Middle Tufted Cells Research Articles

Vasopressin differentially modulates the excitability of rat olfactory bulb neuron subtypes.

Vasopressin (VP) plays a crucial role in social memory even at the level of the olfactory bulb (OB), where OB VP cells are activated during social interactions. However, it remains unclear how VP modulates olfactory processing to enable enhanced discrimination of very similar odors, e.g., rat body odors. Thus far, it has been shown that VP reduces firing rates in mitral cells (MCs) during odor presentation in vivo and decreases the amplitudes of olfactory nerve-evoked excitatory postsynaptic potentials (ON-evoked EPSPs) in external tufted cells in vitro. We performed whole-cell patch-clamp recordings and population Ca2+ imaging on acute rat OB slices. We recorded ON-evoked EPSPs as well as spontaneous inhibitory postsynaptic currents (IPSCs) from two types of projection neurons: middle tufted cells (mTCs) and MCs. VP bath application reduced the amplitudes of ON-evoked EPSPs and the frequencies of spontaneous IPSCs in mTCs but did not change those in MCs. Therefore, we analyzed ON-evoked EPSPs in inhibitory interneurons, i.e., periglomerular cells (PGCs) and granule cells (GCs), to search for the origin of increased inhibition in mTCs. However, VP did not increase the amplitudes of evoked EPSPs in either type of interneurons. We next performed two-photon population Ca2+ imaging in the glomerular layer and the superficial GC layer of responses to stronger ON stimulation than during patch-clamp experiments that should evoke action potentials in the measured cells. We observed that VP application increased ON-evoked Ca2+ influx in juxtaglomerular cells and GC somata. Thus, our findings indicate inhibition by VP on projection neurons via strong ON input-mediated inhibitory interneuron activity. This neural modulation could improve representation of odors, hence, better discriminability of similar odors, e.g., conspecific body odors.

Parallel odor processing by mitral and middle tufted cells in the olfactory bulb

The olfactory bulb (OB) transforms sensory input into spatially and temporally organized patterns of activity in principal mitral (MC) and middle tufted (mTC) cells. Thus far, the mechanisms underlying odor representations in the OB have been mainly investigated in MCs. However, experimental findings suggest that MC and mTC may encode parallel and complementary odor representations. We have analyzed the functional roles of these pathways by using a morphologically and physiologically realistic three-dimensional model to explore the MC and mTC microcircuits in the glomerular layer and deeper plexiform layer. The model makes several predictions. MCs and mTCs are controlled by similar computations in the glomerular layer but are differentially modulated in deeper layers. The intrinsic properties of mTCs promote their synchronization through a common granule cell input. Finally, the MC and mTC pathways can be coordinated through the deep short-axon cells in providing input to the olfactory cortex. The results suggest how these mechanisms can dynamically select the functional network connectivity to create the overall output of the OB and promote the dynamic synchronization of glomerular units for any given odor stimulus.

Localization of α1-2 Fucose Glycan in the Mouse Olfactory Pathway

Glycoconjugates in the olfactory system play critical roles in neuronal formation, and α1-2 fucose (α1-2Fuc) glycan mediates neurite outgrowth and synaptic plasticity. Histochemical findings of α1-2Fuc glycan in the mouse olfactory system detected using Ulex europaeus agglutinin-I (UEA-I) vary. This study histochemically assessed the main olfactory and vomeronasal pathways in male and female ICR and C57BL/6J mice aged 3-4 months using UEA-I. Ulex europaeus agglutinin-I reacted with most receptor cells arranged mainly at the basal region of the olfactory epithelium. The olfactory nerve layer and glomerular layer of the main olfactory bulb were speckled with positive UEA-I staining, and positive fibers were scattered from the glomerular to the internal plexiform layer. The lateral olfactory tract and rostral migratory stream were also positive for UEA-I. We identified superficial short-axon cells, interneurons of the external plexiform layer, external, middle and internal tufted cells, mitral cells and granule cells as the origins of the UEA-I-positive fibers in the main olfactory bulb. The anterior olfactory nucleus, anterior piriform cortex and olfactory tubercle were negative for UEA-I. Most receptor cells in the vomeronasal epithelium and most glomeruli of the accessory olfactory bulb were positive for UEA-I. Our findings indicated that α1-2Fuc glycan is located within the primary and secondary, but not the ternary, pathways of the main olfactory system, in local circuits of the main olfactory bulb and within the primary, but not secondary, pathway of the vomeronasal system.

