Detection of Ureaplasma diversum in the upper airways of Australian feedlot cattle.

Bovine respiratory disease (BRD) exerts a major impact on the beef cattle industry nationally and worldwide, with a range of aetiological factors impacting its pathogenesis. Previous research has focussed on an increasing number of bacteria and viruses that have been shown to play a role in eliciting disease. Recently, additional agents have been emerging as potential contributors to BRD, including the opportunistic pathogen Ureaplasma diversum. To determine if U. diversum was present in Australian feedlot cattle and if that presence was linked to BRD, nasal swabs were collected from a cohort of 34 hospital pen animals and compared to 216 apparently healthy animals sampled contemporaneously at feedlot induction and again after 14 days on feed at an Australian feedlot. All samples were subjected to a de novo polymerase chain reaction (PCR) assay targeting U. diversum in combination with other BRD agents. U. diversum was detected at a low prevalence in cattle at induction (Day 0: 6.9%, Day 14: 9.7%), but in a significantly greater proportion of cattle sampled from the hospital pen (58.8%). When considering the presence of other BRD-associated agents, co-detection of U. diversum and Mycoplasma bovis was most common in hospital pen animals receiving treatment for BRD. These findings suggest that U. diversum may be an opportunistic pathogen involved in the aetiology of BRD in Australian feedlot cattle, in combination with other agents, with further studies are warranted to identify if a causal relationship exists.

Efficiency-corrected PCR quantification for identification of prevalence and load of respiratory disease-causing agents in feedlot cattle.

Bovine respiratory disease (BRD) is the most prevalent disease in feedlot cattle worldwide with Bovine alphaherpesvirus 1 (BoAHV1), Histophilus somni, Mannheimia haemolytica, Mycoplasma bovis, Pasteurella multocida and Trueperella pyogenes accepted to be common etiological agents associated with BRD. Although these agents are common in the upper and lower airways in clinical BRD cases, some also exist as normal flora suggesting their presence in the upper airways alone is not necessarily informative with respect to disease status or risk. To determine the relationship between presence, load and disease status, we investigated the relationship between load in the upper airways at induction and active BRD cases in feedlot cattle using efficiency-corrected PCR quantification. By this approach, we were able to accurately determine the prevalence and load of the key BRD agents in the upper respiratory tract showing that cattle in the hospital pen had a higher prevalence, and load, of these agents both singly and in combination compared to cattle sampled at feedlot induction. A combination of agents was the most accurate indicator of BRD risk with cattle with four or more agents detected in the upper airway more likely to be undergoing treatment for BRD than non-BRD ailments. In addition, M. bovis was rarely detected at feedlot induction but was identified at high prevalence in cattle in the hospital pen. These findings present a potential new technological approach for the investigation, analysis and identification of BRD-associated viral and bacterial agents for Australian feedlot systems as well as for BRD disease management and treatment.

Dependence of juvenile reef fishes on semi-arid hypersaline estuary microhabitats as nurseries.

The differences between fish assemblages in three microhabitat types, in relation to vegetation and sediment characteristics of a hypersaline estuary located in an semi-arid zone in north-eastern Brazil, were investigated. Fishes were collected using a beach seine during the rainy and dry seasons in 2012. A total of 78 species were recorded, with the most common families being Gerreidae, Lutjanidae and Tetraodontidae. The majority of species were represented by juveniles, with Eucinostomus argenteus, Ulaema lefroyi and Sphoeroides greeleyi being the dominant species. The fish assemblage structures differed significantly among microhabitat types, with the narrow intertidal flat adjacent to the mangrove fringe supporting the most diverse fish fauna. In addition, only 27 species were common to all of the microhabitats. The results support the hypothesis that hypersaline estuaries serve as important nursery areas for various reef fish species, due to the structural complexity provided by their macroalgae beds and mangroves.

DNA barcoding and morphological identification of neotropical ichthyoplankton from the Upper Paraná and São Francisco.

