Immimohistochemical localization of corticotropin-releasing factor in selected brain areas of the European starling (Sturnus vulgaris) and the song sparrow (Melospiza melodia)

Summary Corticotropin-releasing factor (CRF) was localized in the brains of two passerine species, the European starling (Sturnus vulgaris) and the song sparrow (Melospiza melodia), by means of immunohistochemistry. The hypothalamic distribution of this peptide in these species includes a complex of immunoreactive perikarya observed in the paraventricular nucleus (PVN), in both its medial and lateral divisions. Nerve fibers were also seen running from these areas to the anterior median eminence (AME) where a terminal field is apparent. A wide variety of extra-hypothalamic nuclei containing CRF-immunoreactive cells and fibers were identified. An apparent CRF terminal field can be visualized in the lateral septum. A dense fiber plexus is present in the nucleus accumbens (Ac) and more caudally in the nucleus of the stria terminalis (nST). In colchicinepretreated animals, it was revealed that these areas also contain CRF-stained perikarya. The pattern of CRF immunoreactivity in the Ac-nST complex is continuous, with no distinction apparent between the nuclei. The medial preoptic area (mPOA) and the adjacent diagonal band of Broca contain CRF-fibers, while cells are apparent in the mPOA. In the mesencephalon, cells were visualized in the midbrain central gray; a terminal field and scattered positively stained perikarya were found in areas more ventral to the central grey that are adjacent to the third cranial nerve. Scattered cells were also seen at the border of the nucleus intercollicularis-nucleus mesencephalicus lateralis, pars dorsalis complex. In contrast to mammalian studies, no immunoreactive nerve fibers or perikarya were observed in telencephalic areas homologous to the mammalian neocortex. These studies confirm the presence of a CRF path-way regulating pituitary function and suggest a broad role played by CRF as a neuromodulator or neurotransmitter in autonomic and possibly behavioral activities in these species.     

Localization of corticotropin-releasing factor immunoreactivity in the brain of the teleost Sparus aurata

The distribution of perikarya and fibers containing corticotropin-releasing factor (CRF) was studied in the brain of the teleost Sparus aurata by immunocytochemistry using the peroxidase-antiperoxidase method. Antisera against rat CRF, arginine vasotocin, and human adrenocorticotropin (ACTH) were used. Most CRF-immunoreactive neurons were located in the nucleus lateralis tuberis, but they were absent from the nucleus preopticus, which only contained arginine vasotocin neurons. Few CRF perikarya were identified in the nucleus preopticus periventricularis and in the mesencephalic tegmentum. A conspicuous bundle of immunoreactive fibers ran along the diencephalic floor and pituitary stalk to end near the cells of the hypophysial pars intermedia. No CRF was seen near the adenohypophysial rostral pars distalis. Our results suggest that, in Sparus aurata, CRF is a releasing factor for melanotropic cells. Its role as a releasing factor for ACTH is discussed.     

Localization of pigment-dispersing hormone (PDH) immunoreactivity in the central nervous system of Carcinus maenas and Orconectes limosus (Crustacea), with reference to FMRFamide immunoreactivity in O. limosus

Summary By use of antisera raised against synthetic pigment-dispersing hormone (PDH) of Uca pugilator and FMRFamide, the distribution of immunoreactive structures in the central nervous system (CNS) of Carcinus maenas and Orconectes limosus was studied by light microscopy. In both species, a total of 10–12 PDH-positive perikarya occur amongst the anterior medial, dorsal lateral and angular somata of the cerebral ganglion (CG). In C. maenas, one PDH-perikaryon was found in each commissural ganglion (COG) and several more in the thoracic ganglion. In O. limosus, only four immunopositive perikarya could be demonstrated in the ventral nerve cord, i.e., two somata in the anterior and two in the posterior region of the suboesophageal ganglion (SOG). PDH-immunoreactive tracts and fiber plexuses were present in all central ganglia of both species, and individual axons were observed in the connectives. FMRFamide-immunoreactivity was studied in O. limosus only. Neurons of different morphological types were found throughout the entire CNS, including numerous perikarya in the anterior medial, anterior olfactory, dorsal lateral and posterior cell groups of the CG. Four perikarya were found in the COG, six large and numerous smaller ones in the SOG, and up to eight cells in each of the thoracic and abdominal ganglia. In each ganglion, the perikarya form fiber plexuses. Axons from neurons belonging to the CG could be traced into the ventral nerve cord; nerve fibers arising from perikarya in the SOG appeared to project to the posterior ganglia. In none of the structures examined colocalization of PDH- and FMRF-amide-immunoreactivity was observed.Dedicated to Prof. K.-E. Wohlfarth-Bottermann on the occasion of his 65th birthday     

