The condition of septic arthritis was treated in 12 foals with 21 affected joints (Group I) and in 27 adult horses. The adult horses were divided into three groups, based on aetiology of the condition: haematogenous (Group II, n = 6), iatrogenic (Group III, n = 6), and perforating trauma (Group IV, n = 15). The treatment consisted of an initial systemic antibiotic that anticipated the microbial agents that were considered most likely per group, repeated through-and-through joint lavages every other day and non-steroidal anti-inflammatory drugs. The antibiotics were adjusted to the results of bacteriological culture and susceptibility tests. Joint lavages were continued until the white blood cell count dropped below 15 G/l and bacteriological culture was negative, after which a single dose of a short-acting corticosteroid was administered intra-articularly. Joint recovery rate in group I was 71%. Patient recovery rate of the foals, however, was lower (42%). Three foals were killed for reasons other than arthritis; one foal because of an arthritis-related problem and three foals because of persistent arthritis. Overall joint recovery rate, equalling patient recovery rate, in the adult horses was 81%. The expected predominance of Streptococcus spp. in haematogenous arthritis in adult horses was not confirmed, indicating that in these cases also, an initial antibiotic treatment with a broad-spectrum combination is preferable. It is concluded that with intensive treatment, the prognosis of septic arthritis in the adult horse can be classified as fair to even good. Results in the foals are not as good, but this seems to be more due to the specific problems surrounding the equine neonate than to unresponsiveness to the treatment.

The energy required to fuse synaptic vesicles with the plasma membrane (‘activation energy’) is considered a major determinant in synaptic efficacy. From reaction rate theory, we predict that a class of modulations exists, which utilize linear modulation of the energy barrier for fusion to achieve supralinear effects on the fusion rate. To test this prediction experimentally, we developed a method to assess the number of releasable vesicles, rate constants for vesicle priming, unpriming, and fusion, and the activation energy for fusion by fitting a vesicle state model to synaptic responses induced by hypertonic solutions. We show that complexinI/II deficiency or phorbol ester stimulation indeed affects responses to hypertonic solution in a supralinear manner. An additive vs multiplicative relationship between activation energy and fusion rate provides a novel explanation for previously observed non-linear effects of genetic/pharmacological perturbations on synaptic transmission and a novel interpretation of the cooperative nature of Ca2+-dependent release.DOI: http://dx.doi.org/10.7554/eLife.05531.001

Synaptic transmission depends critically on the Sec1p/ Munc18 protein Munc18-1, but it is unclear whether Munc18-1 primarily operates as a integral part of the fusion machinery or has a more upstream role in fusion complex assembly. Here, we show that point mutations in Munc18-1 that interfere with binding to the free Syntaxin1a N-terminus and strongly impair binding to assembled SNARE complexes all support normal docking, priming and fusion of synaptic vesicles, and normal synaptic plasticity in munc18-1 null mutant neurons. These data support a prevailing role of Munc18-1 before/ during SNARE-complex assembly, while its continued association to assembled SNARE complexes is dispensable for synaptic transmission.

Highlights d We establish a single-cell model to study synapses in iPSCderived neurons d This platform allows quantitative analysis of synaptic transmission and plasticity d The platform is validated for GABA-or glutamatergic iPSCderived human neurons d The platform is scalable and suitable for compound screening and disease modeling

Presynaptic activation of the diacylglycerol (DAG)/protein kinase C (PKC) pathway is a central event in short-term synaptic plasticity. Two substrates, Munc13-1 and Munc18-1, are essential for DAGinduced potentiation of vesicle priming, but the role of most presynaptic PKC substrates is not understood. Here, we show that a mutation in synaptotagmin-1 (Syt1 T112A ), which prevents its PKCdependent phosphorylation, abolishes DAG-induced potentiation of synaptic transmission in hippocampal neurons. This mutant also reduces potentiation of spontaneous release, but only if alternative Ca 2+ sensors, Doc2A/B proteins, are absent. However, unlike mutations in Munc13-1 or Munc18-1 that prevent DAG-induced potentiation, the synaptotagmin-1 mutation does not affect pairedpulse facilitation. Furthermore, experiments to probe vesicle priming (recovery after train stimulation and dual application of hypertonic solutions) also reveal no abnormalities. Expression of synaptotagmin-2, which lacks a seven amino acid sequence that contains the phosphorylation site in synaptotagmin-1, or a synaptotagmin-1 variant with these seven residues removed (Syt1 Δ109-116 ), supports normal DAG-induced potentiation. These data suggest that this seven residue sequence in synaptotagmin-1 situated in the linker between the transmembrane and C2A domains is inhibitory in the unphosphorylated state and becomes permissive of potentiation upon phosphorylation. We conclude that synaptotagmin-1 phosphorylation is an essential step in PKC-dependent potentiation of synaptic transmission, acting downstream of the two other essential DAG/PKC substrates, Munc13-1 and Munc18-1.P resynaptic strength changes rapidly during repetitive stimulation [short-term plasticity (STP)] and activation of intracellular signal transduction pathways (1, 2). The diacylglycerol (DAG)/protein kinase C (PKC) cascade is one of the most potent pathways at the presynaptic terminal. Its activation leads to 50-100% potentiation of spontaneous and action potential (AP)-evoked release (3-5), and is critical for multiple forms of presynaptic plasticity (6-8). DAG directly activates the vesicle priming factor Munc13-1 (9, 10) and indirectly activates downstream effectors via PKC. Activation of both Munc13-1 and PKC is essential for this pathway to operate (Fig. 1A) (8, 11). We previously identified Munc18-1 as an essential PKC substrate, because a nonphosphorylatable Munc18-1 mutant completely inhibits PKC-dependent STP (8, 12). Importantly, a phosphomimetic mutation of Munc18-1 cannot fully bypass the requirement for PKC activation, indicating that other PKC substrates must contribute to this form of plasticity (8). These substrates have not been identified to date.Among other presynaptic PKC substrates is synaptotagmin-1 (Syt1, ref. 13). Syt1 is the vesicular Ca 2+ sensor that mediates fast AP-evoked release in the hippocampus (14) and drives a large fraction of spontaneous release (15). In the latter case, Syt1 competes with alternative sensors, in particular Doc2s, for SNARE binding ...