Neurogenesis in the ventral CNS of Drosophila was studied using staining with toluidine blue and birth dating of cells monitored by incorporation of bromodeoxyuridine into DNA. The ventral CNS of the larva contains sets of neuronal stem cells (neuroblasts) which are thought to be persistent embryonic neuroblasts. Each thoracic neuromere has at least 47 of these stem cells whereas most abdominal neuromeres possess only 6. They occur in stereotyped locations so that the same neuroblast can be followed from animal to animal. The thoracic neuroblasts begin enlarging at 18-26 hr of larval life, DNA synthesis commences by 31-36 hr, and the first mitoses occur shortly thereafter. Mitotic activity continues through the remainder of larval life with the neuroblasts showing a minimum cell cycle time of less than 55 min during the late third larval instar. By 12 hr after pupariation each neuroblast has produced approximately 100 progeny which are collected with it into a discrete packet. The progeny accumulate in an immature, arrested state and only finish their differentiation into mature neurons with the onset of metamorphosis. Most of the abdominal neuroblasts differ from their thoracic counterparts in their minimum cell cycle time (less than 2 hr) and the duration of proliferation (from about 50 to 90 hr of larval life). Neurons produced during the larval stage account for more than 90% of the cells found in the ventral CNS of the adult.

We examine the role of synaptic activity in the development of identified Drosophila embryonic motorneurons. Synaptic activity was blocked by both pan-neuronal expression of tetanus toxin light chain (TeTxLC) and by reduction of acetylcholine (ACh) using a temperature-sensitive allele of choline acetyltransferase (Cha ts2 ). In the absence of synaptic activity, aCC and RP2 motorneurons develop with an apparently normal morphology and retain their capacity to form synapses. However, blockade of synaptic transmission results in significant changes in the electrical phenotype of these neurons. Specifically, increases are seen in both voltage-gated inward Na ϩ and voltage-gated outward K ϩ currents. Voltage-gated Ca 2ϩ currents do not change. The changes in conductances appear to promote neuron excitability. In the absence of synaptic activity, the number of action potentials fired by a depolarizing ramp (Ϫ60 to ϩ60 mV) is increased and, in addition, the amplitude of the initial action potential fired is also significantly larger. Silencing synaptic input to just aCC, without affecting inputs to other neurons, demonstrates that the capability to respond to changing levels of synaptic excitation is intrinsic to these neurons. The alteration to electrical properties are not permanent, being reversed by restoration of normal synaptic function. Whereas our data suggest that synaptic activity makes little or no contribution to the initial formation of embryonic neural circuits, the electrical development of neurons that constitute these circuits seems to depend on a process that requires synaptic activity.

We have used a retrograde labeling technique to identify motorneurons for each of the 30 body wall muscles of an abdominal hemisegment in the late stage 16 Drosophila embryo. Each motorneuron has a characteristic cell body position, dendritic arborization, and axonal projection. In addition, we have determined the neuroblasts of origin for most of the motorneurons we describe. Some organizational principles for the neuromuscular system have become apparent: (1) There is no obvious topographic relationship between the cell body positions of motorneurons and the position or orientation of the muscles they innervate; (2) motorneurons that innervate muscles of similar position and orientation are often clustered and have overlapping dendritic trees; (3) morphologically similar motorneurons are generally derived from a common neuroblast and innervate operationally related muscles; and (4) neuroblasts can give rise to more than one morphological type of motorneuron.

In this study, we describe the development of electrical properties of Drosophila embryonic central neurons in vivo. Using whole-cell voltage clamp, we describe the onset of expression of specific voltage- and ligand-gated ionic currents and the first appearance of endogenous and synaptic activity. The first currents occur during midembryogenesis [late stage 16, 13-14 hr after egg laying (AEL)] and consist of a delayed outward potassium current (IK) and an acetylcholine-gated inward cation current (IACh). As development proceeds, other voltage-activated currents arise sequentially. An inward calcium current (ICa) is first observed at 15 hr AEL, an inward sodium current (INa) at 16 hr AEL, and a rapidly inactivating outward potassium current (IA) at 17 hr AEL. The inward calcium current is composed of at least two individual and separable components that exhibit small temporal differences in their development. Endogenous activity is first apparent at 15 hr AEL and consists of small events (peak amplitude, 5 pA) that probably result from the random opening of relatively few numbers of ion channels. At 16 hr AEL, discrete (10-15 msec duration) currents that exhibit larger amplitude (25 pA maximum) and rapid activation but slower inactivation first appear. We identify these latter currents as EPSCs, an indication that functional synaptic transmission is occurring. In the neurons from which we record, action potentials first occur at 17 hr AEL. This study is the first to record from Drosophila embryonic central neurons in vivo and makes possible future work to define the factors that shape the electrical properties of neurons during development.

The Drosophila neuromuscular junction has attracted widespread attention as an excellent model system for studying the cellular and molecular mechanisms of synaptic development and neurotransmission. In Drosophila the advantages of invertebrate small systems, where individual cells can be examined with single-cell resolution, are combined with the powerful techniques of patch-clamp analysis and molecular genetics. In this review we examine myogenesis and motoneuron development, the problems of axon outgrowth and target selection, the differentiation of the synapse, and the mechanisms of both synaptic function and plasticity in this model genetic system.