior sub-regions accessory olfactory bulb in mice, as has been reported to occur in other species.

Estradiol masculinizes the posteromedial cortical nucleus of the amygdala in the rat

It has been demonstrated that the posteromedial cortical amygdaloid nucleus (PMCo), is sexually dimorphic. It is shown (Experiment 1) that male orchidectomy on the day of birth (D1) decreases the volume and number of neurons of the PMCo, while a single injection of propionate testosterone to the female on D1 masculinizes the PMCo in this gender. Since male gonadectomy on D1 (Experiment 2) is counteracted by a single injection of estradiol benzoate in males it has been suggested that the masculinization of the PMCo is due to the aromatization of testosterone to estradiol in this structure. These findings support the hypothesis that the development of sex differences in structures that belong to the vomeronasal system are due to the aromatization of testosterone to estradiol shortly after birth.

Zinc-containing afferent projections to the rat corticomedial amygdaloid complex: a retrograde tracing study.

The mammalian amygdaloid complex is densely innervated by zinc-containing neurons. The distribution of the terminals throughout the region has been described, but the origins of these zinc-containing fibers have not. The present work describes the origins of one major component of the zinc-containing innervation of the amygdaloid complex, namely, the component that innervates the corticomedial complex. Selective labeling of zinc-containing axons was accomplished by intracerebral microinfusion of selenium anions (SeO3(2-)), a procedure that produces a ZnSe precipitate in zinc-containing axonal boutons with subsequent retrograde transport to the neurons of origin. After infusions of SeO3(2-) into combinations of cortical, medial, or amygdalohippocampal regions, retrogradely labeled zinc-containing somata were found in all amygdaloid nuclei except for the medial and central nuclei, the bed nucleus of the accessory olfactory tract, the nucleus of the lateral olfactory tract, and the anterior amygdaloid area. Extrinsic zinc-containing projections to the same amygdaloid terminal fields were found to originate from the infralimbic, cingulate, piriform, perirhinal and entorhinal cortices, and from the prosubiculum and CA1. Commissural zinc-containing projections were found to originate from the posterolateral and posteromedial cortical nuclei and from the posterior part of the basomedial nucleus. Zinc-containing neurons have been implicated in the pathophysiology of epilepsy, in cell death after seizure or stroke, and in Alzheimer's disease, all clinical conditions that involve the amygdaloid complex. Identification of the zinc-containing pathways is a prerequisite to the elucidation of zinc's role in these disorders.

Maternal exposure to Δ 9-tetrahydrocannabinol facilitates morphine self-administration behavior and changes regional binding to central μ opioid receptors in adult offspring female rats

Opiates and cannabinoids are among the most widely consumed habit-forming drugs in humans. Several studies have demonstrated the existence of interactions between both kind of drugs in a variety of effects and experimental models. The present study has been focused to determine whether perinatal Δ 9-tetrahydrocannabinol (Δ 9-THC) exposure affects the susceptibility to reinforcing effects of morphine in adulthood and whether these potential changes were accompanied by variations in μ opioid receptor binding in brain regions related to drug reinforcement. Adult female rats born from mothers that were daily treated with Δ 9-THC during gestation and lactation periods, exhibited a statistically significant increase in the rate of acquisition of intravenous morphine self-administration behavior when compared with females born from vehicle-exposed mothers, an effect that did not exist in Δ 9-THC-exposed male offspring. This increase was significantly greater on the last day of acquisition period. There were not significant differences when the subjects were lever pressing for food. In parallel, we have also examined the density of μ opioid receptors in the brain of adult male and female offspring that were exposed to Δ 9-THC during the perinatal period. Collectively, perinatal exposure to Δ 9-THC produced changes in μ opioid receptor binding that differed regionally and that were mostly different as a function of sex. Thus, Δ 9-THC-exposed males exhibited a lower density for these receptors than their respective oil-exposed controls in the caudate-putamen area as well as in the amygdala (posteromedial cortical nucleus). On the contrary, Δ 9-THC-exposed females exhibited higher density of these receptors than their respective oil-exposed controls in the prefrontal cortex, the hippocampus (CA3 area), the amygdala (posteromedial cortical nucleus), the ventral tegmental area and the periaqueductal grey matter, whereas the binding was lower than control females only in the lateral amygdala. These results support the notion that perinatal Δ 9-THC exposure alters the susceptibility to morphine reinforcing effects in adult female offspring, in parallel with changes in μ opioid receptor binding in several brain regions.

