RESUMEN
Basically, all mammalian tissues are constantly exposed to a variety of environmental mechanical signals. Depending on the signal strength, mechanics intervenes in a multitude of cellular processes and is thus capable of inducing simple cellular adaptations but also complex differentiation processes and even apoptosis. The underlying recognition typically depends on mechanosensitive proteins, which most often sense the mechanical signal for the induction of a cellular signaling cascade by changing their protein conformation. However, the fate of mechanosensors after mechanical stress application is still poorly understood, and it remains unclear whether protein degradation pathways affect the mechanosensitivity of cells. Here, we show that cyclic stretch induces autophagosome formation in a time-dependent manner. Formation depends on the cochaperone BAG family molecular chaperone regulator 3 (BAG3) and thus likely involves BAG3-mediated chaperone-assisted selective autophagy. Furthermore, we demonstrate that strain-induced cell reorientation is clearly delayed upon inhibition of autophagy, suggesting a bidirectional cross-talk between mechanotransduction and autophagic degradation. The strength of the observed delay depends on stable adhesion structures and stress fiber formation in a Ras homologue family member A (RhoA)-dependent manner.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas Reguladoras de la Apoptosis/metabolismo , Mecanorreceptores/metabolismo , Animales , Apoptosis/fisiología , Autofagosomas/metabolismo , Autofagia/fisiología , Fenómenos Biomecánicos , Línea Celular , Fibroblastos/citología , Fibroblastos/metabolismo , Mecanorreceptores/citología , Mecanotransducción Celular , Ratones , Músculo Liso/citología , Músculo Liso/metabolismo , Proteolisis , Ratas , Transducción de Señal/fisiología , Factores de Transcripción/metabolismoRESUMEN
Primary mechanosensory neurons play an important role in converting mechanical forces into the sense of touch. In zebrafish, Rohon-Beard (RB) neurons serve this role at embryonic and larval stages of development. Here we examine the morphology and physiology of RBs in larval zebrafish to better understand how mechanosensory stimuli are represented along the spinal cord. We report that the morphology of RB neurons differs along the rostrocaudal body axis. Rostral RB neurons arborize in the skin near the cell body whereas caudal cells arborize at a distance posterior to their cell body. Using a novel electrophysiological approach, we also found longitudinal differences in the mechanosensitivity and physiological properties of RB neurons. Rostral RB neurons respond to mechanical stimulations close to the soma and produce up to three spikes with increasing stimulus intensity, whereas caudal cells respond at more distal locations and can produce four or more spikes when the intensity of the mechanical stimulus increases. The mechanosensory properties of RB neurons are consistent with those of rapidly adapting mechanoreceptors and can signal the onset, offset and intensity of mechanical stimulation. This is the first report of the intensity encoding properties of RB neurons, where an increase in spike number and a decrease in spike latency are observed with increasing stimulation intensity. This study reveals an unappreciated complexity of the larval zebrafish mechanosensory system and demonstrates how differences in the morphological and physiological properties of RBs related to their rostrocaudal location can influence the signals that enter the spinal cord.
Asunto(s)
Mecanorreceptores/citología , Mecanorreceptores/fisiología , Médula Espinal/citología , Médula Espinal/fisiología , Pez Cebra/anatomía & histología , Pez Cebra/fisiología , Animales , Electrofisiología , Procesamiento de Imagen Asistido por ComputadorRESUMEN
Biological cellular structures have inspired many scientific disciplines to design synthetic structures that can mimic their functions. Here, we closely emulate biological cellular structures in a rationally designed synthetic multicellular hybrid ion pump, composed of hydrogen-bonded [EMIM+][TFSI-] ion pairs on the surface of silica microstructures (artificial mechanoreceptor cells) embedded into thermoplastic polyurethane elastomeric matrix (artificial extracellular matrix), to fabricate ionic mechanoreceptor skins. Ionic mechanoreceptors engage in hydrogen bond-triggered reversible pumping of ions under external stimulus. Our ionic mechanoreceptor skin is ultrasensitive (48.1-5.77 kPa-1) over a wide spectrum of pressures (0-135 kPa) at an ultra-low voltage (1 mV) and demonstrates the ability to surpass pressure-sensing capabilities of various natural skin mechanoreceptors (i.e., Merkel cells, Meissner's corpuscles, Pacinian corpuscles). We demonstrate a wearable drone microcontroller by integrating our ionic skin sensor array and flexible printed circuit board, which can control directions and speed simultaneously and selectively in aerial drone flight.
