At fast CNS synapses, the role of asynchronous release following initial synchronous release is poorly understood. We examined the contribution of asynchronous release to GABAergic transmission in the cochlear nucleus across a 40-fold range of electrical stimulus frequencies. Whereas quantal release was highly synchronized at low frequencies, it was largely continuous and desynchronized at high frequencies. Despite the change in release mode, intense and steady inhibitory transmission was virtually maintained. Experimental analyses and modeling studies indicated that this "desynchronization" process was dependent on presynaptic Ca2+ accumulation, facilitation of vesicle release, and short-term depletion of available vesicles. Asynchronous release at high frequencies may help generate a smooth inhibitory "tone" by minimizing the consequences of random timing of presynaptic action potentials.

The firing pattern of neurons is shaped by the convergence of excitation and inhibition, each with finely tuned magnitude and duration. In an auditory brainstem nucleus, glycinergic inhibition features fast decay kinetics, the mechanism of which is unknown. By applying glycine to native or recombinant glycine receptors, we show that response decay times are accelerated by addition of GABA, a weak partial agonist of glycine receptors. Systematic variation in agonist exposure time revealed that fast synaptic time course may be achieved with submillisecond exposures to mixtures of glycine and GABA at physiological concentrations. Accordingly, presynaptic terminals generally contained both transmitters, and depleting terminals of GABA slowed glycinergic synaptic currents. Thus, coreleased GABA accelerates glycinergic transmission by acting directly on glycine receptors, narrowing the time window for effective inhibition. Packaging both weak and strong agonists in vesicles may be a general means by which presynaptic neurons regulate the duration of postsynaptic responses.

_ gradient, measured using whole-cell recording. With gramicidin-perforated patch recording, E GABA was _25 ± 5 mV (mean ± S.D.), and was stable between embryonic day 17 and posthatch day 10. With normal intracellular Cl _ , GABA depolarized neurons by 12 mV.3. In current clamp, repetitive activation of the GABAergic axons reduced the probability of spiking in response to simultaneous stimulation of excitatory axons. However, IPSPs could themselves elicit action potentials, and facilitation of IPSPs by repetitive activation could lead to a characteristic pattern of spiking.4. These data indicate that IPSPs with reversal potentials positive to spike threshold may have dual functions, depending on the context of their activation.

The effect of exogenous CaCl2 on photosystem I and II (PSI and PSII) activities, cyclic electron flow (CEF), and proton motive force of tomato leaves under low night temperature (LNT) was investigated. LNT stress decreased the net photosynthetic rate (Pn), effective quantum yield of PSII [Y(II)], and photochemical quenching (qP), whereas CaCl2 pretreatment improved Pn, Y(II), and qP under LNT stress. LNT stress significantly increased the non-regulatory quantum yield of energy dissipation [Y(NO)], whereas CaCl2 alleviated this increase. Exogenous Ca2+ enhanced stimulation of CEF by LNT stress. Inhibition of oxidized PQ pools caused by LNT stress was alleviated by CaCl2 pretreatment. LNT stress reduced zeaxanthin formation and ATPase activity, but CaCl2 pretreatment reversed both of these effects. LNT stress caused excess formation of a proton gradient across the thylakoid membrane, whereas CaCl2 pretreatment decreased the said factor under LNT. Thus, our results showed that photoinhibition of LNT-stressed plants could be alleviated by CaCl2 pretreatment. Our findings further revealed that this alleviation was mediated in part by improvements in carbon fixation capacity, PQ pools, linear and cyclic electron transports, xanthophyll cycles, and ATPase activity.

The conventional calpains, m- and micro-calpain, are suggested to be involved in apoptosis triggered by many different mechanisms. However, it has not been possible to definitively associate calpain function with apoptosis, largely because of the incomplete selectivity of the cell permeable calpain inhibitors used in previous studies. In the present study, Chinese hamster ovary (CHO) cell lines overexpressing micro-calpain or the highly specific calpain inhibitor protein, calpastatin, have been utilized to explore apoptosis signals that are influenced by calpain content. This approach allows unambiguous alteration of calpain activity in cells. Serum depletion, treatment with the endoplasmic reticulum (ER) calcium ATPase inhibitor thapsigargin, and treatment with calcium ionophore A23187 produced apoptosis in CHO cells, which was increased in calpain overexpressing cells and decreased by induced expression of calpastatin. Inhibition of calpain activity protected beta-spectrin, but not alpha-spectrin, from proteolysis. The calpains seemed not to be involved in apoptosis triggered by a number of other treatments. Calpain protected against TNF-alpha induced apoptosis. In contrast to previous studies, we found no evidence that calpains proteolyze I kappa B-alpha in TNF-alpha-stimulated cells. These studies indicate that the conventional calpains participate in some, but not all, apoptotic signaling mechanisms. In most cases, they contributed to apoptosis, but in at least one case, they were protective.