The identification of fish larvae from two neotropical hydrographic basins using traditional morphological taxonomy and DNA barcoding revealed no conflicting results between the morphological and barcode identification of larvae. A lower rate (25%) of correct morphological identification of eggs as belonging to migratory or non-migratory species was achieved. Accurate identification of ichthyoplankton by DNA barcoding is an important tool for fish reproductive behaviour studies, correct estimation of biodiversity by detecting eggs from rare species, as well as defining environmental and management strategies for fish conservation in the neotropics.

Nutrigenomics: definitions and advances of this new science.

The search for knowledge regarding healthy/adequate food has increased in the last decades among the world population, researchers, nutritionists, and health professionals. Since ancient times, humans have known that environment and food can interfere with an individual's health condition, and have used food and plants as medicines. With the advance of science, especially after the conclusion of the Human Genome Project (HGP), scientists started questioning if the interaction between genes and food bioactive compounds could positively or negatively influence an individual's health. In order to assess this interaction between genes and nutrients, the term "Nutrigenomics" was created. Hence, Nutrigenomics corresponds to the use of biochemistry, physiology, nutrition, genomics, proteomics, metabolomics, transcriptomics, and epigenomics to seek and explain the existing reciprocal interactions between genes and nutrients at a molecular level. The discovery of these interactions (gene-nutrient) will aid the prescription of customized diets according to each individual's genotype. Thus, it will be possible to mitigate the symptoms of existing diseases or to prevent future illnesses, especially in the area of Nontransmissible Chronic Diseases (NTCDs), which are currently considered an important world public health problem.

Allometric growth patterns in hatchery-reared larvae of the catfish Lophiosilurus alexandri (Pisces: Pseudopimelodidae) / Padrões de crescimento alométrico em larvas criadas em cativeiro do pacamã Lophiosilurus alexandri (Pisces: Pseudopimelodidae)

The aim of the present study was to characterize the relationships that occur between morphometric variables of larvae of Lophiosilurus alexandri Steindachner, 1876 through their allometric growth analysis. Total length, head length and body height were correlated with standard length, head height and body height, as well as eye diameter and head height with head length. The results revealed allometric positive relation during the initial development, where the b coefficient varied between 1.10 and 2.81. The variable pre-anal distance in relation to the standard length and the snout length in relation to the head length revealed an allometric negative relation, where the b coefficient was 0.85 and 0.94, respectively.

O objetivo do presente estudo foi caracterizar os tipos de relações que ocorrem entre as variáveis morfométricas das larvas de Lophiosilurus alexandri Steindachner, 1876, por meio da análise de seu crescimento alométrico. As variáveis comprimento total, comprimento da cabeça e altura do corpo foram correlacionadas com o comprimento padrão; a altura da cabeça com a altura do corpo; e o diâmetro do olho e a altura da cabeça com o comprimento da cabeça. Os resultados revelaram relações alométricas positivas durante o desenvolvimento inicial, quando o coeficiente b variou entre 1,10 e 2,81. A variável distância pré-anal em relação ao comprimento padrão e a variável comprimento do focinho em relação ao comprimento da cabeça revelaram relação alométrica negativa quando o coeficiente b foi 0,85 e 0,94, respectivamente.

Anatomical and histological organization of the testes of the inseminating catfish Trachelyopterus striatulus (Steindachner, 1877) (Siluriformes: Auchenipteridae).

