Deep homologies in chordate caudal central nervous systems.

Invertebrate chordates, such as the tunicate , can offer insight into the evolution of the chordate phylum. Anatomical features that are shared between invertebrate chordates and vertebrates may be taken as evidence of their presence in a common chordate ancestor. The central nervous systems of larvae and vertebrates share a similar anatomy despite the CNS having ~180 neurons.

preparation of brains enables high-throughput FIB-SEM connectomics.

Deriving the detailed synaptic connections of an entire nervous system is the unrealized goal of the nascent field of connectomics. For the fruit fly , in particular, we need to dissect the brain, connectives, and ventral nerve cord as a single continuous unit, fix and stain it, and undertake automated segmentation of neuron membranes. To achieve this, we designed a protocol using progressive lowering of temperature dehydration (PLT), a technique routinely used to preserve cellular structure and antigenicity.

Neuronal circuits integrating visual motion information in Drosophila melanogaster.

The detection of visual motion enables sophisticated animal navigation, and studies on flies have provided profound insights into the cellular and circuit bases of this neural computation. The fly's directionally selective T4 and T5 neurons encode ON and OFF motion, respectively. Their axons terminate in one of the four retinotopic layers in the lobula plate, where each layer encodes one of the four directions of motion.

Presynaptic Mitochondrial Volume and Packing Density Scale with Presynaptic Power Demand.

Stable neural function requires an energy supply that can meet the intense episodic power demands of neuronal activity. Neurons have presumably optimized the volume of their bioenergetic machinery to ensure these power demands are met, but the relationship between presynaptic power demands and the volume available to the bioenergetic machinery has never been quantified. Here, we estimated the power demands of six motor nerve terminals in female larvae through direct measurements of neurotransmitter release and Ca entry, and via theoretical estimates of Na entry and power demands at rest.

A connectome is not enough - what is still needed to understand the brain of Drosophila?

Understanding the structure and operation of any nervous system has been a subject of research for well over a century. A near-term opportunity in this quest is to understand the brain of a model species, the fruit fly Drosophila melanogaster. This is an enticing target given its relatively small size (roughly 200,000 neurons), coupled with the behavioral richness that this brain supports, and the wide variety of techniques now available to study both brain and behavior.

Ultrastructural 3D reconstruction of the smallest known insect photoreceptors: The stemmata of a first instar larva of Strepsiptera (Hexapoda).

Stemmata of strepsipteran insects represent the smallest arthropod eyes known, having photoreceptors which form fused rhabdoms measuring an average size of 1.69 × 1.21 × 1.

A connectome and analysis of the adult central brain.

The neural circuits responsible for animal behavior remain largely unknown. We summarize new methods and present the circuitry of a large fraction of the brain of the fruit fly . Improved methods include new procedures to prepare, image, align, segment, find synapses in, and proofread such large data sets.

Neuronal Glutamatergic Synaptic Clefts Alkalinize Rather Than Acidify during Neurotransmission.

The dogma that the synaptic cleft acidifies during neurotransmission is based on the corelease of neurotransmitters and protons from synaptic vesicles, and is supported by direct data from sensory ribbon-type synapses. However, it is unclear whether acidification occurs at non-ribbon-type synapses. Here we used genetically encoded fluorescent pH indicators to examine cleft pH at conventional neuronal synapses.

The Organization of the Second Optic Chiasm of the Optic Lobe.

Visual pathways from the compound eye of an insect relay to four neuropils, successively the lamina, medulla, lobula, and lobula plate in the underlying optic lobe. Among these neuropils, the medulla, lobula, and lobula plate are interconnected by the complex second optic chiasm, through which the anteroposterior axis undergoes an inversion between the medulla and lobula. Given their complex structure, the projection patterns through the second optic chiasm have so far lacked critical analysis.

Novel type of sub-retinal pigment shield in the miniaturized compound eye of Trichogramma evanescens.

Pigment granules, found in different cell types of the retina in insect compound eyes, fulfill important functions. They isolate the individual ommatidia from stray light, regulate the angular sensitivity, and restrict the light that reaches the photoreceptor according to ambient light intensities. Descriptions of pigment cells within the retina are included in ultrastructural eye descriptions, but knowledge of pigment cell types beneath the retina and basal matrix (BM) are relatively limited in insects.

Control of Synaptic Specificity by Establishing a Relative Preference for Synaptic Partners.

The ability of neurons to identify correct synaptic partners is fundamental to the proper assembly and function of neural circuits. Relative to other steps in circuit formation such as axon guidance, our knowledge of how synaptic partner selection is regulated is severely limited. Drosophila Dpr and DIP immunoglobulin superfamily (IgSF) cell-surface proteins bind heterophilically and are expressed in a complementary manner between synaptic partners in the visual system.

The Fly Brain Atlas.

The brain's synaptic networks endow an animal with powerfully adaptive biological behavior. Maps of such synaptic circuits densely reconstructed in those model brains that can be examined and manipulated by genetic means offer the best prospect for understanding the underlying biological bases of behavior. That prospect is now technologically feasible and a scientifically enabling possibility in neurobiology, much as genomics has been in molecular biology and genetics.

Transcriptional Feedback Links Lipid Synthesis to Synaptic Vesicle Pools in Drosophila Photoreceptors.

Neurons can maintain stable synaptic connections across adult life. However, the signals that regulate expression of synaptic proteins in the mature brain are incompletely understood. Here, we describe a transcriptional feedback loop between the biosynthesis and repertoire of specific phospholipids and the synaptic vesicle pool in adult Drosophila photoreceptors.

