Damage-induced signalling mechanisms in the neonatal rat cochlea.

Sound overstimulation and exposure to ototoxic drugs damage cochlear hair cells (HCs) and cause their death. The surrounding support cells maintain an epithelial barrier and the appropriate physiological environment for surviving HCs during pathological conditions. Coordination of this homeostatic process requires cellular signalling. However, the signalling events that are activated during damage in the mammalian cochlea, are poorly understood. Neonatal rat cochlear explants were subjected to mechanical damage or exposed to neomycin - an ototoxin. Mechanical damage triggered the immediate propagation of an intercellular wave of increased intracellular Ca2+ from the lesion site into distinct cochlear regions. The properties of the Ca2+ wave and the source of Ca2+ required were specific to the cochlear region. IP3-mediated release from intracellular stores and influx of extracellular Ca2+ contribute differentially to the rise in intracellular Ca2+. The release of extracellular ATP is crucial for the propagation of the damage-induced Ca2+ wave. Gap junctions or connexin hemichannels also contribute to its formation. A subsequent damage-induced signalling event is the transient phosphorylation of ERK1/2 that arises within minutes of the insult occurring in support cells specifically. Similarly to the formation of the Ca2+ wave, release of extracellular ATP and gap junctions are critical for ERK1/2 activation. UTP-induced activation of ERK1/2 reveals the involvement of P2Y receptors. In addition, a requirement for the influx of extracellular Ca2+ also suggests a role for ion channels - potentially P2X receptors. P2X2,3,4 and P2Y2.4, n receptors were expressed in cochlear explants with P2X2 and P2Y2 being exclusive to support cells. Damage-induced currents were recorded from Deiters' cells in a syncytium during mechanical damage of the cochlea. Finally, when HCs were specifically targeted using neomycin, ERK1/2 activation occurred in support cells surrounding pyknotic HC nuclei. Inhibition of ERK1/2 delayed HC death

PAX6 disease models for aniridia

Aniridia is a pan-ocular genetic developmental eye disorder characterized by complete or partial iris and foveal hypoplasia, for which there is no treatment currently. Progressive sight loss can arise from cataracts, glaucoma, and aniridia-related keratopathy, which can be managed conservatively or through surgical intervention. The vast majority of patients harbor heterozygous mutations involving the PAX6 gene, which is considered the master transcription factor of early eye development. Over the past decades, several disease models have been investigated to gain a better understanding of the molecular pathophysiology, including several mouse and zebrafish strains and, more recently, human-induced pluripotent stem cells (hiPSCs) derived from aniridia patients. The latter provides a more faithful cellular system to study early human eye development. This review outlines the main aniridia-related animal and cellular models used to study aniridia and highlights the key discoveries that are bringing us closer to a therapy for patients

Unique activities of two overlapping PAX6 retinal enhancers

Enhancers play a critical role in development by precisely modulating spatial, temporal, and cell type-specific gene expression. Sequence variants in enhancers have been implicated in diseases; however, establishing the functional consequences of these variants is challenging because of a lack of understanding of precise cell types and developmental stages where the enhancers are normally active. PAX6 is the master regulator of eye development, with a regulatory landscape containing multiple enhancers driving the expression in the eye. Whether these enhancers perform additive, redundant or distinct functions is unknown. Here, we describe the precise cell types and regulatory activity of two PAX6 retinal enhancers, HS5 and NRE. Using a unique combination of live imaging and single-cell RNA sequencing in dual enhancer-reporter zebrafish embryos, we uncover differences in the spatiotemporal activity of these enhancers. Our results show that although overlapping, these enhancers have distinct activities in different cell types and therefore likely nonredundant functions. This work demonstrates that unique cell type-specific activities can be uncovered for apparently similar enhancers when investigated at high resolution in vivo

The lexicocalorimeter: Gauging public health through caloric input and output on social media

We propose and develop a Lexicocalorimeter: an online, interactive instrument for measuring the caloric content of social media and other large-scale texts. We do so by constructing extensive yet improvable tables of food and activity related phrases, and respectively assigning them with sourced estimates of caloric intake and expenditure. We show that for Twitter, our naive measures of caloric input , caloric output , and the ratio of these measures are all strong correlates with health and well-being measures for the contiguous United States. Our caloric balance measure in many cases outperforms both its constituent quantities; is tunable to specific health and well-being measures such as diabetes rates; has the capability of providing a real-time signal reflecting a population\u27s health; and has the potential to be used alongside traditional survey data in the development of public policy and collective self-awareness. Because our Lexicocalorimeter is a linear superposition of principled phrase scores, we also show we can move beyond correlations to explore what people talk about in collective detail, and assist in the understanding and explanation of how population-scale conditions vary, a capacity unavailable to black-box type methods