Correlated firing in tufted cells of mouse olfactory bulb

Temporally correlated spike discharges are proposed to be important for the coding of olfactory stimuli. In the olfactory bulb, correlated spiking is known in two classes of output neurons, the mitral cells and external tufted cells. We studied a third major class of bulb output neurons, the middle tufted cells, analyzing their bursting and spike timing correlations, and their relation to mitral cells. Using patch-clamp and fluorescent tracing, we recorded spontaneous spiking from tufted-tufted or mitral-tufted cell pairs with visualized dendritic projections in mouse olfactory bulb slices. We found peaks in spike cross-correlograms indicating correlated activity on both fast (peak width 1–50 ms) and slow (peak width>50 ms) time scales, only in pairs with convergent glomerular projections. Coupling appeared tighter in tufted-tufted pairs, which showed correlated firing patterns and smaller mean width and lag of narrow peaks. Some narrow peaks resolved into 2–3 sub-peaks (width 1–12 ms), indicating multiple modes of fast correlation. Slow correlations were related to bursting activity, while fast correlations were independent of slow correlations, occurring in both bursting and non-bursting cells. The AMPA receptor antagonist NBQX (20 μM) failed to abolish broad or narrow peaks in either tufted-tufted or mitral-tufted pairs, and changes of peak height and width in NBQX were not significantly different from spontaneous drift. Thus, AMPA-receptors are not required for fast and slow spike correlations. Electrical coupling was observed in all convergent tufted-tufted and mitral-tufted pairs tested, suggesting a potential role for gap junctions in concerted firing. Glomerulus-specific correlation of spiking offers a useful mechanism for binding the output signals of diverse neurons processing and transmitting different sensory information encoded by common olfactory receptors.

Mitral and tufted cells differ in the decoding manner of odor maps in the rat olfactory bulb.

Mitral and tufted cells in the mammalian olfactory bulb are principal neurons, each type having distinct projection pattern of their dendrites and axons. The morphological difference suggests that mitral and tufted cells are functionally distinct and may process different aspects of olfactory information. To examine this possibility, we recorded odorant-evoked spike responses from mitral and middle tufted cells in the aliphatic acid- and aldehyde-responsive cluster at the dorsomedial part of the rat olfactory bulb. Homologous series of aliphatic acids and aldehydes were used for odorant stimulation. In response to adequate odorants, mitral cells showed spike responses with relatively low firing rates, whereas middle tufted cells responded with higher firing rates. Examination of the molecular receptive range (MRR) indicated that most mitral cells exhibited a robust inhibitory MRR, whereas a majority of middle tufted cells showed no or only a weak inhibitory MRR. In addition, structurally different odorants that activated neighboring clusters inhibited the spike activity of mitral cells, whereas they caused no or only a weak inhibition in the middle tufted cells. Furthermore, responses of mitral cells to an adequate excitatory odorant were greatly inhibited by mixing the odorant with other odorants that activated neighboring glomeruli. In contrast, odorants that activated neighboring glomeruli did not significantly inhibit the responses of middle tufted cells to the adequate excitatory odorant. These results indicate a clear difference between mitral and middle tufted cells in the manner of decoding the glomerular odor maps.