Asunto(s)
Técnicas Biosensibles/instrumentación , Electroquímica/instrumentación , Enlace de Hidrógeno , Mecanorreceptores/fisiología , Mecanotransducción Celular/fisiología , Fenómenos Fisiológicos de la Piel , Adulto , Biomimética/instrumentación , Técnicas Biosensibles/métodos , Humanos , Mecanorreceptores/química , Mecanorreceptores/citología , Células de Merkel/metabolismo , Estimulación Física , Poliuretanos , Presión , Gel de Sílice , Piel/citología , Tacto/fisiologíaRESUMEN
Differential coordination of growth and patterning across metazoans gives rise to a diversity of sizes and shapes at tissue, organ and organismal levels. Although tissue size and tissue function can be interdependent1-5, mechanisms that coordinate size and function remain poorly understood. Planarians are regenerative flatworms that bidirectionally scale their adult body size6,7 and reproduce asexually, via transverse fission, in a size-dependent manner8-10. This model offers a robust context to address the gap in knowledge that underlies the link between size and function. Here, by generating an optimized planarian fission protocol in Schmidtea mediterranea, we show that progeny number and the frequency of fission initiation are correlated with parent size. Fission progeny size is fixed by previously unidentified mechanically vulnerable planes spaced at an absolute distance along the anterior-posterior axis. An RNA interference screen of genes for anterior-posterior patterning uncovered components of the TGFß and Wnt signalling pathways as regulators of the frequency of fission initiation rather than the position of fission planes. Finally, inhibition of Wnt and TGFß signalling during growth altered the patterning of mechanosensory neurons-a neural subpopulation that is distributed in accordance with worm size and modulates fission behaviour. Our study identifies a role for TGFß and Wnt in regulating size-dependent behaviour, and uncovers an interdependence between patterning, growth and neurological function.
Asunto(s)
Tipificación del Cuerpo/fisiología , Tamaño Corporal/fisiología , Planarias/crecimiento & desarrollo , Planarias/fisiología , Factor de Crecimiento Transformador beta/metabolismo , Vía de Señalización Wnt/fisiología , Animales , Tipificación del Cuerpo/genética , Tamaño Corporal/genética , Sistema Nervioso Central/citología , Mecanorreceptores/citología , Mecanorreceptores/fisiología , Planarias/anatomía & histología , Planarias/citología , Interferencia de ARN , Reproducción Asexuada/fisiología , Vía de Señalización Wnt/genéticaRESUMEN
How the somatosensory cortex (S1) encodes complex patterns of touch, such as those that occur during tactile exploration, is poorly understood. In the mouse whisker S1, temporally dense stimulation of local whisker pairs revealed that most neurons are not classical single-whisker feature detectors, but instead are strongly tuned to two-whisker sequences that involve the columnar whisker (CW) and one specific surround whisker (SW), usually in a SW-leading-CW order. Tuning was spatiotemporally precise and diverse across cells, generating a rate code for local motion vectors defined by SW-CW combinations. Spatially asymmetric, sublinear suppression for suboptimal combinations and near-linearity for preferred combinations sharpened combination tuning relative to linearly predicted tuning. This resembles computation of motion direction selectivity in vision. SW-tuned neurons, misplaced in the classical whisker map, had the strongest combination tuning. Thus, each S1 column contains a rate code for local motion sequences involving the CW, thus providing a basis for higher-order feature extraction.
Asunto(s)
Mecanorreceptores/citología , Corteza Somatosensorial/citología , Percepción del Tacto/fisiología , Vibrisas/inervación , Animales , Ratones , Tacto/fisiologíaRESUMEN
Sensory hair cells are mechanoreceptors required for hearing and balance functions. From embryonic development, hair cells acquire apical stereociliary bundles for mechanosensation, basolateral ion channels that shape receptor potential, and synaptic contacts for conveying information centrally. These key maturation steps are sequential and presumed coupled; however, whether hair cells emerging postnatally mature similarly is unknown. Here, we show that in vivo postnatally generated and regenerated hair cells in the utricle, a vestibular organ detecting linear acceleration, acquired some mature somatic features but hair bundles appeared nonfunctional and short. The utricle consists of two hair cell subtypes with distinct morphological, electrophysiological and synaptic features. In both the undamaged and damaged utricle, fate-mapping and electrophysiology experiments showed that Plp1+ supporting cells took on type II hair cell properties based on molecular markers, basolateral conductances and synaptic properties yet stereociliary bundles were absent, or small and nonfunctional. By contrast, Lgr5+ supporting cells regenerated hair cells with type I and II properties, representing a distinct hair cell precursor subtype. Lastly, direct physiological measurements showed that utricular function abolished by damage was partially regained during regeneration. Together, our data reveal a previously unrecognized aberrant maturation program for hair cells generated and regenerated postnatally and may have broad implications for inner ear regenerative therapies.
