Distribution of proneuropeptide Y-derived peptides in the brain of Rana esculenta and Xenopus laevis

The distribution ofproneuropeptide Y-containing perikarya and nerve fibers in the brain of Rana esculenta and Xenopus lavis was determined with antisera directed toward neuropeptide Y and the carboxyl terminal flanking peptide. The distribution of proneuropeptide Y-like immunoreactivity was similar in both anurans. In the telencephalon, immunoreactive perikarya were found in the olfactory bulb, all subdivisions of the pallium, the septum, pars lateralis of the amygdala, the nucleus accumbens, and the anterior preoptic area. In the diencephalon, labelled perikarya were detected in the ventromedial, ventrolateral and central thalamic nuclei, the magnocellular preoptic nucleus, the suprachiasmatic nucleus, the posterior tuberculum, and the infundibulum. Amacrine-like cells were stained in the retina. In the pretectal area, posterior thalamic neurons showed intense, Golgi-like immunostaining. In the mesencephalon, immunoreactive cells were found in the reticular nucleus, the anteroventral tegmental nucleus, the optic tectum, the interpeduncular nucleus, and the torus semicircularis. In the rhombencephalon, labelled perikarya were detected in the secondary visceral nucleus, the central gray, the nucleus of the solitary tract, the dorsal column nuclei, and the spinal nucleus of the trigeminal nerve. Immunoreactive nerve fibers were observed in all areas of the brain that contained labelled perikarya. The densest accumulations were found in the accessory olfactory bulb, pars lateralis of the amygdala, the ventral habenula, the posterior pituitary, the optic tectum, the interpeduncular nucleus, and the saccular nucleus. The distribution of proneuropeptide Y-like immunoreactivity in the anuran brain showed many similarities to the distribution described for the amniote brain. © 1993 Wiley-Liss, Inc.     

Distribution of galanin-like immunoreactivity in the brain of Rana esculenta and Xenopus laevis.

The immunocytochemical distribution of galanin-containing perikarya and nerve terminals in the brain of Rana esculenta and Xenopus laevis was determined with antisera directed toward either porcine or rat galanin. The pattern of galanin-like immunoreactivity appeared to be identical with antisera directed toward either target antigen. The distribution of galanin-like immunoreactivity was similar in Rana esculenta and Xenopus laevis except for the absence of a distinct laminar distribution of immunoreactivity in the optic tectum of Xenopus laevis. Galanin-containing perikarya were located in all major subdivisions of the brain except the metencephalon. In the telencephalon, immunoreactive perikarya were detected in the pars medialis of the amygdala and the preoptic area. In the diencephalon, immunoreactive perikarya were detected in the caudal half of the suprachiasmatic nucleus, the nucleus of the periventricular organ, the ventral hypothalamus, and the median eminence. In the mesencephalon, immunoreactive perikarya were detected near the midline of the rostroventral tegmentum, in the torus semicircularis and, occasionally, in lamina A and layer 6 of the optic tectum. In the myelencephalon, labelled perikarya were detected only in the caudal half of the nucleus of the solitary tract. Immunoreactive nerve fibers of varying density were observed in all subdivisions of the brain with the densest accumulations of fibers occurring in the pars lateralis of the amygdala and the preoptic area. Dense accumulations of nerve fibers were also found in the lateral septum, the medial forebrain bundle, the periventricular region of the diencephalon, the ventral hypothalamus, the median eminence, the mesencephalic central gray, the laminar nucleus of the torus semicircularis, several laminae of the optic tectum, the interpeduncular nucleus, the isthmic nucleus, the central gray of the rhombencephalon, and the dorsolateral caudal medulla. The extensive system of galanin-containing perikarya and nerve fibers in the brain of representatives of two families of anurans showed many similarities to the distribution of galanin-containing perikarya and nerve fibers previously described for the mammalian brain.     

