J.R. Alonso

Nitric oxide synthase in the brain of a urodele amphibian (Pleurodeles waltl) and its relation to catecholaminergic neuronal structures

The neuronal structures with NADPH-diaphorase activity and nitric oxide synthase (NOS) immunoreactivity have been studied in the brain of the urodele amphibian Pleurodeles waltl by means of histochemical and immunocytochemical techniques. Both approaches resulted in the selective labeling of the same neurons and fiber tracts in the brain, except for the primary olfactory fibers that did not stain for NOS but were positive for NADPH-diaphorase. NOS-containing neurons were found in the olfactory bulbs, pallial regions, septum, caudal striatum, amygdala and preoptic area. Only a few diencephalic cells were labeled in the posterior tubercle and ventral hypothalamus. In the brainstem, abundant cells were labeled in the tectum, mesencephalic tegmentum and isthmic region. The most conspicuous cell population was found in the isthmic-pretrigeminal region. Particularly well stained cells were distributed throughout the rhombencephalon in areas related to the descending trigeminal tract, solitary tract, raphe nucleus and the mid-caudal reticular formation. In the cervical spinal cord, NOS-containing cells were present in the dorsal, intermediate and ventral grey fields. Cells in the preoptic, postotic and dorsal root ganglia were also labeled. Double labeling techniques revealed an extensive codistribution of neurons with NOS and catecholamines in the urodele brain but actual colocalization in the same cells was never observed. The organization of the central systems in urodeles with NOS appears to share many features not only with other anamniotes but also with amniotes.

Distribution of neuropeptide Y-like immunoreactive cell bodies and fibers in the brain stem of the cat

By using intratissue injections of colchicine and an indirect immunoperoxidase technique, we studied the distribution of cell bodies and fibers containing neuropeptide Y-like immunoreactivity in the brain stem of the cat. The densest clusters of immunoreactive perikarya were observed in the following nuclei: anteroventral cochlear, lateral reticular (internal and external divisions), dorsal tegmental, inferior colliculus and dorsal nucleus of the lateral lemniscus. By contrast, the nuclei abducens, the nucleus of the trapezoid body, preolivary, interpeduncularis, infratrigeminal, gigantocellular tegmental field, coeruleus and dorsal motor nucleus of the vagus had the lowest density. Finally, a moderate density of neuropeptide Y-like immunoreactive cell bodies was found in the nuclei: lateral tegmental field, laminar spinal trigeminal, praepositus hypoglossi, superior colliculus, lateral vestibular and motor trigeminal. In addition, a mapping of the neuropeptide Y-like immunoreactive fibers was carried out. Thus, the densest network of immunoreactive fibers was observed in the laminar spinal trigeminal nucleus. The nuclei periaqueductal gray, inferior central, praepositus hypoglossi, postpyramidal raphe, dorsal raphe, incertus and medial vestibular contained a moderate density of immunoreactive fibers, whereas the nuclei interpeduncularis, inferior colliculus, superior central, gracile, retrorubral, Kölliker-Fuse, dorsal tegmental, ambiguus and alaminar spinal trigeminal had the lowest density of neuropeptide Y-like immunoreactive fibers. The anatomical location of neuropeptide Y-like immunoreactivity suggests that the peptide could play an important role in several physiological functions, e.g., those involved in cardiovascular, auditory, motor, visual, nociceptive and somatosensory mechanisms.

Calretinin-like immunoreactivity in the optic tectum of the tench (Tinca tinca L.)

The distribution of calretinin-like immunopositive cells and fibers in the optic tectum of the tench (Tinca tinca) was studied by using a polyclonal antibody and the avidin-biotin-peroxidase technique. A clear laminated pattern of calretinin-like immunoreactivity was observed. The stratum periventriculare demonstrated a large number of strongly labeled cells whereas in the strata album centrale and griseum centrale, and at the boundary between the strata griseum centrale and fibrosum et griseum superficiale, some scarce, weakly immunostained cells were observed. No immunoreactive cells were seen in the strata fibrosum et griseum superficiale, opticum and marginale. Cells belonging to neuronal types X and XIV, previously characterized using Golgi impregnation, were found to be calretinin-like immunoreactive. Most calretinin-like immunopositive fibers were found in the strata fibrosum et griseum superficiale and opticum with a distribution pattern similar to retinotectal axons in these layers. In agreement with previous biochemical studies, our data suggest that, by contrast to all other classes of vertebrates, instead of calretinin and calbindin D-28k, only one protein is present in teleosts. Nevertheless, the calretinin-like immunostaining pattern in the teleost optic tectum was more complex than that previously described for calbindin D-28k. When compared to the calretinin-immunostaining in the rat superior colliculus, it is evident the presence in both amniotes and anamniotes of calretinin-immunopositive retinotectal axons. However, the distribution patterns of intrinsic calretinin-immunoreactive cells were different.(ABSTRACT TRUNCATED AT 250 WORDS)

Distribution of parvalbumin immunoreactivity in the rat septal area

The distribution of parvalbumin (PV)-containing neurons and processes in the septal area of the rat brain was studied using a monoclonal antibody and the avidin-biotin immunoperoxidase method. PV-immunoreactive neurons were mainly located in the medial septum/diagonal band complex and in the horizontal limb of the diagonal band of Broca, showing a high density of heavily immunostained neurons and fibers. Nonimmunoreactive cells surrounded by PV-positive cells and processes were observed in the same region, but no pericellular basket-like arrangements were found. On the contrary, the dorsal, intermediate, and ventral nuclei of the lateral septum were practically devoid of PV-positive neurons and processes. Thus, in these nuclei only a very low density of isolated neurons was labeled; these were specially scattered in the ventrolateral septal nucleus and in the dorsolateral septal nucleus just below the corpus callosum. Delicate PV-positive axonal plexuses were also observed in the dorsal and intermediate nuclei of the lateral septum. The immunopositive neurons displayed very different sizes and morphologies among the various septal nuclei and inside each of them, indicating that they do not belong to a single morphological class of neurons. Finally, the distribution of PV in the rat septal area is not directly related to cholinergic and GABAergic septal neurons.

