{ "paper": "artiushin-2026-spider-atlas", "claims": 17, "panels": { "total": [ "fig1a", "fig1b", "fig2a", "fig2b", "fig2c", "fig2d", "fig3a", "fig3b", "fig3c", "fig3d", "fig4a", "fig4b", "fig4c", "fig4d", "fig4e", "fig5a", "fig5b", "fig5c", "fig6a", "fig6b", "fig6c", "fig6d", "fig7a", "fig7b", "fig7c", "fig7d", "fig7e", "fig7f", "fig7g", "fig7h", "fig7i", "fig8a", "fig8b", "fig9a", "fig9b", "fig9c", "fig10a", "fig10b", "fig11a", "fig11b", "fig11c", "fig11d", "fig11e", "fig11f", "fig11g", "fig12", "fig13a", "fig13b", "fig13c", "inlinetable1" ], "orphan": [ "fig1a", "fig1b", "fig2a", "fig2b", "fig2c", "fig2d", "fig3a", "fig3b", "fig3c", "fig3d", "fig4a", "fig4b", "fig4c", "fig4d", "fig4e", "fig5a", "fig5b", "fig5c", "fig6a", "fig6b", "fig6c", "fig6d", "fig7c", "fig7d", "fig7f", "fig7g", "fig7i", "fig8a", "fig8b", "fig9a", "fig9b", "fig9c", "fig10a", "fig10b", "fig11a", "fig11b", "fig11e", "fig11f", "fig11g", "fig13a", "fig13b", "fig13c", "inlinetable1" ], "figure_level": [ "fig1", "fig2" ], "phantom": [], "pct": 14.0 }, "statistics": { "total": 0, "orphan": [], "pct": 100.0 }, "spans": { "segmented": 582, "obligations": 202, "textual": 380, "accounted": 74, "pct": 36.6, "orphans": [ { "uid": "results-002", "section": "results", "text": "Residing within the prosoma, the central nervous system, or synganglion, as it has been called ( Steinhoff et al., 2017 ), is comprised of two major divisions named in reference to the esophageal passage traveling between them \u2013 the subesophageal mass, comprised primarily of motor and sensory interneurons and comparable to the ventral nerve cord in insects, and the supraesophageal mass, containing the higher-order integration centers ( Figure 1A ).", "stats": [], "panels": [ "fig1a" ] }, { "uid": "results-004", "section": "results", "text": "Consistent with general arthropod nervous system morphology, the neuronal somata are found superficially ( Figure 1B ), while the internal structure of the brain is comprised primarily of neuropil.", "stats": [], "panels": [ "fig1b" ] }, { "uid": "results-006", "section": "results", "text": "( A ) 3D rendering of U. diversus (female) synganglion from averaged \u03b1-synapsin volume, composed of 909 z -planes, oblique posterior\u2013lateral (left) and oblique anterior\u2013lateral (right) views.", "stats": [], "panels": [ "fig1a" ] }, { "uid": "results-007", "section": "results", "text": "( B ) 3D rendering of \u03b1-synapsin (green) and DAPI stained (blue) synganglion, posterior, lateral, and anterior views.", "stats": [], "panels": [ "fig1b" ] }, { "uid": "results-025", "section": "results", "text": "( A ) 3D rendering of leg neuropils as annotated from averaged synapsin volume with posterior oblique, anterior oblique, and dorsal views, left to right.", "stats": [], "panels": [ "fig2a" ] }, { "uid": "results-026", "section": "results", "text": "( B ) Maximum intensity projections of z -planes 160\u2013190 for leg neuropil expression of synapsin (\u03b1-synapsin, gray), ( i ) serotonergic (\u03b1-5-HT, green), (ii) octopaminergic/tyraminergic (\u03b1-TDC2, magenta), (iii) cholinergic (\u03b1-ChAT, cyan), (iv) proctolin (\u03b1-Proctolin, yellow), ( v ) FMRFamide (\u03b1-FMRFamide, red), (vi) cardioactive peptide (\u03b1-CCAP, cyan), and (vii) allatostatin A (\u03b1-AstA, green) immunoreactivity in the standard brain.", "stats": [], "panels": [ "fig2b" ] }, { "uid": "results-027", "section": "results", "text": "Dotted perimeter marks the boundary of Leg Neuropil 1. ( C ) Maximum intensity projections of z -planes 230\u2013260. Dotted perimeter marks the boundary of Leg Neuropil 1. ( D ) Maximum intensity projection showing dopaminergic (\u03b1-TH, green) immunoreactivity, dotted perimeter marks the boundary of Leg Neuropil 1. Dopaminergic immunoreactivity is shown in a separate panel because this volume, unlike the other stains, is not aligned to the standard atlas.", "stats": [], "panels": [ "fig2c", "fig2d" ] }, { "uid": "results-034", "section": "results", "text": "Dopaminergic signal (anti-TH) is more expansive, evenly filling each LN with a mesh-like network of TH+ varicosities ( Figure 2D ).", "stats": [], "panels": [ "fig2d" ] }, { "uid": "results-041", "section": "results", "text": "CCAP immunoreactivity within the LNs is predominantly in the posterior halves, where sparse puncta are evenly distributed ( Figure 2B, Cvi ).", "stats": [], "panels": [ "fig2b" ] }, { "uid": "results-042", "section": "results", "text": "FMRFamide immunoreactivity is also evident in the LNs ( Figure 2B, Cv ).", "stats": [], "panels": [ "fig2b" ] }, { "uid": "results-043", "section": "results", "text": "Opisthosomal neuropil The OpN is found at the posterior and dorsal-most extension of the subesophageal mass ( Figure 3A ; Figure 3\u2014videos 1\u20133 ).", "stats": [], "panels": [ "fig3a", "fig3" ] }, { "uid": "results-045", "section": "results", "text": "Within the OpN, there are a number of antero-posterior longitudinal tracts, while a ladder-like appearance of medio-laterally running tracts in the \u237a-synapsin channel can also be observed ( Figure 3B, C ).", "stats": [], "panels": [ "fig3b", "fig3c" ] }, { "uid": "results-046", "section": "results", "text": "Posteriorly traveling tracts also diverge laterally to follow the circumference of the OpN ( Figure 3B\u2013D ).", "stats": [], "panels": [ "fig3b", "fig3c", "fig3d" ] }, { "uid": "results-047", "section": "results", "text": "Figure 3. Opisthosomal neuropil.", "stats": [], "panels": [ "fig3" ] }, { "uid": "results-048", "section": "results", "text": "( A ) 3D rendering of the opisthosomal neuropil as annotated from averaged synapsin volume with posterior oblique, anterior oblique, and dorsal views, left to right.", "stats": [], "panels": [ "fig3a" ] }, { "uid": "results-049", "section": "results", "text": "( B ) Maximum intensity projections of z -planes 415\u2013425 for opisthosomal neuropil expression of synapsin (\u03b1-synapsin, gray), ( i ) serotonergic (\u03b1-5-HT, green), (ii) octopaminergic/tyraminergic (\u03b1-TDC2, magenta), (iii) cholinergic (\u03b1-ChAT, cyan), (iv) proctolin (\u03b1-Proctolin, yellow), ( v ) FMRFamide (\u03b1-FMRFamide, red), (vi) cardioactive peptide (\u03b1-CCAP, cyan), and (vii) allatostatin A (\u03b1-AstA, green) immunoreactivity in the standard brain.", "stats": [], "panels": [ "fig3b" ] }, { "uid": "results-052", "section": "results", "text": "( C ) Maximum intensity projections of z -planes 448\u2013465. Longitudinal and lateral projections form a ladder-like structure in the \u03b1-TDC2 subpanel (brace).", "stats": [], "panels": [ "fig3c" ] }, { "uid": "results-054", "section": "results", "text": "( D ) Maximum intensity projection showing dopaminergic (\u03b1-TH, green) immunoreactivity in the opisthosomal neuropil.", "stats": [], "panels": [ "fig3d" ] }, { "uid": "results-056", "section": "results", "text": "Figure 3\u2014video 1. Opisthosomal 3D volume.", "stats": [], "panels": [ "fig3" ] }, { "uid": "results-057", "section": "results", "text": "Figure 3\u2014video 2. Neurotransmitter immunostains in the opisthosomal neuropil.", "stats": [], "panels": [ "fig3" ] }, { "uid": "results-058", "section": "results", "text": "Figure 3\u2014video 3. Neuropeptide immunostains in the opisthosomal neuropil.", "stats": [], "panels": [ "fig3" ] }, { "uid": "results-061", "section": "results", "text": "The lateral perimeter tracts are likewise revealed by ChAT and proctolin immunoreactivity ( Figure 3B, Ciii, iv ) as well as by fibers from dopaminergic populations (anti-TH) ( Figure 3D ).", "stats": [], "panels": [ "fig3b", "fig3d" ] }, { "uid": "results-065", "section": "results", "text": "Intense boutons line a tract running parallel to the midline, while also giving rise dorsally and laterally to a ladder-like structure of projections in the anterior\u2013posterior direction, which can best be seen with octopaminergic/tyraminergic innervation (TDC2+) ( Figure 3Cii \u2013 brace) and dopaminergic signal (TH+) ( Figure 3D ).", "stats": [], "panels": [ "fig3d" ] }, { "uid": "results-067", "section": "results", "text": "Ample FMRFamide signal is seen within the opisthosomal neuromere, but its signal is more uniform than the other targets investigated ( Figure 3B, Cv ).", "stats": [], "panels": [ "fig3b" ] }, { "uid": "results-068", "section": "results", "text": "Pedipalpal neuropil As the LNs begin to diminish in the dorsal region of the subesophageal mass, a smaller antero-lateral synaptic density becomes apparent, representing the pedipalpal neuropil (PdN) ( Figure 4A ; Figure 4\u2014videos 1 and 2 ), which serves the anterior-facing pedipalp appendages.", "stats": [], "panels": [ "fig4a", "fig4" ] }, { "uid": "results-074", "section": "results", "text": "Figure 4. Pedipalp, Blumenthal, and cheliceral neuropil.", "stats": [], "panels": [ "fig4" ] }, { "uid": "results-075", "section": "results", "text": "( A ) 3D rendering of the pedipalp neuropil as annotated from averaged synapsin volume with posterior oblique and anterior oblique views, left to right, and dorsal view, lower.", "stats": [], "panels": [ "fig4a" ] }, { "uid": "results-076", "section": "results", "text": "( B ) Optical slices (z275) from the standard brain, containing a cross-section of the pedipalp neuropil (dashed line boundary), with expression of (i) synapsin (\u03b1-synapsin, gray), (ii) octopaminergic/tyraminergic (\u03b1-TDC2, magenta), (iii) cholinergic (\u03b1-ChAT, cyan), and (iv) proctolin (\u03b1-Proctolin, yellow) immunoreactivity.", "stats": [], "panels": [ "fig4b" ] }, { "uid": "results-078", "section": "results", "text": "( C ) Optical slices (z360) from the standard brain, with a cropped and enlarged selection showing the Blumenthal neuropil (brace), with expression of (i) synapsin (\u03b1-synapsin, gray) and proctolin (\u03b1-Proctolin, yellow) immunoreactivity above, and only (ii) proctolin below.", "stats": [], "panels": [ "fig4c" ] }, { "uid": "results-080", "section": "results", "text": "( D ) 3D rendering of the cheliceral neuropil as annotated from averaged synapsin volume with posterior oblique and anterior oblique views, left to right, and dorsal view, lower.", "stats": [], "panels": [ "fig4d" ] }, { "uid": "results-081", "section": "results", "text": "( E ) Optical slices (z400) from the standard brain, containing a cross-section of the cheliceral neuropil (dashed line boundary), with expression of (i) synapsin (\u03b1-synapsin, gray), (ii) octopaminergic/tyraminergic (\u03b1-TDC2, magenta), (iii) allatostatin A (\u03b1-AstA, green), (iv) proctolin (\u03b1-Proctolin, yellow), (v) serotonergic (\u03b1-5-HT, green), and (vi) cholinergic (\u03b1-ChAT, cyan) immunoreactivity.", "stats": [], "panels": [ "fig4e" ] }, { "uid": "results-084", "section": "results", "text": "Figure 4\u2014video 1. Pedipalp neuropil 3D volume.", "stats": [], "panels": [ "fig4" ] }, { "uid": "results-085", "section": "results", "text": "Figure 4\u2014video 2. Neurotransmitter and neuropeptide immunostains in the pedipalp neuropil.", "stats": [], "panels": [ "fig4" ] }, { "uid": "results-086", "section": "results", "text": "Figure 4\u2014video 3. Cheliceral 3D volume.", "stats": [], "panels": [ "fig4" ] }, { "uid": "results-087", "section": "results", "text": "Figure 4\u2014video 4. Neurotransmitter immunostains in the cheliceral neuropil.", "stats": [], "panels": [ "fig4" ] }, { "uid": "results-088", "section": "results", "text": "Figure 4\u2014video 5. Neuropeptide immunostains in the cheliceral neuropil.", "stats": [], "panels": [ "fig4" ] }, { "uid": "results-089", "section": "results", "text": "Figure 4\u2014video 6. Immunostains for \u03b1-synapsin and \u03b1-proctolin in the blumenthal neuropil.", "stats": [], "panels": [ "fig4" ] }, { "uid": "results-091", "section": "results", "text": "Although we also see a paired, synapsin-dense entity close to the midline ( Figure 4C \u2013 brace), in the approximate anterio-ventral subesophageal location as described for C. salei , we cannot be certain that this is the same structure \u2013 a question which will benefit from tracing techniques.", "stats": [], "panels": [ "fig4c" ] }, { "uid": "results-093", "section": "results", "text": "In dorsal planes, this immunoreactivity morphs into laterally moving tracts Figure 4\u2014video 6 , becoming difficult to trace.", "stats": [], "panels": [ "fig4" ] }, { "uid": "results-095", "section": "results", "text": "Deutocerebral features Cheliceral neuropil At the level of the esophageal passage, an anterior\u2013lateral neuropil begins, wrapping medially to become the ChN ( Figure 4D ; Figure 4\u2014videos 3\u20135 ).", "stats": [], "panels": [ "fig4d", "fig4" ] }, { "uid": "results-103", "section": "results", "text": "Supraesophageal (Protocerebral) neuropils Ventral features The supraesophageal mass begins dorsally to the closure of the esophageal passage, and in its ventral-most planes, can be subdivided anteriorly and posteriorly into two sectors of slightly differing anti-synapsin immunoreactivity ( Figure 5A\u2013C \u2013 dashed perimeter, anti-synapsin).", "stats": [], "panels": [ "fig5a", "fig5b", "fig5c" ] }, { "uid": "results-106", "section": "results", "text": "Figure 5. Ventral supraesophageal features.", "stats": [], "panels": [ "fig5" ] }, { "uid": "results-107", "section": "results", "text": "Posterior and anterior stalk region expression of synapsin (\u03b1-synapsin, gray), ( i ) serotonergic (\u03b1-5-HT, green), (ii) cholinergic (\u03b1-ChAT, cyan), (iii) octopaminergic/tyraminergic (\u03b1-TDC2, magenta), (iv) proctolin (\u03b1-Proctolin, yellow), ( v ) allatostatin A (\u03b1-AstA, green), and (vi) cardioactive peptide (\u03b1-CCAP, cyan) immunoreactivity in the standard brain, for: ( A ) z -plane 461. Arrow in the \u03b1-TDC2 subpanel