Development of parvalbumin immunoreactivity in the chick Edinger Westphal nucleus.

To determine when the calcium-binding protein parvalbumin appears during development, neurons in the chick Edinger Westphal nucleus were examined for parvalbumin immunoreactivity at a variety of embryonic stages. Parvalbumin immunoreactivity appeared on embryonic day 14 (E14, Hamburger and Hamilton stage 40) in predominantly lateral Edinger Westphal neurons. Cytochrome oxidase activity within the nucleus was examined throughout development, as an indicator of physiological activity, and expression of cytochrome oxidase was compared with that of parvalbumin. Cytochrome oxidase activity was found to be uniformly high in all parts of the Edinger Westphal nucleus throughout development. Either the Edinger Westphal nucleus in physiologically active quite early in its development or other energy demands mask the correlation of cytochrome oxidase with electrical activity. Cytochrome oxidase was expressed well before parvalbumin immunoreactivity appeared. Voltage-activated calcium currents were characterized in E12 Edinger Westphal neurons. In both amplitude and composition, E12 calcium currents resemble those of E16 neurons, excluding the possibility that calcium currents appear de novo during or just prior to the appearance of parvalbumin. Both cytochrome oxidase activity and calcium currents are observed in Edinger Westphal neurons well before the appearance of parvalbumin during development. These findings do not exclude the possibility that physiological activity affects the expression of parvalbumin since other factors such as changing patterns of synaptic activity or the appearance of calcium conducting NMDA receptors have yet to be examined. However, they raise the possibility that additional factors such as an intrinsic developmental program or a change in the neuron's basal intracellular calcium requirements may also be involved.

Multiple subtypes of voltage-gated calcium currents in the Edinger-Westphal nucleus.

Avian Edinger-Westphal (EW) neurons provide a unique opportunity to compare electrophysiologically the membranes of cell bodies and terminals in the same population of neurons. Axons that originate from neurons in the lateral region of the EW nucleus form a morphologically distinct presynaptic terminal, known as a calyx, on ciliary ganglion neurons. Several studies have shown that calyciform terminals in the ciliary ganglion exhibit predominantly N-type, high-voltage-activated (HVA) calcium channels. The goal of this study was to characterize and compare the calcium currents expressed in EW cell somas with those reported in the terminals. Whole-cell patch-clamp techniques were used to record from cell bodies in the lateral EW nucleus in slice preparations. Slices were obtained from embryonic day 16 chicks, matching the age of the embryos in which calyces were studied. Recordings were localized to the lateral region of the EW nucleus using Lucifer yellow fills. Voltage-step commands from -70 to 0 mV produced calcium currents with both a sustained and an inactivating component. Depolarization steps to 0 mV from a holding potential of -40 mV eliminated the inactivating component. These recordings suggested the presence of both LVA and HVA calcium currents. Application of 0.1 mM NiCl2 produced a reversible decrease in the amplitude of the whole-cell calcium current, preferentially affecting the inactivating component. The Ni2+(-)sensitive current activated and inactivated rapidly in a voltage-dependent manner. Treatment with 0.1 mM cadmium chloride caused a reversible reduction in the amplitude of the calcium current.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcium-binding proteins in the chick Edinger Westphal nucleus.

It has been suggested that the calcium-binding proteins, parvalbumin and calbindin-D28K, are involved in the control of intracellular calcium levels but their exact functions are unknown. Immunoreactivity for parvalbumin has been associated with rapidly firing cells, while calbindin has been implicated in protecting neurons from excitotoxicity. Since the chick Edinger Westphal nucleus contains two populations of neurons with different firing patterns, parvalbumin and calbindin immunoreactivities were examined in the Edinger Westphal nuclei of posthatch chicks to determine whether a particular subpopulation of neurons is associated with either protein. Moderate levels of parvalbumin immunoreactivity were found consistently associated with repetitively firing neurons in the lateral Edinger Westphal nucleus. In contrast, medial neurons expressed much lower levels of parvalbumin immunoreactivity. Many medial neurons were negative for parvalbumin although occasionally a few medial neurons stained as strongly as lateral neurons. Definitive calbindin immunoreactivity was absent from Edinger Westphal nuclei despite robust staining of cells in other parts of the brainstem and in control sections of cerebellum.