Journal: Neuron

Article Title: Multimodal Single-Cell Analysis Reveals Physiological Maturation in the Developing Human Neocortex

doi: 10.1016/j.neuron.2019.01.027

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Biocytin was detected by Streptavidin Dylight 549 (1:500, Vector Labs SA-5549).

Techniques: Plasmid Preparation, RNAscope 2.5 HD Assay, RNA Sequencing Assay, Software

Journal: Frontiers in Neuroanatomy

Article Title: Registry of Compartmental Ephrin-B3 Guidance Patterns With Respect to Emerging Multimodal Midbrain Maps

doi: 10.3389/fnana.2021.649478

Figure Lengend Snippet: Developmental progression of GAD67-GFP and ephrin-B3 expression patterns in the neonatal lateral cortex of the inferior colliculus (LCIC). Transient ephrin-B3 expression (red) with respect to developing glutamic acid decarboxylase (GAD)-positive modules (green) at P0 (A–C) , P4 (D–F) , P8 (G–I) , and P12 (J–L) . At birth, GAD-positive modules are in early development and are not readily identifiable, and ephrin-B3, while present in the LCIC, does not yet exhibit any clear compartmental organization. By P4, modules are distinguishable (dashed contours) and become more evident with age. Over this critical period for emerging LCIC compartments, the ephrin-B3 expression is complementary to that of GAD and preferentially restricted to the surrounding LCIC matrix. Note discrete layer 2 modular voids at P4 and P8 ( E,H , arrowheads), encompassed by concentrated ephrin-B3 expression. With the modular-extramodular framework and afferent patterns largely in place by P12 ( see Lamb-Echegaray et al., ), LCIC ephrin-B3 expression appears strikingly downregulated (K,L) . Scale bars = 100 μm.

Article Snippet: For double-labeling studies, a directly conjugated secondary was used for CR visualization (Alexa Fluor 350 donkey anti-rabbit IgG, 1:25, A10039, Thermo Fisher Scientific, RRID: AB_2534015 ), paired with a biotinylated-streptavidin detection system for ephrin-B3 (biotinylated horse anti-goat IgG, 1:600, BA-9500, Vector Laboratories, RRID: AB_2336123 ; Dylight 549 streptavidin, 1:200, SA-5549, Vector Laboratories).

Techniques: Expressing

Journal: Frontiers in Neuroanatomy

Article Title: Registry of Compartmental Ephrin-B3 Guidance Patterns With Respect to Emerging Multimodal Midbrain Maps

doi: 10.3389/fnana.2021.649478

Figure Lengend Snippet: Quantification of LCIC ephrin-B3 expression patterns relative to GAD modular expression over the early critical period. The negatively sloped linear regression of cross-correlation y-intercept values with age (A) supports the observed developmental trend that ephrin-B3 patterns become increasingly extramodular and offset from emerging GAD-positive layer 2 modules. The absence of any prominent values in P0 cross-correlation functions [representative plot shown in panel (B) ] indicates the lack of any significantly similar pattern features between ephrin-B3 and GAD-positive signals at birth. In contrast, by P8 [representative plot shown in panel (C) ] pronounced negative cross-correlation y-intercepts and smooth transitions to prominent positive cross-correlation values at shifts of approximately ±200–250 μm indicate a significant mismatch between signal features at zero relative signal shift and again at regular intervals in keeping with the discontinuous LCIC compartmental arrangement at this age.

Article Snippet: For double-labeling studies, a directly conjugated secondary was used for CR visualization (Alexa Fluor 350 donkey anti-rabbit IgG, 1:25, A10039, Thermo Fisher Scientific, RRID: AB_2534015 ), paired with a biotinylated-streptavidin detection system for ephrin-B3 (biotinylated horse anti-goat IgG, 1:600, BA-9500, Vector Laboratories, RRID: AB_2336123 ; Dylight 549 streptavidin, 1:200, SA-5549, Vector Laboratories).

Techniques: Expressing

Journal: Frontiers in Neuroanatomy

Article Title: Registry of Compartmental Ephrin-B3 Guidance Patterns With Respect to Emerging Multimodal Midbrain Maps

doi: 10.3389/fnana.2021.649478

Figure Lengend Snippet: Ephrin-B3 LCIC expression with respect to GAD (modular) and calretinin (CR; matrix) labeling at P8. Digital merges from a coronal series (A–C) through the modular extent of the LCIC at P8 [ephrin-B3 = red, GAD = green, calretinin (CR) = blue]. Dashed contours indicate the change in the appearance of GAD modules along the caudal-to-rostral dimension of the LCIC (in keeping with that previously described, see Dillingham et al., ; Gay et al., ). The ephrin-B3 expression is strong throughout the coronal series, exhibiting patterns complementary to GAD-positive modules that overlap the calretinin-defined matrix. Within layers 1 and 3 and intermodular bridges that together comprise extramodular regions, many calretinin-positive cells are also positive for ephrin-B3 (purple). Scale bars = 100 μm.

Article Snippet: For double-labeling studies, a directly conjugated secondary was used for CR visualization (Alexa Fluor 350 donkey anti-rabbit IgG, 1:25, A10039, Thermo Fisher Scientific, RRID: AB_2534015 ), paired with a biotinylated-streptavidin detection system for ephrin-B3 (biotinylated horse anti-goat IgG, 1:600, BA-9500, Vector Laboratories, RRID: AB_2336123 ; Dylight 549 streptavidin, 1:200, SA-5549, Vector Laboratories).

Techniques: Expressing, Labeling

Journal: Frontiers in Neuroanatomy

Article Title: Registry of Compartmental Ephrin-B3 Guidance Patterns With Respect to Emerging Multimodal Midbrain Maps

doi: 10.3389/fnana.2021.649478

Figure Lengend Snippet: LCIC sampling and corresponding brightness profiles of extracted GAD (green), ephrin-B3 (red), and calretinin (blue) channels in a representative P8 mouse. Consistent ROI sampling curved contour in panel (A) bisecting LCIC layer 2 modular fields facilitates comparisons of relative signal fluctuations for each label (B) . Each exhibits a periodic component, in keeping with discontinuous compartments along the ventrolateral-to-dorsomedial extent of the LCIC. Ephrin-B3 and calretinin waveforms are largely in-phase with each other (red and blue arrows, respectively), and out-of-phase with that of GAD oscillations (green arrows). Such reliable alignment (with CR) and offset (with GAD) suggests a high degree of ephrin-B3 expression specificity that corresponds with the emerging LCIC extramodular matrix. Scale bar in (A) = 100 μm.

Article Snippet: For double-labeling studies, a directly conjugated secondary was used for CR visualization (Alexa Fluor 350 donkey anti-rabbit IgG, 1:25, A10039, Thermo Fisher Scientific, RRID: AB_2534015 ), paired with a biotinylated-streptavidin detection system for ephrin-B3 (biotinylated horse anti-goat IgG, 1:600, BA-9500, Vector Laboratories, RRID: AB_2336123 ; Dylight 549 streptavidin, 1:200, SA-5549, Vector Laboratories).

Techniques: Sampling, Expressing