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KIF5C, a novel neuronal kinesin enriched in motor neurons.

Kinesin superfamily proteins (KIFs) are the molecular motors conveying cargos along microtubules. KIF5s, the heavy chains of conventional kinesin (KHC), are originally identified members of KIFs, and neuronal KIF5A and ubiquitous KIF5B have been identified so far. In the present work, we cloned a novel member of KIF5, KIF5C, and generated specific antibodies against three KIF5s to investigate their distribution and functions. KIF5A showed pan-neuronal distribution in the nervous system. KIF5B showed a glial cell distribution pattern in general; however, interestingly, its expression was strongly upregulated in axon-elongating neurons, such as olfactory primary neurons and mossy fibers. KIF5C was also a neuronal KIF5 like KIF5A but was highly expressed in lower motor neurons in 2-week-old or older mice, suggesting its important roles in the maintenance of motor neurons rather than in their formation, such as axonal elongation. Because a large part of KIF5s in adult motor neurons were expected to be KIF5C, we generated mice lacking the kif5C gene to investigate the functions of KIF5C in neurons in living animals. The mutant mice showed smaller brain size but were viable and did not show gross changes in the nervous system. Closer examinations revealed the relative loss of motor neurons to sensory neurons. Because three KIF5s showed high similarity in the amino acid sequence, could rescue the KIF5B mutant cells, and could form heterodimers, we think that there are functional redundancy among the three KIF5s and that KIF5A and KIF5B prevented the KIF5C null mice from the severe phenotype.

Amino Acid Sequence↗

[Quantitative evaluation of populations of neurons expressing dopamine synthesis enzymes in the arcuate nucleus neurons in developing rats].

The ratio of neuron populations expressing either tyrosine hydroxylase or aromatic L-amino acid decarboxylase, which are enzymes of dopamine synthesis, was estimated quantitatively in the arcuate nucleus of male and female rats on 21st day of intrauterine development, the 9th day of postnatal development, and in adult animals. The enzymes in neurons were revealed by double immunocytochemical labeling, followed by identification under a fluorescence microscope. At all the developmental stages, three neuron populations differing in the expression of these enzymes were revealed. By the end of the prenatal period, most of the neurons (99%) contained only one of the enzymes, and the proportion of neurons expressing both enzymes (dopaminergic neurons) did not exceed 1%. During postnatal development, the proportion of neurons with one enzyme proved to decrease, whereas that of dopaminergic neurons increased. However, the latter proportion, even in adult animals, did not exceed 50% of the total number of neurons expressing the enzymes of dopamine synthesis. Thus, the population of neurons expressing both enzymes increases during rat ontogeny, whereas the number of neurons expressing only one enzyme decreases.

Animals↗

[The interaction of neurons characterized by a tonic reaction to sound with other neurons in the cat auditory cortex].

Interaction of neurons with tonic response to sound with adjacent or distant (approximately 400-500 micrograms) cortex neurons was studied in acute experiments on 15 immobilized cats using a method of the cross-correlation analysis. A presence of synchronizing excitatory input common for the cells has been revealed in 26 pairs (72%) on the cross-correlograms. The results of the cross-correlation analysis in five pairs of neurons show mono- or polysynaptic excitatory effect of a tonic neuron to impulse activity of another neuron. Negative correlation indicative of the inhibitory influence of tonic neurons on impulse responses of other neurons of the same or adjacent auditory cortex column is revealed in five pairs of neurons, but the inhibitory influences may be considered as monosynaptic ones only in 3 pairs of these neurons (latency of interaction 1.0-1.5 ms). The data obtained permit concluding that the group of neurons characterized by tonic response to sound is a heterogeneous one in the functional respect. An assumption that some neurons of the tonic type are inhibitory interneurons of the auditory cortex, other excitatory ones is under discussion.

Acoustic Stimulation↗

Cytosine arabinoside kills postmitotic neurons: evidence that deoxycytidine may have a role in neuronal survival that is independent of DNA synthesis.