Distribution of dynorphin and enkephalin peptides in the rat brain

The neuroanatomical distribution of dynorphin B-like immunoreactivity (DYN-B) was studied in the adult male and female albino rat. The distribution of DYN B in colchicine- and noncolchicine-treated animals was also compared to that of another opioid peptide derived from the prodynorphin precursor dynorphin A (1-8) (DYN 1-8), and an opioid peptide derived from the proenkephalin precursor met-enkephalin-arg-gly-leu (MERGL). DYN B cell bodies were present in nonpyramidal cells of neo- and allocortices, medium-sized cells of the caudate-putamen, nucleus accumbens, lateral part of the central nucleus of the amygdala, bed nucleus of the stria terminalis, preoptic area, and in sectors of nearly every hypothalamic nucleus and area, medial pretectal area, and nucleus of the optic tract, periaqueductal gray, raphe nuclei, cuneiform nucleus, sagulum, retrorubral nucleus, peripeduncular nucleus, lateral terminal nucleus, pedunculopontine nucleus, mesencephalic trigeminal nucleus, parabigeminal nucleus, dorsal nucleus of the lateral lemniscus, lateral superior olivary nucleus, superior paraolivary nucleus, medial superior olivary nucleus, ventral nucleus of the trapezoid body, lateral dorsal tegmental nucleus, accessory trigeminal nucleus, solitary nucleus, nucleus ambiguus, paratrigeminal nucleus, area postrema, lateral reticular nucleus, and ventrolateral region of the reticular formation. Fiber systems are present that conform to many of the known output systems of these nuclei, including major descending pathways (e.g., striatonigral, striatopallidal, reticulospinal, hypothalamospinal pathways), short projection systems (e.g., mossy fibers in hippocampus, hypothalamo-hypophyseal pathways), and local circuit pathways (e.g., in cortex, hypothalamus). The distribution of MERGL was, with a few notable exceptions, in the same nuclei as DYN B. From these neuroanatomical data, it appears that the dynorphin and enkephalin peptides are strategically located in brain regions that regulate extrapyramidal motor function, cardiovascular and water balance systems, eating, sensory processing, and pain perception.     

Distribution of neuromedin U-like immunoreactivity in the central nervous system of Rana esculenta

The distribution of perikarya and nerve fibers containing neuromedin U-like immunoreactivity in the brain of Rana esculenta was determined with an antiserum directed toward the carboxyl terminus of the peptide. In the telencephalon, immunoreactive perikarya were found in the olfactory bulb, the medial septum, and the diagonal band. In the diencephalon, labeled perikarya were detected in the anterior and posterior preoptic areas, the dorsal nucleus of the hypothalamus, the caudal part of the infundibulum, and the posterior tuberculum. In the mesencephalon, immunoreactive cell bodies were found only in the laminar nucleus of the torus semicircularis and the anterodorsal tegmental nucleus. In the rhombencephalon, labeled perikarya were detected in the secondary visceral nucleus, the cerebellar nucleus, the central gray, and the nucleus of the solitary tract. Immunoreactive nerve fibers were observed in all areas of the brain that contained labeled perikarya. The densest accumulations were found in the nucleus accumbens; the dorsal part of the lateral septum; the periventricular region of the ventral thalamus; the lateral part of the infundibulum; the anterodorsal, anteroventral, posterodorsal, and posteroventral tegmental nuclei; and the periaqueductal region of the tegmentum. The distribution of neuromedin U-like immunoreactivity in the frog brain was substantially different from the distribution described for the rodent brain. © 1996 Wiley-Liss, Inc.     

Distribution of urocortin-like immunoreactivity in the central nervous system of the frog Rana esculenta