Serotonin-like immunoreactivity in the central nervous system and gonad of the scallop,Patinopecten yessoensis

Summary The localization of neurons containing serotonin in the central nervous system and the gonad of the scallop, Patinopecten yessoensis, was examined immunohistochemically. In the central nervous system a large number of immunoreactive perikarya were observed in the following regions: a part of the anterior lobe of the cerebral ganglion; the posterior lobe of the cerebral ganglion; the pedal ganglion; and the accessory ganglion. No immunoreactive perikarya were found in the visceral ganglion. Numerous immunoreactive fibers were revealed in the neuropil of all central ganglia. In the gonadal region immunoreactive fibers were distributed around the gonoduct and along the germinal epithelium.This work was supported by a grant from the Ministry of Education, Science and Culture, Japan     

The distribution of corticotropin-releasing factor-immunoreactive neurons and nerve fibers in the brain of the snake,Natrix maura

Summary The anatomical distribution of neurons and nerve fibers containing corticotropin-releasing factor (CRF) has been studied in the brain of the snake, Natrix maura, by means of immunocytochemistry using an antiserum against rat CRF. To test the possible coexistence of CRF with the neurohypophysial peptides arginine vasotocin (AVT) and mesotocin (MST) adjacent sections were stained with antisera against the two latter peptides. CRF-immunoreactive (CRF-IR) neurons exist in the paraventricular nucleus (PVN). In some neurons of the PVN, coexistence of CRF with MST or of CRF with AVT has been shown. Numerous CRF-IR fibers run along the hypothalamo-hypophysial tract and end in the outer layer of the median eminence. In addition, some fibers reach the neural lobe of the hypophysis. CRF-IR perikarya have also been identified in the following locations: dorsal cortex, nucleus accumbens, amygdala, subfornical organ, lamina terminalis, nucleus of the paraventricular organ, nucleus of the oculomotor nerve, nucleus of the trigeminal nerve, and reticular formation. In addition to all these locations CRF-IR fibers were also observed in the lateral septum, supraoptic nucleus, habenula, lateral forebrain bundle, paraventricular organ, hypothalamic ventromedial nucleus, raphe and interpeduncular nuclei.     

Immunohistochemical localization of urotensin I and other neuropeptides in the caudal neurosecretory system of three species of teleosts and two species of elasmobranchs

Summary In three species of teleosts — carp Cyprinus carpio; grass carp Ctenopharyngodon idella; and crucian carp Carassius auratus — the caudal neurosecretory system displays small, medium-sized and large neurons. Urotensin I (UI)-immunoreactive and UI-nonreactive neurons were found in all three groups; in general, the number of the latter neurons exceeded that of the former. Noteworthy are: (i) UI-immunoreactive fibers in the caudal spinal cord and (ii) dense accumulations of UI-immunoreactive product around the capillaries of the urophysis. In two species of elasmobranchs — cat shark Heterodontus japonicas and swell shark Cephaloscyllium umbratile — neurosecretory neurons decreased in size in rostro-caudal direction. Most of the neurosecretory perikarya, their axons and the corresponding neurohemal areas were UI-immunoreactive, but a small number of secretory neurons was devoid of immunoreaction. Oxytocin, arginine vasopressin, substance P, somatostatin, neurotensin, vasoactive intestinal polypeptide and gastrin-releasing peptide were not detected in the caudal neurosecretory system of the carp.     