The testicular morphology, spermatogenesis and occurrence of sperm in the ovarian lumen of Trachelyopterus striatulus were studied using anatomical, histological and biometric techniques. A total of 50 catfish (T. striatulus) were captured, measuring 14.9 ± 2.5 cm of standard length, body weight was 81.2 ± 34.5 g and their testes weighed 16.9 ± 6.1 g. The testes of T. striatulus are paired organs, showing two distinct regions: cranial, which shows a compact medial part and with fringes ventrally, and caudal region, which is formed of the seminal vesicle with fringes laterally and two saculiform expansions. The testes presented a length of 35.2 ± 6.9 mm, and the fringes showed a cranial length of 12.1 ± 3.8 mm and caudal length of 6.4 ± 2.6 mm. Histologically, the cranial fringes are spermatogenic and showed cells with significantly different nuclear diameters, ranging from 8.2 ± 1.5 µm (primary spermatogonia) to 1.88 ± 0.3 µm (spermatid). The seminal vesicles and saculiform expansions showed tubules with a simple prismatic secretory epithelium containing spermatozoa and secretion into the lumen. The caudal fringes are exclusively for secretory flow, consisting of tubules with a simple cuboidal epithelium. The common spermatic duct showed a simple cuboidal epithelium and contained spermatozoa with secretion into the lumen. The secretion of the caudal region is acidophilic, with neutral glycoproteins and sialomucin. T. striatulus ovaries showed free spermatozoa or were organized in spermatozeugmata into the ovarian lumen and between the ovuligers lamellae.

Composição química e atividade in vitro de três óleos essenciais sobre Colletotrichum gloeosporioides do maracujazeiro / Chemical composition and in vitro activity of three essential oils on Colletotrichum gloeosporioides from passion fruit

Objetivou-se avaliar a ação de diferentes concentrações de óleos essenciais de três plantas medicinais sobre o crescimento micelial, a produção e a germinação de conídios de Colletotrichum gloeosporioides (Penz.). Utilizou-se os óleos essenciais de Lippia sidoides Cham., Cymbopogon citratus (DC) Stapf.) e Ocimum gratissimum L. em cinco concentrações (0 µL mL-1, 1 µL mL-1, 3 µL mL-1, 5 µL mL-1 e 7 µL mL-1) com 4 repetições no delineamento inteiramente casualizado, em esquema fatorial 5x3. Os óleos foram analisados por cromatografia gasosa acoplada com espectrometria de massas. Para todas as características verificou-se efeito significativo da interação "Óleo" versus "Concentração". O óleo essencial de C. citratus proporcionou a maior inibição do crescimento micelial em todas as concentrações testadas. Os outros óleos não se diferenciaram, exceto a partir da concentração de 5 µL mL-1, onde o óleo de L. sidoides foi superior ao óleo de O. gratissimum. Na produção e germinação de conídios, o óleo de C. citratus, foi superior aos óleos de L. sidoides e de O. gratissimum nas concentrações de 1 µL mL-1 e 3 µL mL-1, inibindo completamente a produção e germinação a partir da concentração de 3 µL mL-1. As concentrações influenciam em todas as variáveis, ajustando-se à equação do modelo linear. Observa-se o decréscimo nessas, à medida em que se aumentam as concentrações dos óleos testados. Os óleos essenciais testados possuem efeito fungitóxico e fungistático.

The objective of this study was to evaluate the action of different concentrations of essential oils of three medicinal plants on the mycelial growth, production and conidial germination of Colletotrichum gloeosporioides (Penz.). We used a completely randomized design, in a 5x3 factorial design, being five concentrations (0 µL mL-1, 1 µL mL-1, 3 µL mL-1, 5 µL mL-1 e 7 µL mL-1) x three oils from Lippia sidoides Cham., Cymbopogon citratus (DC) Stapf. and Ocimum gratissimum L., with four replications. The oils were analyzed by gas chromatography with mass spectrometry. There was a significant interaction of concentration versus species on all analyzed traits. The essential oil from C. citratus yielded the highest mycelial growth inhibition at all concentrations. The other oils were not differentiated, except at the concentration of 5µL mL-1, in which the oil of L. sidoides was more efficient that the oil of O. gratissimum. In conidial production and germination, the oil of C. citratus was superior to the oil of L. sidoides and O. gratissimum at the concentrations of 1 µL mL-1 and 3 µL mL-1, as it completely inhibited the production and germination of conidia at the concentrations of 3 µL mL-1 and above. The concentrations influenced all the variables, adjusting themselves to the linear model equation. We observed a decrease in the variables, as the concentrations of the oils tested are increased. The essential oils used have antifungal and fungicidal effect.

Ovarian structure and oogenesis of catfish Pimelodella vittata (Lütken, 1874) (Siluriformes, Heptapteridae).