Asunto(s)
Diferenciación Celular/fisiología , Células Ciliadas Auditivas/fisiología , Células Ciliadas Vestibulares/fisiología , Mecanorreceptores/fisiología , Regeneración/fisiología , Sáculo y Utrículo/fisiología , Animales , Fenómenos Electrofisiológicos/fisiología , Células Ciliadas Auditivas/citología , Células Ciliadas Vestibulares/citología , Mecanorreceptores/citología , Ratones Transgénicos , Sáculo y Utrículo/citología , Transmisión Sináptica/fisiologíaRESUMEN
We investigated the relationship between whisker mechanoreceptive inputs and the neural responses to optical stimulation in layer 2/upper 3 (L2/U3) of the barrel cortex using optogenetics since, ideally, we should investigate interactions among inputs with spatiotemporal acuity. Sixteen whisker points of a transgenic rat (W-TChR2V4), that expresses channelrhodopsin 2 (ChR2)-Venus conjugate (ChR2V) in the peripheral nerve endings surrounding the whisker follicles, were respectively connected one-by-one with 16 LED-coupled optical fibres, which illuminated the targets according to a certain pattern in order to evaluate interactions among the inputs in L2/U3. We found that the individual L2/U3 neurons frequently received excitatory inputs from multiple whiskers that were arrayed in a row. Although the interactions among major afferent inputs (MAIs) were negligible, negative interactions with the surrounding inputs suggest that the afferent inputs were integrated in the cortical networks to enhance the contrast of an array to its surroundings. With its simplicity, reproducibility and spatiotemporal acuity, the optogenetic approach would provide an alternative way to understand the principles of afferent integration in the cortex and should complement knowledge obtained by experiments using more natural stimulations.
Asunto(s)
Optogenética/métodos , Corteza Somatosensorial/fisiología , Animales , Femenino , Luz , Masculino , Mecanorreceptores/citología , Mecanorreceptores/fisiología , Neuronas Aferentes/citología , Neuronas Aferentes/fisiología , Optogenética/instrumentación , Estimulación Física , Ratas , Ratas Transgénicas , Corteza Somatosensorial/citología , Vibrisas/inervaciónRESUMEN
Low-threshold mechanosensitive C fibers (C-LTMRs) that express the vesicular glutamate transporter VGLUT3 are thought to signal affective touch, and may also play a role in mechanical allodynia. However, the nature of the central termination of C-LTMRs in the dorsal horn remains largely unexplored. Here, we used light and electron microscopy in combination with VGLUT3 immunolabeling as a marker of C-LTMR terminations to investigate this issue. VGLUT3+ C-LTMRs formed central terminals of Type II glomeruli in the inner part of lamina II of the dorsal horn, often establishing multiple asymmetric synapses with postsynaptic dendrites but also participating in synaptic configurations with presynaptic axons and dendrites. Unexpectedly, essentially all VGLUT3+ C-LTMR terminals showed substantial VGLUT1 expression in the rat, whereas such terminals in mice lacked VGLUT1. Most VGLUT3+ C-LTMR terminals exhibited weak-to-moderate VGLUT2 expression. Further, C-LTMR terminals formed numerous synapses with excitatory protein kinase Cγ (PKCγ) interneurons and inhibitory parvalbumin neurons, whereas synapses with calretinin neurons were scarce. C-LTMR terminals rarely if ever established synapses with neurokinin 1 receptor (NK1R)-possessing dendrites traversing lamina II. Thus, VGLUT3+ C-LTMR terminals appear to largely correspond to neurofilament-lacking central terminals of Type II glomeruli in inner lamina II and can thus be identified at the ultrastructural level by morphological criteria. The participation of C-LTMR terminals in Type II glomeruli involving diverse populations of interneuron indicates highly complex modes of integration of C-LTMR mediated signaling in the dorsal horn. Furthermore, differences in VGLUT1 expression indicate distinct species differences in synaptic physiology of C-LTMR terminals.