Corticotropin-releasing factor (CRF), sauvagine, and urotensin I are all members of the so-called CRF neuropeptide family. Urocortin (Ucn), a 40-amino-acid neuropeptide recently isolated from the rat brain, is the newest member of this family. Until now, the distribution of Ucn in the central nervous system (CNS) has been studied only in placental mammals. We used a polyclonal antiserum against rat Ucn to determine the distribution of Ucn-like immunoreactivity in the CNS of the green frog, Rana esculenta. The great majority of Ucn-immunoreactive perikarya was seen in the anterior preoptic area, ventromedial thalamic nucleus, posterior tuberculum, nucleus of the medial longitudinal fasciculus, and Edinger-Westphal nucleus. Urocortin-immunoreactive nerve cells were also observed in the motor nuclei of the trigeminal and facial nerves and in the hypoglossal nucleus. Immunoreactive fibers were found in the medial and lateral septal nuclei, bed nucleus of the stria terminalis, many of the thalamic and hypothalamic nuclei, mesencephalic tectum, tegmental nuclei, torus semicircularis, and dorsal horn and central field of the spinal cord. Only scattered Ucn-immunoreactive axon terminals were observed in the external zone of the medial eminence. The densest accumulations of Ucn-immunoreactive nerve terminals were seen in the granular layer of the cerebellum and cochlear nuclei. Our results suggest that an ortholog of mammalian Ucn occurs in the CNS of the green frog. The distribution of Ucn-like immunoreactivity in Rana esculenta showed many similarities to the distribution in placental mammals. The distribution of Ucn-like immunoreactivity in the anuran CNS was different from that of CRF and sauvagine, so our results suggest that at least three different lineages of the CRF neuropeptide family occur in the anuran CNS.     

Changes in the dynamic state of brain proenkephalin-derived peptides during amygdaloid kindling

The dynamic state of the proenkephalin (PE) gene products during and after development of amygdaloid kindling was assayed by monitoring changes of the accumulation of PE mRNA and changes in proenkephalin-related peptides. A parallel determination of PE mRNA and peptides from the same sample was conducted in this study. Electrical stimulation of the amygdala causes early increases in the PE mRNA content in that structure and in the hippocampus. Other areas related with the amygdaloid complex do not exhibit such an early increase, but this alteration occurs when the kindling process is fully established. Enkephalin content increases early in amygdala and hippocampus presumably owing to an increase in synthesis rate. Also, the enkephalin content of areas connected with amygdala and hippocampus such as the entorhinal cortex, the nucleus accumbens, and the frontal and occipital cortex exhibits an increase. A clear tendency towards normalization is observed after a recovery period of 2-3 months. Rekindling of the animals after this recovery period does not elicit a similar pattern of changes in the dynamic state of enkephalin system, even though the animals rekindle with just one single stimulation. The present data suggest that the enkephalinergic neurons participate in the development and spreading of kindling phenomena after amygdaloid stimulation, but they do not seem to play any role in mediating maintenance of the kindling state.     

The distribution of proenkephalin-derived peptides in the central nervous system of turtles

The present study was carried out to examine if peptides similar to the various opioid peptide products of mammalian proenkephalin are present in the turtle central nervous system and to determine their distribution. Antisera against several enkephalin peptides were used: leucine-enkephalin (LENK), methionine-enkephalin (MENK), methionine-enkephalin-arg6-phe7 (MERF), methionine-enkephalin-arg6-gly7-leu8 (MERGL), Peptide E (PEPE), and BAM22P. Their specificity and cross-reactivity were carefully examined. The results indicated that LENK, MENK, and MERF (or highly similar peptides) are present in the turtle central nervous system, and that a peptide showing immunological similarity to BAM22P and PEPE also appeared to be present. In contrast, MERGL did not appear to be present. The distributions of the immunoreactive labeling for LENK, MENK, MERF, BAM22P, and PEPE were indistinguishable, and double-label studies showed that LENK, MERF, and BAM22P were colocalized within individual neurons and fibers. Although all of the above substances were observed in the same cell groups, there was some regional variation, in terms of which enkephalin peptide appeared to be most abundant. The distributions of these enkephalin peptides were very similar to those previously described in mammals and birds. Enkephalin was more abundant in the basal ganglia than in overlying telencephalic regions. Within the basal ganglia, enkephalin was present in striatal neurons and fibers and in pallidal fibers, thereby suggesting the existence of an enkephalinergic striatopallidal projection. Sensory relay nuclei of the thalamus were generally poor in enkephalinergic fibers, whereas the hypothalamus was rich in enkephalinergic neurons and fibers. Enkephalinergic neurons and fibers were present in the midbrain central gray. As is true of neurons of the nucleus spiriformis lateralis of the avian pretectum, the neurons of the homologous cell group in turtles, the dorsal nucleus of the posterior commissure of the pretectum, were found to contain enkephalin and have an enkephalinergic projection to the deep layers of the ipsilateral tectum. Enkephalinergic neurons and fibers were also abundant in the entry zones of the trigeminal nerve and dorsal root fibers of the spinal cord.(ABSTRACT TRUNCATED AT 400 WORDS)     