marks tracks along perimeter of the opisthosomal neuropil, supplied anteriorly (brace).", "stats": [], "panels": [ "fig5a" ] }, { "uid": "results-110", "section": "results", "text": "( B ) z -plane 490. Further ventrally, a bridging structure is also visible on the posterior end of the ventral supraesophageal, as seen in the \u03b1-Proctolin subpanel (brace).", "stats": [], "panels": [ "fig5b" ] }, { "uid": "results-113", "section": "results", "text": "( C ) z -plane 511. Arrowhead in the \u03b1-ChAT subpanel marks prominent cholinergic immunoreactivity in the PCT, more clearly visible at this plane.", "stats": [], "panels": [ "fig5c" ] }, { "uid": "results-117", "section": "results", "text": "Figure 5\u2014video 1. Neurotransmitter Immunostains in the Ventral Supraesophageal Ganglion.", "stats": [], "panels": [ "fig5" ] }, { "uid": "results-118", "section": "results", "text": "Figure 5\u2014video 2. Neuropeptide immunostains in the ventral supraesophageal ganglion.", "stats": [], "panels": [ "fig5" ] }, { "uid": "results-119", "section": "results", "text": "The esophageal passage is bridged at the anterior side by a region named the stomodeal bridge (StB) ( Steinhoff et al., 2017 ; Figure 5A \u2013 brace).", "stats": [], "panels": [ "fig5a" ] }, { "uid": "results-120", "section": "results", "text": "A bridge structure also exists at the posterior end, where additional undifferentiated synaptic density is flanking ( Figure 5B \u2013 brace with asterisk).", "stats": [], "panels": [ "fig5b" ] }, { "uid": "results-121", "section": "results", "text": "Within these planes, a protocerebral tract (PCT) is essentially parallel to the ventro-dorsal axis and appears as twin, dense nodes rising in the central burgeoning protocerebrum ( Figure 5B, C ). 5-HT immunoreactivity is prominent in the posterior bridging area dorsal to the esophageal passage ( Figure 5B, Ci , Figure 5\u2014video 1 ), as well as the laterally adjacent tissue, and not as apparent in the anterior StB. 5-HT immunoreactivity is otherwise weak within the stalk regions.", "stats": [], "panels": [ "fig5b", "fig5c", "fig5" ] }, { "uid": "results-122", "section": "results", "text": "TDC2 immunoreactivity is prominent in the StB and adjacent areas, and at this plane, two lateral bands of immunoreactivity appear which do not correspond to a clear demarcation in the synapsin channel ( Figure 5Biii \u2013 arrow, Figure 5\u2014video 1 ).", "stats": [], "panels": [ "fig5" ] }, { "uid": "results-124", "section": "results", "text": "Concentrations of TDC2+ immunoreactivity are also apparent centrally, both anterior and lateral to the PCTs ( Figure 5Ciii \u2013 arrows, Figure 5\u2014video 1 ).", "stats": [], "panels": [ "fig5" ] }, { "uid": "results-125", "section": "results", "text": "Similar to what has been described for M. muscosa as the StB ( Steinhoff et al., 2017 ), the area adjacent to the esophagus on the anterior side has immunoreactivity to allatostatin A, although the actual bridge which crosses the midline is modest, with thin representation in the posterior commissure ( Figure 5Av \u2013 arrowhead, Figure 5\u2014video 2 ).", "stats": [], "panels": [ "fig5" ] }, { "uid": "results-127", "section": "results", "text": "The posterior region adjacent to the midline, previously highlighted with 5HT immunoreactivity, also shows partial AstA+ innervation, displaying a unique oxbow type pattern ( Figure 5Cv \u2013 arrow, Figure 5\u2014video 2 ).", "stats": [], "panels": [ "fig5" ] }, { "uid": "results-132", "section": "results", "text": "There is also proctolin immunoreactivity seen centrally, medial to where the PCT is ascending ( Figure 5B, Civ ).", "stats": [], "panels": [ "fig5b" ] }, { "uid": "results-133", "section": "results", "text": "ChAT+ immunoreactivity is diffusely present throughout the stalk regions ( Figure 5Cii ), and unlike the other antisera, co-stains the posterior aspect of the PCT, which is prominently visible with synapsin immunoreactivity ( Figure 5B, Cii \u2013 arrowheads and dashed circles).", "stats": [], "panels": [ "fig5b" ] }, { "uid": "results-134", "section": "results", "text": "Hagstone neuropil and mid-supraesophageal features We will define the hagstone neuropil as a central, midline adjacent, paired structure, whose ventral bounds share the same plane as the appearance of a prominent protocerebral commissure (standard brain, ~ z = 540), and which continues dorsally until the formation of the MB bridge (standard brain, ~ z = 615).", "stats": [ "z = 540", "z = 615" ], "panels": [] }, { "uid": "results-135", "section": "results", "text": "This