RESUMEN
The spatial organization of projections from olfactory receptor neurons to the main olfactory bulb (MOB) was studied in hamsters by using fluorescent stilbene isothiocyanates as retrograde tracers. Injections confined to small sectors of the MOB produce labeling of receptor neurons that is more restricted circumferentially (i.e., with respect to the medial-lateral and dorsal-ventral axes) than longitudinally (i.e., with respect to the rostral-caudal axis) along the mucosal sheet. This restricted labeling is also discontinuous, giving an initial impression that the peripheral input is only crudely organized with respect to the medial-lateral and dorsal-ventral axes of the nasal cavity. However, from analyses of serial sections, it is apparent that each set of mucosal segments shares convergent projections to a circumferential quadrant of the MOB with other segments that are positioned around a common domain of the nasal cavity airspace. The primary afferent projections to the MOB, thus, are organized rhinotopically (i.e., with respect to the three-dimensional position of receptor neurons in olfactory space) rather than mucosotopically.
Asunto(s)
Neuronas/fisiología , Bulbo Olfatorio/fisiología , Vías Olfatorias/fisiología , Células Receptoras Sensoriales/fisiología , Transmisión Sináptica , Ácido 4-Acetamido-4'-isotiocianatostilbeno-2,2'-disulfónico/farmacocinética , Ácido 4-Acetamido-4'-isotiocianatostilbeno-2,2'-disulfónico/farmacología , Animales , Cricetinae , Inyecciones , Masculino , Mesocricetus , Cavidad Nasal/fisiología , Tabique Nasal/fisiología , Fibras Nerviosas/metabolismo , Vías Olfatorias/citologíaRESUMEN
The spatial organization of olfactory receptor surfaces and odorant passageways within the nasal cavity was studied in hamsters through descriptive and morphometric analyses of a complete stereotaxically defined series of coronal, sagittal, and horizontal sections through the snout. These analyses reveal that the caudal two-thirds of each cavity is divided into two longitudinally oriented medial and lateral channels. The olfactory mucosa that lines these two channels projects selectively onto the medial and lateral halves of the main olfactory bulb (MOB), respectively. Moreover, the ethmoturbinates of the caudal recesses create highly convoluted channels, lined by ventrally projecting mucosa, that lie ventral, lateral, and dorsal to a relatively smooth central channel lined by dorsally projecting mucosa. The rhinotopic map makes equivalent representations of medial and lateral olfactory space to the MOB but gives the smooth space lined by dorsally projecting mucosa a disproportionately larger representation on the MOB than the convoluted space lined by the more expansive ventrally projecting mucosa. Recent descriptions of the spatial distribution of probes for odorant receptor proteins conform closely to this organization, giving credence to the idea that rhinotopy is a basis for representing to the MOB the specific molecular features of odorant molecules.
Asunto(s)
Cavidad Nasal/inervación , Odorantes , Vías Olfatorias/anatomía & histología , Células Receptoras Sensoriales/citología , Animales , Cricetinae , Masculino , Mesocricetus , Neuronas/fisiología , Mucosa Olfatoria/inervación , Vías Olfatorias/citología , Células Receptoras Sensoriales/fisiologíaAsunto(s)
Proteínas Asociadas a Microtúbulos/fisiología , Proteínas del Tejido Nervioso/fisiología , Sistema Nervioso/química , Animales , Ácido Aspártico/fisiología , Axones/química , Axones/ultraestructura , Dendritas/química , Dendritas/ultraestructura , Regulación de la Expresión Génica , Glutamatos/fisiología , Ácido Glutámico , Humanos , Proteínas Asociadas a Microtúbulos/análisis , Proteínas Asociadas a Microtúbulos/clasificación , Microtúbulos/fisiología , Microtúbulos/ultraestructura , Factores de Crecimiento Nervioso/fisiología , Proteínas del Tejido Nervioso/análisis , Sistema Nervioso/ultraestructura , Plasticidad Neuronal , Sensación/fisiologíaRESUMEN
Five high-molecular-weight microtubule-associated proteins (MAPs) were identified in brain tissue in previous work from this laboratory (Bloom et al., 1984). These proteins were termed MAP 1A, 1B, 1C, 2A, and 2B. The MAP 1's differed from the MAP 2's, and showed little evidence of interrelationship on the basis of immunological and biochemical comparison. We report here that MAP 1A and MAP 1B are, in fact, related at the level of subunit composition. Immunoprecipitation of the individual MAPs showed that both contained low-molecular-weight subunits of Mr 30,000 and Mr 19,000 (light chains 1 and 3). An additional subunit, light chain 2 (Mr 28,000), was primarily found in preparations of MAP 1A. The light chains co-sedimented with microtubules after chymotryptic digestion