Low light (LL) is one of the main limiting factors that negatively affect tomato growth and yield. Techniques of chemical regulation are effective horticultural methods to improve stress resistance. Strigolactones (SLs), newly discovered phytohormones, are considered as important regulators of physiological responses. We investigated the effects of foliage spray of GR24, a synthesized SLs, on tomato seedlings grown under LL stress conditions. The results showed that application of GR24 effectively mitigated the inhibition of plant growth and increased the fresh and dry weight of tomato plants under LL. Additionally, GR24 also increased the chlorophyll content (Chl a and Chl b ), the net photosynthetic rate (Pn), the photochemical efficiency of photosystem (PS) II (Fv/Fm), and the effective quantum yield of PSII and I [Y(II) and Y(I)], but decreased the excitation pressure of PSII (1-qP), the non-regulatory quantum yield of energy dissipation [Y(NO)] and the donor side limitation of PSI [Y(ND)] under LL. Moreover, application of GR24 to LL-stressed tomato leaves increased the electron transport rate of PSII and PSI [ETR(II) and ETR(I)], the ratio of the quantum yield of cyclic electron flow (CEF) to Y(II) [Y(CEF)/Y(II)], the oxidized plastoquinone (PQ) pool size and the non-photochemical quenching. Besides, GR24 application increased the activity and gene expression of antioxidant enzymes, but it reduced malonaldehyde (MDA) and hydrogen peroxide (H 2 O 2 ) content in LL-stressed plants. These results suggest that exogenous application of GR24 enhances plant tolerance to LL by promoting plant utilization of light energy to alleviate the photosystem injuries induced by excess light energy and ROS, and enhancing photosynthesis efficiency to improve plant growth.

The development of synaptic function was examined at auditory nerve synapses in the rostromedial region of the cochlear nucleus magnocellularis of the chick. EPSCs were studied beginning at embryonic day 12 (E12), when synaptic transmission was first observed, through E19. The amplitude of evoked EPSCs produced by AMPA receptor (AMPA-R) increased 30-fold over this age range, whereas NMDA receptor (NMDA-R)-mediated transmission peaked at E14 and then declined almost completely. At E12, Ͼ80% of the miniature EPSCs exhibited both receptor components, and Ͻ10% were NMDA-R only. With age, the contribution of NMDA-R to miniature EPSCs steadily declined, suggesting that NMDA-R number is gradually reduced at individual postsynaptic sites. Between E12 and E16, the number of axonal inputs to each cell reduced by half. In simultaneous recordings from adjacent neurons, synchronous EPSCs were observed that resulted from spontaneous firing of the same presynaptic fiber. The difference in amplitude of the EPSCs in the two cells was greater in E14 than E12, whereas at E16 synchronous events were no longer observed, suggesting that the weaker input was destined for elimination. The relative amplitude of the NMDA-R component, compared with the AMPA-R component, was larger for the weaker inputs. When elimination was underway, AMPA-R quantal size was much reduced for the weakest terminals. Thus, elimination of auditory nerve terminals and pruning of axonal branches is preceded by a reduction in quantal efficacy.

High temperature and high light intensity is a common environment posing a great risk to organisms. This study aimed to elucidate the effects of sub-high temperature and high light intensity stress (HH, 35°C, 1000 μmol⋅m-2⋅s-1) and recovery on the photosynthetic mechanism, photoinhibiton of photosystem II (PSII) and photosystem I (PSI), and reactive oxygen (ROS) metabolism of tomato seedlings. The results showed that with prolonged stress time, net photosynthetic rate (Pn), Rubisco activity, maximal photochemistry efficiency (Fv/Fm), efficient quantum yield and electron transport of PSII [Y(II) and ETR(II)] and PSI [Y(I) and ETR(I)] decreased significantly whereas yield of non-regulated and regulated energy dissipation of PSII [Y(NO) and Y(NPQ)] increased sharply. The donor side limitation of PSI [Y(ND)] increased but the acceptor side limitation of PSI [Y(NA)] decreased. Content of malondialdehyde (MDA) and hydrogen peroxide (H2O2) were increased while activity of superoxide dismutase (SOD) and peroxidase (POD) were significantly inhibited compared with control. HH exposure affected photosynthetic carbon assimilation, multiple sites in PSII and PSI, ROS accumulation and elimination of Solanum lycopersicum L.