Medial Division Research Articles

Zonal distribution of Purkinje cells in the zebrafish cerebellum: analysis by means of a specific monoclonal antibody.

We have isolated a monoclonal antibody that recognizes a 42-kDa protein from adult zebrafish brain. The antibody stains the typical drop-shaped perikaryon of Purkinje cells and their dendrites. The cerebellum of teleosts has complex features. It is composed of three parts; the valvula cerebelli (Va), the corpus cerebelli (CCe), and the crista cerebellaris (CC). In higher vertebrates, the molecular layer is always found as the most outer layer of the cerebellum, but in teleosts, some of the granular cells are located on the surface of the Va. In higher vertebrates, the boundary between the granular and molecular layers always contains Purkinje cells, but this does not occur in teleosts. The Purkinje cells are found only in a part of the boundary in Va. We have found that the layer containing Purkinje cells forms a continuous zone in the cerebellum in the zebrafish. The complex structure of the cerebellum is more easily understood with the aid of the concept of a "Purkinje zone". The Purkinje zone starts at the caudal end of Val (lateral division of Va), turns at the edge of Va toward Vam (medial division of Va), connects to CCe, and ends at the bottom of CCe. The dendrites are found only on one side of the zone. The dendrites of the Purkinje cells in Vam are planar and are packed regularly, similar to those of higher vertebrates. However, the dendrites in Val and the posterior part of CCe are not planar and are irregularly packed.

Cadherin expression by embryonic divisions and derived gray matter structures in the telencephalon of the chicken

The expression of three cadherins (cadherin-6B, cadherin-7, and R-cadherin) was studied by immunohistochemistry in the telencephalon of chicken embryos at intermediate stages of development (11 and 15 days of incubation). Expression patterns were related to cytoarchitecture and to previously published data on functional connections and on the expression of gene regulatory proteins. Our results indicate that, like in other regions of the embryonic chicken brain, the expression of each cadherin is restricted to parts of embryonic divisions as well as to particular nuclei, areas or their subdivisions. The expression patterns are largely complementary with partial overlap. The regional expression of the cadherins respects the boundary between the pallium and the subpallium as well as between various pallial and subpallial subdivisions. Novel subdivisions were found in several telencephalic areas. For example, subjacent to the hyperstriatum, the neostriatum contains multiple islands of cells with a profile of cadherin expression that differs from the surrounding matrix ("island fields"). Moreover, the expression of each cadherin is apparently associated with parts of intratelencephalic neural circuits and of thalamopallial and basal ganglia pathways. These results support a role for cadherins in the aggregation and differentiation of gray matter structures within embryonic brain divisions. The cadherin immunostaining patterns are interpreted in the context of a recently proposed divisional scheme of the avian pallium that postulates medial, dorsal, lateral, and ventral divisions as complete radial histogenetic units (Puelles et al. [2000]).

The extrusion-capture model for chromosome partitioning in bacteria.

