Static and dynamic properties of synaptic transmission at the cyto-neural junction of frog labyrinth posterior canal

The properties of synaptic transmission have been studied at the cyto-neural junction of the frog labyrinth posterior canal by examining excitatory postsynaptic potential (EPSP) activity recorded intraaxonally from the afferent nerve after abolishing spike firing by tetrodotoxin. The waveform, amplitude, and rate of occurrence of the EPSPs have been evaluated by means of a procedure of fluctuation analysis devised to continuously monitor these parameters, at rest as well as during stimulation of the semicircular canal by sinusoidal rotation at 0.1 Hz, with peak accelerations ranging from 8 to 87 deg.s-2. Responses to excitatory and inhibitory accelerations were quantified in terms of maximum and minimum EPSP rates, respectively, as well as total numbers of EPSPs occurring during the excitatory and inhibitory half cycles. Excitatory responses were systematically larger than inhibitory ones (asymmetry). Excitatory responses were linearly related either to peak acceleration or to its logarithm, and the same occurred for inhibitory responses. In all units examined, the asymmetry of the response yielded nonlinear two-sided input-output intensity functions. Silencing of EPSPs during inhibition (rectification) was never observed. Comparison of activity during the first cycle of rotation with the average response over several cycles indicated that variable degrees of adaptation (up to 48%) characterize the excitatory response, whereas no consistent adaptation was observed in the inhibitory response. All fibers appeared to give responses nearly in phase with angular velocity, at 0.1 Hz, although the peak rates generally anticipated by a few degrees the peak angular velocity. From the data presented it appears that asymmetry, adaptation, and at least part of the phase lead in afferent nerve response are of presynaptic origin, whereas rectification and possible further phase lead arise at the encoder. To confirm these conclusions a simultaneous though limited study of spike firing and EPSP activity has been attempted in a few fibers

FcRn-mediated antibody transport across epithelial cells revealed by electron tomography

The neonatal Fc receptor (FcRn) transports maternal IgG across epithelial barriers, thereby providing the fetus or newborn with humoral immunity before its immune system is fully functional. In newborn rats, FcRn transfers IgG from milk to blood by apical-to-basolateral transcytosis across intestinal epithelial cells. The pH difference between the apical (pH 6.0–6.5) and basolateral (pH 7.4) sides of intestinal epithelial cells facilitates the efficient unidirectional transport of IgG, because FcRn binds IgG at pH 6.0–6.5 but not at pH 7 or more. As milk passes through the neonatal intestine, maternal IgG is removed by FcRn-expressing cells in the proximal small intestine (duodenum and jejunum); remaining proteins are absorbed and degraded by FcRn-negative cells in the distal small intestine (ileum). Here we use electron tomography to make jejunal transcytosis visible directly in space and time, developing new labelling and detection methods to map individual nanogold-labelled Fc within transport vesicles and simultaneously to characterize these vesicles by immunolabelling. Combining electron tomography with a nonperturbing endocytic label allowed us to conclusively identify receptor-bound ligands, resolve interconnecting vesicles, determine whether a vesicle was microtubule-associated, and accurately trace FcRn-mediated transport of IgG. Our results present a complex picture in which Fc moves through networks of entangled tubular and irregular vesicles, only some of which are microtubule-associated, as it migrates to the basolateral surface. New features of transcytosis are elucidated, including transport involving multivesicular body inner vesicles/tubules and exocytosis through clathrin-coated pits. Markers for early, late and recycling endosomes each labelled vesicles in different and overlapping morphological classes, revealing spatial complexity in endo-lysosomal trafficking

The kinetics of nerve-evoked quantal secretion.

Current views on quantal release of neurotransmitters hold that after the vesicle migrates towards release sites (active zones), multiple protein interactions mediate the docking of the vesicle to the presynaptic membrane and the formation of a multimolecular protein complex (the 'fusion machine') which ultimately makes the vesicle competent to release a quantum in response to the action potential. Classical biophysical studies of quantal release have modelled the process by a binomial system where n vesicles (sites) competent for exocytosis release a quantum, with probability p, in response to the action potential. This is likely to be an oversimplified model. Furthermore, statistical and kinetic studies have given results which are difficult to reconcile within this framework. Here, data are presented and discussed which suggest a revision of the biophysical model. Transient silencing of release is shown to occur following the pulse of synchronous transmitter release, which is evoked by the presynaptic action potential. This points to a schema where the vesicle fusion complex assembly is a reversible, stochastic process. Asynchronous exocytosis may occur at several intermediate stages in the process, along paths which may be differentially regulated by divalent cations or other factors. The fusion complex becomes competent for synchronous release (armed vesicles) only at appropriately organized sites. The action potential then triggers (deterministically rather than stochastically) the synchronous discharge of all armed vesicles. The existence of a specific conformation for the fusion complex to be competent for synchronous evoked fusion reconciles statistical and kinetic results during repetitive stimulation and helps explain the specific effects of toxins and genetic manipulation on the synchronization of release in response to an action potential

La cellula come sistema biologico integrato

in "Fisiologia Medica". F. Conti Ed

Controllo del pH dell'organismo

in "Fisiologia Medica". F. Conti Ed