Localization of growth hormone-releasing factor-like immunoreactivity in the hypothalamo-hypophysial system of some teleost species

Summary An antiserum to growth hormone-releasing factor (GRF) 1-44 was applied on brain and pituitary sections of nine teleost species. Immunoreactive (ir) perikarya were demonstrated in parvo- and magnocellular portions of the preoptic nucleus (PON) and occasionally in the nucleus lateralis tuberis. The two tracts originating in the PON ran ventro-laterally toward the optic chiasm and then caudally in the basal hypothalamus. In the pars distalis (PD) of the eel, carp, goldfish and salmonids, GRF-ir fibers did not enter the rostral PD and few fibers passed close to somatotropes. In.Myoxocephalus andMugil, a variable number of ir-fibers passed close to cells of the rostral and proximal PD. In the neurointermediate lobe, GRF-ir fibers were located exclusively in the neural tissue of the eel and trout. In goldfish, carp andMyoxocephalus, GRF-ir fibers entered the intermediate lobe. This antiserum also labeled corticotrops and, to a lesser extent, melanotrops in the pituitary of cyprinids. A variable number of perikarya contained both GRF and vasotocin in the PON of the eel. In all teleost species studied so far, the distribution patterns of GRF are different, and the function of the various adenohypophysial cell types appears to be differently modulated, according to the variable distribution of GRF in the pituitary.     

Immunocytochemical distribution of corticotropin-releasing factor in the brain and hypophysis of larval Bufo arenarum; effect of KClO4 during early development

The maturation of the corticotropin-releasing factor (CRF) neuronal system was evaluated by immunocytochemistry and morphometry in Bufo arenarum, during spontaneous metamorphosis and in tadpoles with inhibited thryroid function. The first appearance of CRF immunoreactive fibers was at the end of premetamorphosis (stage VIII). These fibers were found in small numbers and weakly stained in the median eminence and infundibular stalk. With the advance of larval development, CRF-like material stained intensely and tended to aggregate in the external zone of the median eminence. At climax stages, immunoreactive fibers and perikarya (weakly stained) were identified in the interpeduncular nucleus and in the dorsal infundibular nucleus. Morphometric and immunocytochemical results indicate that the maturation of the CRF neuronal system in Bufo arenarum occurs just before metamorphic climax, coinciding with high levels of thyroid and steroid hormones. We have also found that in larvae with inhibited thyroid function, the CRF neuronal system is able to develop, and that thyroid hormone could exert a negative feedback control on the synthesis of CRF.     

CRF-immunoreactive nerve fibers in the circumventricular organs of the monkey,Macaca fuscata

Summary The occurrence of CRF (corticotropin-releasing factor)-immunoreactive nerve fibers in the circumventricular organs of adult male monkeys, Macaca fuscata, was studied on serially sectioned brains, by means of the peroxidase-antiperoxidase technique in combination with a highly specific and sensitive CRF antiserum. CRF-containing nerve fibers were found in high concentrations in the infundibulum and, in addition, in small numbers in the posterior lobe, organum vasculosum laminae terminalis, subfornical organ, and area postrema; they were missing in the pineal body and the subcommissural organ. The CRF immunoreactive nerve fibers distributed in these organs were located in the proximity of the blood vessels.Supported by a grant (No. 56440022) from the Ministry of Education, Science, and Culture, Japan     

Immuno-enzyme cytochemical demonstration of mesotocinergic nerve fibres in the pars intermedia of the amphibian hypophysis

Summary Immuno-enzyme cytochemical investigations showed that the whole amphibian pars intermedia of the hypophysis is innervated by an intercellular network of peptidergic varicose nerve fibres which contain mesotocin or (and) parts of the mesotocin molecule. The pars intermedia does not contain vasotocinergic fibres. The mesotocinergic fibres are branches of axons leaving the pituitary stalk and the neural lobe. In animals of which the hypothalamic magnocellular neurosecretory preoptic nuclei had been completely removed, the immuno-reactive mesotocinergic fibres of the pars intermedia had totally disappeared. From this result, it is concluded that the mesotocinergic fibres of the pars intermedia of the amphibian hypophysis are axons of neurosecretory perikarya located in the hypothalamic magnocellular neurosecretory preoptic nuclei.Dedicated to Professor Berta Scharrer on the accasion of her 70th birthdayThis investigation was supported by a grant from the Belgian Nationaal Fonds voor Geneeskundig Wetenschappelijk Onderzoek     

Monoamine-containing structures in the intestines of bivalve molluscs and a polychaete annelid revealed by fluorescence histochemistry

Summary Monoamine-containing elements in the intestines of Bivalvia and Polychaeta species have been found by use of histochemical fluorescence methods according to Falck and Furness. Catecholamine-containing perikarya and fibers are seen within the epithelium and subepithelial layers of the midgut of the bivalves Mytilus edulis, Mya arenaria, Arctica islandica, as well as the polychaete Harmothoe imbricata. In addition, intraepithelial cell bodies and fibers containing serotonin-like substance are present in Mytilus edulis. Results obtained with the Furness method, applied earlier to vertebrates, correlate with those obtained with the Falck method.     