Successful cellular reproduction requires accurate duplication and partitioning (segregation) of the genome. Failure to correctly partition the sister genomes results in aneuploidy. The consequences of these errors range from loss of normal cellular function (e.g., the loss of normal growth controls in tumor cells) to cell death. In prokaryotes with a single chromosome, partitioning failures are fatal for at least one of the two daughter cells; a so-called anucleate cell forms when one daughter receives no chromosome and the other daughter receives two chromosomes. Several findings in recent years have fundamentally altered our view of chromosome partitioning in prokaryotes (for review, see Gerdes et al. 2000; Gordon and Wright 2000; Hiraga 2000; Moller-Jensen et al. 2000; Donachie 2001; Sawitzke and Austin 2001). The flurry of new observations was ignited by adaptation of cell biological techniques used in eukaryotes (immunofluorescence, GFP, and fluorescent in situ hybridization) for use in prokaryotes (Harry et al. 1995; Pogliano et al. 1995; Webb et al. 1995; Niki and Hiraga 1998). This review focuses on chromosome partitioning in bacteria that have a single circular chromosome, specifically, the gram-positive organism, Bacillus subtilis, and the gramnegative organisms, Escherichia coli and Caulobacter crescentus. In these bacteria, DNA replication initiates once per cell division cycle from a specific chromosomal locus, oriC, and proceeds bidirectionally to terminate in a defined region opposite the origin, terC (Fig. 1). The basic components of the DNA replication machinery are highly conserved in bacteria (Kornberg and Baker 1992); in fact, these basic components are functionally conserved from bacteria to mammals (Baker and Bell 1998). Fundamental differences exist between chromosome partitioning in eukaryotes and prokaryotes. In eukaryotes, chromosomes are duplicated in S phase, and sister chromosomes remain together during G2. Chromosome partitioning occurs during M phase, when sister chromosomes are lined up on a metaphase plate, separated from each other, and finally segregated in opposite directions by the combined action of the microtubular spindle and mitotic motors. In contrast to the temporal separation of chromosome replication and partitioning in eukaryotes, regions of the bacterial chromosome, starting with the origin, appear to be partitioned soon after duplication, whereas the remainder of the chromosome awaits replication (Glaser et al. 1997; Gordon et al. 1997; Lewis and Errington 1997; Lin et al. 1997; Mohl and Gober 1997; Webb et al. 1997, 1998; Niki and Hiraga 1998; Sharpe and Errington 1998; Teleman et al. 1998; Jensen and Shapiro 1999; Niki et al. 2000). Thus, in bacteria, DNA replication, chromosome refolding, and chromosome partitioning are concurrent. Given these differences, it is not surprising that there is no evidence that bacteria contain eukaryotic-like mitotic spindles or mitotic motors. As discussed below, it appears that bacteria with circular chromosomes probably power chromosome partitioning differently from eukaryotes. This may be possible because of the much smaller distances that chromosomes move in bacteria. Many bacteria, including B. subtilis and E. coli, are capable of dividing in one-half to one-third of the time it takes to duplicate the genome. To accomplish this, new rounds of DNA replication are initiated before a previous round is completed, giving the replication cycle a head start on the division cycle. This results in cells with multiple bidirectional DNA replication forks (so-called multifork replication), multiple copies of oriC (2, 4, or 8), but only a single, unduplicated terminus region (Fig. 2). Soon after duplication, sister origins are each partitioned in opposite directions. Thus, during multifork replication, bacteria contain positional information regarding not only the current medial division site, but also the future division planes (the cell quarters and cell eighths). Other bacteria, for exampleC. crescentus, are not known to be capable of multifork replication and do not have more than two copies of the origin per cell. Recent studies have revealed common mechanisms involved in chromosome partitioning, along with differences in some of the details between organisms. This review summarizes the following recent findings that have contributed new insights into the mechanism by which bacteria accomplish accurate chromosome partitioning. First, the bacterial chromosome has a defined orientation within cells, and individual regions move 3Corresponding author. E-MAIL adg@mit.edu; FAX (617) 253-2643. Article and publication are at http://www.genesdev.org/cgi/doi/10.1101/ gad.913301.

Temporal frequency of whisker movement. I. Representations in brain stem and thalamus.

How does processing of information change the internal representations used in subsequent stages of sensory pathways? To approach this question, we studied the representations of whisker movements in the lemniscal and paralemniscal pathways of the rat vibrissal system. We recently suggested that these two pathways encode movement frequency in different ways. We proposed that paralemniscal thalamocortical circuits, functioning as phase-locked loops (PLLs), translate temporally coded information into a rate code. Here we focus on the two major trigeminal nuclei of the brain stem, nucleus principalis and subnucleus interpolaris, and on their thalamic targets, the ventral posteromedial nucleus (VPM) and the medial division of the posterior nucleus (POm). This is the first study in which these brain stem and thalamic nuclei were explored together in the same animals and using the same stimuli. We studied both single- and multi-unit activity. We moved the whiskers both mechanically and by air puffs; here we present air-puff-induced movements because they are more similar to natural movements than movements induced by mechanical stimulations. We describe the basic properties of the responses in these brain stem and thalamic nuclei. The responses in both brain stem nuclei were similar; responses to air puffs were mostly tonic and followed the trajectory of whisker movement. The responses in the two thalamic nuclei were similar during low-frequency stimulations or during the first pulses of high-frequency stimulations, exhibiting more phasic responses than those of brain stem neurons. However, with frequencies >2 Hz, VPM and POm responses differed, generating different representations of the stimulus frequency. In the VPM, response amplitudes (instantaneous firing rates) and spike counts (total number of spikes per stimulus cycle) decreased as a function of the frequency. In the POm, latencies increased and spike count decreased as a function of the frequency. Having described the basic response properties in the four nuclei, we then focus on a specific test of our PLL hypothesis for coding in the paralemniscal pathway. We used short-duration air puffs, much shorter than whisker movements during natural whisking. The activity in this situation was consistent with the prediction we made on the basis of the PLL hypothesis.

