Bianca Kramer

Developmental axon pruning mediated by BDNF-p75NTR–dependent axon degeneration

The mechanisms that regulate the pruning of mammalian axons are just now being elucidated. Here, we describe a mechanism by which, during developmental sympathetic axon competition, winning axons secrete brain-derived neurotrophic factor (BDNF) in an activity-dependent fashion, which binds to the p75 neurotrophin receptor (p75NTR) on losing axons to cause their degeneration and, ultimately, axon pruning. Specifically, we found that pruning of rat and mouse sympathetic axons that project to the eye requires both activity-dependent BDNF and p75NTR. p75NTR and BDNF are also essential for activity-dependent axon pruning in culture, where they mediate pruning by directly causing axon degeneration. p75NTR, which is enriched in losing axons, causes axonal degeneration by suppressing TrkA-mediated signaling that is essential for axonal maintenance. These data provide a mechanism that explains how active axons can eliminate less-active, competing axons during developmental pruning by directly promoting p75NTR-mediated axonal degeneration.

The localization, trafficking and retrograde transport of BDNF bound to p75NTR in sympathetic neurons

BDNF, through p75NTR, promotes apoptosis and inhibits axonal growth of sympathetic neurons, antagonizing the pro-survival and axon growth-promoting actions of NGF through TrkA. While the trafficking of the TrkA:NGF complex is well characterized, little is known about p75NTR:BDNF trafficking in these neurons. Here we show that BDNF binds to and appears inside sympathetic neurons relatively slowly, although the temperature-sensitive internalization step itself is rapid. P75NTR internalization is partially sensitive to disruption of clathrin- or raft-mediated internalization, while that of TrkA is entirely clathrin-mediated. P75NTR, but not Trk, associates with neurotrophins in lipid rafts and coimmunoprecipitates with the truncated beta-caveolin-1 isoform. Finally, we directly visualize the retrograde transport of p75NTR ligands to cell bodies, which is insensitive to inhibitors of Trk retrograde transport, suggesting mechanistic differences. We postulate that beta-caveolin-1-containing lipid rafts and possibly intracellular endosomes might be compartments to which p75NTR:BDNF complexes are trafficked separately from Trk.

The comparative recall of Google Scholar versus PubMed in identical searches for biomedical systematic reviews: a review of searches used in systematic reviews

BackgroundThe usefulness of Google Scholar (GS) as a bibliographic database for biomedical systematic review (SR) searching is a subject of current interest and debate in research circles. Recent research has suggested GS might even be used alone in SR searching. This assertion is challenged here by testing whether GS can locate all studies included in 21 previously published SRs. Second, it examines the recall of GS, taking into account the maximum number of items that can be viewed, and tests whether more complete searches created by an information specialist will improve recall compared to the searches used in the 21 published SRs.MethodsThe authors identified 21 biomedical SRs that had used GS and PubMed as information sources and reported their use of identical, reproducible search strategies in both databases. These search strategies were rerun in GS and PubMed, and analyzed as to their coverage and recall. Efforts were made to improve searches that underperformed in each database.ResultsGS’ overall coverage was higher than PubMed (98% versus 91%) and overall recall is higher in GS: 80% of the references included in the 21 SRs were returned by the original searches in GS versus 68% in PubMed. Only 72% of the included references could be used as they were listed among the first 1,000 hits (the maximum number shown). Practical precision (the number of included references retrieved in the first 1,000, divided by 1,000) was on average 1.9%, which is only slightly lower than in other published SRs. Improving searches with the lowest recall resulted in an increase in recall from 48% to 66% in GS and, in PubMed, from 60% to 85%.ConclusionsAlthough its coverage and precision are acceptable, GS, because of its incomplete recall, should not be used as a single source in SR searching. A specialized, curated medical database such as PubMed provides experienced searchers with tools and functionality that help improve recall, and numerous options in order to optimize precision. Searches for SRs should be performed by experienced searchers creating searches that maximize recall for as many databases as deemed necessary by the search expert.

Regulation of neurons in the suprachiasmatic nucleus of Xenopus laevis

In the amphibian Xenopus laevis, suprachiasmatic melanotrope-inhibiting neurons (SMINs) play an important role in the regulation of the background adaptation process. In this study, we investigated the innervation of the SMINs at the light- and electron- microscopical level. Immunocytochemistry in combination with confocal laser scanning microscopy revealed co-existence of neuropeptide Y (NPY) and synaptobrevin in spots in the direct vicinity of the SMINs, suggesting the existence of NPY-containing synapses on these cells. At the ultrastructural level, the SMINs showed a high degree of plasticity, containing more electron-dense vesicles and a larger extent of RER in white- than in black-adapted animals. In black-adapted animals, symmetric synapses (Gray type II) were observed on the soma of the SMINs, suggesting an inhibitory input to these cells. The synaptic profiles contained electron-lucent and electron-dense vesicles, indicating the involvement of both a classical neurotransmitter and a neuropeptide (possibly NPY) in this input. In white-adapted animals, synapses were only found at some distance from the SMIN somata. Our findings indicate a striking plasticity of the innervation of the SMINs in relation to background adaptation and support the hypothesis that the SMINs are innervated by NPY-containing interneurons that inhibit SMIN activity in black-adapted animals.

Multiple control and dynamic response of the Xenopus melanotrope cell.

Some amphibian brain-melanotrope cell systems are used to study how neuronal and (neuro)endocrine mechanisms convert environmental signals into physiological responses. Pituitary melanotropes release alpha-melanophore-stimulating hormone (alpha-MSH), which controls skin color in response to background light stimuli. Xenopus laevis suprachiasmatic neurons receive optic input and inhibit melanotrope activity by releasing neuropeptide Y (NPY), dopamine (DA) and gamma-aminobutyric acid (GABA) when animals are placed on a light background. Under this condition, they strengthen their synaptic contacts with the melanotropes and enhance their secretory machinery by upregulating exocytosis-related proteins (e.g. SNAP-25). The inhibitory transmitters converge on the adenylyl cyclase system, regulating Ca(2+) channel activity. Other messengers like thyrotropin-releasing hormone (TRH) and corticotropin-releasing hormone (CRH, from the magnocellular nucleus), noradrenalin (from the locus coeruleus), serotonin (from the raphe nucleus) and acetylcholine (from the melanotropes themselves) stimulate melanotrope activity. Ca(2+) enters the cell and the resulting Ca(2+) oscillations trigger alpha-MSH secretion. These intracellular Ca(2+) dynamics can be described by a mathematical model. The oscillations travel as a wave through the cytoplasm and enter the nucleus where they may induce the expression of genes involved in biosynthesis and processing (7B2, PC2) of pro-opiomelanocortin (POMC) and release (SNAP-25, munc18) of its end-products. We propose that various environmental factors (e.g. light and temperature) act via distinct brain centers in order to release various neuronal messengers that act on the melanotrope to control distinct subcellular events (e.g. hormone biosynthesis, processing and release) by specifically shaping the pattern of melanotrope Ca(2+) oscillations.

p75 nerve growth factor receptor is important for retrograde transport of neurotrophins in adult cholinergic basal forebrain neurons

The role of the p75 nerve growth factor receptor in the retrograde transport of neurotrophins in the adult CNS was investigated by comparing the transport of 125I-labeled neurotrophins by normal and p75 nerve growth factor receptor-deficient cholinergic septohippocampal neurons. In control mice, nerve growth factor was selectively transported from the hippocampal formation to the cholinergic neurons in the septum. Nerve growth factor labeling was found in three to four times as many septal cholinergic neuronal cell bodies than labeling for neurotrophin-3 or neurotrophin-4/5, and transported brain-derived neurotrophic factor was barely detectable. Cells were considered as labeled when the number of grains per cell exceeded five times background. In p75 nerve growth factor receptor-deficient mice, the number of cholinergic neurons labeled with each of the neurotrophins was reduced by 85-95%. Retrograde labeling of septohippocampal neurons with Fluorogold was not obviously reduced in p75 nerve growth factor receptor-deficient mice, suggesting that general transport mechanisms were not impaired. Despite the reduced neurotrophin transport, cholinergic neurons of p75 nerve growth factor receptor-deficient mice were larger than controls and had an apparently normal density of immunostaining for choline acetyltransferase. Since nerve growth factor is reportedly involved in size regulation and choline acetyltransferase expression, this raises the possibility that the retrograde transport itself is not essential for these events. Thus, p75 nerve growth factor receptor plays an important, although not exclusive, role in the transport of neurotrophins by cholinergic basal forebrain neurons, and retrograde transport of nerve growth factor may not be needed for regulating certain cellular processes.

Functional organization of the suprachiasmatic nucleus of Xenopus laevis in relation to background adaptation

The process of background adaptation in the toad Xenopus laevis is controlled by neurons in the suprachiasmatic nucleus (SC) that inhibit the release of α‐melanophore‐stimulating hormone from the neuroendocrine melanotrope cells in the pituitary gland. We have identified the structural and functional organization of different neuropeptide Y (NPY)‐containing cell groups in the Xenopus SC in relation to background adaptation. A ventrolateral, a dorsomedial, and a caudal group were distinguished, differing in location as well as in number, size, and shape of their cells. They also show different degrees of NPY immunoreactivity in response to different background adaptation conditions. In situ hybridization using a Xenopus mRNA probe for the exocytosis protein DOC2 revealed that melanotrope cells of black‐adapted animals have a much higher expression of DOC2‐mRNA than white‐adapted ones. This establishes that the degree of DOC2‐mRNA expression is a good parameter to measure cellular secretory activity in Xenopus. We show that in the ventrolateral SC group, more NPY‐positive neurons express DOC2‐mRNA in white‐ than in black‐adapted animals. In contrast, NPY‐positive neurons in the dorsomedial group have a high secretory activity under the black‐adaptation condition. We propose that in black‐adapted animals, NPY‐positive neurons in the ventrolateral group, known to inhibit the melanotrope cells in white‐adapted animals synaptically, are inhibited by NPY‐containing interneurons in the dorsmedial group. NPY‐positive neurons in the caudal group have similar secretory dynamics as the dorsomedial NPY neurons, indicating that they also play a role in background adaptation, distinct from that exerted by the ventrolateral and dorsomedial group. J. Comp. Neurol. 432:346–355, 2001.

Expression of carp-cdx1, a caudal homolog, in embryos of the carp, Cyprinus carpio.

A carp caudal cDNA of 1.3 kb was cloned after screening an early segmentation stage cDNA library with a probe produced by PCR using conserved homeobox sequences as primers and genomic DNA as template. The homeobox gene was called carp-cdxl. The gene appears highly similar to other vertebrate caudal homologs, especially the zebrafish gene cdx(Zf-cad). The possible relationship to homeobox genes within the Hox-C gene complexes is discussed. A weak expression of the gene, detected by in situ hybridization, was found shortly before gastrulation (at 25% epiboly) in cells likely to have a posterior fate. During gastrulation expression became stronger. At the early segmentation stage, cells of the neural keel in the area of the prospective spinal cord expressed the gene. During the progression of segmentation, expression retracted in a caudal direction. The tailbud expressed the gene throughout, but the somites lost expression shortly after their formation. Only the most lateral mesoderm cells maintained expression in the trunk area. Carp-cdxl was also expressed in the endoderm. At 24 h after fertilization the gene was only expressed in the tailbud. At 48 h, no expression could be detected. The expression pattern suggests a function for carp-cdxl in gastrulation and patterning along the anterior-posterior axis of the embryo.

α-Melanophore-stimulating hormone in the brain, cranial placode derivatives, and retina of Xenopus laevis during development in relation to background adaptation

The amphibian Xenopus laevis can adapt the color of its skin to the light intensity of the background. A key peptide in this adaptation process is α‐melanophore‐stimulating hormone (α‐MSH), which is derived from proopiomelanocortin (POMC) and released by the endocrine melanotrope cells in the pituitary pars intermedia. In this study, the presence of α‐MSH in the brain, cranial placode derivatives, and retina of developing Xenopus laevis was investigated using immunocytochemistry, to test the hypothesis that POMC peptide‐producing neurons and endocrine cells have a common embryonic origin and a common function, i.e., controlling each others activities and/or being involved in the process of physiological adaptation. The presence of α‐MSH‐positive cells in the suprachiasmatic nucleus, ventral hypothalamic nucleus, epiphysis, and endocrine melanotrope and corticotrope cells, which are all involved in regulation of adaptation processes, has been detected from stage 37/38 onward. This is consistent with the presumed common origin of these cells, the anterior neural ridge (ANR) of the neural‐plate‐stage embryo. The olfactory epithelium and the otic and epibranchial ganglia also contain α‐MSH, indicating that these placodal derivatives originate from a common placodal domain continuous with the ANR. Furthermore, we demonstrate the presence of α‐MSH in a subpopulation of retinal ganglion cells (RGCs), which is possibly also derived from the ANR. Immunoreactivity for α‐MSH in RGCs that are located in the dorsal part of the retina is dependent on the background light intensity, suggesting that these cells are involved in the regulation of background adaptation. Taken together, the results support the hypothesis that POMC peptide‐producing cells have a common embryonic origin and are involved in adaptation processes. J. Comp. Neurol. 456:73–83, 2003.

Innovations in scholarly communication - global survey on research tool usage.

Many new websites and online tools have come into existence to support scholarly communication in all phases of the research workflow. To what extent researchers are using these and more traditional tools has been largely unknown. This 2015-2016 survey aimed to fill that gap. Its results may help decision making by stakeholders supporting researchers and may also help researchers wishing to reflect on their own online workflows. In addition, information on tools usage can inform studies of changing research workflows. The online survey employed an open, non-probability sample. A largely self-selected group of 20663 researchers, librarians, editors, publishers and other groups involved in research took the survey, which was available in seven languages. The survey was open from May 10, 2015 to February 10, 2016. It captured information on tool usage for 17 research activities, stance towards open access and open science, and expectations of the most important development in scholarly communication. Respondents’ demographics included research roles, country of affiliation, research discipline and year of first publication.