of the MAPs. This suggested an association of the light chains with the microtubule binding domains of the MAPs, which are identified here as distinct fragments of Mr 60,000 for MAP 1A and 120,000 for MAP 1B. A panel of monoclonal anti-MAP 1A and anti-MAP 1B antibodies, including one that reacts with a common phosphorylated epitope, was used to examine the distribution of these proteins in the developing rat brain and spinal cord. MAP 1B was found to be abundant in the newborn brain and to decrease with development, in contrast to MAP 1A which increased with development. By immunohistochemistry MAP 1B was found to be highly concentrated in developing axonal processes in the cerebellar molecular layer, the corticospinal tract, the mossy fibers in the hippocampus, and the olfactory nerve. Of particular interest, the mossy fiber and olfactory nerve staining persisted in the adult, indicating continued outgrowth of the mossy fibers as well as olfactory nerve axons. MAP 1A staining was, in contrast, weak or absent in developing axonal fibers but moderate in mature axons and intense in developing and mature dendritic processes. Our results indicate that MAP 1A and MAP 1B are structurally related components of the neuronal cytoskeleton with complementary patterns of expression.
Asunto(s)
Sistema Nervioso Central/metabolismo , Proteínas Asociadas a Microtúbulos/metabolismo , Animales , Bovinos , Sistema Nervioso Central/crecimiento & desarrollo , Fenómenos Químicos , Química , Epítopos , Inmunohistoquímica , Proteínas Asociadas a Microtúbulos/inmunología , Fosforilación , Distribución TisularRESUMEN
The effects of castration and testosterone (T) replacement on levels of substance P (SP) and luteinizing hormone-releasing hormone (LHRH) were assessed in discrete areas of the male hamster brain. The animals were either castrated, castrated and given a chronically low or high dose of T by Silastic implant, or sham-operated. Brain tissues and trunk blood were collected 3 weeks after surgery. Plasma T levels were maintained within the normal range by the implants but at significantly lower or higher levels than the mean for sham-operated males. Levels of SP and LHRH were quantified in the olfactory bulbs, rostral basal forebrain, anterior hypothalamic and preoptic area, medial basal hypothalamic area, medial basal hypothalamic area and median eminence, and brain stem. In general, castration and T replacement effected opposite changes in levels of SP and LHRH. In the medial basal hypothalamic area and median eminence SP levels were found to be inversely related to the chronic T levels, whereas the LHRH levels were directly correlated. In the anterior hypothalamic and preoptic area, castration reduced levels of SP. Conversely, castration elevated levels of LHRH in this area. This inverse dynamic relationship between changing peptide levels was also observed in the rostral basal forebrain but not in the olfactory bulbs. In most of these forebrain regions, the dose-response curves for the experimental groups could not incorporate the peptide levels in the sham-operated control group. SP levels in the brain stem showed a monotonic inverse relationship to circulating T levels which did include the control group values.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Química Encefálica , Hormona Liberadora de Gonadotropina/metabolismo , Sustancia P/metabolismo , Testosterona/sangre , Animales , Tronco Encefálico/metabolismo , Cricetinae , Diencéfalo/metabolismo , Masculino , Bulbo Olfatorio/metabolismo , OrquiectomíaRESUMEN
Following placement into a test cage filled with pine shavings, a litter of 7-8 golden hamster pups (aged 3-18 days postnatal: P3-18) initially displays a period of locomotion which ends reliably in huddling. The latency to establish a huddle (i.e., the duration of locomotion) is significantly longer in the presence of novel odors (fresh or lemon shavings) than more familiar odors (slightly soiled fresh or lemon shavings) but only in pups aged P12 or older. Pups aged P9 or younger do not locomote differentially in the presence of novel or familiar odors. This age difference represents the emergence of olfactory exploration in hamsters between P9 and P12. Exploration of novel odors interferes with initial attempts to establish a single huddle site by a litter, but does not preclude the ultimate aggregation of all pups at a single site as guided by conspecific odors and possibly thermotactile cues as well. Such shifts in the control of behavior by non-nest and nest-related, conspecific stimuli correspond well with the first occurrence of nest exits at P11-12 (e.g., Dieterlen, 1959) coupled with the persistent return of hamster pups to the maternal nest for as long as it is maintained (Rowell, 1961).