Projections from the nociceptive area of the central nucleus of the amygdala to the forebrain: a PHA-L study in the rat.

The lateral capsular division (CeLC) of the central nucleus (Ce) of the amygdala, in the rat, has been shown to be the main terminal area of a spino(trigemino)-parabrachio-amygdaloid nociceptive pathway [Bernard & Besson (1990) J. Neurophysiol. 63, 473-490; Bernard et al. (1992) J. Neurophysiol. 68, 551-569; Bernard et al. (1993) J. Comp. Neurol. 329, 201-229]. The projections to the forebrain from the CeLC and adjacent regions were studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin (PHA-L) restricted in subdivisions of the Ce and the basolateral amygdaloid nucleus anterior (BLA). Our data showed that the entire CeLC projects primarily and extensively to the substantia innominata dorsalis (SId). The terminal labelling is especially dense in the caudal aspect of the SId. The other projections of the CeLC in the forebrain were dramatically less dense. They terminate in the bed nucleus of the stria terminalis (BST) and the posterior hypothalamus (pLH). No (or only scarce) other projections were found in the remaining forebrain areas. The Ce lateral division (CeL) and the Ce medial division (CeM), adjacent to the CeLC, also project to the SId with slightly lower density labelling. However, contrary to the case of the CeLC, both the CeL and the CeM extensively project to the ventrolateral subnucleus of the BST (BSTvl) with a few additional terminals found in other regions of the lateral BST. Only the CeM projects densely to both the interstitial nucleus of the posterior limb of the anterior commissure and the caudal most portion of the pLH. The projections of the BLA are totally different from those of the Ce as they terminate in the dorsal striatum, the accumbens nucleus, the olfactory tubercle, the nucleus of olfactory tract and the rostral pole of the cingulate/frontal cortex. This study demonstrates that the major output of the nociceptive spino(trigemino)-parabrachio-CeLC pathway is to the SId. It is suggested that the CeLC-SId pathway could have an important role in anxiety, aversion and genesis of fear in response to noxious stimuli.

Basic organization of projections from the oval and fusiform nuclei of the bed nuclei of the stria terminalis in adult rat brain.

The organization of axonal projections from the oval and fusiform nuclei of the bed nuclei of the stria terminalis (BST) was characterized with the Phaseolus vulgaris-leucoagglutinin (PHAL) anterograde tracing method in adult male rats. Within the BST, the oval nucleus (BSTov) projects very densely to the fusiform nucleus (BSTfu) and also innervates the caudal anterolateral area, anterodorsal area, rhomboid nucleus, and subcommissural zone. Outside the BST, its heaviest inputs are to the caudal substantia innominata and adjacent central amygdalar nucleus, retrorubral area, and lateral parabrachial nucleus. It generates moderate inputs to the caudal nucleus accumbens, parasubthalamic nucleus, and medial and ventrolateral divisions of the periaqueductal gray, and it sends a light input to the anterior parvicellular part of the hypothalamic paraventricular nucleus and nucleus of the solitary tract. The BSTfu displays a much more complex projection pattern. Within the BST, it densely innervates the anterodorsal area, dorsomedial nucleus, and caudal anterolateral area, and it moderately innervates the BSTov, subcommissural zone, and rhomboid nucleus. Outside the BST, the BSTfu provides dense inputs to the nucleus accumbens, caudal substantia innominata and central amygdalar nucleus, thalamic paraventricular nucleus, hypothalamic paraventricular and periventricular nuclei, hypothalamic dorsomedial nucleus, perifornical lateral hypothalamic area, and lateral tegmental nucleus. Moderately dense inputs are found in the parastrial, tuberal, dorsal raphé, and parabrachial nuclei and in the retrorubral area, ventrolateral division of the periaqueductal gray, and pontine central gray. Light projections end in the olfactory tubercle, lateral septal nucleus, posterior basolateral amygdalar nucleus, supramammillary nucleus, and nucleus of the solitary tract. These and other results suggest that the BSTov and BSTfu are basal telencephalic parts of a circuit that coordinates autonomic, neuroendocrine, and ingestive behavioral responses during stress.