Cholecystokinin peptides in the brain and pituitary of the bullfrog Rana catesbeiana: distribution and characterization

The distribution of CCK peptides in the bullfrog brain was determined with a CCK radioimmunoassay. Frog brain CCK distribution resembles rat porcine and human brain in that CCK concentration is moderate to high in hypothalamus, diencephalon, and medulla (3 18.4 ng/mg protein) and low in cerebellum (0.6 ng/mg protein). However, unlike all mammalian species examined, the CCK content of frog cerebral cortex, hippocampus and olfactory lobe is quite low (0.03 0.23 ng/mg protein). The elution of CCK-like peptides in frog brain extracts was determined on two HPLC systems. On both systems the bulk of the CCK-like material eluted with CCK 8 sulfate and separated from gastrin and other CCK peptides. These data suggest that though the chemical structure of CCK appears to be the same in the brains of frogs and mammals, the distribution of CCK in the brain appears to have shifted during the course of evolution, becoming a cortical, hippocampal, and olfactory system peptide only in more evolved organisms.     

Topological analysis of the brain stem of the frogs Rana esculenta and Rana catesbeiana.

The ventricular sulcal pattern and the cytoarchitectonic organization of the brain stem of the frogs Rana esculenta and Rana catesbeiana have been studied in transversely cut, Nissl stained serial sections. Four longitudinal sulci, the sulcus medianus inferior, the sulcus intermedius ventralis, the sulcus limitans and the sulcus medianus superior could be distinguished in both species. A fifth longitudinal groove, the sulcus intermedius dorsalis, was found only in Rana esculenta. With the aid of the usual cytoarchitectonic criteria 25 cell masses have been delineated in Rana esculenta and 27 in Rana catesbeiana. These cell masses can be distributed over the following categories (numbers added in brackets for Rana catesbeiana, if different from those in Rana esculenta): primary efferent or motor, 8; primary afferent or sensory, 4(6); "relay" centers, 7. Contrary to statements in the literature the reticular formation can be divided into six separate cell groups. The majority of the nuclei form part of the central gray, which constitutes a rather wide zone in anurans; three reticular nuclei lie partly within the stratum griseum and partly within the stratum album; six nuclei are entirely embedded in the stratum album. The morphological pattern of the cell masses and their relationship to the ventricular sulci were studied with the aid of a graphical reconstruction procedure termed topological analysis (cf. Nieuwenhuys, '74 and figs. 15, 16). This analysis yielded the following results: The sulcus limitans extends throughout the rhombencephalon, dividing this brain part into a basal plate and an alar plate. The cell masses in the basal plate fit into two longitudinal zones, a medial area ventralis and a lateral area intermedioventralis. The area ventralis contains three somatic motor nuclei (IV, VI and XII) and the rhombencephalic medial reticular zone. The latter may be primarily considered as a somatic motor coordinating center. The area intermedioventralis contains the visceral motor nuclei of V, VII, IX and X. However, the basal plate also contains a number of non-motor centers, for example the superior olive. The alar plate contains visceral sensory, general somatic sensory and special somatic sensory centers. Two cell masses, the nucleusfasciculi solitarii and the nucleus visceralis secundarius, represent together a discontinuous visceral sensory zone. Both of these nuclei are situated immediately dorsal to the sulcus limitans. The special somatic sensory area, i.e., the area of termination of the eighth nerve, occupies a considerable part of the alar plate. This area comprises, apart from a large zone of diffuse gray, three distinct cell masses...     

