Systematic comparison of adeno‐associated virus and biotinylated dextran amine reveals equivalent sensitivity between tracers and novel projection targets in the mouse brain

As an anterograde neuronal tracer, recombinant adeno‐associated virus (AAV) has distinct advantages over the widely used biotinylated dextran amine (BDA). However, the sensitivity and selectivity of AAV remain uncharacterized for many brain regions and species. To validate this tracing method further, AAV (serotype 1) was systematically compared with BDA as an anterograde tracer by injecting both tracers into three cortical and 15 subcortical regions in C57BL/6J mice. Identical parameters were used for our sequential iontophoretic injections, producing injections of AAV that were more robust in size and in density of neurons infected compared with those of BDA. However, these differences did not preclude further comparison between the tracers, because the pairs of injections were suitably colocalized and contained some percentage of double‐labeled neurons. A qualitative analysis of projection patterns showed that the two tracers behave very similarly when injection sites are well matched. Additionally, a quantitative analysis of relative projection intensity for cases targeting primary motor cortex (MOp), primary somatosensory cortex (SSp), and caudoputamen (CP) showed strong agreement in the ranked order of projection intensities between the two tracers. A detailed analysis of the projections of two brain regions (SSp and MOp) revealed many targets that have not previously been described in the mouse or rat. Minor retrograde labeling of neurons was observed in all cases examined, for both AAV and BDA. Our results show that AAV has actions equivalent to those of BDA as an anterograde tracer and is suitable for analysis of neural circuitry throughout the mouse brain. J. Comp. Neurol. 522:1989–2012, 2014. © 2014 Wiley Periodicals, Inc.     

Selective anterograde tracing of the individual serotonergic and nonserotonergic components of the dorsal raphe nucleus projection to the vestibular nuclei

It is well known that the dorsal raphe nucleus (DRN) sends serotonergic and nonserotonergic projections to target regions in the brain stem and forebrain, including the vestibular nuclei. Although retrograde tracing studies have reported consistently that there are differences in the relative innervation of different target regions by serotonergic and nonserotonergic DRN neurons, the relative termination patterns of these two projections have not been compared using anterograde tracing methods. The object of the present investigation was to trace anterogradely the individual serotonergic and nonserotonergic components of the projection from DRN to the vestibular nuclei in rats. To trace nonserotonergic DRN projections, animals were pretreated with the serotonergic neurotoxin 5,7-dihydroxytryptamine (5,7-DHT), and then, after 7 days, the anterograde tracer biotinylated dextran amine (BDA) was iontophoretically injected into the DRN. In animals treated with 5,7-DHT, nonserotonergic BDA-labeled fibers were found to descend exclusively within the ventricular plexus and to terminate predominantly within the periventricular aspect of the vestibular nuclei. Serotonergic DRN projections were traced by injecting 5,7-DHT directly into DRN, and amino-cupric-silver staining was used to visualize the resulting pattern of terminal degeneration. Eighteen hours after microinjection of 5,7-DHT into the DRN, fine-caliber degenerating serotonergic terminals were found within the region of the medial vestibular nucleus (MVN) that borders the fourth ventricle, and a mixture of fine- and heavier-caliber degenerating serotonergic terminals was located further laterally within the vestibular nuclear complex. These findings indicate that fine-caliber projections from serotonergic and nonserotonergic DRN neurons primarily innervate the periventricular regions of MVN, whereas heavier-caliber projections from serotonergic DRN neurons innervate terminal fields located in more lateral regions of the vestibular nuclei. Thus, serotonergic and nonserotonergic DRN axons target distinct but partially overlapping terminal fields within the vestibular nuclear complex, raising the possibility that these two DRN projection systems are organized in a manner that permits regionally-specialized regulation of processing within the vestibular nuclei.     

Balance control interferes with the tracing performance of a pattern with mirror-reversed vision in older persons

GeroScience - When tracing a template with mirror-reversed vision (or distorted vision), the sensory information arising from the movement does not match the expected sensory consequences. In such...     