Neurocalcin immunoreactivity in the rat main olfactory bulb

The morphological characteristics and distribution of neurocalcin (NC)-immunoreactive elements were studied in the rat main olfactory bulb (OB) using a polyclonal antibody and the avidin–biotin immunoperoxidase method. NC-positive elements were abundant in the glomerular layer (GL), where numerous immunostained external tufted cells and periglomerular cells were detected. Other less abundant NC-immunolabeled populations included middle and internal tufted cells, Van Gehuchten cells, horizontal cells, vertical cells of Cajal, deep short-axon cells and granule cells. This study demonstrates the presence of NC immunoreactivity in subsets of different neuronal types in the rat main OB. This calcium-binding protein has been found in interneurons, and no evidence of immunoreactivity to NC is detected in projecting neurons. Despite the large population of labeled external tufted cells, most of them belong according to morphological criteria to the local circuit group and some others to those with interbulbar and/or intrabulbar connections. The identification of neuronal subpopulations expressing NC provides a further characterization and shows the existence of biochemical differences within morphologically identical neurons. Thus, this marker may be a useful tool in unravelling the circuitries of the rodent OB in both normal and experimental conditions. The exact physiological function of NC in the olfactory system remains unknown. On the basis of similarities to recoverin, it could be involved in mechanisms responsible for sensory adaptation. Additionally, its calcium-binding abilities may contribute to improve the temporal precision of stimuli transmission, or be concerned with general calcium-related events occurring in specific interneuronal groups.

The structural localization of galanin, and its function in modulating acetylcholine release in the olfactory bulb of adult rat.

The localization of galanin immunoreactivity was analyzed within the olfactory bulb of adult rats. Galanin-positive neurons were differentially distributed among the bulb layers. The density of stained neurons was highest in the glomerular and external plexiform layers. According to morphology, size, location and arrangement, a large proportion of galanin-immunoreactive neurons corresponds to external tufted cells and short-axon neurons in the superficial part of the external plexiform and glomerular layers. A smaller number were middle tufted cells and short-axons neurons while only a few short-axon neurons were labeled in the granule cell layer. Galanin-stained nerve fibers had different structures (thick fibers with or without varicosities, and thin fibers with or without varicosities). Among them were afferent immunoreactive nerve fibers entering the bulb through the olfactory nerve layer, but penetrating superficial layers. Correspondingly, a large number of galanin-positive axons (with or without varicosities) were observed in the olfactory nerve layer. A number of galanin-positive nerve fibers was also present in the glomerular and internal plexiform layers, while these fibers were scarce in the granule cell layer, their density was lowest in the external plexiform layer. These results suggest that galanin-positive axons present in the olfactory bulb originate from at least four different sources. From the periphery axon bundles enter the bulb together with olfactory nerve fibers from the rostral direction and with a fiber bundle from the ventral posterior surface, i.e. at the border between the olfactory tract and the main olfactory bulb along a large blood vessel. Central sources are local interneurons in the olfactory bulb and some extrabulbar brain regions. Double-labeling experiments combining acetylcholinesterase histochemistry with galanin immunocytochemistry did not show any co-localization of acetylcholinesterase and galanin in nerve cell perikarya or nerve fibers. Synthetic porcine galanin (1-29) promoted acetylcholine release in olfactory bulb tissue slices, suggesting that galanin can effectively modulate cholinergic transmission and perhaps other forms of neuronal transmission. It is concluded that galanin may be significantly involved in olfactory processing at cellular and synaptic levels.