Melanin-concentrating hormone system in the brain of the lungfish Protopterus annectens

The neurochemical anatomy of the lungfish brain is of particular interest, because many features in these animals might be representative of the common ancestor of land vertebrates. In the present study, we have investigated the localization and biochemical characteristics of melanin-concentrating hormone (MCH)-immunoreactive material in the central nervous system of the African lungfish, Protopterus annectens. The most prominent group of MCH-immunoreactive cell bodies was found in the dorsal hypothalamus. Additional groups of MCH-immunoreactive perikarya were detected in the telencephalon within the medial and dorsal pallium, the medial subpallium, and the ventral part of the lateral subpallium. Brightly immunofluorescent nerve fibers were seen in the anterior olfactory nucleus, the ventral part of the medial pallium, the medial subpallium, and the anterior preoptic area. In the diencephalon, the hypothalamus and the medial region of the dorsal thalamus exhibited a dense accumulation of fibers. MCH-immunoreactive fibers were also found in the tectum and the tegmentum of the mesencephalon and within the reticular formation of the rhombencephalon. In the pituitary, several small groups of cells of the intermediate lobe showed a bright fluorescence. Reversed-phase high-performance liquid chromatography (HPLC) analysis of diencephalon and pituitary extracts resolved a major MCH-immunoreactive peak that coeluted with synthetic salmon MCH. The distribution of MCH in the brain of P. annectens suggests that, in lungfishes, this peptide may exert neuromodulator or neurotransmitter functions. The presence of MCH-like immunoreactivity in the intermediate lobe of the pituitary indicates that, in dipnoans, MCH may also act as a typical pituitary hormone. J. Comp. Neurol. 390:41–51, 1998. © 1998 Wiley-Liss, Inc.     

Distribution of melanin-concentrating hormone-like immunoreactivity in the central nervous system of Rana esculenta

The distribution of salmon and rat melanin-concentrating hormone (MCH)-like and neuropeptide glutamate-isoleucine (NEI)-like immunoreactivity in the brain and spinal cord of the frog Rana esculenta was studied with immunohistochemistry. In the telencephalon, only fibers showed immunoreactivity in the olfactory bulb, lateral pallium, diagonal band, septum, and the amygdala. Immunoreactive fibers were abundant in all diencephalic structures, except the optic tract, the visual neuropils, and the habenula. Several cells in the central thalamic nucleus and a few in the suprachiasmatic nucleus were stained with the MCH antisera. Cells and their processes were intensely stained in the dorsal hypothalamus with the MCH and NEI antisera. Immunoreactive fibers were found in all tegmental nuclei and the white matter of the mesencephalon. They formed terminal plexuses in the deep layers of the optic tectum and the laminar nucleus of the torus semicircularis. Immunoreactive fibers were sparse in the rhombencephalon and the spinal cord.     

Distribution of vasoactive intestinal peptide-like immunoreactivity in the brain and pituitary of the frog (Rana esculenta) during development

The localization of vasoactive intestinal peptide (VIP)-like immunoreactive (ir) elements was investigated in the brain of the anuran amphibian, Rana esculenta, during development. Using an antiserum raised against the porcine VIP, ir cell bodies and fibers were observed in the forebrain of tadpoles a few days after hatching. During early premetamorphosis, ir perikarya were distributed in the ventral infundibular nucleus of the hypothalamus and in the posterocentral nucleus of the thalamus. Labeled fibers were detected in the olfactory bulbs and in the hypothalamus. In these larvae, furthermore, several VIP-ir cells were found in the pars distalis of the pituitary and there were ir fibers in the pars nervosa. In tadpoles at stages VIII-IX, a new group of VIP-labeled neurons was observed in the dorsal part of the infundibular nucleus. In other brain regions, the distribution of the immunoreactivity was similar to that described in the earliest stages, i.e., IV-VII. During mid-premetamorphosis, stages X-XII of development, an additional set of ir perikarya appeared in the ventrolateral area of the thalamus. During late premetamorphosis, stages XIII-XVIII, the organization of VIP-like immunoreactivity was more complex and its distribution more widespread. Two new groups of ir cell bodies appeared, one in the preoptic nucleus and another in the anteroventral area of the thalamus, and for the first time, VIP immunoreactivity was observed in the median eminence. This distribution pattern persisted through to the prometamorphic, four-limb stage. Strikingly, no VIP-ir elements were observed anywhere in the mid- and hindbrain. The present results indicate that a VIP-like ir peptide may be involved in the processing of olfactory information or may act as a neurohormone, hypophysiotropic factor, and neuromodulator in the brain of R. esculenta during development.     