Central 5-HT 3 receptors in P and in AA alcohol-preferring rats: An autoradiographic study

Considerable evidence exists for an involvement of serotonergic mechanisms in the control of alcohol consumption. In the present study, an extensive 5-hydroxytryptamine (5-HT 3) receptor autoradiographical investigation was performed using two genetically selected rat strains, alcohol preferring (P) and Alko alcohol (AA) alcohol-preferring rats, as well as the corresponding alcohol nonpreferring (NP) and Alko nonalcohol (ANA) alcohol-nonpreferring rats. The aim was to determine if there are any differences in 5-HT 3 binding levels that may illuminate mechanisms of alcohol preference in these animals. For quantitating 5-HT 3 binding sites, [ 3H]S(−)zacopride (0.5 nM) was used. Non-specific binding was measured in the presence of granisetron 10 −6 M. The [ 3H]S(−)zacopride binding density was measured in two subregions of the amygdaloid nucleus, frontal cortex, piriform cortex, cingulate laminae, parietal anterior cortex, parietal medial cortex, hippocampus CA1, hippocampus CA3, and entorhinal cortex. In all the brain areas investigated, the results showed no differences between AA and ANA rats. In P rats, compared to NP controls, there was a 30% lower 5-HT 3 binding level in the lateral nucleus and the posteromedial cortical nucleus of the amygdala. These findings suggest that the expression of high alcohol preference in genetically selected P and AA rats is not associated with a general alteration of central 5-HT 3 receptors, although a lower 5-HT 3 receptor level in the amygdala of P rats may contribute to the phenotype of this strain of animals.

Central Connections of the Ovine Olfactory Bulb Formation Identified Using Wheat Germ Agglutinin-Conjugated Horseradish Peroxidase

Pheromonal stimuli elicit rapid behavioral and reproductive endocrine changes in the ewe. The neural pathways responsible for these effects in sheep are unknown, in part, because the olfactory bulb projections have not been examined in this species. Using the anterograde and retrograde neuronal tracer, wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP), we describe the afferent and efferent olfactory bulb connections of the Suffolk ewe. Injections of WGA-HRP limited to the main olfactory bulb resulted in retrograde labeling of cells in numerous telencephalic, diencephalic, and metencephalic regions. Terminal labeling was limited to layer Ia of ipsilateral cortical structures extending rostrally from the anterior olfactory nucleus (AON), piriform cortex, anterior-, and posterolateral-cortical amygdaloid nuclei to lateral entorhinal cortex caudally. Injections involving the accessory olfactory bulb and AON produced additional labeling of cells within the bed nucleus of the stria terminalis (BNST), medial nucleus of the amygdala, and a few cells in the posteromedial cortical nucleus of the amygdala. Terminal labeling included a small dorsomedial quadrant of BNST and also extended to the far lateral portions of the supraoptic nucleus. A clearly defined accessory olfactory tract and nucleus was not evident, perhaps due to limitations in the sensitivity of the method. With this possible exception, the afferent and efferent olfactory connections in the sheep appear similar to those reported for other species.