approximately kidney-bean-shaped structure is pierced by a circular spot lacking synapsin immunoreactivity ( Figure 6A, C, D \u2013 dashed outline, Figure 6\u2014videos 1\u20133 ) \u2013 which may be indicative of a space occupied by a fiber tract, tracheal passageway, and/or potentially glia.", "stats": [], "panels": [ "fig6a", "fig6c", "fig6d", "fig6" ] }, { "uid": "results-136", "section": "results", "text": "Figure 6. Hagstone neuropil and mid-supraesophageal features.", "stats": [], "panels": [ "fig6" ] }, { "uid": "results-137", "section": "results", "text": "( A ) 3D rendering of the hagstone neuropil as annotated from averaged synapsin volume with posterior oblique, anterior oblique, and dorsal views, top to bottom.", "stats": [], "panels": [ "fig6a" ] }, { "uid": "results-138", "section": "results", "text": "( B\u2013D ) Hagstone neuropil and posterior feature expression of synapsin (\u03b1-synapsin, gray), ( i ) serotonergic (\u03b1-5-HT, green), (ii) proctolin (\u03b1-Proctolin, yellow), (iii) octopaminergic/tyraminergic (\u03b1-TDC2, magenta), and (iv) allatostatin A (\u03b1-AstA, green) immunoreactivity in the standard brain, fo: ( B ) z -plane 540. A circular tract pattern distinctive to the \u03b1-5-HT stain is visible in the posterior mid-supraesophageal (brace, \u03b1-5-HT subpanel).", "stats": [], "panels": [ "fig6b", "fig6c", "fig6d" ] }, { "uid": "results-139", "section": "results", "text": "( C ) z -planes 565, with the boundary of hagstone neuropil (dashed perimeter).", "stats": [], "panels": [ "fig6c" ] }, { "uid": "results-141", "section": "results", "text": "( D ) z -plane 585, with the boundary of hagstone neuropil (dashed perimeter).", "stats": [], "panels": [ "fig6d" ] }, { "uid": "results-145", "section": "results", "text": "Figure 6\u2014video 1. Hagstone 3D volume.", "stats": [], "panels": [ "fig6" ] }, { "uid": "results-146", "section": "results", "text": "Figure 6\u2014video 2. Neurotransmitter immunostains in the hagstone neuropil.", "stats": [], "panels": [ "fig6" ] }, { "uid": "results-147", "section": "results", "text": "Figure 6\u2014video 3. Neuropeptide immunostains in the hagstone neuropil.", "stats": [], "panels": [ "fig6" ] }, { "uid": "results-150", "section": "results", "text": "The semi-circular tracts bend medially ( Figure 6Bi ), before a fusion of seemingly all three directions is seen immediately dorsal ( Figure 6C ).", "stats": [], "panels": [ "fig6c" ] }, { "uid": "results-151", "section": "results", "text": "The hagstone neuropil is essentially completely filled with serotonergic immunoreactivity, matching the outline defined in the anti-synapsin channel ( Figure 6C, Di ).", "stats": [], "panels": [ "fig6c" ] }, { "uid": "results-152", "section": "results", "text": "On the posterior end is a diffuse, arching band of varicosities, with fiber tracts at the midline, resembling an umbrella-like form ( Figure 6C, Di \u2013 brace, anti-5-HT).", "stats": [], "panels": [ "fig6c" ] }, { "uid": "results-154", "section": "results", "text": "TDC2 immunoreactivity is also present in the umbrella-like posterior region innervated by 5-HT ( Figure 6Ciii \u2013 brace, Figure 6\u2014video 2 ), and sparser puncta within the bounds of the hagstone neuropil.", "stats": [], "panels": [ "fig6" ] }, { "uid": "results-165", "section": "results", "text": "( C ) Optical sections of the supraesophageal ganglion from an averaged \u03b1-synapsin volume (ventral (top) to dorsal (bottom)).", "stats": [], "panels": [ "fig7c" ] }, { "uid": "results-167", "section": "results", "text": "( D ) \u03b1-\u03b2Tubulin3 (magenta) immunoreactivity aligned with \u03b1-Synapsin volume (gray) (compare to bottom portion of previous subfigure) showing the arching form of the mid-line spanning MB bridge.", "stats": [], "panels": [ "fig7d" ] }, { "uid": "results-169", "section": "results", "text": "( F ) \u03b1-\u03b2Tubulin3 (magenta) and \u03b1-Synapsin (green) immunoreactivity in the supraesophageal ganglion at the plane where the MB hafts appear (round, intensely immunoreactive).", "stats": [], "panels": [ "fig7f" ] }, { "uid": "results-171", "section": "results", "text": "( G ) Tripart tract entering at the MB head to fuse with the tract descending through the MB.", "stats": [], "panels": [ "fig7g" ] }, { "uid": "results-177", "section": "results", "text": "Synapsin immunoreactivity is modest within the bridge region, whose true thickness is better visualized with staining for \u03b2Tubulin3 ( Figure 7D ).", "stats": [], "panels": [ "fig7d" ] }, { "uid": "results-182", "section": "results", "text": "Although