Functional and structural study comparing the C-terminal amidated ß-neurotoxin Ts1 with its isoform Ts1-G isolated from Tityus serrulatus venom.

Mature Ts1, the main neurotoxin from Tityus serrulatus venom, has its C-terminal Cys amidated, while the isolated isoform of Ts1, named Ts1-G, keeps the non-amidated Gly residue at the C-terminal region, allowing the study of the comparative functional importance of amidation at the C-terminal between these two native toxins. Voltage dependent sodium current measurements showed that the affinity of Ts1-G for sodium channels is smaller than that of the mature Ts1, confirming the important role played by the C-terminal amidation in determining Ts1 activity.

Índices de eficiência reprodutiva de búfalos da Amazônia Oriental do Brasil / Indices of reproductive efficiency of buffaloes of the Brazilian Eastern Amazon

Foram avaliados registros de manejo reprodutivo de búfalos das raças Jafarabadi, Murrah, Mediterrâneo, Carabao e Tipo Baio, bem como seus mestiços, no período de 1983 a 2005. Estudaram-se as características reprodutivas: idade à primeira cria (média de 1052,52±120,45 dias), intervalo de partos (média de 399,69±23,78 dias), eficiência reprodutiva (média de 91,09±1,89%), fertilidade real adaptada (média de 29,30±4,40 quilogramas de bezerro parido por ano), produtividade ao primeiro parto adaptada (média de 33,75±6,89 quilogramas) e produtividade acumulada adaptada (média de 22,86±6,55 quilogramas de bezerro parido por ano). Foram verificadas influências do ano de parto e o grau de sangue da fêmea sobre as características estudadas, o que indica que tais efeitos devem ser incluídos na avaliação genética e na seleção dos animais. Os índices produtivos utilizados mostraram-se de grande valia para a seleção, devendo ser mais estudadas suas interações relativas ao desempenho geral do rebanho.

We evaluated records of reproductive management Jaffarabadi buffalo breeds, Murrah, Mediterranean, Carabao and Baio types, and their crosses, in the period from 1983 to 2005. We studied the following reproductive characteristics: Age at first calving (mean 1052.52±120.45 days), calving interval (399.69±23.78 mean days), Reproductive Efficiency (average 91.09±1, 89%), Real Adapted Fertility (average 29.30±4.40kg calf calved each year), the First Labor Productivity Adapted (mean 33.75±6.89kg) and Accumulated Productivity Adapted (mean 22.86±6.55kg calf calved each year). Influences were observed in the year of delivery and degree of blood on the female traits, suggesting that such effects should be included in genetic evaluation and selection of animals. The production indices used were of great value for the selection, and should be well studied for their interactions with the overall performance of the herd.

Nitric oxide modulates the firing rate of the rat supraoptic magnocellular neurons.

In vitro, nitric oxide (NO) inhibits the firing rate of magnocellular neurosecretory cells (MNCs) of hypothalamic supraoptic and paraventricular nuclei and this effect has been attributed to GABAergic activation. However, little is known about the direct effects of NO in MNCs. We used the patch-clamp technique to verify the effect of L-arginine, a precursor for NO synthesis, and N(omega)-nitro-L-arginine methyl ester hydrochloride (L-NAME), an inhibitor of NOS, on spontaneous electrical activity of MNCs after glutamatergic and GABAergic blockade in Wistar rat brain slices. 6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX) (10 microM) and dl-2-amino-5-phosphonovaleric acid (dl-AP5) (30 microM) were used to block postsynaptic glutamatergic currents, and picrotoxin (30 microM) and saclofen (30 microM) to block ionotropic and metabotropic postsynaptic GABAergic currents. Under these conditions, 500 microM L-arginine decreased the firing rate from 3.7+/-0.6 Hz to 1.3+/-0.3 Hz. Conversely, 100 microM L-NAME increased the firing rate from 3.0+/-0.3 Hz to 5.8+/-0.4 Hz. All points histogram analysis showed changes in resting potential from -58.1+/-0.8 mV to -62.2+/-1.1 mV in the presence of L-arginine and from -59.8+/-0.7 mV to -56.9+/-0.8 mV by L-NAME. Despite the nitrergic modulator effect on firing rate, some MNCs had no significant changes in their resting potential. In those neurons, hyperpolarizing after-potential (HAP) amplitude increased from 12.4+/-1.2 mV to 16.8+/-0.7 mV by L-arginine, but without significant changes by L-NAME treatment. To our knowledge, this is the first demonstration that NO can inhibit MNCs independent of GABAergic inputs. Further, our results point to HAP as a potential site for nitrergic modulation.

Neurotensin modulates synaptic transmission in the nucleus of the solitary tract of the rat.