Asunto(s)
Sistemas de Transporte de Aminoácidos Acídicos/metabolismo , Mecanorreceptores/citología , Fibras Nerviosas Amielínicas/metabolismo , Médula Espinal/citología , Sinapsis/metabolismo , Proteínas de Transporte Vesicular de Glutamato/metabolismo , Animales , Femenino , Expresión Génica , Proteínas de Andamiaje Homer/metabolismo , Masculino , Mecanorreceptores/metabolismo , Ratones Endogámicos C57BL , Ratas Sprague-Dawley , Médula Espinal/metabolismo , Proteína 1 de Transporte Vesicular de Glutamato/metabolismo , Proteína 2 de Transporte Vesicular de Glutamato/metabolismoRESUMEN
Loss of one sensory modality can cause other types to become more perceptive (cross-modal plasticity). To test the hypothesis that the loss of vision changes the perceptual threshold in the somatosensory system, we applied optogenetics to directly manipulate the afferent inputs involved in the whisker-barrel system using a transgenic rat (W-TChR2V4) that expresses channelrhodopsin-2 (ChR2) selectively in the large mechanoreceptive neurons in the trigeminal ganglion (TG) and their peripheral nerve terminals. The licking behavior of W-TChR2V4 rat was conditioned to a blue LED light cue on the whisker area while the magnitude and duration of light pulses were varied. The perceptual threshold was thus quantitatively determined for each rat according to the relationship between the magnitude/duration of light and the reaction time between the LED light cue and the first licking event after it. We found that the perceptual threshold was more significantly reduced than the control non-deprived rats when the rats were visually deprived at postnatal 26-30 days (P26-30, early VD group), but not at P58-66 (late VD group). However, the sensory threshold of a late VD animal was similar to that of a control. Our results suggest the presence of cross-modal plasticity by which the loss of vision at the juvenile period increased the sensitivity of the somatosensory system involved in the touch of whiskers.
Asunto(s)
Plasticidad Neuronal/fisiología , Umbral Sensorial/fisiología , Corteza Somatosensorial/fisiología , Tacto/fisiología , Vibrisas/fisiología , Visión Ocular/fisiología , Animales , Condicionamiento Psicológico , Femenino , Aseo Animal/fisiología , Luz , Masculino , Mecanorreceptores/citología , Mecanorreceptores/fisiología , Optogenética/métodos , Nervios Periféricos/citología , Nervios Periféricos/fisiología , Ratas , Ratas Transgénicas , Corteza Somatosensorial/citología , Factores de Tiempo , Ganglio del Trigémino/citología , Ganglio del Trigémino/fisiología , Vibrisas/citologíaRESUMEN
Tunicates, the sister group of vertebrates, possess a mechanoreceptor organ, the coronal organ, which is considered the best candidate to address the controversial issue of vertebrate hair cell evolution. The organ, located at the base of the oral siphon, controls the flow of seawater into the organism and can drive the "squirting" reaction, i.e., the rapid body muscle contraction used to eject dangerous particles during filtration. Coronal sensory cells are secondary mechanoreceptors and share morphological, developmental, and molecular traits with vertebrate hair cells. In the colonial tunicate Botryllus schlosseri, we described coronal organ differentiation during asexual development. Moreover, we showed that the ototoxic aminoglycoside gentamicin caused morphological and mechanosensorial impairment in coronal cells. Finally, fenofibrate had a strong protective effect on coronal sensory cells due to gentamicin-induced toxicity, as occurs in vertebrate hair cells. Our results reinforce the hypothesis of homology between vertebrate hair cells and tunicate coronal sensory cells.
Asunto(s)
Diferenciación Celular , Mecanorreceptores/fisiología , Urocordados/fisiología , Animales , Evolución Biológica , Mecanorreceptores/citología , Urocordados/citologíaRESUMEN
Meissner's and Pacinian corpuscles are cutaneous mechanoreceptors responsible for different modalities of touch. The development of these sensory formations in humans is poorly known, especially regarding the acquisition of the typical immunohistochemical profile related to their full functional maturity. Here we used a panel of antibodies (to specifically label the main corpuscular components: axon, Schwann-related cells and endoneurial-perineurial-related cells) to investigate the development of digital Meissner's and Pacinian corpuscles in a representative sample covering from 11 weeks of estimated gestational age (wega) to adulthood. Development of Pacinian corpuscles starts at 13 wega, and it is completed at 4 months of life, although their basic structure and immunohistochemical characteristics are reached at 36 wega. During development, around the axon, a complex network of S100 positive Schwann-related processes is progressively compacted to form the inner core, while the surrounding mesenchyme is organized and forms the outer core and the capsule. Meissner's corpuscles start to develop at 22 wega and complete their typical morphology and immunohistochemical profile at 8 months of life. In developing Meissner's corpuscles, the axons establish complex relationships with the epidermis and are progressively covered by Schwann-like cells until they complete the mature arrangement late in postnatal life. The present results demonstrate an asynchronous development of the Meissner's and Pacini's corpuscles and show that there is not a total correlation between morphological and immunohistochemical maturation. The correlation of the present results with touch-induced cortical activity in developing humans is discussed.