Comparisons between the ON- and OFF-edge motion pathways in the brain.

Understanding the circuit mechanisms behind motion detection is a long-standing question in visual neuroscience. In , recently discovered synapse-level connectomes in the optic lobe, particularly in ON-pathway (T4) receptive-field circuits, in concert with physiological studies, suggest a motion model that is increasingly intricate when compared with the ubiquitous Hassenstein-Reichardt model. By contrast, our knowledge of OFF-pathway (T5) has been incomplete.

Three-dimensional ultrastructural organization of the ommatidium of the minute parasitoid wasp Trichogramma evanescens.

Existing information on insect compound eyes is mainly limited to two-dimensional information derived from histological or ultrathin sections. These allow a basic description of eye morphology, but are limited in z-axis resolution because of the section thickness or intervals between sections, so that accurate volumetric information cannot be generated. Here we use serial-sectioning transmission electron microscopy to present a 3-D reconstruction at ultrastructural level of a complete ommatidium of a miniaturized insect compound eye.

Neuronal identity: the neuron types of a simple chordate sibling, the tadpole larva of Ciona intestinalis.

Neurons of the sparsely populated nervous system of the tadpole larva in the tunicate Ciona intestinalis, a chordate sibling, are known from sporadic previous studies but especially two recent reports that document the connectome of both the central and peripheral nervous systems at EM level. About 330 CNS cells comprise mostly ciliated ependymal cells, with ∼180 neurons that constitute about 50 morphologically distinguishable types. The neurons reveal various chordate characters amid many features that are idiosyncratic.

A resource for the antennal lobe provided by the connectome of glomerulus VA1v.

Using FIB-SEM we report the entire synaptic connectome of glomerulus VA1v of the right antennal lobe in . Within the glomerulus we densely reconstructed all neurons, including hitherto elusive local interneurons. The -positive, sexually dimorphic VA1v included >11,140 presynaptic sites with ~38,050 postsynaptic dendrites.

Location and functions of Inebriated in the eye.

Histamine (HA) is a neurotransmitter in arthropod photoreceptors. It is recycled via conjugation to β-alanine to form β-alanylhistamine (carcinine). Conjugation occurs in epithelial glia that surround photoreceptor terminals in the first optic neuropil, and carcinine (CA) is then transported back to photoreceptors and cleaved to liberate HA and β-alanine.

The world of the identified or digital neuron.

In general, neurons in insects and many other invertebrate groups are individually recognizable, enabling us to assign an index number to specific neurons in a manner which is rarely possible in a vertebrate brain. This endows many studies on insect nervous systems with the opportunity to document neurons with great precision, so that in favourable cases we can return to the same neuron or neuron type repeatedly so as to recognize many separate morphological classes. The visual system of the fly's compound eye particularly provides clear examples of the accuracy of neuron wiring, allowing numerical comparisons between representatives of the same cell type, and estimates of the accuracy of their wiring.

Of what use is connectomics? A personal perspective on the connectome.

The brain is a network of neurons and its biological output is behaviour. This is an exciting age, with a growing acknowledgement that the comprehensive compilation of synaptic circuits densely reconstructed in the brains of model species is now both technologically feasible and a scientifically enabling possibility in neurobiology, much as 30 years ago genomics was in molecular biology and genetics. Implemented by huge advances in electron microscope technology, especially focused ion beam-scanning electron microscope (FIB-SEM) milling (see Glossary), image capture and alignment, and computer-aided reconstruction of neuron morphologies, enormous progress has been made in the last decade in the detailed knowledge of the actual synaptic circuits formed by real neurons, in various brain regions of the fly It is useful to distinguish synaptic pathways that are major, with 100 or more presynaptic contacts, from those that are minor, with fewer than about 10; most neurites are both presynaptic and postsynaptic, and all synaptic sites have multiple postsynaptic dendrites.

Sidekick is required in developing photoreceptors to enable visual motion detection.

The assembly of functional neuronal circuits requires growth cones to extend in defined directions and recognize the correct synaptic partners. Homophilic adhesion between vertebrate Sidekick proteins promotes synapse formation between retinal neurons involved in visual motion detection. We show here that Sidekick accumulates in specific synaptic layers of the developing motion detection circuit and is necessary for normal optomotor behavior.

From two to three dimensions: The importance of the third dimension for evaluating the limits to neuronal miniaturization in insects.

Most studies dealing with the limits to miniaturization in insect brains have until now relied on information based on data collected in two dimensions: either histological sections imaged by light microscopy, or electron micrographs of single ultrathin sections imaged by transmission electron microscopy (TEM). To test the validity of transferring information gained from two-dimensional images to the third dimension, we examined a 3D image stack from serial-section TEM (ssTEM) of the optic neuropiles of the miniature parasitic wasp Trichogramma brassicae (Bezdenko, 1968). We reinvestigated the proposed lower limit of 2 µm for the diameters of neuronal somata and found average volumes of 6.

The peripheral nervous system of the ascidian tadpole larva: Types of neurons and their synaptic networks.

Physical and chemical cues from the environment are used to direct animal behavior through a complex network of connections originating in exteroceptors. In chordates, mechanosensory and chemosensory neurons of the peripheral nervous system (PNS) must signal to the motor circuits of the central nervous system (CNS) through a series of pathways that integrate and regulate the output to motor neurons (MN); ultimately these drive contraction of the tail and limb muscles. We used serial-section electron microscopy to reconstruct PNS neurons and their hitherto unknown synaptic networks in the tadpole larva of a sibling chordate, the ascidian, Ciona intestinalis.