difficult to trace the source, it appears the hafts are innervated from the posterior side ( Figure 7F \u2013 arrows).", "stats": [], "panels": [ "fig7f" ] }, { "uid": "results-183", "section": "results", "text": "By \u03b2Tubulin3 immunoreactivity, we observe two tracts which straddle the MB hafts as they descend from the dorsal somata layer ( Figure 7G ).", "stats": [], "panels": [ "fig7g" ] }, { "uid": "results-192", "section": "results", "text": "By \u03b2Tubulin3 staining, we also observed a trident of tracts feeding into the dorsal aspect of the MB head ( Figure 7G ).", "stats": [], "panels": [ "fig7g" ] }, { "uid": "results-201", "section": "results", "text": "The optic neuropils in U. diversus tended to show weaker synapsin immunoreactivity but were clearly seen with antisera to HRP ( Figure 8A ).", "stats": [], "panels": [ "fig8a" ] }, { "uid": "results-202", "section": "results", "text": "Figure 8. Visual pathways.", "stats": [], "panels": [ "fig8" ] }, { "uid": "results-203", "section": "results", "text": "( A ) Immunostaining for \u03b1-HRP (magenta) for neuropil and use of DAPI (blue) for nuclei, arrows show the primary (1\u00b0) and secondary (2\u00b0) visual pathway extensions from the bulk of the supraesophageal tissue.", "stats": [], "panels": [ "fig8a" ] }, { "uid": "results-204", "section": "results", "text": "( B ) 3D renderings of synapsin immunoreactivity in the dorsal supraesophageal ganglion, with tissue of the primary (1\u00b0) and secondary (2\u00b0) visual pathway visible.", "stats": [], "panels": [ "fig8b" ] }, { "uid": "results-207", "section": "results", "text": "As in other species, the secondary pathway is larger ( Figure 8B ), lifting away anteriodorsally from the zone of the MB heads.", "stats": [], "panels": [ "fig8b" ] }, { "uid": "results-208", "section": "results", "text": "This continuity can be seen from the sliced three-dimensional maximum intensity projection of synapsin ( Figure 8B \u2013 brace).", "stats": [], "panels": [ "fig8b" ] }, { "uid": "results-209", "section": "results", "text": "The primary pathway is diminutive in U. diversus and emerges as a bulbous shape at the dorsal-most end of the brain through a field of somata ( Figure 8A ).", "stats": [], "panels": [ "fig8a" ] }, { "uid": "results-217", "section": "results", "text": "Between the two halves, at the midline, is a furrow which is negative for synapsin immunoreactivity, giving this neuropil, in conjunction with the synapsin-negative zone, a likeness to tonsils when viewed from the horizontal optical planes ( Figure 9A, C \u2013 \u03b1-synapsin, Figure 9\u2014videos 1\u20133 ) \u2013 hence our referral to this structure as the tonsillar neuropil.", "stats": [], "panels": [ "fig9a", "fig9c", "fig9" ] }, { "uid": "results-218", "section": "results", "text": "Figure 9. Tonsillar neuropil.", "stats": [], "panels": [ "fig9" ] }, { "uid": "results-219", "section": "results", "text": "( A ) 3D rendering of tonsillar neuropil as annotated from averaged synapsin volume with posterior oblique, anterior oblique, and dorsal views, left to right.", "stats": [], "panels": [ "fig9a" ] }, { "uid": "results-220", "section": "results", "text": "( B ) Oblique horizontal optical section of supraesophageal ganglion with \u03b1-Synapsin (green) and \u03b1-\u03b2Tubulin3 (magenta) immunoreactivity.", "stats": [], "panels": [ "fig9b" ] }, { "uid": "results-222", "section": "results", "text": "( C ) Ventral (z682) and dorsal (z700) views of the tonsillar neuropil, demarcated by dashed lines, from the standard brain, showing expression of synapsin (gray), (i, vi) serotonergic (\u03b1-5-HT, green), (ii, vii) octopaminergic/tyraminergic (\u03b1-TDC2, magenta), (iii, viii) proctolin (\u03b1-Proctolin, yellow), (iv, ix) allatostatin A (\u03b1-AstA, green) and (v, x ) FMRFamide (\u03b1-FMRFamide, red) immunoreactivity.", "stats": [], "panels": [ "fig9c" ] }, { "uid": "results-224", "section": "results", "text": "Figure 9\u2014video 1. Tonsillar neuropil 3D volume.", "stats": [], "panels": [ "fig9" ] }, { "uid": "results-225", "section": "results", "text": "Figure 9\u2014video 2. Neurotransmitter immunostains in the tonsillar neuropil.", "stats": [], "panels": [ "fig9" ] }, { "uid": "results-226", "section": "results", "text": "Figure 9\u2014video 3. Neuropeptide immunostains in the tonsillar neuropil.", "stats": [], "panels": [ "fig9" ] }, { "uid": "results-228", "section": "results", "text": "By tubulin immunoreactivity, it appears to bifurcate the structure below the bridge in the dorsal portion ( Figure 9B ).", "stats": [], "panels": [ "fig9b" ] }, { "uid": "results-229", "section": "results", "text": "A subset of antisera for specific neuronal populations is instrumental in confirming this neuropil, as their immunoreactivity is circumscribed by its boundaries, with little neighboring signal to obscure the distinction ( Figure 9C ).", "stats": [], "panels": [ "fig9c" ] }, { "uid": "results-236", "section": "results", "text": "This is the dorsal-most neuropil seen in the interior of the protocerebrum before reaching the dense cap of somata ( Figure 10 ; Figure 10\u2014videos 1\u20133 ).", "stats": [], "panels": [ "fig10" ] }, { "uid": "results-237", "section": "results", "text": "Figure 10. Protocerebral bridge (PCB) neuropil.", "stats": [], "panels": [ "fig10" ] }, { "uid": "results-238", "section": "results", "text": "( A ) 3D rendering of PCB neuropil as annotated from averaged synapsin volume, with posterior oblique, anterior oblique, and dorsal views, left to right.", "stats": [], "panels": [ "fig10a" ] }, { "uid": "results-239", "section": "results", "text": "( B ) Ventral (z722) and dorsal (z730) views of the PCB, as demarcated by dashed lines, from the standard brain, showing expression of synapsin (gray), (i, vi) GABAergic (\u03b1-GAD, red), (ii, vii) octopaminergic/tyraminergic (\u03b1-TDC2, magenta), (iii, viii) proctolin (\u03b1-Proctolin, yellow), (iv, ix) cholinergic (\u03b1-ChAT, cyan), and (v, x) allatostatin A (\u03b1-AstA, green) immunoreactivity.", "stats": [], "panels": [ "fig10b" ] }, { "uid": "results-242", "section": "results", "text": "Figure 10\u2014video 1. Protocerebral bridge 3D volume.", "stats": [], "panels": [ "fig10" ] }, { "uid": "results-243", "section": "results", "text": "Figure 10\u2014video 2. Neurotransmitter immunostains in the protocerebral bridge.", "stats": [], "panels": [ "fig10" ] }, { "uid": "results-244", "section": "results", "text": "Figure 10\u2014video 3. Neuropeptide immunostains in the protocerebral bridge.", "stats": [], "panels": [ "fig10" ] }, { "uid": "results-253", "section": "results", "text": "We refer to this as the dorsal protocerebral commissure ( Figure 10B \u2013 arrows) and the neuronal subtype populations which show PCB expression tend to also innervate this commissure.", "stats": [], "panels": [ "fig10b" ] }, { "uid": "results-254", "section": "results", "text": "The strongest of these are octopaminergic/tyraminergic neurons (anti-TDC2) and proctolin+ neurons (anti-proctolin) ( Figure 10B \u2013 \u03b1-TDC2, \u03b1-proctolin).", "stats": [], "panels": [ "fig10b" ] }, { "uid": "results-256", "section": "results", "text": "A similar pattern is also seen for allatostatin A ( Figure 10B \u2013 \u03b1-AstA).", "stats": [], "panels": [ "fig10b" ] }, { "uid": "results-257", "section": "results", "text": "Cholinergic immunoreactivity is also apparent in the posterior arch, but is more subtle and in a single layer in the anterior domain, less comparable to that of TDC2, proctolin, and allatostatin A. Arcuate body The arcuate body (AB) is a prominent neuropil structure found in all spider species whose central nervous system anatomy has been examined closely to date.", "stats": [], "panels": [ "fig10a", "fig10b" ] }, { "uid": "results-262", "section": "results", "text": "( A ) 3D rendering of arcuate body neuropil as annotated from averaged \u03b1-synapsin volume, posterior oblique, posterior, and anterior oblique views, left to right, respectively ( B ) Individual 3D rendering of the ventral arcuate body lobe (ABv, dark green) and dorsal arcuate body lobe (ABd, light green), with magenta envelope representing space which would be occupied by the missing lobe in each image.", "stats": [], "panels": [ "fig11a", "fig11b" ] }, { "uid": "results-268", "section": "results", "text": "( E ) \u03b1-\u03b2Tubulin3 (magenta) and \u03b1-Synapsin (green) immunoreactivity in the arcuate body (ventral to dorsal as top to bottom, respectively), with arrows marking where pronounced fiber tracts pass through the arcuate body layers.", "stats": [], "panels": [ "fig11e" ] }, { "uid": "results-269", "section": "results", "text": "( F ) Dorsal view of arcuate body showing layering in ABv and ABd (brace), for proctolin (\u03b1-Proctolin, yellow) and octopaminergic/tyraminergic (\u03b1-TDC2, magenta) immunoreactivity which have overlapping but distinct innervation patterns in the anterior ABd.", "stats": [], "panels": [ "fig11f" ] }, { "uid": "results-270", "section": "results", "text": "( G ) Arrowheads marking patterning of individual units of \u03b1-AstA immunoreactivity in the arcuate body, suggestive of a columnar