Asunto(s)
Animales Recién Nacidos/crecimiento & desarrollo , Conducta Exploratoria/fisiología , Comportamiento de Nidificación/fisiología , Olfato/fisiología , Factores de Edad , Animales , Cricetinae , Femenino , Masculino , Mesocricetus , OdorantesRESUMEN
Antisera generated to substance P-Gly (SP-G) and substance P-Gly-Lys (SP-G-K), the likely unamidated COOH-terminally extended forms of substance P, were used to quantify and localize substance P precursor forms in hamster brain stem and spinal cord. The precursor determinant SP-G-K was liberated from larger heterogeneous forms by mild trypsinization of tissue extracts and was converted into the second precursor determinant, SP-G, by subsequent treatment with carboxypeptidase B. The basal levels of SP-G-K in brain stem and spinal cord were approximately equal to 0.5 pg/mg of tissue and rose 43- to 64-fold after trypsinization. Basal levels of SP-G were comparable to those of SP-G-K and rose 10- to 29-fold after combined enzyme treatments. Immunohistochemical labeling of axons and somata with anti-SP-G-K increased dramatically after trypsinization. This labeling was eliminated by preadsorption with authentic SP-G-K but not substance P or SP-G. Gel-permeation chromatography revealed SP-G-K-like immunoreactivity in fractions corresponding to considerably higher molecular weight than mature substance P. Collectively, these results support the hypothesis that substance P is synthesized from larger precursors and demonstrate that extended precursor forms are normally present in the axons and somata of neural systems that synthesize substance P.
Asunto(s)
Tronco Encefálico/análisis , Precursores de Proteínas/análisis , Médula Espinal/análisis , Sustancia P/análogos & derivados , Sustancia P/análisis , Animales , Tronco Encefálico/citología , Cromatografía Líquida de Alta Presión , Cricetinae , Sueros Inmunes , Masculino , Mesocricetus , Radioinmunoensayo , Médula Espinal/citologíaRESUMEN
The organization of intrinsic axonal projections of principal neurons in the main olfactory bulb (MOB) was studied in hamsters by using wheat germ agglutinin-horseradish peroxidase (WGA-HRP) and fluorescent dyes. Punctate injections of either WGA-HRP or fast blue (FB) that are restricted to small sectors on one side of the MOB produce comparably restricted fields of retrograde labeling on the opposite side. Label is found predominantly in superficially situated (middle and external) tufted cells that lie near and at the border between the external plexiform and glomerular layers. Few of the deeper middle tufted, internal tufted, or mitral cells and no external tufted cells that lie in the superficial two-thirds of the glomerular layer are labeled in regions remote to the injection site. Anterograde transport of WGA-HRP from the injection site labels axons that travel dorsally and ventrally in restricted bands through the internal plexiform layer and then terminate within this layer in the punctate sector on the opposite side that contains retrogradely labeled neurons. Such reciprocal projections between opposing regions of the medial and lateral sides of the MOB are found at all rostrocaudal and dorsoventral levels. When punctate injections of FB into the MOB are paired with restricted injections of a second fluorescent tracer (nuclear yellow or diamidino yellow dihydrochloride) into the appropriate sector of pars externa (pE) of the anterior olfactory nucleus, the punctate region of remote retrogradely labeled principal neurons is embedded within a topographically restricted longitudinal wedge of retrogradely labeled mitral and tufted cells that project extrinsically to or through pE. However, extremely few of these neurons are double-retrogradely labeled. The results reveal the existence of an intrabulbar associational system in which principal neurons engage in point-to-point, reciprocal projections between opposing regions of the medial and lateral MOB. Moreover, the results indicate that this associational system largely arises from superficially situated tufted cells distinct from those that support bulbofugal projections into the topographically organized interbulbar commissural system via pE.