Medial Geniculate, Amygdalar and Cingulate Cortical Training-Induced Neuronal Activity during Discriminative Avoidance Learning in Rabbits with Auditory Cortical Lesions

This study addressed the neural mediation of discriminative avoidance learning, wherein rabbits step in a wheel apparatus in response to an acoustic conditional stimulus, the CS+, to avoid a foot shock, and they learn to ignore a different stimulus, the CS-, not followed by foot shock. Previously, muscimol-induced inactivation of the amygdala in the first session of training prevented learning during the inactivation and permanently blocked the development of discriminative training-induced neuronal activity (TIA) in the medial division of the medial geniculate nucleus (MGm). These results suggested that amygdalar neurons induce discriminative TIA in the MGm via basolateral (BL) amygdalar axonal projections to the auditory cortex. To test this hypothesis, the activity of neurons in the MGm was recorded during learning in rabbits with lesions of the auditory cortex. Recordings were also made in the lateral and BL amygdalar nuclei and in the cingulate cortex. In support of the hypothesis, discriminative learning in rabbits with lesions was impaired significantly during early training sessions 1-4; in these same sessions, discriminative TIA was abolished in the MGm, the BL nucleus, and the anterior cingulate cortex. The lesions also blocked posterior cingulate cortical discriminative TIA in training sessions 1-2 but spared TIA in sessions 3-7. Lateral amygdalar neurons showed gradual development of discrimination that was not significantly affected by the lesions. The results demonstrate a critical role of auditory cortex in early discriminative learning and in the production of early discriminative TIA in multiple areas.

Cardiovascular responses to subseptic doses of endotoxin contribute to differential neuronal activation in rat brain

The contribution of cardiovascular activity in the early central responses to systemic inflammation was assessed in rats following intravenous administration of subseptic doses of lipopolysaccharide (LPS). LPS at 12.5 μg/kg increased heart rate (HR) but did not alter mean arterial pressure (MAP), and induced interleukin-1β (IL-1β) gene expression at 1 h in circumventricular organs (CVOs), choroid plexus, meninges, blood vessels, and pituitary gland. IL-1β mRNA levels were attenuated at 2 h in most regions studied. LPS at 50 μg/kg caused a biphasic change in MAP, increased HR, increased levels of arginine vasopressin heteronuclear RNA in the hypothalamic paraventricular nucleus (PVN), and induced IL-1β gene expression in the nucleus of the solitary tract (NTS) at 1 h. LPS (both doses) induced Fos-like immunoreactivity (FLI) in the area postrema, organum vasculosum of the lamina terminalis, NTS, preoptic area, supraoptic nucleus, and PVN at 1 h. In the PVN, neurons with FLI were found primarily in the dorsal and dorsal medial parvocellular divisions after 12.5 μg/kg of LPS whereas neurons with FLI were found throughout the PVN after 50 μg/kg of LPS. After 2 h, FLI was widespread throughout the brain. Plasma ACTH levels were elevated at 1 and 2 h in response to both doses of LPS, and levels of CRF mRNA were increased after 2 h in the parvocellular PVN. Our results reveal that central responses to increasing doses of LPS show different patterns which are related to activation of distinct immune and viscerosensory pathways, and that cardiovascular responses contribute to early neuronal activation as LPS concentrations are increased.

Projections from the amygdalo-piriform transition area to the amygdaloid complex: a PHA-l study in rat.

The amygdalo-piriform transition area is a poorly defined region in the temporal lobe that is heavily connected with the olfactory system. As part of an ongoing project aimed at understanding the neuronal pathways that provide sensory information to the amygdala, we investigated the cytoarchitectonic and chemoarchitectonic features of the amygdalo-piriform transition area and its connections to the amygdaloid complex in 13 rats by using the anterograde tracer, Phaseolus vulgaris-leucoagglutinin. Our analysis indicates that the amygdalo-piriform transition area has medial (rostral and caudal portions) and lateral parts. The rostromedial part projects heavily to the intermediate and lateral divisions of the central nucleus, whereas the caudomedial part projects mainly to the medial division. The lateral part of the amygdalo-piriform transition area projects heavily to the capsular and lateral divisions of the central nucleus. Electron microscopic analysis revealed that the projection to the lateral division of the central nucleus forms asymmetric contacts with the spines and shafts of postsynaptic neurons and, therefore, is assumed to be excitatory. The amygdalo-piriform transition area also projects moderately to other amygdaloid nuclei, including the parvicellular division of the basal nucleus, the anterior cortical nucleus, and the nucleus of the lateral olfactory tract. The lateral and medial parts of the amygdalo-piriform transition area also project to the distal temporal CA1 and distal temporal subiculum, respectively. Unlike the adjacent entorhinal cortex, the amygdalo-piriform transition area does not project to the dentate gyrus. These data suggest that the amygdalo-piriform transition area is a region that influences both emotional and memory processing in parallel by means of pathways to the amygdala and the hippocampus, respectively.