The morphology of the ovaries and oogenesis of Pimelodella vittata were studied using anatomical and histological techniques to provide information of its reproductive biology. Eighty adult females were captured trimonthly during the period November 2005 to October 2006. The ovaries are paired, saculiform organs, which are coated with tunica albuginea and contain ovigerous lamellae, where the oocytes develop before being released into the ovarian lumen and following the ovarian duct until reaching the genital papilla. Oogenesis was divided into stages based on the alterations to the nucleus, ooplasm and surrounding follicular layers. Oogonia form groups from the germinal epithelium have asynchronous development and differentiate into initial perinucleolar oocytes. The formation of the zona pellucida is initiated in the advanced perinucleolar oocytes reaching a thickness of 1.46±0.58 µm in the vitellogenic oocytes. The follicular cells are squamous in perinucleolar oocytes, become cubical in the pre-vitellogenic oocytes and prismatic in the vitellogenic oocytes with a height of 11.20±4.74 µm. The histochemical reactions indicate that zona pellucida, cortical alveoli and yolk globules contain neutral glycoproteins and the follicular cells contain neutral glycoproteins in association with carboxylated and sulphated glycoconjugates. Statistical analyses showed significant differences in the diameter of the oocytes and follicular cells height as oocytes matured. This study represents the first data about the ovarian structure and oogenesis of this species.

Influence of subphase parameters on the nanostructures of 5,10,15,20-tetraphenyl-21H,23H-porphine films at air-water interface.

In this work, we have evaluated the influence of concentration of 5,10,15,20-tetraphenyl-21H,23H-porphine solutions on pi-A isotherms, varying the pH of subphase. The obtained pi-A isotherms indicated the existence of different aggregation states and the dependence of area/molecule with the pH was established. There is an increase in the area/molecule for both pH = 5 and pH = 8 in low concentrations with a marked increase for the acid pH. Such trend has given such clearly evidence that proton (H+) effect in the chromophore macrocycle is potentially indicated for the pH-sensitive gas sensors.

Effectiveness of a non-surgical alternative to the Mules operation in sheep.

OBJECTIVE: To measure changes to the perineal bare area, local tissue reaction and healing responses of young sheep, following intradermal administration of cetrimide and polyvinylpyrrolidone (PVP), with and without ethanol, to the breech and tail. METHOD: A needle-less injector was used to deposit formulations containing 40 g/L cetrimide and 30 g/L PVP (group 2) or 20 g/L cetrimide, 30 g/L PVP and 15 g/L ethanol (group 3), within the dermis of the tail and the region surrounding the perineal bare breech area of groups (N = 8) of Merino weaner sheep. The dimensions of the perineal bare area (length, width and diagonal distances left and right) and tail width were recorded before and at intervals after treatment for 60 days. Observations of swelling and bruising and scab formation at the treatment sites were recorded for up to 35 days after treatment. Rectal temperatures were monitored for up to 35 days after treatment and bodyweight for up to 60 days after treatment. An untreated control group (group 1) was included. RESULTS: Comparison of day -3 and day 35 measurement data showed that both treated groups had significantly (P < 0.05) wider breech bare areas compared to the untreated controls and that group 2 sheep had significantly (P < 0.05) longer breech bare areas compared to group 3 sheep or to the untreated controls, which were not significantly different. At this time scabs were still firmly in place on many treated sheep. At day 35 there was no increase in tail bare area caused by either treatment. By day 60 there was no significant difference between the treated and control groups in either the breech or tail regions indicating that the changes present at day 35, were not permanent. Mean weight gain in the groups throughout the 60-day interval was unaffected by treatment. Intradermal treatment was associated with a significant elevation in body temperature. This effect lasted for 3 days and was associated with signs of discomfort and depressed appearance in at least some of the treated sheep. Bruising was mild to severe in all treated sheep within two days of treatment but was not evident in any sheep by day 21. Mild to moderate swelling was also associated with treatment but was not uniform across sheep in the groups. The tail of one sheep was severely swollen for several days. Swelling remained obvious in most treated sheep until day 14 but was not present at day 21. CONCLUSION: Under the conditions of this study intradermal injection of cetrimide had no permanent effect on bare area measurements on the breech or the amount of wool-bearing skin on the tail. It also caused signs of discomfort and pain that raise welfare concerns.