Localization of corticotropin-releasing hormone (CRF)-like immunoreactivity in the central nervous system of the elasmobranch fish,Scyliorhinus canicula

Summary The occurrence and localization of immunoreactive corticotropin-releasing factor (CRF) in the brain and pituitary of the elasmobranch fish Scyliorhinus canicula, were studied by means of specific radioimmunoassay and immunohistochemistry using the indirect immunofluorescence method. Brain and pituitary extracts showed a good cross-reactivity with the ovine CRF antiserum, but serial dilutions of tissue samples did not completely parallel the standard curve. Relatively high concentrations of CRF-like material were found within the pituitary, diencephalon, and telencephalon. CRF-like immunoreactive perikarya were observed in the preoptic nucleus and in the nucleus lateralis tuberis. Numerous immunoreactive cells appeared to be of the CSF-contacting type. CRF-like immunopositive fibers were seen to run through the hypothalamus within the ventro-medial floor of the infundibular region. A dense plexus of immunoreactive nerve endings terminated in the median eminence and the neurointermediate lobe of the pituitary. These results indicate that a neurosecretory system containing CRF-like immunoreactivity exists in the brain of elasmobranchs, a group of vertebrates which has diverged early from the evolutionary line leading to mammals. In addition, our data support the notion that a CRF-like molecule is involved in the regulation of corticotropic and melanotropic cell activity in this primitive species of fish.     

The Lethal Fungus Batrachochytrium dendrobatidis Is Present in Lowland Tropical Forests of Far Eastern Panamá

The fungal disease chytridiomycosis, caused by Batrachochytrium dendrobatidis (Bd), is one of the main causes of amphibian population declines and extinctions all over the world. In the Neotropics, this fungal disease has caused catastrophic declines in the highlands as it has spread throughout Central America down to Panamá. In this study, we determined the prevalence and intensity of Bd infection in three species of frogs in one highland and four lowland tropical forests, including two lowland regions in eastern Panamá in which the pathogen had not been detected previously. Bd was present in all the sites sampled with a prevalence ranging from 15–34%, similar to other Neotropical lowland sites. The intensity of Bd infection on individual frogs was low, ranging from average values of 0.11–24 zoospore equivalents per site. Our work indicates that Bd is present in anuran communities in lowland Panamá, including the Darién province, and that the intensity of the infection may vary among species from different habitats and with different life histories. The population-level consequences of Bd infection in amphibian communities from the lowlands remain to be determined. Detailed studies of amphibian species from the lowlands will be essential to determine the reason why these species are persisting despite the presence of the pathogen.     

Factors determining the structure and diversity of parasitoid complexes in tephritid fruit flies

Summary The relative importance of phylogenetic affinity of hosts versus their ecological characteristics in determining the composition of their parasitoid complexes was examined using the parasitoid complexes of six species of frugivorous fruit flies from Central Europe. The hosts were four Rhagoletis and two other trypetine species, ranging in their relatedness from host races to members of different genera. They also differed in ecological characteristics, utilizing host plants of three different families, and developing either as pulp- or seedfeeders inside the host fruit. These features made it feasible to test the following pair of hypotheses. The ecological hypothesis predicts that ecological traits such as host-plant and fruit fly phenologies and host-fruit texture should be more important for the composition of parasitoid complexes than the taxonomic relatedness of the fly species. Assuming that ecological relationships do not parallel phylogenetic ones, the alternative phylogenetic hypothesis predicts the opposite. In fruit and soil samples, taken between 1983 and 1989, three guilds of parasitoids comprising 20 species were found: guild 1 — koinobiotic larval parasitoids (e.g. Opius spp., which attack the host larvae but develop inside the host puparia); guild 2 — idiobiotic larval parasitoids (e.g. Pteromalus spp., which consume the host larvae at once); and guild 3 — idiobiotic puparium parasitoids (e.g. Phygadeuon spp.). Although some results support the phylogenetic hypothesis, the majority of results support the ecological hypothesis.     