Cytosine arabinoside (ARA C), a competitive inhibitor of the incorporation of 2'-deoxycytidine into DNA in other cell types, caused a concentration-dependent inhibition of KCl- and insulin-stimulated survival of postmitotic ciliary parasympathetic ganglion neurons, and the nerve growth factor (NGF)-stimulated survival of postmitotic dorsal root ganglion (DRG) sensory neurons in vitro. The IC50 for survival was 2 x 10(-8) M for both types of neurons after 4 d under the culture conditions used. The inhibition of DRG survival by ARA C in the presence of varying concentrations of NGF indicated that ARA C acted as an apparent noncompetitive antagonist of NGF. This cytotoxic effect of ARA C was blocked by 2'-deoxycytidine, but not by cytosine, 2'-deoxyadenosine, 2'-deoxyguanosine, or 2'-deoxythymine, indicating that ARA C was interfering with a deoxycytidine-specific survival process. Cytidine could block ARA C toxicity, but it was 40 times less potent than 2'-deoxycytidine. The blockade of the cytotoxic effect of ARA C by 2'-deoxycytidine indicated that 2'-deoxycytidine was an apparent competitive antagonist of ARA C toxicity. 2'-Deoxycytidine, by itself, was not survival-promoting. Other antimitotic agents, such as adenine arabinoside, thymine arabinoside, 5-fluorodeoxyuridine, 5-bromodeoxycytidine, 5-azadeoxycytidine, and aphidicolin had no effect on neuronal survival at a concentration 5000 times the EC50 of ARA C, indicating that inhibition of DNA synthesis or repair was probably not the mechanism by which ARA C inhibited neuronal survival and that other 2'-deoxynucleosides were not involved in the survival-promoting process. Nitrobenzylthioinosine, an inhibitor of 2'-deoxycytidine and ARA C membrane transport in other cell types, inhibited the cytotoxic effect of ARA C in neurons, suggesting that ARA C entered the neurons through a similar transport mechanism and that ARA C needed to gain access to the inside of the neuron to be effective. These results indicate that ARA C, in addition to being an antimitotic agent for dividing cells, is also cytotoxic for postmitotic neurons. This inhibition of neuronal survival by ARA C is hypothesized to be due to inhibition of a 2'-deoxycytidine-dependent process that is independent of DNA synthesis or repair. Thus, 2'-deoxycytidine may have an important and previously unrecognized role in cellular function that in the case of neurons is critical for survival.

Animals↗

Lead neurotoxicity: neuronal and non-neuronal cell survival in fetal and adult DRG cell cultures.

In the present study, neural cell cultures of fetal and adult mouse dorsal root ganglia (DRG) were prepared, exposed to various concentrations of lead (Pb) for two different schedules, and dose survival curves obtained. For the late exposure (LE) to Pb, Pb was applied from 8 DIV to 20 DIV (12 day exposure) while for the early exposure (EE), Pb was applied from 2 DIV to 20 DIV (18 day exposure). For LE there was evidence for increased vulnerability of adult compared to fetal neurons and the LD50 for both was greater than 100 microM. For both LE and EE and for both adult and fetal tissue, the non-neuronal cells were much more sensitive to the toxic Pb effects than neurons. For the LE, the adult and the fetal non-neuronal cells had LD50s of 28 microM and 58 microM respectively. For EE the LD50 for adult neurons was 35 microM. The differential effect of Pb on fetal and adult non-neuronal cells observed for LE was reversed in direction for EE; the LD50 for adult non-neuronal cells being 5.5 microM and the LD50 for fetal non-neuronal cells being only 1.7 microM, making the fetal cells about three times more sensitive than adult non-neuronal cells. An explanation of the differential effects of Pb on neurons and non-neuronal cells and on adult and fetal cells was proposed which incorporated known cell dynamics in vitro.

Age Factors↗

Neuronal and non-neuronal aromatase in primary cultures of developing zebra finch telencephalon.