Germinação, vigor e sanidade de sementes de fava d'anta (Dimorphandra mollis Benth. ) obtidas de frutos coletados no solo e na planta / Germination, vigor and sanity of "fava d'anta" (Dimorphandra mollis Benth. ) seeds obtained from fruits collected both in the soil and in the plant

O objetivo do trabalho foi avaliar a qualidade fisiológica e sanitária das sementes obtidas a partir de frutos maduros coletados no solo e nas plantas de fava d'anta, espécie medicinal nativa do Cerrado, ameaçada de extinção, cujos frutos são ricos em rutina. O delineamento experimental empregado foi o inteiramente casualizado, com 10 repetições. Cada repetição foi constituída de uma caixa gerbox contendo 15 sementes cada. A qualidade fisiológica foi determinada por meio de testes de germinação e vigor, avaliando-se a porcentagem de germinação, o índice de velocidade de germinação (IVG) e a massa seca das raízes primárias. A qualidade sanitária foi determinada pela identificação dos microorganismos patogênicos associados às sementes, bem como pela porcentagem de ocorrência destes em cada parcela. Os fungos identificados nas sementes pertencem aos gêneros Fusarium, Alternaria, Aspergillus, Cladosporium, Colletrotrichum, Helminthosporium, Nigrospora, Penicillum, Phoma e Phomopsis. Sementes oriundas de frutos coletados no solo foram mais contaminadas por agentes patogênicos e apresentaram germinação (56,67 por cento), vigor e sanidade inferiores às sementes obtidas de frutos coletados na planta.

The aim of this work was to evaluate the physiological and sanitary quality of seeds obtained from mature fruits collected in the soil and in "fava d'anta" plants. Dimorphandra mollis constitutes an endangered medicinal species native to cerrado and presents fruits rich in rutin. The experimental design was completely randomized, with 10 replicates. Each replicate consisted of a gerbox containing 15 seeds each. Physiological quality was assessed through germination and vigor tests by evaluating germination percentage, germination velocity index (GVI), and primary root dry matter. Sanitary quality was evaluated by identifying pathogenic microorganisms associated with the seeds, as well as their occurrence percentage in each plot. The identified fungi belonged to the genera Fusarium, Alternaria, Aspergillus, Cladosporium, Colletotrichum, Helminthosporium, Nigrospora, Penicillium, Phoma and Phomopsis. Seeds from fruits collected in the soil were more contaminated by pathogenic agents and presented lower germination (56.67 percent), vigor and health than seeds from fruits collected in the plant.

Resistance to benzoylphenyl urea insecticides in Australian populations of the sheep body louse.

A laboratory bioassay based on the failure of eggs laid by diflubenzuron-treated sheep body lice Bovicola ovis (Schrank) (Phthiraptera; Trichodectidae) to produce nymphs is described. The test is capable of detecting benzoylphenyl urea-resistant populations. A susceptible discriminating dose (topical treatment of female lice with 0.10 microg diflubenzuron) suitable for screening suspect resistant populations is suggested but testing of further populations that are naïve to these insect growth regulator insecticides is warranted.

What can we learn from the oral intake of prions by sheep?