structure.", "stats": [], "panels": [ "fig11g" ] }, { "uid": "results-296", "section": "results", "text": "Neurons of the PCL send their projections anteriorly through the ventral arcuate layers, as revealed by immunostaining for \u03b2Tubulin3 in conjunction with synapsin ( Figure 11E ).", "stats": [], "panels": [ "fig11e" ] }, { "uid": "results-300", "section": "results", "text": "An example is the dorsal arcuate body (ABd), where TDC2 immunoreactivity shows a prominent columnar, flagstone innervation, while proctolin has a sparse field of fine puncta in the same layer ( Figure 11F ).", "stats": [], "panels": [ "fig11f" ] }, { "uid": "results-302", "section": "results", "text": "For example, a series of discernible units of allatostatin A+ immunoreactivity are seen in the posABv layer, suggesting the columnar organization which is present, but generally obscured by the density of staining ( Figure 11G ).", "stats": [], "panels": [ "fig11g" ] }, { "uid": "discussion-005", "section": "discussion", "text": "Given the utility of specific stains for understanding of neuropil structures ( Figure 13 , Figure 13\u2014video 1 ), tracts, and other features, this atlas provides a rich source for comparative anatomy in an orb-weaving spider, U. diversus , while also extending knowledge of a number of different neurosignaling pathways for spiders at large.", "stats": [], "panels": [ "fig13" ] }, { "uid": "discussion-006", "section": "discussion", "text": "Figure 13. Summary of U . diversus neuropils.", "stats": [], "panels": [ "fig13" ] }, { "uid": "discussion-007", "section": "discussion", "text": "( A ) Posterior oblique, dorsal, and anterior oblique views of 3D renderings of the standard U. diversus brain with annotations of major established and novel neuropils throughout the subesophageal and supraesophageal masses.", "stats": [], "panels": [ "fig13a" ] }, { "uid": "discussion-008", "section": "discussion", "text": "( B ) 3D renderings of the standard U. diversus brain with annotations of potential central complex constituents in shades of green (protocerebral bridge, arcuate body ventral and dorsal lobes, tonsillar neuropil), also showing the mushroom body (purple).", "stats": [], "panels": [ "fig13b" ] }, { "uid": "discussion-009", "section": "discussion", "text": "( C ) 3D neuropil renderings from neuropils of the central complex as found in the insects Rhyparabia maderae , Scarabaeus lamarcki , and Manduca sexta (images from https://insectbraindb.org/app/ ).", "stats": [], "panels": [ "fig13c" ] }, { "uid": "discussion-011", "section": "discussion", "text": "Figure 13\u2014video 1. 3D rendering of all identified brain regions.", "stats": [], "panels": [ "fig13" ] }, { "uid": "discussion-081", "section": "discussion", "text": "Based on gross morphology, it is tempting to speculate that these novel neuropils, when considered along with each individual lobe of the arcuate body, may form an equivalent to a central complex in U. diversus ( Figure 13 ).", "stats": [], "panels": [ "fig13" ] }, { "uid": "captions-010", "section": "captions", "text": "[panels detected: a, b, c, d] === Figure 3 === Figure 3. Opisthosomal neuropil.", "stats": [], "panels": [ "fig3" ] }, { "uid": "captions-019", "section": "captions", "text": "[panels detected: a, b, c, d] === Figure 4 === Figure 4. Pedipalp, Blumenthal, and cheliceral neuropil.", "stats": [], "panels": [ "fig4" ] }, { "uid": "captions-029", "section": "captions", "text": "[panels detected: a, b, c, d, e] === Figure 5 === Figure 5. Ventral supraesophageal features.", "stats": [], "panels": [ "fig5" ] }, { "uid": "captions-040", "section": "captions", "text": "[panels detected: a, b, c] === Figure 6 === Figure 6. Hagstone neuropil and mid-supraesophageal features.", "stats": [], "panels": [ "fig6" ] }, { "uid": "captions-062", "section": "captions", "text": "[panels detected: a, b, c, d, e, f, g, h, i] === Figure 8 === Figure 8. Visual pathways.", "stats": [], "panels": [ "fig8" ] }, { "uid": "captions-067", "section": "captions", "text": "[panels detected: a, b] === Figure 9 === Figure 9. Tonsillar neuropil.", "stats": [], "panels": [ "fig9" ] }, { "uid": "captions-073", "section": "captions", "text": "[panels detected: a, b, c] === Figure 10 === Figure 10. Protocerebral bridge (PCB) neuropil.", "stats": [], "panels": [ "fig10" ] }, { "uid": "captions-105", "section": "captions", "text": "Compass abbreviations: A = anterior, P = posterior, D = dorsal, V = ventral, L = left, R = right. === Figure 13 === Figure 13. Summary of U . diversus neuropils.", "stats": [], "panels": [ "fig13" ] } ] } }