Estrogenic metabolites of circulating androgens have important effects on the organization and activation of neural circuits controlling reproductive behavior and physiology in males of many vertebrate species. Previous studies indicate that aromatase, the enzyme that converts androgens to estrogens, is expressed most abundantly in neurons in limbic brain regions. Songbirds are unique in that aromatase is expressed at unusually high levels throughout the telencephalon of both males and females. We assume that estrogens formed in the telencephalon itself masculinize neural circuits controlling song, since the brain is a major source of circulating estrogens in adult males. However, the cellular localization of telencephalic aromatase in songbirds remains unknown. We have established primary cultures from telencephalons of developing zebra finches and found aromatase activity (conversion of 3H-androstenedione or 3H-testosterone to 3H-estrone plus 3H-estradiol) at some of the highest levels reported for brain tissue of any species. Both neurons and glia were identified in these cultures based on cell morphology and labeling by specific immunohistochemical markers. However, when culture conditions were manipulated to reduce the incidence of either neurons or glia by varying the age of cultures or their plating density, treating with the neurotoxin kainic acid, physically shaking off loosely attached neurons, or preparing cultures in media that encouraged enrichment of neurons, high levels of aromatase persisted. Furthermore, Northern blot analysis of total RNA extracted from enriched neuronal or glial cultures indicated the presence of aromatase mRNA in both cell preparations. In situ hybridization with a zebra finch aromatase cDNA probe conjugated to digoxigenin showed the cultures contained darkly labeled neurons and lightly labeled non-neuronal cells, presumably astrocytes. We conclude that aromatase is expressed in both neuronal and non-neuronal cells in these cultures, suggesting that both cell types may also express the enzyme in vivo. The presence of aromatase outside of neurons suggests that glia may be targets of estrogen action or that glia may supply some estrogen to the estrogen-sensitive neural circuits in this species.

Aging↗

Role of neuron-glial junctional domain proteins in the maintenance and termination of neuronal migration across the embryonic cerebral wall.

To identify glial membrane proteins that contribute to the process of neuronal migration in the developing brain, we developed a polyclonal antiserum (D4) and a monoclonal antibody (NJPA1: neuron-glial junctional polypeptide antibody) that recognize membrane proteins localized to the plasmalemmal junction between migrating neurons and adjacent radial glial fibers (Cameron and Rakic, 1994). Here, we show that in the developing cerebral cortex, immunoreactivity for these junctional polypeptides is present throughout the neuronal migratory pathway but becomes minimal or absent where radial glial cell processes enter the marginal zone region, the barrier at which newly arrived neurons normally stop their migration and detach from their glial fiber substrate. We thus tested, using imprints of embryonic cerebral wall and slice preparations, whether the junctional membrane proteins detected by our antibodies contribute to the regulation of neuronal migration in the cerebral cortex. The rate of neuronal migration on glial cell substrates was reduced significantly in the presence of D4 or NJPA1 antibodies. Antibody exposure typically led to the withdrawal of leading processes, changes in microtubular organization and, in some instances, to detachment of neurons from their glial cell substrates. These results suggest that the polypeptides recognized by the D4 and NJPA1 antibodies are essential for the maintenance of normal neuronal migration. Dismantling of neuron-glial cell junctional domains formed by these membrane proteins may underlie neuronal cell detachment from glial migratory substrates at the interface between cortical plate and marginal zone in the developing cerebral wall.

Animals↗

Response of postmitotic neurons to X-irradiation: implications for the role of DNA damage in neuronal apoptosis.

The molecular changes responsible for inducing neuronal apoptosis are unknown. Rat cortical neurons were treated with x-irradiation 7 d after isolation to test for the role of DNA damage in neuronal death. The response of neurons to x-irradiation was compared with that of astrocytes that had been isolated 3 weeks earlier from newborn rats. At the time of irradiation, the neurons appeared well differentiated morphologically and were predominantly (90-95%) noncycling, based on flow cytometric analysis. There was a similar, linear increase in DNA double-strand breaks with increasing radiation dose in neurons and astrocytes. However, whereas doses as low as 2 Gy induced typical apoptotic changes in neurons, including nuclear fragmentation and/or internucleosomal DNA fragmentation, doses as high as 32 Gy caused little or no apoptosis in astrocytes. Radiation-induced apoptosis of neurons started 4-8 hr after irradiation, was maximal at 12 hr, and was dependent on dose up to 16 Gy. It was prevented when cycloheximide, a protein synthesis inhibitor, was added up to 6 hr after irradiation. In addition to their distinct apoptotic response, neurons rejoined radiation-induced DNA double-strand breaks more slowly than astrocytes. Treatment with benzamide to inhibit ADP-ribosylation and strand break repair increased apoptosis; splitting the dose of radiation to allow increased time for DNA repair decreased apoptosis. These data suggest that DNA damage may induce neuronal apoptosis, that the extent of damage may determine the degree of apoptosis induced, and that slow repair of damage may play a role in the susceptibility of neurons to apoptosis.

