Biology subjects

Blagburn, J. M.

Publications and source records attributed to Blagburn, J. M..

5 recordsLinked to original sources

bioRxiv2023

Somatotopic organization among parallel sensory pathways that promote a grooming sequence in Drosophila

Mechanosensory neurons located across the body surface respond to tactile stimuli and elicit diverse behavioral responses, from relatively simple stimulus location-aimed movements to complex movement sequences. How mechanosensory neurons and their postsynaptic circuits influence such diverse behaviors remains unclear. We previously discovered that Drosophila perform a body location-prioritized grooming sequence when mechanosensory neurons at different locations on the head and body are simultaneously stimulated by dust (Hampel et al., 2017; Seeds et al., 2014). Here, we identify nearly all mechanosensory neurons on the Drosophila head that individually elicit aimed grooming of specific head locations, while collectively eliciting a whole head grooming sequence. Different tracing methods were used to reconstruct the projections of these neurons from different locations on the head to their distinct arborizations in the brain. This provides the first synaptic resolution somatotopic map of a head, and defines the parallel-projecting mechanosensory pathways that elicit head grooming.

neuroscience↗

bioRxiv2021

Retinoic acid treatment recruits macrophages and increases axonal regeneration after optic nerve injury in the frog Rana pipiens

Retinoic acid (RA) plays major roles during nervous system development, and during regeneration of the adult nervous system. We have previously shown that components of the RA signaling pathway are upregulated after optic nerve injury, and that exogenous application of all-trans retinoic acid (ATRA) greatly increases the survival of axotomized retinal ganglion cells (RGCs). The objective of the present study is to investigate the effects of ATRA application on the macrophages in the optic nerve after injury, and to determine whether this affects axonal regeneration. The optic nerve was crushed and treated with PBS, ATRA and/or clodronate-loaded liposomes. Nerves were examined at one and two weeks after axotomy with light microscopy, immunocytochemistry and electron microscopy. ATRA application to the optic nerve caused transient increases in the number of macrophages and microglia one week after injury. The macrophages are consistently labeled with M2-type markers, and have considerable phagocytic activity. ATRA increased ultrastructural features of ongoing phagocytic activity in macrophages at one and two weeks. ATRA treatment also significantly increased the numbers of regenerating GAP-43-labeled axons. Clodronate liposome treatment depleted macrophage numbers by 80%, completely eliminated the ATRA-mediated increase in axonal regeneration, and clodronate treatment alone decreased axonal numbers by 30%. These results suggest that the success of axon regeneration is partially dependent on the presence of debris-phagocytosing macrophages, and that the increases in regeneration caused by ATRA are in part due to their increased numbers. Further studies will examine whether macrophage depletion affects RGC survival.

neuroscience↗

bioRxiv2019

A new method of recording from the giant fiber of Drosophila melanogaster shows that the strength of its auditory inputs remains constant with age

There have been relatively few studies of how central synapses age in adult Drosophila melanogaster. In this study we investigate the aging of the synaptic inputs to the Giant Fiber (GF) from the Johnstons Organ neurons (JONs). In previously published experiments an indirect assay of this synaptic connection was used; here we describe a new, more direct assay, which allows reliable detection of the GF action potential in the neck connective, and long term recording of its responses to sound. Genetic ablation using diphtheria toxin expressed in the GF with R68A06-GAL4 was used to show that these signals indeed arose from the GF and not from other descending neurons. As before, the sound-evoked action potentials (SEPs) in the antennal nerve were recorded via an electrode inserted at the base of the antenna. We then used this technique to quantify the response of the JONs to a high frequency sound pulse, and also the strength of the JON-GF synapse, in males and female of different ages. At no age was there any significant difference between males and females, for any of the parameters measured. Sensitivity of the JONs increased between 1 d and 10 d, with the sound intensity that elicited a half-maximal SEP decreasing by 40%. This measure almost doubled by 20 d and had increased 3-fold by 50 d compared to 10 d. Thus, JONs are most responsive around the period when most matings are taking place. The strength of the JON-GF synaptic connection itself was quite variable and did not change significantly with age. As a result, the GFs sensitivity to sound approximately followed the JONs sensitivity, but with greater variability.

neuroscience↗

bioRxiv2018

Application of CNTF or FGF-2 increases the number of M2-like macrophages after optic nerve injury in adult Rana pipiens

We have previously shown that a single application of the growth factors ciliary neurotrophic factor (CNTF) or fibroblast growth factor 2 (FGF-2) to the crushed optic nerve of the frog, Rana pipiens, increases the numbers and elongation rate of regenerating retinal ganglion cell axons. Here we investigate the effects of these factors on the numbers and types of macrophages that invade the regeneration zone. In control PBS-treated nerves, many macrophage-like cells are present 100 m distal to the crush site at 1 week after injury; their numbers halve by 2 weeks. A single application of CNTF at the time of injury triples the numbers of macrophages at 1 week, with this increase compared to control being maintained at 2 weeks. Application of FGF-2 is equally effective at 1 week, but the macrophage numbers have fallen to control levels at 2 weeks. Immunostaining with a pan-macrophage marker, ED1, and a marker for M2-like macrophages, Arg-1, showed that the proportion of the putative M2 phenotype remained at approximately 80% with all treatments. Electron microscopy of the macrophage-like cells at 1 week shows strong phagocytic activity with all treatments, with many vacuoles containing axon fragments and membrane debris. At 2 weeks with PBS or FGF-2 treatment the remaining macrophage-like cells are less phagocytically active, containing mainly lipid inclusions. With CNTF treatment, at 2 weeks many of the more numerous macrophages are still phagocytosing axonal debris, although they also contain lipid inclusions. We conclude that the increase in macrophage influx seen after growth factor application is beneficial for the regenerating axons, probably due to more extensive removal of degenerating distal axons, but also perhaps to secretion of growth-promoting substances.

neuroscience↗

bioRxiv2018

Effects of gap junction misexpression on synapses between auditory sensory neurons and the giant fiber of Drosophila melanogaster.

The synapse between auditory Johnstons Organ neurons (JONs) and the giant fiber (GF) of Drosophila is structurally mixed, being composed of cholinergic chemical synapses and Neurobiotin-(NB) permeable gap junctions, which consist of the innexin Shaking-B (ShakB). Misexpression of one ShakB isoform, ShakB(N+16), in a subset of JONs that do not normally form gap junctions, results in their de novo dye coupling to the GF. This is similar to the effect of misexpression of the transcription factor Engrailed (En) in these same neurons, which also causes the formation of additional chemical synapses. In order to test the hypothesis that ShakB misexpression would similarly affect the distribution of chemical synapses, fluorescently-labeled presynaptic active zone protein (Brp) was expressed in JONs and the changes in its distribution were assayed with confocal microscopy. Both ShakB(N+16) and En increased the dye-coupling of JONs with the GF, indicating the formation of ectopic gap junctions. Conversely, expression of the incorrect isoform, ShakB(N) abolishes dye coupling. However, while En misexpression increased the chemical contacts with the GF and the amount of GF medial branching, ShakB misexpression did not. ShakB immunocytochemistry showed that misexpression of ShakB(N+16) increases gap junctional plaques in JON axons but ShakB(N) does not. We conclude that both subsets of JON form chemical synapses onto the GF dendrites but only one population forms gap junctions, comprised of ShakB(N+16). Misexpression of this isoform in all JONs does not result in the formation of new mixed synapses but in the insertion of gap junctions, presumably at the sites of existing chemical synaptic contacts with the GF.

neuroscience↗