Hedonic contrast and the short-term stimulation of appetite

Hedonic contrast describes how liking for one item is influenced by the recent experience of other items which differ in hedonic valence. In the context of food stimuli, there is abundant evidence that hedonic contrast alters liking, but limited information on its impact on intake, and the aim here was to further clarify how hedonic impact modifies intake. Participants (96 female volunteers) rated and consumed ad libitum a sequence of four bowls of a snack (potato crisps) in one of three conditions. In the Palatable (salted crisps) and Bland (unsalted crisps) conditions, all four bowls were the same. In the Contrast condition participants alternated between salted and unsalted crisps. In total, significantly more was consumed in the Palatable (35.0 ± 2.6 g) than Bland (26.6 ± 2.4 g) condition, but most was consumed in the Contrast condition (37.0 ± 1.6 g). The impact of hedonic contrast was seen in the third serving, where those in the Contrast condition consumed the most of any serving, and significantly more than in Palatable or Bland conditions, and at the final serving, when those in the Contrast condition consumed significantly less than in Bland or Palatable conditions. Rated liking for the foods showed a similar pattern, with liking decreasing across servings in Palatable and Bland conditions. However, liking was influenced by the preceding serving in the Contrast condition, and the change in liking produced by contrast predicted subsequent intake. Overall, these data provide clear evidence that hedonic contrast can influence consumption, with intake driven by this adjusted liking

A novel mutation (Cys308Phe) of the LDL receptor gene in families from the South-Eastern part of Poland

The purpose of this investigation was to characterize a new mutation in the LDL-receptor (LDLR) gene in three families with clinically diagnosed familial hypercholesterolemia (FH) from the South-Eastern part of Poland. Mutational screening with exon by exon sequencing analysis was performed in all probands. The novel mutation c986G>T (Cys308Phe) in the exon 7 of LDLR gene was found in three apparently unrelated probands with FH. Analysis of the receptor activity of peripheral blood lymphocytes by binding and uptake of DiL-LDL showed a significant reduction (by 24% versus healthy control) of the fluorescent label in the lymphocytes of patients heterozygous for this mutation. Concentrations of serum LDL-C in probands before treatment were between 9.5 and 10.5 mmol/l. All patients had corneal arcus and tendon xanthoma. Clinically, families were characterized by premature coronary artery disease. This mutation occurred relatively frequently in our group of patients with FH, but this could be explained by a founder effect since we demonstrated their common ancestors

ATP-Evoked Intracellular Ca Signaling of Different Supporting Cells in the Hearing Mouse Hemicochlea

Hearing and its protection is regulated by ATP-evoked Ca2+ signaling in the supporting cells of the organ of Corti, however, the unique anatomy of the cochlea hampers observing these mechanisms. For the first time, we have performed functional ratiometric Ca2+ imaging (fura-2) in three different supporting cell types in the hemicochlea preparation of hearing mice to measure purinergic receptor-mediated Ca2+ signaling in pillar, Deiters' and Hensen's cells. Their resting [Ca2+]i was determined and compared in the same type of preparation. ATP evoked reversible, repeatable and dose-dependent Ca2+ transients in all three cell types, showing desensitization. Inhibiting the Ca2+ signaling of the ionotropic P2X (omission of extracellular Ca2+) and metabotropic P2Y purinergic receptors (depletion of intracellular Ca2+ stores) revealed the involvement of both receptor types. Detection of P2X2,3,4,6,7 and P2Y1,2,6,12,14 receptor mRNAs by RT-PCR supported this finding and antagonism by PPADS suggested different functional purinergic receptor population in pillar versus Deiters' and Hensen's cells. The sum of the extra- and intracellular Ca2+-dependent components of the response was about equal with the control ATP response (linear additivity) in pillar cells, and showed supralinearity in Deiters' and Hensen's cells. Calcium-induced calcium release might explain this synergistic interaction. The more pronounced Ca2+ leak from the endoplasmic reticulum in Deiters' and Hensen's cells, unmasked by cyclopiazonic acid, may also suggests the higher activity of the internal stores in Ca2+ signaling in these cells. Differences in Ca2+ homeostasis and ATP-induced Ca2+ signaling might reflect the distinct roles these cells play in cochlear function and pathophysiology