NADPH‐diaphorase activity in the olfactory system of the hamster and rat

A comparative analysis of nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase activity in the olfactory bulb was conducted in the hamster and rat. The distribution and morphological features of NADPH-stained neurons were compared to those of glutamic acid decarboxylase-like (GAD-LI) and tyrosine hydroxylase-like (TH-LI) immunoreactive somata in order to relate NADPH-staining to neuronal classes with specific biochemical properties. Intense NADPH-staining was located in primary nerve fibers of the accessory and main olfactory systems, producing dense staining of individual glomeruli. The entire vomeronasal nerve and all glomeruli were stained in the accessory olfactory bulb, but olfactory nerve and glomerular staining were restricted to the dorsal half of the main olfactory bulb. The glomerular layer of the main olfactory bulb of both animals contained numerous small NADPH-stained neurons. The range of somal areas of these neurons was relatively narrow and averaged about 60 microns2 (ca. 8 x 11 microns). Most neurons possessed ovoid somata and monoglomerular intraglomerular dendrites. Previous Golgi studies indicate that such features characterize periglomerular cells. The somal areas of GAD-LI somata in the glomerular layer overlapped that of the NADPH-stained neurons, providing additional evidence that these neurons are probably periglomerular cells. The range of somal areas of TH-LI somata in the glomerular layer was broader and included both small and large neurons that usually possessed intraglomerular dendritic tufts. The smaller TH-LI somata corresponded in size to both the NADPH-stained and GAD-LI somata, suggesting an interrelationship among periglomerular cells, GAD-LI, TH-LI, and NADPH-diaphorase activity. The larger TH-LI somata were probably external tufted cells. In the external plexiform layer of the hamster, oriented NADPH-stained neurons were observed that possessed an intraglomerular dendrite. These neurons appeared to be middle tufted cells. Lightly stained and smaller neurons were occasionally seen in the mitral body and internal plexiform layers, corresponding in somal area and morphological features to those of type III granule cells. No internal tufted or mitral cells were stained. The largest NADPH-stained neurons were located in the inner half of the granule cell layer and were classified as Golgi cells. Their somata averaged 125 microns2 (ca. 10 x 17 microns). Many NADPH-stained neurons were observed in all subdivisions of the anterior olfactory nucleus, the anterior hippocampal rudiment, anterior and posterior levels of the piriform cortex, and the vertical and horizontal limbs of the diagonal band of Broca, all of which are known to provide centrifugal inputs to the olfactory bulb.(ABSTRACT TRUNCATED AT 400 WORDS)

Substance Pimmunoreactivfty in the superficial laminae of the hamster olfactory bulb

The direct application of colchicine to the hamster olfactory bulb has engendered the appearance of a broader distribution of substance P-like immunoreactivity (SP-LI) than has been previously reported. In the glomerular layer, both external tufted and periglomerular cells were strongly immunopositive. These two classes could be identified by somal size and the presence and branching pattern of intraglomerular dendrites. SP-LI neurons were also identified in the external plexiform layer and many were similar to middle tufted cells. These findings demonstrate that substance P expression is not uniquely associated with classes of projection neurons, but is also found in periglomerular cells, a class of inhibitory local circuit neurons.

Structure of the olfactory bulb of the hedgehog (Erinaceus europaeus): A Golgi study of the intrinsic organization of the superficial layers

The intrinsic organization of the mitral cell, external plexiform, and glomerular layers of the main olfactory bulb of the insectivore hedgehog were studied with the Golgi method. This study completes our previous description of the cell types in the granule cell layer in the same subject (López-Mascaraque et al., J. Comp. Neurol. 253:135-152, '86). In the present contribution the morphology of mitral, internal, and middle tufted cells is described with particular interest in the formation and arrangement of glomerular tufts by the primary dendrites, and the disposition of the secondary dendrites. Three types of intrinsic cells were found in the external plexiform layer: Van Gehuchten cells, satellite cells, and horizontal cells. All these cells display unusual patterns of branching processes that were difficult to classify as dendritic or axonal. The close relationship between some processes of the satellite cells with the mitral or tufted cell dendrites is noted, suggesting that these cells may be inhibitory in nature. The external tufted cells are described, and several aspects pertaining to their morphology were considered in order to distinguish them from periglomerular cells and from the remaining tufted cells. External tufted and periglomerular cells appear to be intrinsic neurons, having axons distributed in the periglomerular region, most probably devoted to relating different olfactory glomeruli. The arrangement of glomeruli in the glomerular layer and distinctive characteristics with respect to other mammals were considered from a comparative point of view.

