High Speed Two-Photon Imaging of Calcium Dynamics in Dendritic Spines: Consequences for Spine Calcium Kinetics and Buffer Capacity

Rapid calcium concentration changes in postsynaptic structures are crucial for synaptic plasticity. Thus far, the determinants of postsynaptic calcium dynamics have been studied predominantly based on the decay kinetics of calcium transients. Calcium rise times in spines in response to single action potentials (AP) are almost never measured due to technical limitations, but they could be crucial for synaptic plasticity. With high-speed, precisely-targeted, two-photon point imaging we measured both calcium rise and decay kinetics in spines and secondary dendrites in neocortical pyramidal neurons. We found that both rise and decay kinetics of changes in calcium-indicator fluorescence are about twice as fast in spines. During AP trains, spine calcium changes follow each AP, but not in dendrites. Apart from the higher surface-to-volume ratio (SVR), we observed that neocortical dendritic spines have a markedly smaller endogenous buffer capacity with respect to their parental dendrites. Calcium influx time course and calcium extrusion rate were both in the same range for spines and dendrites when fitted with a dynamic multi-compartment model that included calcium binding kinetics and diffusion. In a subsequent analysis we used this model to investigate which parameters are critical determinants in spine calcium dynamics. The model confirmed the experimental findings: a higher SVR is not sufficient by itself to explain the faster rise time kinetics in spines, but only when paired with a lower buffer capacity in spines. Simulations at zero calcium-dye conditions show that calmodulin is more efficiently activated in spines, which indicates that spine morphology and buffering conditions in neocortical spines favor synaptic plasticity

Produção de maracujazeiro-amarelo sob diferentes densidades de plantio

O adensamento de plantas em cultivos do maracujazeiro-amarelo é importante por propiciar maior rentabilidade em menor área. O objetivo deste trabalho foi avaliar o efeito de diferentes densidades de plantio na produção, qualidade dos frutos e rentabilidade do maracujazeiro-amarelo. O experimento foi instalado e conduzido em pomar comercial da Fazenda Sant'ana, Município de São Tiago, MG. O delineamento experimental utilizado foi o de blocos casualizados com cinco tratamentos e quatro repetições. Os tratamentos constituíram-se de diferentes espaçamentos na linha de plantio: T1, 1,0 m (3.330 plantas/ha); T2, 1,5 m (2.220 plantas/ha); T3, 2,0 m (1.660 plantas/ha); T4, 3,0 m (1.100 plantas/ha) e T5, 4,0 m (830 plantas/ha). O espaçamento entre linhas foi de 3,0 m em todos os tratamentos. Cada parcela foi constituída de 12 m de comprimento por 3,0 m de largura (36 m²). O plantio foi realizado em outubro de 2001, e a colheita, a partir de abril, estendendo-se até agosto de 2002. A maior produtividade foi estimada em 11,9 t/ha na densidade de 1.841 plantas/ha. O adensamento não altera a qualidade do fruto. A máxima eficiência econômica foi alcançada na densidade de 1.340 plantas/ha, com rentabilidade de R$ 1.321,92/ha

Draft genomes of gammaproteobacterial methanotrophs isolated from terrestrial ecosystems

Genome sequences of Methylobacter luteus, Methylobacter whittenburyi, Methylosarcina fibrata, Methylomicrobium agile, and Methylovulum miyakonense were generated. The strains represent aerobic methanotrophs typically isolated from various terrestrial ecosystems

All classes of calcium channel couple with equal efficiency to exocytosis in rat melanotropes, inducing linear stimulus-secretion coupling

1. The contribution of low voltage-activated (LVA) T-type Ca(2+) channels and four different types of high voltage-activated (HVA) Ca(2+) channel to exocytosis, and the relationship between calcium influx and exocytosis during action potentials (APs) were studied in pituitary melanotropes. 2. Selective HVA Ca(2+) channel blockers reduced exocytosis, monitored by membrane capacitance measurements, proportional to the reduction in Ca(2+) influx. The efficacy of Ca(2+) in stimulating exocytosis did not change in the presence of the Ca(2+) channel blockers, indicating that all HVA Ca(2+) channels act together in stimulating exocytosis. 3. The relationship between Ca(2+) influx and exocytosis during the AP was examined using APs recorded from spontaneously active melanotropes as command templates under voltage clamp. Under voltage clamp, multiphasic Ca(2+) currents were activated over the entire duration of the APs, i.e. during the rising phase as well as the plateau phase. The maximum amplitude of the Ca(2+) current coincided with the peak of the AP. 4. The relationship between Ca(2+) entry and exocytosis was linear for the different phases of the AP. Also, the influx of Ca(2+) through LVA T-type channels stimulated exocytosis with the same efficacy as through the HVA channels. 5. APs of increasing duration (∼50 to ∼300 ms) evoked increasing amounts of exocytosis. The number of entering Ca(2+) ions and the capacitance change were linearly related to AP duration, resulting in a fixed relationship between Ca(2+) entry and exocytosis. 6. The results show that Ca(2+) ions, entering a melanotrope, couple with equal strength to exocytosis regardless of the channel type involved. We suggest that the linear relationship between Ca(2+) entry and secretion observed under physiological conditions (during APs), results from the equal strength with which LVA and HVA channels in melanotropes couple to exocytosis. This guarantees that secretion takes place over the entire duration of the AP

Frequency matching of vocalizations to inner-ear sensitivity along an altitudinal gradient in the coqui frog

Acoustic communication involves both the generation and the detection of a signal. In the coqui frog (Eleutherodactylus coqui ), it is known that the spectral contents of its calls systematically change with altitude above sea level. Here, distortion product otoacoustic emissions are used to assess the frequency range over which the inner ear is sensitive. It is found that both the spectral contents of the calls and the innerear sensitivity change in a similar fashion along an altitudinal gradient. As a result, the call frequencies and the auditory tuning are closely matched at all altitudes. We suggest that the animal's body size determines the frequency particulars of the call apparatus and the inner ear

Self-taken pharyngeal and rectal swabs are appropriate for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae in asymptomatic men who have sex with men

Introduction: Self-taken specimens from men who have sex with men (MSM) could be important in reducing high levels of demand on sexual health services. The performance of self-taken specimens for the detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC) from both pharyngeal and rectal sites in asymptomatic MSM was assessed. Methods: MSM were examined according to clinic protocol: a rectal and pharyngeal swab for GC culture and a rectal swab for the CT strand displacement assay. An extra set of nurse-taken and self-taken pharyngeal and rectal specimens were also requested and were tested using the Aptima Combo 2 assay and the result compared with the routine clinic result, which was considered the gold standard. Results: A total of 272 MSM was recruited and the sensitivity and specificity of nurse-taken and patient-taken swabs, respectively, was as follows: rectal GC: 94.9% and 90.1% (nurse); 92.3% and 87.9% (patient); pharyngeal GC: 88.2% and 91.8% (nurse); 100% and 87.8% (patient); rectal CT: 80.0% and 99.6% (nurse); 91.4% and 98.2% (patient). No significant difference in sensitivity or specificity was observed between the nurse-taken and the patient-taken rectal swabs for either GC or CT. For the detection of GC from the pharynx, comparable sensitivities were achieved between nurse-taken and patient-taken swabs (p?=?0.5); however, a significant difference in specificity was observed (p?=?0.006). This was due to a higher number of false GC-positive self-taken pharyngeal swabs from patients with high rates (90.9%; 10/11) of confirmed concurrent GC infection in different anatomical sites. Conclusions: MSM are able to collect self-taken rectal and pharyngeal swabs that are comparable to those taken by clinicians. <br/