Distribution of neurons of origin of zinc-containing projections in the amygdala of the rat

The present study describes the distribution of neurons of origin of zinc-containing pathways in the amygdaloid complex of the rat, using the selenium method for simultaneous retrograde labeling of all zinc-containing neurons. With this method, vesicular ionic zinc is precipitated intravitally with selenium compounds and transported retrogradely to the parent neurons, where it can be visualized by silver amplification. Neurons labeled retrogradely with silver-amplified precipitate were observed in all amygdaloid nuclei except for the lateral olfactory tract nucleus, the accessory olfactory tract nucleus and the central nucleus. Very few labeled cell bodies were seen in the anterior amygdaloid area and the medial nucleus. The amygdalo-hippocampal area and the amygdalo-piriform transition area both showed a substantial number of labeled somata throughout their rostrocaudal extent. In the anterior cortical nucleus, very few labeled cell bodies were found in the rostral pole, whereas they were abundant in the caudal quarter of the nucleus. In the posterolateral cortical nucleus, the number of labeled cell bodies increased gradually; there were none in the rostral pole, but most of the neurons in the caudal part were labeled. The posteromedial cortical nucleus contained a great number of labeled somata, but with some variation in the rostrocaudal extent of the nucleus. Considerable numbers of labeled neurons were observed throughout the lateral nucleus. In the basolateral nucleus, a small number of labeled cell bodies was present in the rostral half, but a gradual increase was observed in the caudal direction. Finally, in the basomedial nucleus, very few labeled cell bodies were present in the rostral two-thirds, whilst a considerable number was encountered in the caudal one-third. Possible functional implications of neuronal zinc are considered. The distribution of neurons of origin of zinc-containing projections has been compared with previously described intrinsic connections of the rat amygdala, and tracts that may possibly be zinc-containing are outlined and discussed. It is concluded that in all probability a substantial proportion of the intrinsic connectivity of the rat amygdaloid complex is zinc-containing.

An iodinated vasopressin (V 1) antagonist blocks flank marking and selectively labels neural binding sites in golden hamsters

An arginine-vasopressin (AVP) derivative, [d(CH 2) 5,Sar 7]AVP (SAVP), has been characterized as an antagonist to vasopressin V 1 receptors. Using AVP-dependent flank-marking behavior as a bioassay, it was possible to verify that iodinated SAVP (I-SAVP) retains biological activity within the central nervous system, as the antagonist blocked the behavior. Furthermore, 125I-SAVP was used to localize specific V 1 binding sites in the brain. The resulting binding was localized to discrete anatomical sites, and highly specific to V 1-like receptors. While we confirmed previous findings using 3H-AVP in golden hamsters, we also identified binding in many areas previously unreported (e.g., arcuate and paraventricular nuclei of the hypothalamus, tenia tecta, posteromedial cortical nucleus of the amygdala, and zona incerta), suggesting that 125I-SAVP provides a greater level of resolution. In addition, specific binding was observed in the lateral septum, anterior hypothalamus, and midbrain central gray, areas that have previously been shown to trigger flank marking in response to AVP microinjection. The presence of AVP binding sites in limbic and mesencephalic areas involved in the regulation of flank marking suggests that this neuropeptide may play an important role as a neurotransmitter at multiple levels in the neural circuits controlling this behavior.

Convergence of main and accessory olfactory pathways onto single neurons in the hamster amygdala.

Chemoreceptor pathways from the vomeronasal organ (VNO), and main olfactory system are known to be separate as they pass into the brain, at least until the level of the amygdala. In the amygdala, vomeronasal pathways project to the posteromedial cortical nucleus (PMCN), and medial nucleus (MN). The main olfactory pathways have terminations in the posterolateral cortical nucleus (PLCN), and anterior cortical nucleus (ACN), both of which project to the PMCN and MN. The anatomy thus suggests that the PMCN and MN are sites for convergence of input from the main and accessory olfactory pathways. We have recorded single units in the amygdala and found that electrical stimulation of either the main olfactory bulbs or the VNO could drive some of the same units in the PMCN. Units were also found that were driven by one system but not the other, and units in which activity driven by one system was suppressed by stimulation of the other system.