Processing of Thyrotropin-Releasing Hormone (TRH) Prohormone in the Rat Olfactory Bulb Generates Novel TRH-Related Peptides*

Based on the deduced amino acid sequence of rat TRH prohormone (pro-TRH), proteolytic processing of this polyprotein precursor is expected to produce, beside TRH, several other novel peptides. These peptides should correspond to connecting segments flanking the repeated TRH progenitor sequence and to various C- and/or N-terminally extended forms of TRH. The profile of the endogenous products of the TRH system was studied in rat brain using multiple RIAs coupled to molecular sieve filtration and HPLC separations. In extracts from the rat hypothalamus, TRH and two pro-TRH-connecting peptides, prepro-TRH-(160-169) and prepro-TRH-(178-199) were detected in molar ratios corresponding to those expected for a nearly complete processing of the prohormone molecule. In the olfactory bulb, pro-TRH is processed differently, since peptides containing TRH at their N-termini, [pGlu172] prepro-TRH-(172-199) and [pGlu154]prepro-TRH-(154-169), were found to be major end products along with prepro-TRH-(160-169) and prepro-TRH-(178-199). The dissimilarity in tissue content suggests that differential processing of TRH precursor by various enzymatic pathways may act as a regulating mechanism for TRH and TRH-related activities. The cellular localization of C-terminally extended forms of TRH in rat olfactory bulb was examined by the indirect immunoperoxidase method, using antisera directed against prepro-TRH-(160-169) and pre-pro-TRH-(178-199). Cell bodies and nerve fibers were detected in the glomerular and external plexiform layers of the main olfactory bulb. The presence of extended forms of TRH in interneurons and middle tufted cells of the main olfactory bulb suggests that in light of the recent biological properties described for prepro-TRH-(160-169), these peptides may act as neuromodulators for olfactory epithelium inputs or neurotransmitters within more rostrally located olfactory areas in the forebrain.

Cytochrome oxidase staining marks dendritic zones of the rat olfactory bulb external plexiform layer.

Cytochrome oxidase staining of the rat olfactory bulb external plexiform layer (EPL) produces a darkly stained intermediate zone bordered by lightly stained superficial and deep zones. Similar zonal staining was seen in cats, rabbits, and hamsters. These zones vary in relative thickness around the circumference of the olfactory bulb; the deep zone is proportionally thicker in the most dorsal and ventral parts of the bulb. Tufted cell somata are unevenly distributed within the EPL; the outer part of the EPL has more somata. The distribution of the cytochrome oxidase reaction product shows that the darkly stained intermediate zone is not produced by staining of tufted cell somata. Zones of cytochrome oxidase staining correspond to the sublaminar distribution of mitral and tufted cell basal dendrites. This was demonstrated by labeling mitral and tufted cells with small extracellular horseradish peroxidase injections and processing alternate sections for horseradish peroxidase and for cytochrome oxidase. Because there was cross-reaction of the cytochrome oxidase procedure with horseradish peroxidase, it was possible to trace many neurons through both series of sections. Middle tufted cells of the superficial EPL have basal dendrites confined to the superficial zone of light cytochrome oxidase staining. Internal tufted cells and middle tufted cells of the intermediate zone send their basal dendrites into the intermediate zone. One group of mitral cells (type I) has basal dendrites confined to the deep zone of lighter cytochrome oxidase staining. A second group of mitral cells (type II) and tufted cells of the intermediate EPL has basal dendrites primarily confined to the intermediate zone of dark cytochrome oxidase staining. The correlation of the enzyme staining with the dendritic laminar patterns supports the existence of three relatively distinct sublaminae in the EPL and supports the designation of two types of mitral cells. The staining pattern also provides an independent method for evaluating the sublaminae of the EPL without the necessity of labeling individual groups of cells. Finally, the staining pattern suggests that the intermediate zone of the EPL may be subjected to more tonic synaptic input, causing it to have an increased level of metabolic activity.

Distribution of cholecystokinin-like immunoreactivity in the rat main olfactory bulb.