Adenosine Diphosphate Ribose↗

Loss of primary sensory neurons in the very old rat: neuron number estimates using the disector method and confocal optical sectioning.

Loss of neurons has been considered to be a prime cause of nervous disturbances that occur with advancing age. However, the notion of a constitutive aging-related loss of neurons has been challenged recently in several studies that used up-to-date methods for counting neurons. In this study, we have applied stereological techniques with the objective of obtaining quantitative data on total neuron numbers and the distribution of neuron cross-sectional areas in the fifth cervical (C5) and fourth lumbar (L4) dorsal root ganglion (DRG) of 3- and 30-month-old Sprague-Dawley rats. Tissue data were recorded on a confocal laser-scanning microscope with the use of the optical-disector technique and random, systematic sampling. Aged rats of both sexes disclosed only a small decrease (approximately 12%) in the number of cervical and lumbar DRG neurons. Furthermore, there was no significant correlation between the degree of neuron loss and the extent of behavioral deficits among the aged individuals. The DRG neurons of aged rats had a smaller mean cross-sectional area (approximately 15%; P < 0.001) at both DRG levels. Further analysis of the male cohorts was carried out by using isolectin B4 and neurofilament subunit (phosphorylated 200 kDa; RT97) immunoreactivity (IR) as selective markers for unmyelinated and myelinated axons, respectively, and disclosed no significant change in the relative frequencies of immunoreactive neuron profiles in the old rats. However, RT97-IR DRG neurons of the aged rats had significantly smaller cross-sectional areas (approximately 9% in C5; approximately 16% in L4; P < 0.001) than the young adult rats, indicating a selective cell body atrophy among myelinated primary afferents during aging. The results indicate that loss of primary sensory neurons cannot exclusively explain the functional deficits in sensory perception among senescent individuals. It seems likely that other factors at the subcellular level and/or target interaction(s) contribute substantially to the sensory impairments observed with advancing age.

Animals↗

Quantitative analyses of neuronal development in the lateral motor column of mouse spinal cord. III. Generation and settling patterns of large and small neurons.

The generation and settling patterns of large and small lateral motor column (LMC) neurons were compared in the spinal cords of three inbred strains of mice by means of tritiated thymidine autoradiography. No significant strain differences were observed for the number of large LMC cells (presumptive alpha motor neurons) that were heavily labeled on each injection day, although there were significant strain variations for this measure with regard to small LMC neurons (presumed gamma motor neurons and interneurons). The generation of both large and small LMC neurons began at the same time, but peak production of large cells preceded that of the small neurons. There were no strain differences observed for this relationship between the large and small cells. These findings indicate that the LMC, from the time of its initial formation, contains cells destined to become large and small neurons. The positions of large and small neurons within the adult LMC relative to their times of origin (settling patterns) were analyzed statistically. A significant ventrodorsal sequence for early-to-late generated cells was observed for both large and small LMC neurons. No significant strain differences were found in the analysis of settling patterns. A ventrodorsal settling pattern also has been described for amphibia (Prestige, '73) and, in conjunction with the proximodistal sequence of limb development described by other investigators, the ventrodorsal sequence could play a key role in the development of motor neuronal somatotopic organization.

Animals↗

S100 is present in developing chicken neurons and Schwann cells and promotes motor neuron survival in vivo.