Distribution of choline acetyltransferase immunoreactivity in the pigeon brain

We have investigated the distribution of cholinergic perikarya and fibers in the brain of the pigeon (Columba livia). With this aim, pigeon brain sections were processed immunohistochemi-cally by using an antiserum specific for chicken choline acetyltransferase. Our results show cholinergic neurons in the pigeon basal telencephalon, the hypothalamus, the habenula, the pretectum, the midbrain tectum, the dorsal isthmus, the isthmic tegmentum, and the cranial nerve motor nuclei. Cholinergic fibers were prominent in the dorsal telencephalon, the striatum, the thalamus, the tectum, and the interpeduncular nucleus. Comparison of our results with previous studies in birds suggests some major cholinergic pathways in the avian brain and clarifies the possible origin of the cholinergic innervation of some parts of the avian brain. In addition, comparison of our results in birds with those in other vertebrate species shows that the organization of the cholinergic systems in many regions of the avian brain (such as the basal forebrain, the epithalamus, the isthmus, and the hindbrain) is much like that in reptiles and mammals. In contrast, however, birds appear largely to lack intrinsic cholinergic neurons in the dorsal (“neocortex-like”) parts of the telencephalon. © 1994 Wiley-Liss, Inc.     

Distribution of pro-opiomelanocortin-derived peptides and enkephalins in the rat central nucleus of the amygdala

The distribution and origin of beta-endorphin (BE) and alpha-melanocyte stimulating hormone (alpha-MSH) terminals was studied in the central nucleus of the rat amygdala (CNA). The distributions of BE and alpha-MSH within the CNA were compared to that of Met- and Leu-enkephalin (ENK). BE and a-MSH terminals were found mainly in the medial subdivision of the CNA, and originated, at least in part, from the arcuate nucleus region of the hypothalamus. ENK terminals were found in both the medial and lateral subdivisions of the CNA, although a considerably higher density of terminals was seen within the lateral subdivision. This differential distribution of proopiomelanocortins and ENKs may reflect different functions for these endogenous opiates in the CNA.     

Distribution and origin of serotoninergic afferents to guinea pig cochlear nucleus

The distribution of serotoninergic fibers in the guinea pig cochlear nucleus was studied with serotonin immunohistochemistry. In addition, the origin of the serotoninergic fibers was determined by combining the retrograde transport of wheat germ agglutinin-apohorseradish peroxidase (gold conjugated) with serotonin immunohistochemistry. Immunoreactivity was present in varicose and nonvaricose fibers that were unevenly distributed throughout the cochlear nucleus. The fibers were most prominent in the superficial layers of the dorsal cochlear nucleus and the anterior spherical cell area of the anteroventral cochlear nucleus. Although less prominent, serotonin-positive fibers were also present in the remaining part of the anteroventral cochlear nucleus and the posteroventral cochlear nucleus. A few positive fibers were present in the auditory nerve root and the dorsal and intermediate acoustic stiae. Double-labeled cells were found throughout the rostral- caudal extent of the serotoninergic system from the caudal linear nucleus to the nucleus raphe pallidus. However, most were confined to the dorsal (52%) and median (18%) raphe nuclei. Some serotoninergic cell groups contained retrogradely labeled cells that were not serotonin immunoreactive, indicating nonauditory afferents to cochlear nucleus containing other neurotransmitter substances. Serotonin may tonically modulate auditory processing within the cochlear nucleus as well as influence certain ascending auditory pathways. Most of the serotonin in the cochlear nucleus comes from superior raphe nuclei that also project to basal ganglia motor systems and limbic strctures. Therefore, the effect of serotonin on the cochlear nucleus may be related to level of arousal or behavioral state. © 1995 Willy-Liss, Inc.     