The anatomical localization of cholecystokinin-like immunoreactivity (CCK-I) within the rat main olfactory bulb was analyzed by using the peroxidase-antiperoxidase immunocytochemical technique. Neurons or neuronal processes containing CCK-I were localized within all laminae of the olfactory bulb except the olfactory nerve fiber layer. A large population of CCK-I neurons, with morphology, size, and distribution corresponding to that of the middle and external tufted cells, was observed within a zone extending from the deep periglomerular region through the superficial one-half to one-third of the external plexiform layer. A smaller number of immunoreactive perikarya were found in the deep external plexiform layer, the glomerular layer, and rarely within the inner plexiform layer. These CCK-I neurons appeared to correspond to internal tufted cells, periglomerular cells, and deep short-axon cells, respectively. Dense CCK-I staining of fibers and terminals was present within the internal plexiform layer and, less densely, within the neuropil of the granule cell layer. In addition, terminal-like CCK-I was localized within layer 1A of the anterior olfactory nucleus, the olfactory tubercle, and the most rostral piriform cortex. This observation provides corroboration for the identification of the principal CCK-I neuron in the rat olfactory bulb as the centrally projecting middle tufted cell. The present results, demonstrating the localization of CCK-I to both local circuit and projection neurons of the olfactory bulb and to terminal-like puncta in the internal plexiform and granule cell layers, suggest that CCK may be significantly involved in olfactory processing at several levels.

Distribution of local axon collaterals of mitral, displaced mitral, and tufted cells in the rabbit olfactory bulb.

To determine the projection fields of intrabulbar axon collaterals, mitral, displaced mitral, and middle tufted cells in the rabbit olfactory bulb were stained by intracellular injection of HRP. The axon collaterals of mitral cells and displaced mitral cells were distributed exclusively within the granule cell layer (GrL). Those of middle tufted cells were distributed mostly in the GrL and on rare occasions in the mitral cell layer. None of these collaterals entered the external plexiform layer. Axon collaterals of mitral cells, emitted at the depths of the GrL, were distributed widely from the deep portion to the most superficial portion of the GrL. Collaterals of displaced mitral cells were also emitted in the deep part, but they tended to be distributed more superficially in the GrL than mitral cells. Collaterals of middle tufted cells were released and distributed superficially in the GrL. The axon collaterals of these principal cells typically extended for a longer distance than the secondary dendrites, and they sometimes formed bushlike terminal arborizations. The results indicate that the projection fields of the axon collaterals of principal cells are spatially separated from the dendritic projection fields. This suggests that the output of these principal cells through the collaterals has a functionally different role from the output through the dendrites. The observation that the three types of principal cells differ in the distribution pattern of their axon collaterals in the GrL suggests that there is a functional separation of the sublayers in the GrL.

Distribution of dendrites of mitral, displaced mitral, tufted, and granule cells in the rabbit olfactory bulb.

To determine the dendritic fields, mitral, displaced mitral, middle tufted, and granule cells in the rabbit olfactory bulb were stained by intracellular injection of HRP. The secondary dendrites of mitral cells were distributed mostly in the inner half of the external plexiform layer (EPL). Those of displaced mitral cells extended mainly into the middle and superficial sublayers in the EPL. The secondary dendrites of middle tufted cells were distributed mostly in the superficial portion of the EPL. Mitral cells extended their secondary dendrites in virtually all directions within a plane tangential to the mitral cell layer (MCL) and thus had a disklike projection field with a radius of about 850 microns. Displaced mitral cells had similar dendritic projection fields in the tangential plane but with somewhat distorted shapes. The secondary dendrites of middle tufted cells had a tendency to extend in particular directions. From the projection pattern of the gemmules on the peripheral processes, granule cells were classified into three types. Type I granule cells had gemmules both in the superficial and in the deep sublayers of the EPL. The peripheral processes of Type II granule cells were confined to the deep half of the EPL. The gemmules of Type III granule cells ere distributed in the superficial half of the EPL. The differing dendritic ramification among mitral, displaced mitral, and middle tufted cells suggests the separation of the dendrodendritic synaptic interactions with granule cells in different sublayers in the EPL. It also suggests a functional separation of the sublayers of the EPL.