Vomeronasal and olfactory pathways to the amygdala controlling male hamster sexual behavior: Autoradiographic and behavioral analyses

Previous studies suggest that the rostral corticomedial amygdala (CMA), particularly the medial nucleus, is an important site where vomeronasal and olfactory stimuli critical to male hamster copulatory behavior are processed. To test the possibility that mating deficits seen after lesions of the rostrally-placed medial nucleus may be due to the interruption of chemosensory afferents to more caudal areas, we injected tritiated amino acids into the accessory and main olfactory bulbs of male hamsters in which we had first produced bilateral electrolytic lesions or sham lesions in either the rostral CMA or basolateral amygdala, and then observed mating behavior. Autoradiographic analysis of ‘vomeronasal’ projections from the accessory olfactory bulb and ‘olfactory’ projections from the main bulb, revealed that rostral CMA lesions which damaged the medial nucleus and extended to the ventral surface of the brain (ventral lesions) interrupted vomeronasal input to the more caudally-placed posteromedial cortical nucleus, but spared olfactory inputs to adjacent caidal areas of the amygdala and piriform lobe. In contrast, lesions which damaged a major portion of the medial nucleus but left its ventral surface intact (dorsal lesions) spared both vomeronasal and olfactory inputs to more caudal areas. Animals with both dorsal and ventral lesions failed to mate posteperatively, whereas animals bearing sham lesions of basolateral amygdaloid lesions, which, like dorsal lesions, spared caudally-directed chemosensory afferents, continued to mate normally. We conclude that mating deficits seen after rostral CMA lesions are due primarily to destruction of the medial nucleus.

Connections of the corticomedial amygdala in the golden hamster. II. Efferents of the "olfactory amygdala".

The anterior cortical (C1) and posterolateral cortical (C2) nuclei of the amygdala are designated the "olfactory amygdala" because they each receive direct projections from the main olfactory bulb. The efferents of these nuclei were traced after stereotaxic placement of 1-5 muCi tritiated proline in the corticomedial amygdala of the male golden hamsters. Following survival times of 12, 24, or 48 hours, 20 micron frozen sections of the brains were processed for light microscopic autoradiography. Efferents from C2 terminate in layers II and III of the olfactory tubercle and in layer Ib of pars ventralis and pars medialis of the anterior olfactory nucleus. Fibers from this nucleus also project to layers I and II of the infralimbic cortex and to the molecular layer of the agranular insular cortex. More posteriorly, fibers from C2 terminate in layer I of the dorsolateral entorhinal cortex, and in the endopiriform nucleus. From C1, efferent fibers travel in the stria terminalis and terminate in the precommissural bed nucleus of the stria terminalis and in the mediobasal hypothalamus. Efferents from C1 also innervate the molecular layer of C2, the amygdalo-hippocampal area, and the adjacent piriform cortex. Neurons in both C1 and C2 project to the molecular layer of the medial amygdaloid nucleus and the posteromedial cortical nucleus of the amygdala, the plexiform layer of the ventral subiculum, and the molecular layer of the lateral entorhinal cortex.

Quantitative 3H-thymidine radiographic analyses of neurogenesis in the rat amygdala.