Asunto(s)
Bulbo Olfatorio/anatomía & histología , Animales , Asociación , Axones , Cricetinae , Colorantes Fluorescentes , Peroxidasa de Rábano Silvestre , Lectinas , Masculino , Mesocricetus , Vías Nerviosas/anatomía & histología , Bulbo Olfatorio/citología , Bulbo Olfatorio/fisiología , Coloración y Etiquetado/métodos , Aglutininas del Germen de TrigoRESUMEN
The organization of connections between the main olfactory bulb (MOB) and pars externa (pE) of the anterior olfactory nucleus was studied in hamsters by using wheat germ agglutinin-horseradish peroxidase as both an anterograde and a retrograde neuronal tracer. Bulbar efferents of pE project exclusively to the contralateral MOB. A topographic organization is evident in these efferents, such that distinct sectors of pE project predominantly to certain sectors of the contralateral MOB and lightly to other sectors. The predominant projection to any bulbar sector in the coronal plane is repeated at nearly all rostral-caudal levels, i.e., the pE efferents to the contralateral MOB terminate within long strips or wedges that show a sector-to-sector topographic organization with respect to the medial-lateral and dorsal-ventral axes but not the rostral-caudal axis of the MOB. Afferents to pE arising in the ipsilateral MOB also show a sector-to-sector topographic organization. Injections into restricted sectors along the circumference of pE label all classes of output neurons (mitral cells and internal, middle, and external tufted cells) in restricted sectors of the ipsilateral MOB, and the sectors that have retrograde neuronal labeling are homotopic to those in the contralateral MOB that have dense anterograde terminal labeling. External tufted cells are not labeled and the other classes of MOB output neurons do not have prominent topographic patterns of labeling in cases with injections caudal to pE. The somata of external tufted cell that project to pE are predominantly in the deep part of the glomerular layer; most of the external tufted cells that lie more superficially in the glomerular layer do not appear to have projections extrinsic to the MOB. These results indicate that both the afferent and efferent connections of pE with the MOB are topographically organized and provide a short synaptic pathway between homotopic sectors of the two main olfactory bulbs.
Asunto(s)
Sistema Límbico/anatomía & histología , Bulbo Olfatorio/anatomía & histología , Animales , Mapeo Encefálico , Cricetinae , Masculino , Mesocricetus , Vías Nerviosas/anatomía & histología , Vías Olfatorias/anatomía & histologíaRESUMEN
We prepared a monoclonal antibody to microtubule-associated protein 1 (MAP 1), one of the two major high molecular weight MAP found in microtubules isolated from brain tissue. We found that MAP 1 can be resolved by SDS PAGE into three electrophoretic bands, which we have designated MAP 1A, MAP 1B, and MAP 1C in order of increasing electrophoretic mobility. Our antibody recognized exclusively MAP 1A, the most abundant and largest MAP 1 polypeptide. To determine the distribution of MAP 1A in nervous system tissues and cells, we examined tissue sections from rat brain and spinal cord, as well as primary cultures of newborn rat brain by immunofluorescence microscopy. Anti-MAP 1A stained white matter and gray matter regions, while a polyclonal anti-MAP 2 antibody previously prepared in this laboratory stained only gray matter. This confirmed our earlier biochemical results, which indicated that MAP 1 is more uniformly distributed in brain tissue than MAP 2 (Vallee, R.B., 1982, J. Cell Biol., 92:435-442). To determine the identity of cells and cellular processes immunoreactive with anti-MAP 1A, we examined a variety of brain and spinal cord regions. Fibrous staining of white matter by anti-MAP 1A was generally observed. This was due in part to immunoreactivity of axons, as judged by examination of axonal fiber tracts in the cerebral cortex and of large myelinated axons in the spinal cord and in spinal nerve roots. Cells with the morphology of oligodendrocytes were brightly labeled in white matter. Intense staining of Purkinje cell dendrites in the cerebellar cortex and of the apical dendrites of pyramidal cells in the cerebral cortex was observed. By double-labeling with antibodies to MAP 1A and MAP 2, the presence of both MAP in identical dendrites and neuronal perikarya was found. In primary brain cell cultures anti-MAP 2 stained predominantly cells of neuronal morphology. In contrast, anti-MAP 1A stained nearly all cells. Included among these were neurons, oligodendrocytes and astrocytes as determined by double-labeling with anti-MAP 1A in combination with antibody to MAP 2, myelin basic protein or glial fibrillary acidic protein, respectively. These results indicate that in contrast to MAP 2, which is specifically enriched in dendrites and perikarya of neurons, MAP 1A is widely distributed in the nervous system.