The Amygdala Is Essential for the Development of Neuronal Plasticity in the Medial Geniculate Nucleus during Auditory Fear Conditioning in Rats

The medial geniculate nucleus of the thalamus (MGN) and the basolateral complex of the amygdala (BLA) are critical components of the neural circuit that mediates auditory fear conditioning. Several studies indicate that neurons in both the MGN and BLA exhibit associative plasticity of spike firing during auditory fear conditioning. In the present study, we examined whether the development of plasticity in the MGN requires the BLA. Single units were recorded from chronic multichannel electrodes implanted in the medial division of the MGN of conscious and freely moving rats. Rats received auditory fear conditioning trials, which consisted of a white-noise conditional stimulus (CS) and a co-terminating footshock unconditional stimulus (US). Unpaired (sensitization) controls received the same number of trials as paired animals, but the CS and US were explicitly unpaired. Before fear conditioning, rats received either an intra-amygdala infusion of muscimol, a GABA(A) receptor agonist, to inactivate BLA neurons or an infusion of the saline vehicle. Auditory fear conditioning produced a substantial increase in both CS-elicited spike firing in the MGN and conditional freezing behavior in vehicle-treated rats receiving paired training. Muscimol inactivation of the BLA severely attenuated the development of both conditioning-related increases in CS-elicited spike firing in the MGN and conditional freezing to the auditory CS. Unpaired training did not yield increases in either CS-elicited spike firing or freezing to the tone CS. These results reveal that the BLA is essential to the development of plasticity in the auditory thalamus during fear conditioning.

Glycogen phosphorylase reactivity in the amygdala and bed nucleus of the stria terminalis

The present study examines the reactivity of the glial metabolic enzyme, glycogen phosphorylase, within the amygdala and bed nucleus of the stria terminalis. Reactivity for phosphorylase a, the active form of glycogen phosphorylase, was higher in all parts of the medial amygdaloid nucleus, in the medial division of the central amygdaloid nucelus, in the anterior amygdaloid area and in the bed nucleus of stria terminalis than in all parts of the lateral amygdaloid nucleus, the anterior cortical amygdaloid nucleus, the posteromedial and posterolateral cortical amygdaloid nuclei, the intercalated nucleus of the amygdala, main part and the intercalated nuclei. A greater degree of phosphorylase a reactivity was also observed in the basolateral amygdaloid nucleus, anterior and posterior parts, and in the basomedial amygdaloid nucleus, anterior part, while other parts of these nuclei were less reactive. Reactivity attributed to total glycogen phosphorylase enzyme, phosphorylase a+phosphorylase b activated by AMP, was higher and homogeneous across the amygdala. Phosphorylase a patterns are likely to reflect differences in the contribution of glycogenolysis to the metabolic support of cells in the amygdala and bed nucleus of the stria terminalis. Possible relationships to local neuronal activity and to differences in glycogenolytic neuromodulatory input are discussed.

Interconnectivity between the amygdaloid complex and the amygdalostriatal transition area: a PHA-L study in rat.