We used polyclonal antisera recognizing S100, a small acidic protein highly enriched in nervous tissue, to stain sections of embryonic chicken lumbosacral spinal cord and hindlimb. S100 immunoreactivity was detected in developing sensory neurons of the dorsal root ganglia (DRG) and motor neurons of the ventral spinal cord as early as embryonic day (E) 5, and staining persisted through hatching. In contrast, expression of S100 first became apparent in Schwann cells at E13, just before myelination, and was not detected in developing skin or muscle. Since S100 beta was present in motor and sensory neurons and is known to promote neuronal survival and neurite extension in vitro (Winningham-Major, Staecker, Barger, Coats, and Van Eldik, 1989), we tested the ability of S100 to promote neuron survival in an in ovo survival assay. Addition of S100 to chick embryos in ovo during the period of naturally occurring motor neuron cell death resulted in a significant increase in motor neuron survival, but had no effect on the in vivo survival of sensory neurons in the DRG. The findings that S100 is present in spinal motor neurons and that the addition of S100 enhances the survival of these cells in vivo are consistent with the possibility that S100 may act as a naturally occurring neuron survival factor during development.

Animals↗

Anatomy and physiology of saccadic burst neurons in the alert squirrel monkey. I. Excitatory burst neurons.

Saccadic burst neurons in the pontine reticular formation have been implicated in the generation of saccades in the horizontal plane on the basis of lesion and extracellular recording studies in the cat and monkey. In the present study, saccadic burst neurons were anatomically and physiologically characterized with intraaxonal recording and injection of horseradish peroxidase in the alert squirrel monkey. A population of burst neurons were found that appear analogous to the excitatory burst neurons (EBNs) described previously in the cat. All neurons are located in the caudal pontine reticular formation and have a major axonal projection to the ipsilateral abducens nucleus. Additional projections were found to the medial vestibular nucleus, the nucleus prepositus, and regions of the pontine and medullary reticular formation rostral, ventral, and caudal to the abducens. All neurons fire exclusively during saccades and have a discharge pattern similar to that of medium-lead burst neurons described previously in the cat and monkey. In most neurons the saccadic burst begins 5-15 msec before saccade onset. Linear relationships exist between burst duration and saccade duration, number of spikes in the burst and saccade amplitude, and firing frequency and instantaneous velocity. Physiological activity of each neuron shows the closest relationship with the amplitude of the saccade component in a particular direction. For all neurons, this on-direction is in the ipsilateral hemifield and is predominantly horizontal, but may have either an upward or downward vertical component. These results support a major role for the EBNs in the monkey in generating the saccadic burst in abducens motoneurons, as well as in contributing to the oculomotor activity in other classes of premotor neurons.

Abducens Nerve↗

Trauma-induced Golgi-like staining of neurons: a new approach to neuronal organization and response to injury.

A new esterification-silver approach to the directed staining of the dendritic trees of traumatized neurons is described. Stained neurons compare favorably to those labeled with silver chromate Golgi impregnations in the visualization of dendritic arbors. Cells in all parts of the brain, including the hypothalamus, hippocampus, cerebral cortex, cerebellum, striatum, spinal cord, thalamus, and olfactory bulb, can be detected after focal trauma to that region. Selection of neurons to be stained is made by increasing their affinity for silver with many different types of directed neuronal trauma, including micropipette wounds, pressure, surgical incision, chemical cytotoxicity, and impact trauma. Trauma to one area of the brain results in dendritic arbors staining only in the injured area; other areas of the brain are free of dendritic staining. Injury can be produced either in vivo or in vitro. In vitro injury to neurons allows a high degree of localization and facilitates the analysis of neuronal response to trauma in the absence of complicating factors such as blood flow and secondary injury. The selective affinity for silver staining in this approach is increased very rapidly, allowing detection of traumatized cells fixed minutes after injury. Brains of all mammals used appear to stain similarly, including the rat, mouse, pig, and human. Axons, although labeled less frequently than dendritic arbors, are induced to stain just as rapidly as dendrites. The ability to visualize a large part of the dendritic tree after trauma allows the segregation of neuronal subtypes on the basis of their differential response to injury. Subpopulations of cells in the same area of the brain appear to respond differently to trauma. Differential response of neurons to trauma can easily be detected. For instance, slight variations in trauma can be used to label selectively the major subpopulations of neurons in the hippocampus, including pyramidal cells, interneurons near the pyramidal cell layer, or granule cells. Similarly, neurons of the hypothalamic paraventricular and arcuate nuclei respond to compression trauma much more dramatically than other cell types in the same region of the hypothalamus. Hypothalamic neurons were studied extensively, particularly in regions that are difficult to routinely stain with Golgi impregnations, including the arcuate, paraventricular, supraoptic, and suprachiasmatic nuclei. Trauma to these areas was made in vitro after removal of the brain from the skull, allowing easy access to the ventral surface of the brain.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Distribution of substance P-like immunoreactive neurons in the human medulla oblongata: co-localization with monoamine-synthesizing neurons.