Distribution of somatostatin receptors in the brain of the frog Rana ridibunda: correlation with the localization of somatostatin-containing neurons

The biochemical characterization and anatomical distribution of somatostatin binding sites were examined in the brain of the frog Rana ridibunda, and the distribution of the receptors was compared with the location of somatostatin immunoreactive neurons. The pharmacological profile of somatostatin receptors was determined in the frog brain by means of an iodinated superagonist of somatostatin, [125I-Tyr0,DTrp8]S-14. Membrane-enriched preparations from frog brain homogenates were shown to contain high-affinity receptors (KD = 0.78 +/- 0.34 nM; Bmax = 103 + 12.7 fmoles/mg protein) with pharmacological specificity for [DTrp] substituted S14 and S28 analogs. The distribution of somatostatin-binding sites was studied by autoradiography on coronal sections of frog brain. Various densities of somatostatin receptors were detected in discrete areas of the brain. The highest concentration of binding sites was observed in the olfactory bulb, in the pallium, and in the superficial tectum. Moderate binding was observed in the striatum, amygdaloid complex, preoptic area, and cerebellum. Immunocytochemical studies of the distribution of somatostatin-28 (S28) related peptides were also conducted in the frog brain. Two antisera that recognize distinct epitopes of the somatostatin molecule have been used for immunohistochemical mapping of the peptide. Antiserum SS9 recognizes both S28 and somatostatin-14 (S14) and allowed the labelling of perikarya. Antiserum S320 recognizes the N-terminal fragment (1-12) resulting from enzymatic cleavage of S28. This latter antiserum, which does not cross-react with S28, stained mainly neuronal processes. At the infundibular level, however, both antisera stained cell bodies and fibers. Immunoreactive somatostatin-related peptides were detected in many areas of the frog brain. In the diencephalon, a heavy accumulation of perikarya and fibers was seen in the preoptic nucleus, the dorsal and ventral infundibular nuclei, and the median eminence. Immunoreactive perikarya were also observed in the telencephalon, especially in the pallium and in thalamic nuclei. Immunostained processes were detected in many telencephalic areas and in the tectum. There was good correlation between the distribution of somatostatin-immunoreactive elements and the location of somatostatin-binding sites in several areas of the brain, in particular in the median pallium, the tectum, and the interpeduncular nucleus.(ABSTRACT TRUNCATED AT 400 WORDS)     

Differential changes in the cellular composition of the developing marsupial brain

Throughout development both the body and the brain change at remarkable rates. Specifically, the number of cells in the brain undergoes dramatic nonlinear changes, first exponentially increasing in cell number and then decreasing in cell number. Different cell types, such as neurons and glia, undergo these changes at different stages of development. The current investigation used the isotropic fractionator method to examine the changes in cellular composition at multiple developmental milestones in the short‐tailed opossum, Monodelphis domestica. Here we report several novel findings concerning marsupial brain development and organization. First, during the later stages of neurogenesis (P18), neurons make up most of the cells in the neocortex, although the total number of neurons remains the same throughout the life span. In contrast, in the subcortical regions, the number of neurons decreases dramatically after P18, and a converse relationship is observed for nonneuronal cells. In the cerebellum, the total number of cells gradually increases until P180 and then remains constant, and then the number of neurons is consistent across the developmental ages examined. For the three major structures examined, neuronal density and the percentage of neurons within a structure are highest during neurogenesis and then decrease after this point. Finally, the total number of neurons in the opossum brain is relatively low compared with other small‐brained mammals such as mice. The relatively low number of neurons and correspondingly high number of nonneurons suggests that in the marsupial brain nonneurons may play a significant role in signal processing. J. Comp. Neurol. 521:2602–2620, 2013. © 2013 Wiley Periodicals, Inc.     

Ultrastructural analysis and cholinesterase activity in the brain capillaries of Bufo bufo and Rana esculenta

We have investigated the structure and the cholinesterase features of the brain capillaries in two adult Amphibians (Rana esculenta and Bufo bufo). We found that brain capillaries are un-fenestrated and the endothelial cell edges are joined by tight junctions. The brain capillaries in both species are characterized by high levels of AChE. This enzyme is only localized in the basal membrane, whereas we have found the reaction product neither in the endoplasmatic reticulum nor in the Golgi apparatus. On the contrary the brain capillaries are deprived of BuChE. Only in one experiment traces of reaction product were found. The significance of this datum and the non-nervous role of cholinesterase is discussed.