The central nervous system is the ultimate target of prions, the agents responsible for fatal neurodegenerative diseases known as transmissible spongiform encephalopathies (TSEs). The neuro-invasive phase and its related clinical signs take place after a long incubation period. During this asymptomatic phase, however, active transport and replication of the infectious agent take place in peripheral sites. The oral infection route has been extensively studied because of its implication in the recent epidemic of bovine spongiform encephalopathy (BSE) in cattle and of the resulting human cases of variant Creutzfeldt-Jakob disease (vCJD). Rodent models have been useful in studying some aspects of this pathogenesis. Now, new data on the initial steps of oral infection have been obtained in sheep. This species is naturally infected with scrapie by horizontal transmission and there is strong evidence implicating the oral route. Furthermore, the existence of resistant and susceptible genotypes offers the possibility of comparative studies. The data were obtained using surgical and biochemical procedures to modulate the efficiency of oral infection and show that, in sheep, the abnormal prion protein (PrP) associated with the infectious agent crosses the intact intestinal barrier at the level of the enterocytes and then passes rapidly into lymph. These steps are identical in susceptible and resistant sheep. Thereafter, replication takes place in lymphoid structures. Other results in the same study indicate that alimentary fluids almost completely degrade the PrP of the inoculum. Though not directly transposable to human diseases, in which it is not possible to study these early stages, these data allow the elaboration of a simplified concept for the pathogenesis of TSEs. They also suggest that human contamination at the level of the oral cavity might be more important than previously suspected.

Tissue distribution of bovine spongiform encephalopathy agent in primates after intravenous or oral infection.

BACKGROUND: The disease-associated form of prion protein (PrP(res)) has been noted in lymphoreticular tissues in patients with variant Creutzfeldt-Jakob disease (vCJD). Thus, the disease could be transmitted iatrogenically by surgery or use of blood products. We aimed to assess transmissibility of the bovine spongiform encephalopathy (BSE) agent to primates by the intravenous route and study its tissue distribution compared with infection by the oral route. METHODS: Cynomolgus macaques were infected either intravenously or orally with brain homogenates from first-passage animals with BSE. They were clinically monitored for occurrence of neurological signs and killed humanely at the terminal stage of the disease. Brain, lymphoreticular tissues, digestive tract, and peripheral nerves were obtained and analysed by sandwich ELISA and immunohistochemistry for quantitative and qualitative assessment of their PrP(res) content. FINDINGS: Incubation periods after intravenous transmission of BSE were much shorter than after oral infection. We noted that PrP(res) was present in lymphoreticular tissues such as spleen and tonsils and in the entire gut from the duodenum to the rectum. In the gut, PrP(res) was present in Peyer's patches and in the enteric nervous system and nerve fibres of intestinal mucosa. Furthermore, PrP(res) was found in locomotor peripheral nerves and the autonomic nervous system. Amount of PrP(res) ranged from 0.02% to more than 10% of that recorded in brain. Distribution of PrP(res) was similar in animals infected by the intravenous or oral route. INTERPRETATION: Our findings suggest that the possible risk of vCJD linked to endoscopic procedures might be currently underestimated. Human iatrogenic vCJD cases infected intravenously raise the same public-health concerns as primary cases and need the same precautionary measures with respect to blood and tissue donations and surgical procedures.

Evaluation of quinacrine treatment for prion diseases.

Based on in vitro observations in scrapie-infected neuroblastoma cells, quinacrine has recently been proposed as a treatment for Creutzfeldt-Jakob disease (CJD), including a new variant CJD which is linked to contamination of food by the bovine spongiform encephalopathy (BSE) agent. The present study investigated possible mechanisms of action of quinacrine on prions. The ability of quinacrine to interact with and to reduce the protease resistance of PrP peptide aggregates and PrPres of human and animal origin were analyzed, together with its ability to inhibit the in vitro conversion of the normal prion protein (PrPc) to the abnormal form (PrPres). Furthermore, the efficiencies of quinacrine and chlorpromazine, another tricyclic compound, were examined in different in vitro models and in an experimental murine model of BSE. Quinacrine efficiently hampered de novo generation of fibrillogenic prion protein and PrPres accumulation in ScN2a cells. However, it was unable to affect the protease resistance of preexisting PrP fibrils and PrPres from brain homogenates, and a "curing" effect was obtained in ScGT1 cells only after lengthy treatment. In vivo, no detectable effect was observed in the animal model used, consistent with other recent studies and preliminary observations in humans. Despite its ability to cross the blood-brain barrier, the use of quinacrine for the treatment of CJD is questionable, at least as a monotherapy. The multistep experimental approach employed here could be used to test new therapeutic regimes before their use in human trials.