The raphe nuclei also contained SP-like immunoreactivity (up to 30%) while few monoamine-synthesizing neurons in the lateral and dorsomedial medulla contained SP-like immunoreactivity (approximately 5% of presumed serotonin-, noradrenaline-, and adren- the adult human. The majority of SP-like immunoreactive neurons were found in four main regions: the lateral medulla, the dorsomedial medulla, the spinal trigeminal nucleus, and the raphe nuclei. The morphology of immunoreactive cells varied according to the region in which they were found. In contrast to previous studies, we found large numbers (90,000) of SP-like immunoreactive neurons throughout the adult human medulla oblongata. The distribution of these SP-like immunoreactive neurons appears to be significantly different from those described in the rat and cat. These results were compared to the distributions of monoamine-synthesizing and neuropeptide Y (NPY)-like immunoreactive neurons in the human medulla previously reported (Halliday et al.: Neuroscience, in press, 1988a; J. Comp. Neurol., in press, 1988b). Colocalization studies revealed that many presumed serotonin-synthesizing neurons in the raphe nuclei also contained SP-like immunoreactivity (up to 30%) while few monoamine-synthesizing neurons in the lateral and dorsomedial medulla contained SP-like immunoreactivity (approximately 5% of presumed serotonin-, noradrenaline-, and adrenaline-synthesizing neurons). The distributions of SP- and NPY-like immunoreactive neurons were similar, although SP-like immunoreactive neurons were concentrated in the lateral regions of the same structures. We have found that the distributions of monoamine-synthesizing, NPY-, and SP-like immunoreactive neurons significantly overlap, particularly in the lateral medulla of the adult human. There is a large increase in the number of these cells in this region compared to other species, emphasizing the neuroanatomical differences between humans and other species.

Aged↗

Disease-specific patterns of neuronal loss in the spinal ventral horn in amyotrophic lateral sclerosis, multiple system atrophy and X-linked recessive bulbospinal neuronopathy, with special reference to the loss of small neurons in the intermediate zone.

The ventral horn cells of the fourth lumbar segment were morphometrically analysed in six cases of amyotrophic lateral sclerosis (ALS; there common forms and three pseudopolyneuritic forms), six of multiple system atrophy (MSA) with autonomic failure, four of X-linked recessive bulbospinal neuronopathy (X-BSNP), and seven age-matched autopsy cases of non-neurological disorders. In the common form of ALS, large and medium-sized neurons of the medial and lateral nuclei were markedly lost; small neurons in the intermediate zone were slightly diminished but fairly well preserved. In the pseudopolyneuritic form of ALS, marked loss was present in the large and medium-sized neurons, and in the small neurons located in the intermediate zone as well. In the MSA, in contrast to ALS, there was a marked reduction in small neurons in the intermediate zone, and large and medium-sized neurons of the medial and lateral nuclei tended to be preserved. In X-BSNP, large and medium-sized neurons were almost completely lost and small neurons were also markedly depopulated. These findings indicated that the pattern of neuron loss in the ventral horn is distinct among these diseases depending on size, location and function of the ventral horn cell population. These disease-specific patterns of neuron loss suggest a difference in the process of neuronal degeneration of ventral horn cells among the disease examined.

Adult↗

The location of spinal projection neurons in the cerebellar nuclei (cerebellospinal tract neurons) of the cat. A study with the horseradish peroxidase technique.