Sexual dimorphism in neuronal projections from the antennae of silk moths (Bombyx mori,Antheraea polyphemus) and the gypsy moth (Lymantria dispar)

Summary The antennal lobe of both sexes of the silk moth Bombyx mori contains 55–60 ventrally located antennal glomeruli; in addition, that of the male contains a dorsal macroglomerular complex (MGC). A group of identifiable glomeruli consisting of two lateral large glomeruli (LLG) and four medial small glomeruli (MSG) is present in both sexes, but the LLG are greatly enlarged in the female. A MGC is also present in the male gypsy moth Lymantria dispar and male giant silk moth Antheraea polyphemus. The MGC in all of these species is organized into 3–4 distinct levels of glomeruli. Antennal sensory fibers were stained by cobalt backfills in B. mori, A. polyphemus, and L. dispar. Most fibers stained from cut long hairs (sensilla trichodea) projected to MGC in males and LLG in both sexes of B. mori. The distribution of fibers in the MGC of B. mori was topographically biased in that a majority of fibers from anterior branches projected medially in MGC while most fibers from posterior branches projected laterally or anteriorly. Terminal arborizations of single fibers were each restricted to a single glomerular level of the MGC. Fibers projecting to the posterior antennal center were frequently stained in cut-hair and control preparations, apparently by uptake of cobalt through intact sensilla on flagellar branches.     

江西赣江源区域两栖动物的群落组成和物种多样性特征

赣江发源于武夷山脉南段西麓的高山岭间,该区域独特的地理环境和气候特征蕴育了丰富的生物多样性。然而关于该区域两栖动物群落组成和物种多样性调查非常匮乏,不利于赣江源头两栖动物物种多样性的保护。于2016年4-9月期间在江西赣江源区域采用样线法调查了两栖动物的资源状况、群落组成及物种多样性特征,比较了它们在山地森林区、丘陵森林与农田区和盆地农田区3类栖息地之间的差异性。结果表明：赣江源区域分布有8科23种两栖动物,新增记录3个物种,包括尖舌浮蛙、寒露林蛙和九龙棘蛙,泽陆蛙和饰纹姬蛙为优势种。赣江源区域的两栖动物群落组成和物种多样性存在显著的季节间差异和栖息地间差异;两栖动物季节性繁殖是造成前者差异的主要因素,而土地利用类型、栖息地特征、空间距离、生活习性的物种特性等因素成就了后者差异的出现。因此,春、夏季一般有比秋季见到更高的两栖动物物种丰富度和多样性;山地森林区的两栖动物物种丰富度和多样性最高,盆地农田区次之,丘陵森林与农田区的最低。山地森林区应成为赣江源区域两栖动物多样性保护的优先区域。国家Ⅱ级重点保护野生动物虎纹蛙,在3类栖息地中均有分布,且在盆地农田区和山地森林区能见到更多个体,但农田区分布的虎纹蛙遭受到更大的威胁,建议加强对农田区虎纹蛙及其栖息地的保护。     

Immunocytochemical localization of a galanin-like peptidergic system in the brain of two urodele and two anuran species (Amphibia)

Summary Galanin-like immunoreactivity was localized in the brain of Urodela (Ambystoma, Pleurodeles) and Anura (Bufo, Xenopus) by immunocytochemistry with anti-porcine galanin antiserum. In the four species, immunoreactive perikarya were observed in the telencephalon (striatum, amygdala), diencephalon preoptic area mainly along the anterodorsal wall of the preoptic recessus, suprachiasmatic nucleus, lateral hypothalamus, ventral and dorsal infundibular nuclei, paraventricular organ, and rhombencephalon (nucleus of the solitary tract). Galaninergic fibres extended in similar regions and in the medial septum, ventral telencephalon, ventral hypothalamus, median eminence, and various mesencephalic and rhombencephalic regions. Contacts with the cerebrospinal fluid cavity occurred along the preoptic recessus (Ambystoma) and the ventral infundibular wall (all species). Fibres were scarce in the neurohypophysis. The distal and intermediate lobes of the pituitary were virtually devoid of immunoreactivity. The galaninergic system appeared more developed in adult amphibia than in young animals, suggesting the stimulating influence of sex steroids on the expression of galanin as previously described inAnguilla. The extensive distribution of the galanin-like immunoreactive neurons in amphibian brains suggests that this peptide may act as a neuromodulatur and/or neurotransmitter.     