The amygdala orchestrates the formation of behavioral responses to emotionally arousing stimuli. Many of these responses are initiated by the central nucleus, which converges information from other amygdaloid nuclei. Recently, we observed substantial projections from the amygdala to the amygdalostriatal transition area, which is located dorsal to the central nucleus. These projections led us to question whether the amygdalostriatal transition area has a role in the initiation of behavioral responses in emotionally arousing circumstances. To explore this anatomically, we traced the interconnections between the amygdalostriatal transition area and the amygdaloid complex using the anterograde tracer Phaseolus vulgaris-leucoagglutinin. The lateral (the medial division and the caudal portion of the dorsolateral division) and the accessory basal nuclei (the parvicellular division) provide moderate-to-heavy projections to the amygdalostriatal transition area. Projections back to the amygdala are light and are composed of thin, faintly stained varicose fibers that resemble the labeling of cholinergic terminals. The extra-amygdaloid outputs of the amygdalostriatal transition area are sparse and include moderate projections to the caudoventral globus pallidus, the ansa lenticularis, and the substantia nigra pars lateralis. These data suggest that the amygdalostriatal transition area is one of the major targets for projections originating in the lateral and accessory basal nuclei of the amygdala. Via these pathways, emotionally significant stimuli can evoke behavioral responses that are different from those initiated via projections from the amygdala to the central nucleus. One such candidate response is the orienting response (i.e., saccadic eye movements and head direction) in a pathway that includes a projection from the lateral/accessory basal nucleus of the amygdala to the amygdalostriatal transition area, and from there to the substantia nigra, pars lateralis.

Interconnectivity between the amygdaloid complex and the amygdalostriatal transition area: A PHA-L study in rat

The amygdala orchestrates the formation of behavioral responses to emotionally arousing stimuli. Many of these responses are initiated by the central nucleus, which converges information from other amygdaloid nuclei. Recently, we observed substantial projections from the amygdala to the amygdalostriatal transition area, which is located dorsal to the central nucleus. These projections led us to question whether the amygdalostriatal transition area has a role in the initiation of behavioral responses in emotionally arousing circumstances. To explore this anatomically, we traced the interconnections between the amygdalostriatal transition area and the amygdaloid complex using the anterograde tracer Phaseolus vulgaris-leucoagglutinin. The lateral (the medial division and the caudal portion of the dorsolateral division) and the accessory basal nuclei (the parvicellular division) provide moderate-to-heavy projections to the amygdalostriatal transition area. Projections back to the amygdala are light and are composed of thin, faintly stained varicose fibers that resemble the labeling of cholinergic terminals. The extra-amygdaloid outputs of the amygdalostriatal transition area are sparse and include moderate projections to the caudoventral globus pallidus, the ansa lenticularis, and the substantia nigra pars lateralis. These data suggest that the amygdalostriatal transition area is one of the major targets for projections originating in the lateral and accessory basal nuclei of the amygdala. Via these pathways, emotionally significant stimuli can evoke behavioral responses that are different from those initiated via projections from the amygdala to the central nucleus. One such candidate response is the orienting response (i.e., saccadic eye movements and head direction) in a pathway that includes a projection from the lateral/accessory basal nucleus of the amygdala to the amygdalostriatal transition area, and from there to the substantia nigra, pars lateralis.

Secretion of Bax-Like Protein into Systemic Circulation from the Posterior Pituitary in the Rat Hypothalamo-Posterior Pituitary System.

To investigate the possibility of the neurosecretion of Bax-like protein in the adult rat, we examined its distribution in the hypothalamo-posterior pituitary system using light and electron microscopic immunohistochemistry. We also measured the serum Bax-like protein using an enzyme immunoassay. Neurons in the paraventricular nucleus (PVN) and supraoptic nucleus of the hypothalamus exhibited intense Bax-like immunoreactivity (Bax-IR). Bax-like immunoreactive axons projected to the posterior pituitary through the internal layer of the median eminence. By immunoelectron microscopy, Bax-IR was observed in the granulated vesicles and in the membrane of the roughsurfaced endoplasmic reticulum in the neuronal perikarya in the medial magnocellular division of PVN. It was also detected in the secretory vesicles in the nerve terminals of the posterior lobe of the pituitary gland. Double-labeling histochemistry revealed that almost all Bax-like immunoreactive substances were present in the oxytocin neurons, and many Bax-like immunoreactive neurons were nitro-oxidergic neurons. The enzyme immunoassay revealed that the average Bax-like protein concentration in the serum was 39.0±9.6 pmol/ml in the adult male rat. Colchicine treatment significantly increased the densities of Bax-IR in the PVN and decreased the serum Bax-like protein levels. From these results, Bax-like protein is considered to be produced in the hypothalamic oxytocin neurons, transported through axoplasmic flow and secreted into systemic circulation at the posterior pituitary as a neurohormone

A practical three-dimensional reconstruction method to measure the volume of the sexually-dimorphic central nucleus of the medial preoptic area (MPOC) of the rat hypothalamus