Asunto(s)
Proteínas del Tejido Nervioso/metabolismo , Proteínas/metabolismo , Animales , Anticuerpos Monoclonales , Axones/metabolismo , Encéfalo/metabolismo , Femenino , Técnica del Anticuerpo Fluorescente , Proteínas Asociadas a Microtúbulos , Oligodendroglía/metabolismo , Proteínas/inmunología , Ratas , Distribución TisularAsunto(s)
Sistema Nervioso Central/crecimiento & desarrollo , Neuroglía/fisiología , Bulbo Olfatorio/crecimiento & desarrollo , Vías Olfatorias/crecimiento & desarrollo , Envejecimiento , Animales , Animales Recién Nacidos , Transporte Axonal , Cricetinae , Peroxidasa de Rábano Silvestre , Mesocricetus , Neuroglía/citología , Bulbo Olfatorio/citología , Vías Olfatorias/citologíaRESUMEN
Discretely localized lesions were made in the amygdala to examine how specifically they might alter various measures of feeding behavior in male rats. Behavioral tests included spontaneous intake and body weight regulation, reactivity to saccharin and quinine solutions, conditioned taste aversion, the feeding response to food deprivation, the response to glucose gavage, and teh response to dietary amino acid imbalance. Lesions in virtually all regions of the amygdala disrupted feeding behavior in some respect, but alterations in specific tasks were associated only with highly circumscribed brain damage. Body weight regulation, spontaneous food and water intake, and the responses to glucose gavage and long-term food deprivation were not altered by lesions in the amygdala. The results provide evidence that, in the rat, the amygdala may play a greater role in appetite than in hunger or safety. In particular, amygdaloid nuclei may participate in maintaining a negative bias in the reactivity to all appetitive stimuli.
Asunto(s)
Amígdala del Cerebelo/fisiología , Apetito/fisiología , Hambre/fisiología , Saciedad/fisiología , Animales , Peso Corporal , Cuerpo Estriado/fisiología , Ingestión de Líquidos , Ingestión de Alimentos , Masculino , Ratas , Ratas Endogámicas , Núcleos Talámicos/fisiologíaAsunto(s)
Mapeo Encefálico/métodos , Encéfalo/fisiología , Modelos Neurológicos , Animales , Conducta Animal/fisiología , Circulación Cerebrovascular , Degeneración Nerviosa , Regeneración Nerviosa , Neurotransmisores/metabolismo , Investigación , Coloración y Etiquetado , Técnicas Estereotáxicas , Transmisión SinápticaRESUMEN
The availability and concentration of dietary protein was varied in an examination of the nature of the day-to-day intake of protein solutions by 60-day-old male rats. It was found that the rats consumed a remarkably constant absolute amount of protein each day, adjusting overall caloric intake to maintain protein at a roughly constant proportion of total calories. Factors such as the time of access to a protein source or the extent of prior experience with a protein source were seen to influence the overall constancy of protein intake, whereas daily shifts in preference between two available concentrations of protein did not interfere with such short-term control. The mechanism for this behavioral control is likely to be different from mechanisms mediating the conditioned response to dietary protein, as in the response to dietary amino acid imbalance.