Inheritance of diflubenzuron resistance and monooxygenase activities in a laboratory-selected strain of Lucilia cuprina (Diptera: Calliphoridae).

Inheritance of the high-level diflubenzuron resistance shown by a laboratory-selected strain of Lucilia cuprina (Wiedemann) was examined in matings with a susceptible reference strain. Progeny of reciprocal crosses between resistant females and susceptible males showed higher LC50 values than the alternate reciprocal cross, indicating some maternal influence on inheritance of resistance. Resistance was inherited in a codominant (S male x R female) or incompletely recessive (R male x S female) manner. Monooxygenase activities (aldrin epoxidation) of the F1 generations were also intermediate between the levels shown by the parental lines, however, inheritance of enzyme activities showed greater degrees of dominance than for resistance levels. There was also some maternal influence on inheritance of monooxygenase activities. Backcrosses of F1 generations to both susceptible and resistant parents did not fit the expected patterns for a major sex-linked resistance locus, indicating that the maternal influence on resistance inheritance was not associated with sex-linkage of a major resistance gene. The backcross data also failed to fit the model for a single major autosomal gene, suggesting that the resistance in the diflubenzuron-selected strain is polygenic, involving mechanisms additional to monooxygenases.

Immunolocalization of the cellular prion protein in normal brain.

We examined the localization of PrP(c) in normal brain using free-floating section immunohistochemistry and monclonal antibody 3F4. In the mature hamster and baboon brain, PrP(c) is localized to the neuropil with a synaptic distribution and the PrP(c) immunoreactivity is denser in regions known for ongoing plasticity. Cell bodies and major fiber tracts have little or no PrP(c) immunoreactivity. At the electron microscopic level, PrP(c) immunoreactivity decorates synaptic profiles, both pre- and postsynaptically. Results obtained with two additional antibodies, 3B5 and Pri-304, showed similar patterns of PrP(c) bands on Western blots, although Pri-304 was less sensitive. On sections through the adult hamster hippocampus, 3B5 and Pri-304 both stained the synaptic neuropil while cell bodies in the pyramidal and dentate granule cell layers were not immunoreactive. Pri-304 differentiated between synaptic layers in the hippocampus and closely resembled the pattern of staining obtained with 3F4. Preliminary results of developing brain showed that PrP(c) is initially localized along fiber tracts in the neonate brain. These results show that PrP(c) has a synaptic distribution in the adult brain and suggest that there are important changes in its distribution during brain development. These results also characterize two additional reagents for studies of PrP(c) localization.

Discrete localizations of hedgehog signalling components in the developing and adult rat nervous system.

Sonic hedgehog (Shh), a morphogen molecule implicated in embryonic tissue patterning, displays inductive, proliferative, neurotrophic and neuroprotective activities on various neural cells. Shh might exert its biological functions through binding to patched (Ptc) associated with smoothened (Smo), leading to downstream activation of target genes such as the transcription factor Gli1. We have performed a detailed localization of cells expressing transcripts of Shh, Ptc, Smo and Gli1 in brain and spinal cord of the adult rat as well as in the developing cerebellum. In the adult, Shh-positive cells were mainly observed in forebrain structures, in the Purkinje cells of the cerebellum and in motor neurons. Ptc-positive cells were frequently observed in brain areas devoid of any Shh transcripts, except in the median eminence or the facial nucleus, suggesting local Shh signalling. Interestingly, Smo transcripts were predominantly present within circumventricular organs, in granular cells of the dentate gyrus and in neurons of the reticular thalamic nucleus. The presence of Shh, Ptc and Smo transcripts in hypothalamic areas may indicate a role of Shh signalling in the modulation of neuroendocrine functions. The expression pattern of these three genes as well as of Gli1, and their developmental regulation in the cerebellum, suggest a possible role for Hedgehog signalling in the control of various cell populations within the cerebellum, particularly in granule cell proliferation and/or differentiation that might be impaired in proliferative states such as medulloblastomas.