Whole-cell patch clamp recordings were made from neurons of the rat subpostremal region of the nucleus tractus solitarius (NTS) in transverse brainstem slices. Neurotensin (NT) enhanced the firing rate of action potentials from 0.8 +/- 0.4 Hz in control to 1.9 +/- 1.3 Hz (n = 9) and increased their decay time. The peak amplitude of the after-hyperpolarization was decreased by 34+/-5% (n = 9). These effects were associated with a depolarization of 4 +/- 1 mV (n = 10) in the resting membrane potential and an increase in the input resistance (from 768 +/- 220 MOmega to 986+/-220 MOmega; n = 5) and were compensated by manually hyperpolarizing the cell to control values. In voltage clamp experiments NT decreased an outward current (from 488 +/- 161 to 340 +/- 96 pA at +40 mV; n = 5) which reversed near the potassium equilibrium potential. In addition, NT increased the frequency of both excitatory and inhibitory spontaneous synaptic currents, an effect blocked by tetrodotoxin, and did not change the evoked excitatory or inhibitory postsynaptic currents. The selective NTR1 receptor antagonist SR48692 reversibly blocked the effects of NT on both action potentials and spontaneous synaptic currents. Our results suggest that NTR1 receptors can modulate post-synaptic responses in neurons of the subpostremal NTS by increasing cell excitability as a result of blockade of a potassium conductance.

Electrophysiological evidence for the presence of NR2C subunits of N-methyl-D-aspartate receptors in rat neurons of the nucleus tractus solitarius.

The nucleus tractus solitarius (NTS) plays an important role in the control of autonomic reflex functions. Glutamate, acting on N-methyl-D-aspartate (NMDA) and non-NMDA ionotropic receptors, is the major neurotransmitter in this nucleus, and the relative contribution of each receptor to signal transmission is unclear. We have examined NMDA excitatory postsynaptic currents (NMDA-EPSCs) in the subpostremal NTS using the whole cell patch clamp technique on a transverse brainstem slice preparation. The NMDA-EPSCs were evoked by stimulation of the solitary tract over a range of membrane potentials. The NMDA-EPSCs, isolated pharmacologically, presented the characteristic outward rectification and were completely blocked by 50 microM DL-2-amino-5-phosphonopentanoic acid. The I-V relationship of the NMDA response shows that current, with a mean (+/- SEM) amplitude of -41.2 +/- 5.5 pA, is present even at a holding potential of -60 mV, suggesting that the NMDA receptors are weakly blocked by extracellular Mg2+ at near resting membrane potentials. This weak block can also be inferred from the value of 0.67 +/- 0.17 for parameter delta obtained from a fit of the Woodhull equation to the I-V relationship. The maximal inward current measured on the I-V relationship was at -38.7 +/- 4.2 mV. The decay phase of the NMDA currents was fitted with one exponential function with a decay time constant of 239 +/- 51 and 418 +/- 80 ms at a holding potential of -60 and +50 mV, respectively, which became slower with depolarization (e-fold per 145 mV). The biophysical properties of the NMDA receptors observed in the present study suggest that these receptors in the NTS contain NR2C subunits and may contribute to the synaptic signal integration.

Electrophysiological evidence for the presence of NR2C subunits of N-methyl-D-aspartate receptors in rat neurons of the nucleus tractus solitarius

The nucleus tractus solitarius (NTS) plays an important role in the control of autonomic reflex functions. Glutamate, acting on N-methyl-D-aspartate (NMDA) and non-NMDA ionotropic receptors, is the major neurotransmitter in this nucleus, and the relative contribution of each receptor to signal transmission is unclear. We have examined NMDA excitatory postsynaptic currents (NMDA-EPSCs) in the subpostremal NTS using the whole cell patch clamp technique on a transverse brainstem slice preparation. The NMDA-EPSCs were evoked by stimulation of the solitary tract over a range of membrane potentials. The NMDA-EPSCs, isolated pharmacologically, presented the characteristic outward rectification and were completely blocked by 50 æM DL-2-amino-5-phosphonopentanoic acid. The I-V relationship of the NMDA response shows that current, with a mean (± SEM) amplitude of -41.2 ± 5.5 pA, is present even at a holding potential of -60 mV, suggesting that the NMDA receptors are weakly blocked by extracellular Mg2+ at near resting membrane potentials. This weak block can also be inferred from the value of 0.67 ± 0.17 for parameter delta obtained from a fit of the Woodhull equation to the I-V relationship. The maximal inward current measured on the I-V relationship was at -38.7 ± 4.2 mV. The decay phase of the NMDA currents was fitted with one exponential function with a decay time constant of 239 ± 51 and 418 ± 80 ms at a holding potential of -60 and +50 mV, respectively, which became slower with depolarization (e-fold per 145 mV). The biophysical properties of the NMDA receptors observed in the present study suggest that these receptors in the NTS contain NR2C subunits and may contribute to the synaptic signal integration.