Cross-linking evidence for motional constraints within chemoreceptor trimers of dimers.

Chemotactic behavior in bacteria relies on the sensing ability of large chemoreceptor clusters that are usually located at the cell pole. In Escherichia coli, chemoreceptors exhibit higher-order interactions within those clusters based on a trimer-of-dimers organization. This architecture is conserved in a variety of other bacteria and archaea, implying that receptors in many microorganisms form trimer-of-dimer signaling teams. To gain further insight into the assembly and dynamic behavior of receptor trimers of dimers, we used in vivo cross-linking targeted to cysteine residues at various positions that define six different levels along the cytoplasmic signaling domains of the aspartate and serine chemoreceptors, Tar and Tsr, respectively. We found that the cytoplasmic domains of these receptors are close to each other near the trimer contact region at the cytoplasmic tip and lie farther apart as the receptor dimers approach the cytoplasmic membrane. Tar and Tsr reporter sites within the same or closely adjacent levels readily formed mixed cross-links, whereas reporters located different distances from the tip did not. These findings indicate that there are no significant vertical displacements of one dimer with respect to the others within the trimer unit. Attractant stimuli had no discernible effect on the cross-linking efficiency of any of the reporters tested, but a strong osmotic stimulus reproducibly enhanced cross-linking at most of the reporter sites, indicating that individual dimers may move closer together under this condition.

Spatial choice is biased by chemical cues from conspecifics in the speckeled worm eel Myrophis punctatus

The speckeld worm eel Myrophis punctatus lives in high-densities assemblages, and usually digs through, or lies on the substrate. These behaviours could lead to chemical marks on the substrate and could modulate the spatial distribution in this species. We tested the hypothesis that the spatial choice of the speckled worm eel is modulated by the presence of conspecific odour on the substrate. Here, we showed that the speckled worm eel avoids the substrate area containing the conspecific odour, indicating that this chemical cue modulates the eel's spatial decision. The eels clearly detected the conspecific's odour. This perception might indicate the presence of conspecifics into the substrate. Since the eels avoided an area containing conspecific odour, we suggest this may be a response that avoids the consequences of invading a resident-animal's territory.

A enguia mirongo-mirim Myrophis punctatus vive em agrupamentos de alta densidade populacional e comumente se enterra ou permanece sob o substrato. Esses comportamentos podem levar a marcas químicas no subtrato e podem, portanto, modular o uso do espaço nessa espécie. Neste estudo, testamos a hipótese de que a preferência espacial da enguia mirongo-mirim é influenciada pela presença de odor do animal coespecífico no subtrato. Mostramos que as enguias evitam a área que contém tal odor, indicando que as decisões de ocupação espacial podem ser influenciadas por pistas químicas de coespecíficos. As enguias claramente detectaram o odor de um animal coespecífico e essa percepção poderia ser um indicativo da presença de um coespecífico enterrado no substrato. Visto que elas evitam uma área contendo tal odor, sugerimos que isso poderia ser uma resposta para evitar invadir o território de um animal residente.

Spatial choice is biased by chemical cues from conspecifics in the speckeled worm eel Myrophis punctatus

The speckeld worm eel Myrophis punctatus lives in high-densities assemblages, and usually digs through, or lies on the substrate. These behaviours could lead to chemical marks on the substrate and could modulate the spatial distribution in this species. We tested the hypothesis that the spatial choice of the speckled worm eel is modulated by the presence of conspecific odour on the substrate. Here, we showed that the speckled worm eel avoids the substrate area containing the conspecific odour, indicating that this chemical cue modulates the eel's spatial decision. The eels clearly detected the conspecific's odour. This perception might indicate the presence of conspecifics into the substrate. Since the eels avoided an area containing conspecific odour, we suggest this may be a response that avoids the consequences of invading a resident-animal's territory.(AU)

A enguia mirongo-mirim Myrophis punctatus vive em agrupamentos de alta densidade populacional e comumente se enterra ou permanece sob o substrato. Esses comportamentos podem levar a marcas químicas no subtrato e podem, portanto, modular o uso do espaço nessa espécie. Neste estudo, testamos a hipótese de que a preferência espacial da enguia mirongo-mirim é influenciada pela presença de odor do animal coespecífico no subtrato. Mostramos que as enguias evitam a área que contém tal odor, indicando que as decisões de ocupação espacial podem ser influenciadas por pistas químicas de coespecíficos. As enguias claramente detectaram o odor de um animal coespecífico e essa percepção poderia ser um indicativo da presença de um coespecífico enterrado no substrato. Visto que elas evitam uma área contendo tal odor, sugerimos que isso poderia ser uma resposta para evitar invadir o território de um animal residente.(AU)

In vivo crosslinking methods for analyzing the assembly and architecture of chemoreceptor arrays.

The chemoreceptor molecules that mediate chemotactic responses in bacteria and archaea are physically clustered and operate as highly cooperative arrays. Few experimental approaches are able to investigate the structure-function organization of these chemoreceptor networks in living cells. This chapter describes chemical crosslinking methods that can be applied under normal physiological conditions to explore physical interactions between chemoreceptors and their underlying genetic and structural basis. Most of these crosslinking approaches are based on available atomic structures for chemoreceptor homodimers, the fundamental building block for higher-order networks. However, the general logic of our in vivo crosslinking approaches is readily applicable to other protein-protein interactions and other organisms, even when high-resolution structural information is not available.

In vitro approach to the chemical drive of breathing.

Since its introduction two decades ago, the isolated brain stem-spinal cord preparation of neonatal rodents has been the preferred method used to reveal the mystery underlying the genesis of the respiratory rhythm. Little research using this in vitro approach has focused on the study of the central respiratory chemosensitivity. Some unexpected findings obtained with the brain stem-spinal cord preparation have added new questions that challenge our previous theoretic framework. Some of these findings are addressed here.