Asunto(s)
Dedos/anatomía & histología , Mecanorreceptores/fisiología , Corpúsculos de Pacini/crecimiento & desarrollo , Adolescente , Adulto , Anciano , Animales , Anticuerpos/inmunología , Axones/fisiología , Colágeno Tipo IV/análisis , Femenino , Dedos/embriología , Técnica del Anticuerpo Fluorescente , Edad Gestacional , Humanos , Inmunohistoquímica , Lactante , Recién Nacido , Masculino , Mecanorreceptores/citología , Ratones , Persona de Mediana Edad , Corpúsculos de Pacini/embriología , Embarazo , Conejos , Piel/anatomía & histología , Piel/embriología , Piel/crecimiento & desarrolloRESUMEN
BACKGROUND: The human elbow maintains its stability mainly through its bony structure. Stability is enhanced by ligamentous structures. To allow the ligamento-muscular reflex, which protects against strain and stress, mechanoreceptors are embedded in the ligament. This report describes the existence and the distribution of the elbow medial collateral ligaments (MCLs) mechanoreceptors. HYPOTHESIS: The bony attachment site has the highest density of mechanoreceptors, and the anterior part has the highest density of mechanoreceptors. MATERIALS AND METHODS: Eight MCLs of elbow from fresh frozen cadavers were used. The MCLs were harvested deep to the periosteum from the medial epicondyle to the ulna. The fan-shaped ligaments were divided into six regions of interest (ROI) and stained with modified gold chloride stain. Specimens were evaluated under a light microscope. Golgi, Ruffini, and Pacinian corpuscles were found in every specimen. The number and the distribution of each mechanoreceptor in each ROI were recorded. The density of each mechanoreceptor was calculated in regards to its volume. RESULTS: Golgi, Ruffini, and Pacinian corpuscles were seen in the ligament with small nerve fibers. Ruffini corpuscles had the highest median density of all three corpuscles. The median corpuscle density was higher in the anterior than in the posterior part and higher in the bony attachment than in the mid-substance site except for Golgi corpuscle. CONCLUSION: The three typical types of mechanoreceptors were identified in human MCL with the anterior part and bony attachment as the dominant distribution site. LEVEL OF EVIDENCE: Basic Science Study.
Asunto(s)
Ligamentos Colaterales/citología , Codo , Mecanorreceptores/citología , Anciano , Cadáver , Colorantes , Femenino , Compuestos de Oro , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Within the enteric nervous system, the neurons in charge to control motility of the gastrointestinal tract reside in a particular location nestled between two perpendicular muscle layers which contract and relax. We used primary cultured myenteric neurons of male guinea pigs to study mechanosensitivity of enteric neurons in isolation. Ultrafast Neuroimaging with a voltage-sensitive dye technique was used to record neuronal activity in response to shear stress and strain. Strain was induced by locally deforming the elastic cell culture substrate next to a neuron. Measurements showed that substrate strain was mostly elongating cells. Shear stress was exerted by hydrodynamic forces in a microchannel. Both stimuli induced excitatory responses. Strain activated 14% of the stimulated myenteric neurons that responded with a spike frequency of 1.9 (0.7/3.2) Hz, whereas shear stress excited only a few neurons (5.6%) with a very low spike frequency of 0 (0/0.6) Hz. Thus, shear stress does not seem to be an adequate stimulus for mechanosensitive enteric neurons (MEN) while strain activates enteric neurons in a relevant manner. Analyzing the adaptation behavior of MEN showed that shear stress activated rapidly/slowly/ultraslowly adapting MEN (2/62/36%) whereas strain only slowly (46%) and ultraslowly (54%) MEN. Paired experiments with strain and normal stress revealed three mechanosensitive enteric neuronal populations: one strain-sensitive (37%), one normal stress-sensitive (17%) and one strain- and stress-sensitive (46%). These results indicate that shear stress does not play a role in the neuronal control of motility but normal stress and strain.