Published estimates of the volume of the sexually-dimorphic central nucleus of the medial preoptic area (MPOC) have been quite variable both within and between laboratories. To obtain MPOC volume, most experimenters began with a two-dimensional (2-D) approach. They outlined the MPOC on each of several individual sections; then they added up the area contained on each section and multiplied the total by the section thickness. A 3-D reconstruction approach, although promising, has been somewhat impractical until recently due to the requirements for highly specialized software and massive computing support. Here, we describe the application of commercially-available PC-based software to measure MPOC volume by 3-D reconstruction. Male and female Sprague–Dawley rats, 24 or 50 days of age, were perfusion-fixed with 10% neutral phosphate-buffered formaldehyde. Following processing and embedding, a series of 20-μm sagittal paraffin sections were cut and mounted onto slides. After staining with cresyl violet, they were digitized using a microscope-mounted video camera connected to a frame-grabber in a Pentium-class computer (MCID-M5+). In addition to the MPOC, the anterior commissure, fornix, paraventricular nucleus, medial division of the bed nucleus of the stria terminalis, third ventricle and the bed nucleus of the anterior commissure were identified on the screen image and outlined using a computer mouse. These outlines were then aligned and rendered in 3-D with a solid overlay. The additional areas, such as anterior commissure, form landmarks within 3-D space to improve the accuracy with which the MPOC may be located and outlined. The reconstruction provides a striking illustration of the geometric relations between the structures of the anterior hypothalamus in the male and female rat. Moreover, the volumes determined from the overlays were reproducible between repeated studies in our laboratory. Our volume measurements confirm the sexual dimorphism previously reported for MPOC volumes, and provide a relatively quick, accurate and reliable protocol that should be useful in future experimental studies of environmental estrogenic compounds.

Steroid implants in the medial preoptic area or ventromedial nucleus of the hypothalamus activate female sexual behaviour in the musk shrew.

Female musk shrews are induced ovulators that do not exhibit a spontaneous behavioural oestrous cycle. Testosterone produced by the ovaries and adrenal glands, is the major steroid hormone in circulation at times of mating, and as such, regulates sexual behaviour. In the first experiment, we identified the neural site(s) of action for testosterone. Hormone implants were placed in one of three targeted brain regions. The neural sites selected were the medial anterior division of the bed nucleus of the stria terminalis (BNSTMA), medial preoptic area (mPOA) and the ventromedial nucleus of the hypothalamus (VMN). Ovariectomized females who received a unilateral testosterone propionate implant in either the mPOA or VMN, were significantly more likely to display sexual behaviour as compared to females who received an implant in the BNSTMA or any other hypothalamic nucleus. In experiments 2 and 3, we investigated whether the behavioural effects of testosterone propionate were mediated by an oestrogen receptor or the androgen receptor. Ovariectomized females that received oestradiol (E2) implants in either the mPOA or VMN were more likely to display receptivity, and had significantly shorter behavioural latencies, as compared to females implanted with either dihydrotestosterone or cholesterol. These data show that neural aromatization of testosterone to E2 in the mPOA or VMN is necessary for optimal activation of female musk shrew sexual behaviour. This finding implies a degree of neural redundancy in the networks that control the expression of sexual receptivity.

Differential projection of the posterior paralaminar thalamic nuclei to the amygdaloid complex in the rat.

The thalamic paralaminar nuclei that border the medial and ventral edges of the medial geniculate body, viz. the suprageniculate nucleus (SG), the posterior intralaminar nucleus (PIN), the medial division of the medial geniculate nucleus (MGm), and the peripeduncular nucleus (PP), are regarded as important extralemniscal relay nuclei for sensory stimuli and as an important link for the direct transmission of sensory stimuli to the amygdala. Each of these thalamic nuclei receives a unique pattern of afferent input but an unresolved question is, how each of these thalamic nuclei project to the amygdala and whether there are zones of convergence and/or non-overlapping regions within amygdaloid target nuclei. Small injections of PHA-L or Miniruby, which were made into single thalamic nuclei at different rostrocaudal levels, revealed a non-uniform distribution of anterogradely labeled axons within the amygdaloid complex. Injections into the SG, MGm, and rostral PIN predominantly labeled axons in the laterodorsal and lateroventral portions of the lateral nucleus of the amygdala (LA). Axons from the MGm were located rather in the dorsal part of the LA, whereas SG-derived axons were concentrated in the ventrolateral part of the LA. Injections into the PP labeled axons predominantly in the medial part of the LA, whereas after injections into the caudal PIN axons were seen in the entire LA. In addition, the PIN projects heavily to the anterior basomedial nucleus and medial division of the central nucleus, whereas this projection is virtually absent from the other thalamic nuclei. The lateral part of the central nucleus and the basal nucleus of the amygdala are spared by axons from the thalamic paralaminar nuclei. The present results suggest that, despite a considerable degree of convergence of the thalamoamygdaloid projection in the lateral nucleus, each thalamic nucleus plays a unique role in the transmission of sensory stimuli to the amygdala and in the modulation of intraamygdaloid circuits.