Tyrosine hydroxylase immunoreactivity in a subpopulation of granule cells in the olfactory bulb of teleost fish

The distribution of tyrosine hydroxylase (TH)-like immunoreactivity in the olfactory bulb was studied in three species of teleosts, the tench Tinca tinca, the Mediterranean barbel Barbus meridionalis and the rainbow trout Salmo gairdneri, by using an indirect immunoperoxidase method. The antiserum used displayed a characteristic pattern of immunostaining in the three species, and four main conclusions can be drawn: (1) there is a large population of TH-like positive cell bodies and fibers in the olfactory bulb in fish, mainly in the granule cell and plexiform layers; (2) the immunolabeled cells are identified as granule cells, but only one group of granule cells is positive; (3) specific quantitative variations exist in the pattern of TH immunoreactivity, with use of the same fixative, antibody and localization method, among the three species studied, and (4) the pattern of TH immunoreactivity in the olfactory bulb in teleosts is completely different from that described previously in amniotes.

Immunocytochemical study of angiotensin-II fibres and cell bodies in the brainstem respiratory areas of the cat

The distribution of fibres and cell bodies containing angiotensin-II in brainstem respiratory nuclei was studied using an indirect immunoperoxidase technique. In order to visualize immunoreactive perikarya, intracerebral injections of colchicine were carried out. The richest cluster of immunoreactive perikarya was localized in the Bötzinger complex and Kölliker-Fuse areas, whereas in the nucleus ambiguus and nucleus retroambiguus a moderate density of cell bodies was observed. The nucleus tractus solitarius (ventrolateral portion) and the nucleus parabrachialis medialis had the lowest number of immunoreactive neurons. Angiotensin-II containing fibres were abundant in the nucleus parabrachialis medialis, Bötzinger complex and Kölliker-Fuse area, and scarce in the nuclei tractus solitarius (ventrolateral part), ambiguus and retroambiguus. Moreover, in the dorsal motor nucleus of the vagus a dense network of immunoreactive fibres and a large number of cell bodies were observed. The presence of angiotensin-II in respiratory areas suggests a role for this octapeptide in controlling respiration.

Segregated distribution of nitric oxide synthase-positive cells in the periglomerular region of typical and atypical olfactory glomeruli☆

Using double-staining techniques, the distribution of NADPH-diaphorase (ND)- and nitric oxide synthase (NOS)-positive cells was compared in the periglomerular region of typical and atypical rat olfactory glomeruli. Dorsomedial and ventrolateral atypical glomeruli contained similar number of ND/NOS-positive periglomerular cells. The number of ND/NOS-stained periglomerular cells was much higher (P < 0.001) in typical than in atypical glomeruli. The present results indicate that, in addition to described neurochemical and ultrastructural differences between the neuropile of typical and atypical glomeruli, there are also marked differences in the phenotype of periglomerular interneurons of both glomerular subsets.

Distribution of vasoactive intestinal polypeptide-like immunoreactivity in the olfactory bulb of the rainbow trout (Salmo gairdneri)

The distribution of vasoactive intestinal polypeptide-like structures in the olfactory bulb of the rainbow trout was studied using an indirect-immunoperoxidase technique. Olfactory fibres were very strongly labelled, whereas the fibres or cell bodies in the remaining strata of the olfactory bulb showed no immunoreactivity. In addition, the olfactory nerve fibres were not immunoreactive for methionine- and leucine-enkephalins, motilin, neuropeptide Y, substance P, cholecystokinin-8 and tyrosine-hydroxylase.

Calbindin D-28k immunoreactivity in the rat accessory olfactory bulb.

The distribution pattern and the morphology of calbindin D-28k-immunoreactive neurons were studied in the accessory olfactory bulb of the rat using a monoclonal antibody and the avidin-biotin-immunoperoxidase method. Positive neurons were observed in all layers but the vomeronasal nerve layer. Scarce mono-dendritic periglomerular neurons were calbindin D-28k-immunoreactive. Different morphological types of short-axon cells were calbindin D-28k-immunostained, with different degrees of intensity, in the boundary between the internal and external plexiform layer. In addition, deep short-axon cells located in the granule cell layer were calbindin D-28k-immunopositive. By contrast, previous studies described all cells in the rat accessory olfactory bulb as calbindin D-28k-immunonegative. The staining pattern in the rat accessory olfactory bulb showed both similarities and differences with the distribution pattern of the same calcium-binding protein in the main olfactory bulb.

Substance P-like immunoreactivity in the ganglion cells of the tench terminal nerve.

The distribution of substance P (SP) in the olfactory bulb of the tench Tinca tinca was studied by using an indirect immunoperoxidase technique. Many perikarya and processes of the ganglion cells of the nervus terminalis (NT) were strongly labeled. In addition, SP-like immunopositive fibers were observed in the proximity of these neurons and extending along the olfactory nerves and the olfactory tracts. The ganglion cells of the NT were not immunoreactive for methionine- and leucine-enkephalin, motilin, vasoactive intestinal polypeptide, neuropeptide Y, cholecystokinin-8, and tyrosine hydroxylase.