Optic nerve regeneration: A long view.

The optic nerve conveys information about the outside world from the retina to multiple subcortical relay centers. Until recently, the optic nerve was widely believed to be incapable of re-growing if injured, with dire consequences for victims of traumatic, ischemic, or neurodegenerative diseases of this pathway. Over the past 10-20 years, research from our lab and others has made considerable progress in defining factors that normally suppress axon regeneration and the ability of retinal ganglion cells, the projection neurons of the retina, to survive after nerve injury. Here we describe research from our lab on the role of inflammation-derived growth factors, suppression of inter-cellular signals among diverse retinal cell types, and combinatorial therapies, along with related studies from other labs, that enable animals with optic nerve injury to regenerate damaged retinal axons back to the brain. These studies raise the possibility that vision might one day be restored to people with optic nerve damage.

Isoflurane modulates activation and inactivation gating of the prokaryotic Na+ channel NaChBac.

Voltage-gated Na+ channels (Nav) have emerged as important presynaptic targets for volatile anesthetic (VA) effects on synaptic transmission. However, the detailed biophysical mechanisms by which VAs modulate Nav function remain unclear. VAs alter macroscopic activation and inactivation of the prokaryotic Na+ channel, NaChBac, which provides a useful structural and functional model of mammalian Nav Here, we study the effects of the common general anesthetic isoflurane on NaChBac function by analyzing macroscopic Na+ currents (INa) in wild-type (WT) channels and mutants with impaired (G229A) or enhanced (G219A) inactivation. We use a previously described six-state Markov model to analyze empirical WT and mutant NaChBac channel gating data. The model reproduces the mean empirical gating manifest in INa time courses and optimally estimates microscopic rate constants, valences (z), and fractional electrical distances (x) of forward and backward transitions. The model also reproduces gating observed for all three channels in the absence or presence of isoflurane, providing further validation. We show using this model that isoflurane increases forward activation and inactivation rate constants at 0 mV, which are associated with estimated chemical free energy changes of approximately -0.2 and -0.7 kcal/mol, respectively. Activation is voltage dependent (z ≈ 2e0, x ≈ 0.3), inactivation shows little voltage dependence, and isoflurane has no significant effect on either. Forward inactivation rate constants are more than 20-fold greater than backward rate constants in the absence or presence of isoflurane. These results indicate that isoflurane modulates NaChBac gating primarily by increasing forward activation and inactivation rate constants. These findings support accumulating evidence for multiple sites of anesthetic interaction with the channel.

jShaw1, a low-threshold, fast-activating K(v)3 from the hydrozoan jellyfish Polyorchis penicillatus.

Voltage-gated potassium (K(v)) channels work in concert with other ion channels to determine the frequency and duration of action potentials in excitable cells. Little is known about K(v)3 channels from invertebrates, but those that have been characterized generally display slow kinetics. Here, we report the cloning and characterization of jShaw1, the first K(v)3 isolated from a cnidarian, the jellyfish Polyorchis penicillatus, in comparison with mouse K(v)3.1 and K(v)3.2. Using a two-electrode voltage clamp on Xenopus laevis oocytes expressing the channels, we compared steady-state and kinetic properties of macroscopic currents. jShaw1 is fast activating, and opens at potentials approximately 40 mV more hyperpolarized than the mouse K(v)3 channels. There is an inverse relationship between the number of positive charges on the voltage sensor and the half-activation voltage of the channel, contrary to what would be expected with the simplest model of voltage sensitivity. jShaw1 has kinetic characteristics that are substantially different from the mammalian K(v)3 channels, including a much lower sensitivity of early activation rates to incremental voltage changes, and a much faster voltage-dependent transition in the last stages of opening. jShaw1 opening kinetics were affected little by pre-depolarization voltage, in contrast to both mouse channels. Similar to the mouse channels, jShaw1 was half-blocked by 0.7 mmol l(-1) tetraethyl ammonium and 5 mmol l(-1) 4-aminopyridine. Comparison of sequence and functional properties of jShaw1 with the mouse and other reported K(v)3 channels helps to illuminate the general relationship between amino acid sequence and electrophysiological activity in this channel family.