AbstractNeurogenesis in the rat amygdala was examined with 3H‐thymidine radiography. The animals were the offspring of pregnant females given two injections of 3H‐thymidine on consecutive days in an overlapping series: Embryonic day (E) 12 + E13, + E14. E21 + E22. On 60 days of age, the percentage of labelled cells and the proportion of cells added during each day of formation were determined at several anatomical levels within the following components of the amygdala: anterior amygdaloid area, bed nuclei of the lateral and accessory olfactory tracts, central, medial, anterior cortical, posterolateral cortical, posteromedial cortical, basomedial, basolateral, and lateral nuclei, the amygdalo‐hippocampal area, and the intercalated masses.All large and many small neurons originate in most nuclei between E13 and E17, those in the intercalated masses between E15 and E19, those in the amygdalo‐hippocampal area between E16 and E19. The anterior amygdaloid area, intercalated masses, central, medial, posterolateral cortical, posteromedial cortical, basomedial, basolateral, and lateral nuclei have strong rostral‐to‐caudal intranuclear gradients. There are five additional intranuclear gradients: 1) medial to lateral in the central nucleus, anterior amygdaloid area, and anterior intercalated masses; 2) lateral to medial in the bed nucleus of the lateral olfactory tract and basolateral nucleus; 3) superficial to deep in the amygdalo‐hippocampal area, posterolateral, and posteromedial cortical nuclei; 4) ventral to dorsal in the medial nucleus; and 5) dorsal to ventral between the small and large‐celled parts of the lateral nucleus. Only the bed nucleus of the accessory olfactory tract and the anterior cortical nucleus do not have intranuclear gradients. Between 10 and 15% of the total cell population in most nuclei are very small neurons and/or glia which originate simultaneously between E18 and E20. This population is absent in the ventral part of the medial, anterior cortical, and anterior basomedial nuclei; these contiguous areas may form a distinct subunit in the amygdala.In contrast to the pronounced intranuclear gradient, internuclear gradients are weak. Neurogenesis in the central nucleus and corticomedial and basolateral complexes appears to take place both concurrently and independently. There are groups of early‐originating neurons in the central, medial, and basolateral nuclei located near the periphery of the amygdala. Each of these groups is surrounded by younger neurons farther within the interior. The youngest calls are in the centrally placed intercalated masses. These settling patterns suggest that cells in the amygdala arise simultaneously from more than one neuroepithelial source during morphogenesis.The chronology of neurogenesis in the amygdala can be related to some of its anatomical connections. Rostral‐to‐caudal gradients in the corticomedial complex may be timed to coincide with early vs. late arrival of olfactory fibers. The subdivisions bases on intranuclear gradients in the central, medial, and basolateral nuclei match the subdivisions based on patterns of anatomical connections.

Olfactory relationships of the telencephalon and diencephalon in the rabbit. I. An autoradiographic study of the efferent connections of the main and accessory olfactory bulbs.

The efferent connections of the main and accessory olfactory bulbs in the female albino rabbit have been studied using the autoradiographic method for tracing axonal pathways. Following unilateral injections of 3H-proline or 3H-leucine into the main olfactory bulb, radioactively labeled material transported intraaxonally by axoplasmic flow in an anterograde direction from soma to axon terminal is present ipsilaterally in the superficial half of the plexiform layer (IA) of: the entire circumference of the olfactory peduncle, the tenia tecta, the full mediolateral extent of the olfactory tubercle, the entire length of the prepyriform cortex, a transition area between the prepyriform cortex and the horizontal limb of the nucleus of the diagonal band, the nucleus of the lateral olfactory tract, the anterior cortical and posterolateral cortical amygdaloid nuclei (periamygdaloid areas 1, rostral half of 2, 5 of Rose, '31), and the ventrolateral entorhinal cortex (entorhinal areas 1, 2, 4, 5, 7 of Rose, '31). No subcortical or contralateral projection of main bulb efferents was found. After a unilateral injection of 3H-leucine into the accessory olfactory bulb, transported material could be followed caudally along the dorsal surface of the ipsilateral lateral olfactory tract. This heavily labeled projection is distinct from the unlabeled lateral olfactory tract and has been termed the accessory olfactory tract. Beginning at the level of the caudal third of the olfactory tubercle and extending caudally to the nucleus of the lateral olfactory tract is a group of small neurons intimately associated with the accessory olfactory tract. This cell group is referred to as the bed nucleus of the accessory olfactory tract. Projection sites of the accessory bulb include the bed nucleus of the accessory olfactory tract and layer IA of the medial nucleus and the posteromedial cortical nucleus of the amygdala (periamygdaloid areas 3, 4, PAM, caudal half of 2, 6 of Rose, '31). An additional accessory bulb efferent projection was found to enter the stria terminalis at the level of the medial amygdaloid nucleus and could be traced to a posterior segment of the bed nucleus of the stria terminalis. The autoradiographic findings indicate that the accessory olfactory bulb connects with portions of the amygdala that do not receive afferent input from the main olfactory bulb and provide evidence for the existence of two distinct and separate olfactory systems.