The distribution of spinal projection neurons was studied in the cerebellar nuclei of the cat following injections of horseradish peroxidase (HRP) into the cervical, thoracic and lumbar cord. HRP-positive (labeled) neurons were found in the medial (fastigial) and the posterior interpositus nuclei on the side contralateral to the cervical injection, being most numberous in cases with injections between the C2 and the C3 segments. In the medial nucleus (M) labeled neurons were distributed in the central to the caudal portions, and there was a conspicuous group of labeled small neurons extending from the ventrolateral part to the intermediate zone between the M and the anterior interpositus nucleus. With an increasing number of medium-sized neurons, this neuronal group persisted caudally in a similar position, ventromedial to the posterior interpositus nucleus (IP). Labeled large neurons were seen in the medial third of the IP. In the two cases labeled neurons of medium and small sizes were equal in number, and the neurons of the IP constituted about 10% of the total number of the spinal projection neurons. The present study suggests that the neurons of the M and the IP, including those of the intermediate group located between the two, project the bulk of the crossed descending fibers as far caudally as the C2 and the C3 segments.

Afferent Pathways↗

Crossed projection neurons are generated prior to uncrossed projection neurons in the lateral superior olive of the rat.

The present study examined in the lateral superior olive (LSO) of the rat whether LSO neurons projecting to the ipsilateral inferior colliculus (IC) might be generated later than those projecting to the contralateral IC. Rat fetuses were exposed in utero to 5-bromodeoxyuridine (BrdU), a thymidine analogue, to label neurons proliferating at different embryonic stages from day E11 through to E20. Upon reaching adulthood, the rats were given unilateral injections of fluoro-gold (FG), a retrograde fluorescent tracer, into the IC. Subsequently, the tissue sections of the brains obtained from the rats were immunostained for BrdU to simultaneously detect neurons that were BrdU-positive and/or FG-positive. BrdU-positive LSO neurons were found in the rats which had been exposed to BrdU during E12-E16. In E12 and E13 BrdU-exposure cases, the vast majority of doubled-labeled (BrdU-positive and FG-positive) neurons were seen on the contralateral side to the FG injection. In E14, E15 and E16 BrdU-exposure cases, in contrast, all double-labeled neurons were found on the ipsilateral side to the FG injection. The distribution of these double-labeled neurons within the nucleus was diffuse in all the BrdU-exposure cases. Thus, the results indicate that LSO neurons are generated during E12-E16, that the crossed projection neurons are generated 1-4 days earlier than the uncrossed projection neurons, and that no topographical relationships exist between the early- and the late-generated populations of the LSO neurons.

Animals↗

Neuronal interactions between galanin-like-peptide- and orexin- or melanin-concentrating hormone-containing neurons.

Galanin-like peptide (GALP) is a novel orexigenic neuropeptide that is recently isolated from the porcine hypothalamus. GALP-containing neurons predominantly locate in the hypothalamic arcuate nucleus (ARC). The expression of GALP mRNA within the ARC is increased after the administration of leptin. GALP-containing neurons express leptin receptor and contain alpha-melanocyte-stimulating hormone. We have recently reported that neuropeptide Y (NPY)- and orexin-containing axon terminals are in close apposition with GALP-containing neurons in the ARC. In addition, GALP-containing neurons express orexin-1 receptor (OX1-R). Thus, GALP may function under the influence of leptin and orexin. However, the target neurons of GALP have not yet been clarified. To clarify the neuronal interaction between GALP-containing and other feeding regulating neurons, double-immunostaining method using antibodies against GALP- and orexin- or melanin-concentrating hormone (MCH) was performed in the rat lateral hypothalamus (LH). GALP-immunoreactive fibers appeared to project to the LH around the fornix. They were also found from the rostral to the caudal part of the ARC, paraventricular nucleus (PVH), stria terminalis (BST), medial preoptic area (MPA), and lateral septal nucleus (LSV). Moreover, GALP-like immunoreactive nerve fibers were directly contacted with orexin- and melanin-concentrating hormone (MCH)-like immunoreactive neurons in the LH. Our findings strongly suggest that GALP-containing neurons interact with orexin- and/or MCH-containing neurons in the lateral hypothalamus and that it participates in the regulation of feeding behavior in harmony with other feeding-regulating neurons in the hypothalamus.

Animals↗