Asunto(s)
Mecanorreceptores/fisiología , Plexo Mientérico/fisiología , Potenciales de Acción , Animales , Fenómenos Biomecánicos , Células Cultivadas , Cobayas , Hidrodinámica , Intestino Delgado , Masculino , Mecanorreceptores/citología , Plexo Mientérico/citología , Estimulación Física , Estrés Mecánico , Estrés Fisiológico/fisiología , Imagen de Colorante Sensible al VoltajeRESUMEN
Tactile-foraging ducks are specialist birds known for their touch-dependent feeding behavior. They use dabbling, straining, and filtering to find edible matter in murky water, relying on the sense of touch in their bill. Here, we present the molecular characterization of embryonic duck bill, which we show contains a high density of mechanosensory corpuscles innervated by functional rapidly adapting trigeminal afferents. In contrast to chicken, a visually foraging bird, the majority of duck trigeminal neurons are mechanoreceptors that express the Piezo2 ion channel and produce slowly inactivating mechano-current before hatching. Furthermore, duck neurons have a significantly reduced mechano-activation threshold and elevated mechano-current amplitude. Cloning and electrophysiological characterization of duck Piezo2 in a heterologous expression system shows that duck Piezo2 is functionally similar to the mouse ortholog but with prolonged inactivation kinetics, particularly at positive potentials. Knockdown of Piezo2 in duck trigeminal neurons attenuates mechano current with intermediate and slow inactivation kinetics. This suggests that Piezo2 is capable of contributing to a larger range of mechano-activated currents in duck trigeminal ganglia than in mouse trigeminal ganglia. Our results provide insights into the molecular basis of mechanotransduction in a tactile-specialist vertebrate.
Asunto(s)
Proteínas Aviares/genética , Pico/fisiología , Patos/fisiología , Mecanorreceptores/metabolismo , Percepción del Tacto/fisiología , Tacto/fisiología , Secuencia de Aminoácidos , Animales , Proteínas Aviares/antagonistas & inhibidores , Proteínas Aviares/metabolismo , Pico/citología , Pico/inervación , Pollos , Clonación Molecular , Embrión no Mamífero , Expresión Génica , Vectores Genéticos/genética , Vectores Genéticos/metabolismo , Células HEK293 , Humanos , Canales Iónicos/antagonistas & inhibidores , Canales Iónicos/genética , Canales Iónicos/metabolismo , Cinética , Mecanorreceptores/citología , Mecanotransducción Celular , Ratones , Técnicas de Placa-Clamp , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Homología de Secuencia de Aminoácido , Especificidad de la Especie , Ganglio del Trigémino/citología , Ganglio del Trigémino/metabolismoRESUMEN
We review the development and evolution of the ear neurosensory cells, the aggregation of neurosensory cells into an otic placode, the evolution of novel neurosensory structures dedicated to hearing and the evolution of novel nuclei in the brain and their input dedicated to processing those novel auditory stimuli. The evolution of the apparently novel auditory system lies in duplication and diversification of cell fate transcription regulation that allows variation at the cellular level [transforming a single neurosensory cell into a sensory cell connected to its targets by a sensory neuron as well as diversifying hair cells], organ level [duplication of organ development followed by diversification and novel stimulus acquisition] and brain nuclear level [multiplication of transcription factors to regulate various neuron and neuron aggregate fate to transform the spinal cord into the unique hindbrain organization]. Tying cell fate changes driven by bHLH and other transcription factors into cell and organ changes is at the moment tentative as not all relevant factors are known and their gene regulatory network is only rudimentary understood. Future research can use the blueprint proposed here to provide both the deeper molecular evolutionary understanding as well as a more detailed appreciation of developmental networks. This understanding can reveal how an auditory system evolved through transformation of existing cell fate determining networks and thus how neurosensory evolution occurred through molecular changes affecting cell fate decision processes. Appreciating the evolutionary cascade of developmental program changes could allow identifying essential steps needed to restore cells and organs in the future.
