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Figure 5—figure supplement 4. Influence of pH on PBP-lpo binding affinity. ; Representative binding curves of PBP – Lpo interaction assays at pH 6.9 and 8.2 (indicated to the left). The concentration of Lpo protein injected is plotted against the response its specific binding elicited at equilibrium (Req). Non-linear regression assuming one-site saturation was used to calculate the dissociation constant, KD. The KD is the mean ± SD of three replicates.

2019

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Figure 2—figure supplement 1. Representative images of neurons innervating VTA-projecting LHb neurons. ; (A) Schematics of procedure for viral tracer experiments. (B) Representative pictures of VTA-projecting TVA expressing cells (red label) and first-order rabies infected cells (green label) in the LHb. (C–F) Representative pictures of second-order rabies infected cells in the VP, LPO, rEPN, LH, and VTA after Cav2-Cre injection into VTA.

2019

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Figure 1. LHb-projecting neurons in the rEPN and the LH are significantly activated by repeated footshocks. ; (A) Representative photos of CTb injection site in LHb (brown immunoreactive product). Representative photos of CTb immunostaining (brown) and shock-induced cFos (black) in the rEPN (B, C) and the LH (D–F). Black squares: areas for cFos counting. Red squares: areas enlarged in representative images. f: fornix; opt: optic tract; MHb: medial habenula. (G) Proportion of CTb-labeled neurons expressing cFos in the VTA, rEPN, dlLH, vlLH, LPO, and VP, showing the largest shock-induced increases in the rEPN and LH. (E) Proportion of cFos neurons expressing CTb in these same regions. Scalebars 250 µm (A), 500 µm (B, D), 100 µm (C, E and F).

2019

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Figure 2. rEPN neurons innervate LHb neurons projecting to the RMTg. ; (A, B) Schematics of procedure in which retrogradely transported virus expressing Cre-recombinase was injected into the RMTg or VTA and viruses expressing flox-stopped TVA and RG were injected into the LHb. EnvA-ΔG-rabies virus was injected into the LHb 3 weeks later. Animals were perfused at the end of the forth week. (C) Neurons in the rEPN and the VTA preferentially projected to RMTg-projecting LHb neurons over VTA-projecting LHb neurons. (D) