COST-ATP pretends to establish the role of intravesicular ATP in excitatory and inhibitory synapses of the central nervous system (CNS). Although, several laboratories have characterized the crucial interaction between ATP and catecholamines to permit its large accumulation in secretory vesicles of chromaffin cells, this crucial mechanism has not been studied in synaptic vesicles where high concentrations of neurotransmitters are needed for neuronal communication in the CNS. COST-ATP will combine the experience of the host laboratory in ATP as an accumulator of neurotransmitters in chromaffin cells and the TIRFM technology, and the ample experience of the researcher in cutting edge electrophysiological techniques in hippocampal neurons. The project will use electrophysiology, TIRFM, molecular biology and pharmacological tools to first discern the packagingbrole of vesicular ATP from its actions as neurotransmitter in central synapses using autaptic cultures of mouse hippocampal neurons. COST-ATP wants to explain why ATP is present inside of synaptic vesicles of almost all neurons. The main advantage of our approach is that we can modify the vesicular ATP by acting on the specific vesicular nucleotide carrier (VNUT) without affecting its cellular functions as the molecular energy. The consideration of the ATP, by its colligative properties, as a regulator of the neurotransmission, opens a new door in the neuronal communication. Given that its accumulation is mediated by VNUT a regulation of its activity could constitute a new pharmacological target for the treatment of neurological, psychiatric and cardiovascular diseases without involving membrane receptors.

In neurons, sites of Ca2+ influx and Ca2+ sensors are located within 20-50 nm, in subcellular “Ca2+ nanodomains”. Such tight coupling is crucial for the functional properties of synapses and neuronal excitability. Two key players act together in nanodomains, coupling Ca2+ signal to membrane potential: the voltage-dependent Ca2+ channels (VDCC) and the large conductance Ca2+ and voltage-gated K+ channels (BK). BK channels are characterized by synergistic activation by Ca2+ and membrane depolarization, but the complex molecular mechanism underlying channel function is not adequately understood. Information about the pore region, voltage sensing domain or isolated intracellular domains has been obtained separately using electrophysiology, biochemistry and crystallography. Nevertheless, the specialized behavior of this channel must be studied in the whole protein complex at the membrane in order to determine the complete range of structures and movements critical to its in vivo function. Using a combination of genetics, electrophysiology and spectroscopy, our group has measured for the first time the structural rearrangements accompanying whole BK channel activation at the membrane. From this unique position, our first goal is to fully determine the real time structural dynamics underlying the molecular coupling of Ca2+, voltage and activation of BK channels in the membrane environment, its regulation by accessory subunits and channel effectors. BK subcellular localization and role in Ca2+ nanodomains make these channels perfect candidates as reporters of local changes in [Ca2+] restricted to specific nanodomains close to the neuronal membrane. In our laboratory we have created fluorescent variants of the channel that report BK activity induced by Ca2+ binding, or Ca2+ binding and voltage. Our second aim in this proposal is to optimize and deploy this novel optoelectrical reporters to study physiologically relevant Ca2+-induced processes both in cellular and animal mode

Island ecosystems host a significant proportion of global biodiversity, but their rich insular biotas are also more vulnerable and less resistant to novel pressures than continental biotas. Human colonization has been a relatively recent event on most islands globally, and has nearly always resulted in devastating losses of biodiversity. The Canary Islands were no exception; these oceanic islands were colonized by aborigines more than 2000 years ago and later by Europeans, and have been transformed since this time. Despite being one of the most biodiverse regions within Europe and a target for EU biodiversity policies, there is a key knowledge gap about the pre-human state and natural variability of unmodified Canarian ecosystems that could help planners and managers to guide their strategies. This makes the Canaries the perfect place to reconstruct baseline ecosystem states and the long-term impacts of human activities. ISLANDPALECO will assess the timing and extent of human impact on Canary Island ecosystems by providing detailed reconstructions of past environments, combining leading edge and conventional palaeoecological tools and integrating the results in conservation ecology and management strategies. The project will provide new expertise in Canary Island palaeoenvironmental reconstructions through training the ER in the latest palaeoenvironmental DNA analysis (PalEnDNA) at the Landcare Research Long-Term Ecology Lab, one of the few labs in the world where this rapidly emerging field in palaeoecology is being applied to conservation on island settings. ISLANDPALECO will provide the ER with a new skill set in ancient molecular research that will be transferred to the University of La Laguna where they currently do not exist. This will be the first time that PalEnDNA analysis will be implemented in the Canaries and probably one of the few cases on islands, becoming a valuable benchmark example of the applicability of PalEnDNA analysis in other island regions.

Who were the Neanderthals and what caused their demise? To answer these questions, the classic approach in archaeology relies on the analysis of the Neanderthals' stone-tool assemblages and the mineralized bone remains of their dietary intake. Although this approach has yielded a great deal of important information about the Neanderthals’ fate, it is also limited in the sense that the only evidence that is considered is in-organic in nature. In the current proposal, we attempt to answer these questions by considering microscopic and molecular evidence that is organic in nature. By studying the organic sedimentary record at such fine scales, we are able to extract information about, for example, the fat contents of the Neanderthal food, the way they made fire, the arrangements of their living spaces, their surrounding vegetation and the climatic conditions where they lived. By combining these different sources of information we aim to provide a more complete picture of the Neanderthals and the reason of their disappearance. Specifically, the PALEOCHAR project examines how Neanderthal diet, fire technology, settlement patterns, and surrounding vegetation were affected by changing climatic conditions. To do so, the project will integrate methodologies from micromorphology and organic geochemistry. A key and innovative aspect of the proposal is the consideration of microscopic and molecular evidence that is both organic and charred in nature. Climatic changes and behavioural responses will be examined at two Iberian sites which represent two key points along the Neanderthal time-line. The results of this project will make important contributions to the development of new methods for archaeological research, train a new generation of skilled geoarchaeologists knowledgeable in microstratigraphy and applied chemistry, and yield new insights into the Neanderthals and their demise.