Daniel J. Bumbarger

Understanding the evolution of the bilaterian brain requires a detailed exploration of the precise nature of cellular and subcellular differences between related species. We undertook an electron micrographic reconstruction of the brain of the predatory nematode Pristionchus pacificus and compared the results with the brain of Caenorhabditis elegans , which diverged at least 100 million years ago. We revealed changes in neuronal cell death, neuronal cell position, axodendritic projection patterns, and synaptic connectivity of homologous neurons that display no obvious changes in overall neurite morphology and projection patterns. These multiscale patterns of evolutionary changes show no bias to specific brain regions or neuron types.

1 In 1949, Donald Hebb proposed that groups of neurons that activate stereotypically form the organizational building blocks of perception, cognition, and behavior. Finding the structural underpinning of such assemblies has been technically challenging, due to a lack of large-scale structure-activity maps. Here, we analyze this relation using a novel dataset that links in vivo optical physiology to connectivity using postmortem elec-tron microscopy (EM). From the fluorescence traces, we extract neural assemblies from higher-order correlations in neural activity. Physiologically, we show that these assemblies exhibit properties consistent with Hebb’s theory, including more reliable responses to repeated natural movie inputs than size-matched random ensembles and superior decoding of visual stimuli. Structurally, we find that neurons that participate in assemblies are significantly more integrated into the structural network than those that do not. Contrary to Hebb’s original prediction, we do not observe a marked increase in the strength of monosynaptic excitatory connections between cells participating in the same assembly. However, we find significantly stronger indirect feed-forward inhibitory connections targeting cells in other assemblies. These results show that assemblies can be useful components of perception, and, surprisingly, they are delineated by mutual inhibition.

Neural circuit function is shaped both by the cell types that comprise the circuit and the connections between those cell types ¹ . Neural cell types have previously been defined by morphology ^{2, 3} , electrophysiology ^{4, 5} , transcriptomic expression ^6–8 , connectivity ^9–13 , or even a combination of such modalities ^14–16 . More recently, the Patch-seq technique has enabled the characterization of morphology (M), electrophysiology (E), and transcriptomic (T) properties from individual cells ^17–20 . Using this technique, these properties were integrated to define 28, inhibitory multimodal, MET-types in mouse primary visual cortex ²¹ . It is unknown how these MET-types connect within the broader cortical circuitry however. Here we show that we can predict the MET-type identity of inhibitory cells within a large-scale electron microscopy (EM) dataset and these MET-types have distinct ultrastructural features and synapse connectivity patterns. We found that EM Martinotti cells, a well defined morphological cell type ^{22, 23} known to be Somatostatin positive (Sst+) ^{24, 25} , were successfully predicted to belong to Sst+ MET-types. Each identified MET-type had distinct axon myelination patterns and synapsed onto specific excitatory targets. Our results demonstrate that morphological features can be used to link cell type identities across imaging modalities, which enables further comparison of connectivity in relation to transcriptomic or electrophysiological properties. Furthermore, our results show that MET-types have distinct connectivity patterns, supporting the use of MET-types and connectivity to meaningfully define cell types.

Neurons in the neocortex exhibit astonishing morphological diversity which is critical for properly wiring neural circuits and giving neurons their functional properties. However, the organizational principles underlying this morphological diversity remain an open question. Here, we took a data-driven approach using graph-based machine learning methods to obtain a low-dimensional morphological “bar code” describing more than 30,000 excitatory neurons in mouse visual areas V1, AL and RL that were reconstructed from the millimeter scale MICrONS serial-section electron microscopy volume. Contrary to previous classifications into discrete morphological types (m-types), our data-driven approach suggests that the morphological landscape of cortical excitatory neurons is better described as a continuum, with a few notable exceptions in layers 5 and 6. Dendritic morphologies in layers 2–3 exhibited a trend towards a decreasing width of the dendritic arbor and a smaller tuft with increasing cortical depth. Inter-area differences were most evident in layer 4, where V1 contained more atufted neurons than higher visual areas. Moreover, we discovered neurons in V1 on the border to layer 5 which avoided deeper layers with their dendrites. In summary, we suggest that excitatory neurons’ morphological diversity is better understood by considering axes of variation than using distinct m-types.

Cervidellus sonorensis n. sp. is described from sand near the roots of creosote bush (Larrea tridentata) from Anza-Borrego Desert State Park, CA, USA. It is distinguished from others in the genus by the combination of a greatly distended, guitar-shaped corpus and the morphology of the lips and labial probolae. Distinctive features include a large pharyngeal metacorpus with a double swelling, a knob-like ledge at the base of each labial probola and lips with five filamentous tines, the most apical of which are those closest to the primary axil. One male individual was discovered in this parthenogenetic species. Characters used to diagnose C. sonorensis n. sp. are not definitive with respect to other genera and are probably plesiomorphic or convergent in light of DNA-based phylogenetic hypotheses. The position of C. sonorensis n. sp. and morphologically close congeners in relation to species of Nothacrobeles and Paracrobeles is discussed.

Nematode sensory structures can be divided into two classes; cuticular sensillae, with dendrites ending outside the epidermis, and internal receptors, that typically are single dendrites terminating within the body cavity. Fine structure of the former has been described completely in more than a dozen nematode taxa, while the latter were previously only well understood in the microbial feeder Caenorhabditis elegans. The distantly related nematode Acrobeles complexus has a similar ecology and together the two span a clade representing a large proportion of nematode biodiversity. The cuticular sensillae and internal receptors of A. complexus are here shown to be remarkably similar in number, arrangement, and morphology to those of C. elegans. Several key differences are reported that likely relate to function, and suggest that this nematode has a cuticular sensillum morphology that is closer to that of the common ancestor of the two taxa. Internal sensory receptors have more elaborate termini than those of C. elegans. The existence of a novel form of mechanoreceptor in A. complexus and spatial relationships between sensillum dendrites suggest differences between two classes of sensillae in how a touch‐response behavior may be mediated. J. Morphol., 2007.