Geniculo-collicular descending projections in the gerbil

Axonal projections were labeled using a combination of tract-tracing and intracellular staining to examine the connection between the auditory thalamus and tectum. We report descending projections from the medial and dorsal divisions of the medial geniculate body to the external nucleus of the ipsilateral inferior colliculus and lower brainstem of the gerbil. These were studied in quasi-parasagittal brain slice preparations containing auditory thalamus, tectum and brainstem. Slices were cut in a curved, roughly parasagittal plane from the level of the thalamus to the brainstem. When properly cut, a single thick slice included most of the ipsilateral auditory cell groups and pathways at these levels. The geniculo-collicular projections were revealed after extracellular injections of Biocytin and rhodamine-conjugated biotinylated dextran amine in the medial geniculate body were applied to these slices. Furthermore, cells in the medial geniculate body that had been retrogradely prelabeled with a fluorescent tracer, Fluoro-Gold, injected in the inferior colliculus were intracellularly labeled to confirm the presence of the descending axons. In these preparations, single cells with descending axons were traced through their entire extent to their terminals in the inferior colliculus. Our results clearly demonstrate the geniculo-collicular descending projection and suggest a thalamo-tectal feedback loop.

Projections from the central nucleus of the amygdala to the gastric related area of the dorsal vagal complex: a Phaseolus vulgaris-leucoagglutinin study in rat

Electrophysiological and anatomic studies suggest that the amygdala regulates gastrointestinal motility and gastric acid secretion via projections to the dorsal vagal complex. The topography of these projections is poorly understood. Here, these projections were investigated by injecting anterograde tracer, Phaseolus vulgaris-leucoagglutinin, into the different divisions of the central nucleus of the amygdala in 13 rats. The distribution of immunohistochemically labeled terminals in the different portions of the dorsal vagal complex was analyzed. We found that (1) the dorsal aspect of the medial division of the central nucleus provided moderate projections to the dorsal vagal complex; (2) the heaviest projections terminated in the parvicellular and medial divisions of the nucleus of the solitary tract. These data suggest that via topographically organized projections, the amygdala can modulate the vago-vagal gastrointestinal reflexes in emotional and stressful situations.

The anatomical organization of the macaque pregeniculate complex

Saccade-related activity recorded in the primate pregeniculate nucleus, and its anatomical connections with the pretectal nucleus of the optic tract (NOT) and superior colliculus (SC), suggest that it plays a role in visual-ocular motor integration. To study this role, a clearer understanding of pregeniculate organization is required. Based on its connectivity and neurotransmitter immunocytochemistry, we demonstrate that this nucleus is composed of several subnuclei, suggesting the term, pregeniculate complex (PrGC). The PrGC includes a weakly developed dorsal lamina, rostrally, and a well-developed ventral lamina. The ventral lamina includes the retinorecipient and superior sublayers, rostrally, and the medial division, caudally. A thin lamina of cells lateral to the dorsal lateral geniculate nucleus is contiguous with the PrGC; we term this the lateral division. The PrGC and the lateral division each project to the SC/NOT; the superior sublayer and medial division of the PrGC are connected reciprocally to the SC/NOT. Immunocytochemistry for gamma-aminobutyric acid (GABA) and substance P (SP) further delineate the PrGC subnuclei. The retinorecipient sublayer stains most intensely for GABA and SP. The superior sublayer and medial division also stain strongly for GABA and SP. Essentially all neurons in the lateral division are GABA-positive. The combination of tract tracing and immunocytochemistry demonstrate differences in the connectivity of the PrGC subnuclei and the lateral division with the SC/NOT. This, combined with the differential localization of GABA in the PrGC, provides a basis for further study of its functional role.