Composition of a Neuromere and Its Segmental Diversification under the Control of Hox Genes in the Embryonic CNS of Drosophila

Studies performed at the level of single, identified cells in the fruitfly Drosophila have decisively contributed to our understanding of the mechanisms underlying the development and function of the nervous system. This review highlights some of the work based on single-cell analyses in the embryonic/larval CNS that sheds light on the principles underlying formation and organization of an entire segmental unit and its divergence along the anterior/posterior body axis.     

Comparison of projection neurons in the pontine nuclei and the nucleus reticularis tegmenti pontis of the rat

Dendritic features of identified projection neurons in two precerebellar nuclei, the pontine nuclei (PN) and the nucleus reticularis tegmenti pontis (NRTP) were established by using a combination of retrograde tracing (injection of fluorogold or rhodamine labelled latex microspheres into the cerebellum) with subsequent intracellular filling (lucifer yellow) in fixed slices of pontine brainstem. A multivariate analysis revealed that parameters selected to characterize the dendritic tree such as size of dendritic field, number of branching points, and length of terminal dendrites did not deviate significantly between different regions of the PN and the NRTP. On the other hand, projection neurons in ventral regions of the PN were characterized by an irregular coverage of their distal dendrites by appendages while those in the dorsal PN and the NRTP were virtually devoid of them. The NRTP, dorsal, and medial PN tended to display larger somata and more primary dendrites than ventral regions of the PN. These differences, however, do not allow the differentiation of projection neurons within the PN from those in the NRTP. They rather reflect a dorso-ventral gradient ignoring the border between the nuclei. Accordingly, a cluster analysis did not differentiate distinct types of projection neurons within the total sample. In both nuclei, multiple linear regression analysis revealed that the size of dendritic fields was strongly correlated with the length of terminal dendrites while it did not depend on other parameters of the dendritic field. Thus, larger dendritic fields seem not to be accompanied by a higher complexity but rather may be used to extend the reach of a projection neuron within the arrangement of afferent terminals. We suggest that these similarities within dendritic properties in PN and NRTP projection neurons reflect similar processing of afferent information in both precerebellar nuclei. © 1996 Wiley-Liss, Inc.     

Projections from the nucleus reuniens thalami to the entorhinal cortex,hippocampal field CA1, and the subiculum in the rat arise from different populations of neurons

The entorhinal cortex, CA1, and the subiculum receive a major input from the thalamic midline nucleus reuniens. At present, it is not known whether reuniens projections to these intimately interconnected regions are collateralized or arise from different cell populations. We employed the multiple fluorescent retrograde tracing technique with Fast Blue, Diamidino Yellow, and Fluoro-Gold to examine the possible collateralization of reuniens projections to the entorhinal cortex, CA1, and the subiculum. In addition, we studied the extent of collateralization within each target area. The results indicate that different, yet morphologically indistinguishable, populations of reuniens cells selectively innervate the entorhinal cortex, CA1, or subiculum. Within each of these areas, reuniens fibers display a locally restricted collateralization instead of distributing collaterals throughout the entire target structure. The rostal two-thirds of the nucleus reuniens is the major source of ipsilateral projections to CA1, subiculum, and entorhinal cortex. The perireuniens nucleus selectively projects to the perirhinal cortex. Reuniens projections to CA1 and medial entorhinal cortex originate in the dorsolateral part and throughout the medial one-half of the nucleus, respectively. For these two projections, no topography could be established. However, subicular afferents are topographically organized such that a dorsal-to-ventral gradient in the nucleus reuniens corresponds to a dorsal-to-ventral gradient along the subicular axis. Lateral entorhinal afferents display a subtle topography such that a lateral-to-medial shift of terminal fields in the lateral entorhinal cortex corresponds to a lateral-to-medial shift of projection neurons in the ventral nucleus reuniens. © 1996 Wiley-Liss, Inc.