Using GAL4-Inducible Transgenics to Modulate Rho GTPase Activity in Zebrafish.

Rho GTPases are Ras-family G proteins that regulate many critical cellular functions. Due to their requirement during early embryonic development, investigations into the function of Rho GTPases at a tissue-specific level require inducible and spatially targeted modulation of Rho GTPase activity. Here, we describe the use of ten novel zebrafish transgenics enabling GAL4-specific expression of Rho GTPases to modulate Rho GTPase activity with spatial and temporal control.

A GAL4-inducible transgenic tool kit for the in vivo modulation of Rho GTPase activity in zebrafish.

Background: Rho GTPases are small monomeric G-proteins that play key roles in many cellular processes. Due to Rho GTPases' widespread expression and broad functions, analyses of their function during late development require tissue-specific modulation of activity. The GAL4/UAS system provides an excellent tool for investigating the function of Rho GTPases in vivo.

Role of pH in a nitric oxide-dependent increase in cytosolic Cl- in retinal amacrine cells.

Nitric oxide (NO) synthase-expressing neurons are found throughout the vertebrate retina. Previous work by our laboratory has shown that NO can transiently convert inhibitory GABAergic synapses onto cultured retinal amacrine cells into excitatory synapses by releasing Cl(-) from an internal store in the postsynaptic cell. The mechanism underlying this Cl(-) release is currently unknown.

Expression and localization of CLC chloride transport proteins in the avian retina.

Members of the ubiquitously expressed CLC protein family of chloride channels and transporters play important roles in regulating cellular chloride and pH. The CLCs that function as Cl(-)/H(+) antiporters, ClCs 3-7, are essential in particular for the acidification of endosomal compartments and protein degradation. These proteins are broadly expressed in the nervous system, and mutations that disrupt their expression are responsible for several human genetic diseases.

Sphingosine-1-phosphate elicits receptor-dependent calcium signaling in retinal amacrine cells.

Evidence is emerging indicating that sphingosine-1-phosphate (S1P) participates in signaling in the retina. To determine whether S1P might be involved in signaling in the inner retina specifically, we examine the effects of this sphingolipid on cultured retinal amacrine cells. Whole cell voltage-clamp recordings reveal that S1P activates a cation current that is dependent on signaling through G(i) and phospholipase C.

Local influence of mitochondrial calcium transport in retinal amacrine cells.

Ca2+-dependent synaptic transmission from retinal amacrine cells is thought to be initiated locally at dendritic processes. Hence, understanding the spatial and temporal impact of Ca2+ transport is fundamental to understanding how amacrine cells operate. Here, we provide the first examination of the local effects of mitochondrial Ca2+ transport in neuronal processes.

Nitric oxide transiently converts synaptic inhibition to excitation in retinal amacrine cells.

Nitric oxide (NO) is generated by multiple cell types in the vertebrate retina, including amacrine cells. We investigate the role of NO in the modulation of synaptic function using a culture system containing identified retinal amacrine cells. We find that moderate concentrations of NO alter GABA(A) receptor function to produce an enhancement of the GABA-gated current.