Asunto(s)
Evolución Biológica , Oído Interno/crecimiento & desarrollo , Animales , Vías Auditivas/crecimiento & desarrollo , Vías Auditivas/fisiología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/fisiología , Oído Interno/anatomía & histología , Oído Interno/fisiología , Evolución Molecular , Duplicación de Gen , Células Ciliadas Auditivas/citología , Células Ciliadas Auditivas/fisiología , Audición/genética , Audición/fisiología , Mecanorreceptores/citología , Mecanorreceptores/fisiología , Modelos Biológicos , Células Receptoras Sensoriales/citología , Células Receptoras Sensoriales/fisiologíaRESUMEN
Cell cycle progression and differentiation are highly coordinated during the development of multicellular organisms. The mechanisms by which these processes are coordinated and how their coordination contributes to normal development are not fully understood. Here, we determine the developmental fate of a population of precursor cells in the developing Drosophila melanogaster retina that arrest in G2 phase of the cell cycle and investigate whether cell cycle phase-specific arrest influences the fate of these cells. We demonstrate that retinal precursor cells that arrest in G2 during larval development are selected as sensory organ precursors (SOPs) during pupal development and undergo two cell divisions to generate the four-cell interommatidial mechanosensory bristles. While G2 arrest is not required for bristle development, preventing G2 arrest results in incorrect bristle positioning in the adult eye. We conclude that G2-arrested cells provide a positional cue during development to ensure proper spacing of bristles in the eye. Our results suggest that the control of cell cycle progression refines cell fate decisions and that the relationship between these two processes is not necessarily deterministic.
Asunto(s)
Ojo Compuesto de los Artrópodos/citología , Drosophila melanogaster/citología , Células Epiteliales/citología , Fase G2 , Mecanorreceptores/citología , Animales , Puntos de Control del Ciclo Celular/fisiología , Diferenciación Celular , División Celular , Linaje de la Célula , Ojo Compuesto de los Artrópodos/crecimiento & desarrollo , Ojo Compuesto de los Artrópodos/ultraestructura , Proteínas de Drosophila/fisiología , Drosophila melanogaster/crecimiento & desarrollo , Discos Imaginales/citología , Larva , Mecanorreceptores/ultraestructura , Mecanotransducción Celular , Neuroglía/citología , Células Fotorreceptoras de Invertebrados/citología , Pupa , Células Receptoras Sensoriales/citologíaRESUMEN
In sensory hair cells of auditory and vestibular organs, the ribbon synapse is required for the precise encoding of a wide range of complex stimuli. Hair cells have a unique presynaptic structure, the synaptic ribbon, which organizes both synaptic vesicles and calcium channels at the active zone. Previous work has shown that hair-cell ribbon size is correlated with differences in postsynaptic activity. However, additional variability in postsynapse size presents a challenge to determining the specific role of ribbon size in sensory encoding. To selectively assess the impact of ribbon size on synapse function, we examined hair cells in transgenic zebrafish that have enlarged ribbons, without postsynaptic alterations. Morphologically, we found that enlarged ribbons had more associated vesicles and reduced presynaptic calcium-channel clustering. Functionally, hair cells with enlarged ribbons had larger global and ribbon-localized calcium currents. Afferent neuron recordings revealed that hair cells with enlarged ribbons resulted in reduced spontaneous spike rates. Additionally, despite larger presynaptic calcium signals, we observed fewer evoked spikes with longer latencies from stimulus onset. Together, our work indicates that hair-cell ribbon size influences the spontaneous spiking and the precise encoding of stimulus onset in afferent neurons.SIGNIFICANCE STATEMENT Numerous studies support that hair-cell ribbon size corresponds with functional sensitivity differences in afferent neurons and, in the case of inner hair cells of the cochlea, vulnerability to damage from noise trauma. Yet it is unclear whether ribbon size directly influences sensory encoding. Our study reveals that ribbon enlargement results in increased ribbon-localized calcium signals, yet reduces afferent spontaneous activity and disrupts the timing of stimulus onset, a distinct aspect of auditory and vestibular encoding. These observations suggest that varying ribbon size alone can influence sensory encoding, and give further insight into how hair cells transduce signals that cover a wide dynamic range of stimuli.