Tyrosine hydroxylase immunoreactive neurons and fibers in the olfactory system of the hamster

AbstractTyrosine hydroxylaselike immunoreactivity is associated with several different classes of neurons in the main olfactory bulb of the hamster. Most of the tyrosine hydroxylaselike immunoreactive (THLI) neurons are located in the glomerular layer and in the outer two‐thirds of the external plexiform layer. The majority of THLI neurons in the glomerular layer have somal sizes and dendritic features which correspond to those of external tufted cells. A small population of THLI periglomerular cells also is present. The majority of THLI neurons in the external plexiform layer have the morphological characteristics of middle tufted cells. A small population of THLI neurons which appear to be Van Gehuchten cells is distributed throughout the external plexiform layer and in the mitral body layer. Small populations of THLI deep short‐axon cells are distributed throughout the mitral body, internal plexiform, and granule cell layers. Tyrosine hydroxylase‐positive fibers of central origin also are observed in the main olfactory bulb. They may originate from THLI neurons in the anterior olfactory nucleus, ventral hippocampal rudiment, dorsal peduncular cortex, septal and basal portions of the vertical limb and nucleus of the horizontal limb of the diagonal band, lateral and dorsal hypothalamus, ventral tegmental area and substantia nigra, dorsal and median raphe nuclei, and locus ceruleus. The frontal neocortex, infralimbic and anterior cingulate cortex, and medial amygdaloid nucleus also contain THLI neurons.In contrast to the widespread distribution of tyrosine hydroxylaselike immunoreactivity in the main olfactory bulb, substance P‐like immunoreactivity is restricted to external tufted cells and to centrifugal afferents which may originate in the dorsal and median raphe nuclei. Both types of immunoreactivity appear to be present in centrifugal afferents to the accessory olfactory bulb, but THLI neurons are extremely rare and no neurons with substance P‐like immunoreactivity have been observed in the accessory olfactory bulb. Intraventricular injections of colchicine which were expected to enhance the labeling of neuronal somata did not alter the relative incidences of labeling among the various classes of neurons in the olfactory bulbs.These findings provide immunocytochemical evidence that populations of superficially situated tufted cells in the main olfactory bulb are functionally distinct from the deeper‐lying output neurons and from output neurons in the accessory olfactory bulb, that populations of catecholami‐nergic neurons are present within several olfactory and related cortical structures in addition to the olfactory bulbs, that the medical septum‐diagonal band complex also contains catecholaminergic neurons, and that a substantial population of neurons in the dorsal raphe nucleus is capable of vatechol synthesis.

Electron microscopy of single cells in the olfactory bulb using Golgi impregnation.

A technique for studying the same cell stained by Golgi impregnation with both light and electron microscopy is described. The application of the method, termed Golgi-E.M., to the rat olfactory bulb and its contribution to a wider organizational study of this region is considered. While providing important confirmation of many of the intercellular relationships found by examination of conventional electron microscopic material, it has also allowed certain special features of middle tufted cells to be noted, including reciprocal, axo-dendritic, dendro-axonic synaptic contacts. The advantages and drawbacks of the technique are briefly stated and the role to which it is best suited in organizational study is set out; it is clear that, while unsuitable at present for routine study, this material can provide both important correlations of light and electron microscopy and certain specific types of information, in conjunction with more conventional methods.

Autoradiographic study of histogenesis in the mouse olfactory bulb. I. Time of origin of neurons and neuroglia.

AbstractTime of origin (final cell division) of neurons and neuroglia of the mouse olfactory and accessory olfactory formations was determined by autoradiography. Animals were injected with thymidine‐H3 at various developmental stages and killed at or near maturity. In the olfactory formation mitral cells (the largest neurons) arise first, mainly over the three day period from the eleventh day of gestation (E11) to E13, tufted cells chiefly from E13 to E18, and granule cells (the smallest neurons) mainly from E18 to postnatal day 20. Most of the smaller and more superficial peripheral tufted cells arise later than the deeper and larger middle and internal tufted cells. All three types of granule cells have a time of origin extending well into postnatal life, with internal granule cells arising over a longer and later period than periglomerular cells or granule cells of the mitral cell layer. Neuroglial precursors undergo final cell division chiefly between E17 and P10. In the phylogenetically less evolved accessory olfactory formation, mitral cells originate earlier than their homologues in the olfactory formation; mitral cells principally from E10 to E12 and granule cells chiefly from E12 to E18. The results support the concept that some germinal layers of the central nervous system are programmed to produce a succession of cell types, larger cells before smaller ones.