On the distribution of the genusCousinia (Compositae)

On the basis of new floristic works a distribution map of the genusCousinia is given and discussed. The generic distribution area ofCousinia is nearly identical with the Irano-Turanian Region, but most of the 662 species are concentrated on the Iranian and Turkestanian mountain regions. Here, both in the eastern and in the western parts of the distribution area, four centres of diversity with high numbers of—mostly endemic—species are found: The western Tienshan (61 species), Pamiro-Alaj (169), NE-Afghanistan (80), NW-Afghanistan (44); Kopetdagh (66 in Iran, 33 in Turkmenia), Elburz (66), the northern part of Zagros (44), Azerbaijan (36). The outlines of the generic area are formed by genetic-historical and by climatic factors. The distribution patterns ofCousinia species confirm the close connections between the Iranian and the Turkestanian mountains as centres of origin and conservation of palaeo-xeromorphic mountain floras, contrasting with the Turanian lowland flora.Dedicated to Prof.K. H. Rechinger on the occasion of his 80th birthday.     

Seasonal patterns of activity and community structure in an amphibian assemblage at a pond network with variable hydrology

We studied community structure and seasonal activity patterns in a system of four ponds with seasonally-variable hydrology at a Mediterranean area in central Italy. We used a set of field methods to assess species presence and relative frequency of observation. The network of ponds was inhabited by six species of amphibians, two salamanders and four frogs. The breeding phenology of the six species did not vary remarkably among ponds, but there were significant differences among species in use of ponds. Factorial analysis of pond similarity drawn from percentage composition of the amphibian fauna, revealed that each of the four ponds was treatable as independent units, with no influence of relative inter-pond distance. PCA analysis allowed us to spatially arrange the amphibian species into three main groups: two were monospecific groups (i.e., Triturus vulgaris and Bufo bufo) and the third consisted of those species that selected not only the largest-deepest ponds, but also the ephemeral ones (i.e., Triturus carnifex, Hyla intermedia, the green frogs and Rana dalmatina). Our results suggest that the inter-pond differences in riparian vegetation, water depth, aquatic vegetation structure/abundance, and soil composition may produce differences among pond ecological characteristics (i.e., water turbidity and temperature, shelter availability, abundance of oviposition micro-sites), which may in turn influence different patterns of use by amphibians. To our knowledge, this is the first study emphasizing the potential role of heterochrony in the maintenance of a high species richness in Mediterranean amphibian communities. Preservation of freshwater vertebrate biodiversity requires management and protection not only of the main ponds and water bodies but also the temporary and ephemeral shallow ponds.     

Molecular phylogeny ofForsythia (Oleaceae) based on chloroplast DNA variation

Phylogenetic relationships of ten wild species and several cultivars ofForsythia were reconstructed based on the chloroplast (cp) DNA variation. A total of 216 cpDNA variants, 44 of which were potentially phylogenetically informative, was detected using 24 restriction endonucleases. Phylogenetic analysis usingFontanesia andAbeliophyllum as outgroups revealed four well defined species groups in the genus: 1)F. suspensa, 2)F. europaea — F. giraldiana, 3)F. ovata — F. japonica — F. viridissima, and 4)F. koreana — F. manshurica — F. saxatilis. The amount of support for each monophyletic group was evaluated by various methods including character number, decay analysis, parsimony bootstrapping, Neighbour-Joining (NJ) — bootstrapping, NJ-jackknifing, and the topology-dependent permutation tail probability (T-PTP) test. The data do not support the hybrid origin ofF. intermedia fromF. suspensa andF. viridissima. The disjunctly distributed European species,F. europaea, was identified as a sister species of the ChineseF. giraldiana and it was probably derived through recent long distance dispersal.