Asunto(s)
Potenciales de Acción/fisiología , Señalización del Calcio/fisiología , Mecanorreceptores/citología , Mecanorreceptores/fisiología , Tiempo de Reacción/fisiología , Pez Cebra/fisiología , Animales , Animales Modificados Genéticamente , Tamaño de la Célula , Sistema de la Línea Lateral/citología , Sistema de la Línea Lateral/fisiología , Inhibición Neural/fisiología , Pez Cebra/anatomía & histologíaRESUMEN
In a type of short-term plasticity that is observed in a number of systems, synaptic transmission is potentiated by depolarizing changes in the membrane potential of the presynaptic neuron before spike initiation. This digital-analog form of plasticity is graded. The more depolarized the neuron, the greater the increase in the efficacy of synaptic transmission. In a number of systems, including the system presently under investigation, this type of modulation is calcium dependent, and its graded nature is presumably a consequence of a direct relationship between the intracellular calcium concentration ([Ca2+]i) and the effect on synaptic transmission. It is therefore of interest to identify factors that determine the magnitude of this type of calcium signal. We studied a synapse in Aplysia and demonstrate that there can be a contribution from currents activated during spiking. When neurons spike, there are localized increases in [Ca2+]i that directly trigger neurotransmitter release. Additionally, spiking can lead to global increases in [Ca2+]i that are reminiscent of those induced by subthreshold depolarization. We demonstrate that these spike-induced increases in [Ca2+]i result from the activation of a current not activated by subthreshold depolarization. Importantly, they decay with a relatively slow time constant. Consequently, with repeated spiking, even at a low frequency, they readily summate to become larger than increases in [Ca2+]i induced by subthreshold depolarization alone. When this occurs, global increases in [Ca2+]i induced by spiking play the predominant role in determining the efficacy of synaptic transmission.NEW & NOTEWORTHY We demonstrate that spiking can induce global increases in the intracellular calcium concentration ([Ca2+]i) that decay with a relatively long time constant. Consequently, summation of the calcium signal occurs even at low firing frequencies. As a result there is significant, persistent potentiation of synaptic transmission.
Asunto(s)
Calcio/metabolismo , Espacio Intracelular/metabolismo , Plasticidad Neuronal/fisiología , Sinapsis/metabolismo , Transmisión Sináptica/fisiología , Análisis de Varianza , Animales , Aplysia , Cationes Bivalentes/metabolismo , Femenino , Ganglios de Invertebrados/citología , Ganglios de Invertebrados/metabolismo , Masculino , Mecanorreceptores/citología , Mecanorreceptores/metabolismo , Microelectrodos , Técnicas de Cultivo de Tejidos , Imagen de Colorante Sensible al VoltajeRESUMEN
New tools for applying force to animals, tissues, and cells are critically needed in order to advance the field of mechanobiology, as few existing tools enable simultaneous imaging of tissue and cell deformation as well as cellular activity in live animals. Here, we introduce a novel microfluidic device that enables high-resolution optical imaging of cellular deformations and activity while applying precise mechanical stimuli to the surface of the worm's cuticle with a pneumatic pressure reservoir. To evaluate device performance, we compared analytical and numerical simulations conducted during the design process to empirical measurements made with fabricated devices. Leveraging the well-characterized touch receptor neurons (TRNs) with an optogenetic calcium indicator as a model mechanoreceptor neuron, we established that individual neurons can be stimulated and that the device can effectively deliver steps as well as more complex stimulus patterns. This microfluidic device is therefore a valuable platform for investigating the mechanobiology of living animals and their mechanosensitive neurons.
Asunto(s)
Dispositivos Laboratorio en un Chip , Mecanorreceptores , Técnicas Analíticas Microfluídicas , Animales , Animales Modificados Genéticamente , Caenorhabditis elegans , Calcio/metabolismo , Diseño de Equipo , Mecanorreceptores/química , Mecanorreceptores/citología , Mecanorreceptores/fisiología , Técnicas Analíticas Microfluídicas/instrumentación , Técnicas Analíticas Microfluídicas/métodos , Imagen Óptica , Optogenética , Estimulación Física/instrumentación , Estimulación Física/métodosRESUMEN
OBJECTIVE: Many skin flaps have been described for fingertip reconstruction; however, they have not been compared histologically. The aim of this study is to compare the histological features of common insensate flaps that are used for fingertip reconstruction. METHOD: Skin from fingertips and common flap donor sites on the hand and forearm of cadavers were harvested. This study investigated four histological characteristics, namely thickness of the epidermis and dermis; the ratio of collagen to elastic fibres (C/E ratio) in subdermal tissues, and distribution densities of Merkel cells and Meissner's corpuscles. It then compared the values obtained to determine which flap donor site best matched the fingertip. RESULTS: Epidermal thickness of the reverse digital artery island flap, thenar flap, and hypothenar flap was similar to that of fingertip tissue; dermal thickness of the hypothenar flap was similar to that of fingertip tissue. The C/E ratio of the reverse digital artery island flap was similar to that of fingertip tissue. Merkel cells were abundant in the reverse digital artery island flap, but Meissner's corpuscles were few in each of the flaps compared with fingertip tissue. CONCLUSION: The flap donor site with histological properties most similar to fingertip tissue was the palmar lateral aspect at the finger base, representative of the reverse digital artery island flap with respect to epidermal thickness, C/E ratio, and presence of Merkel cells. The thenar and hypothenar flaps also showed similar properties.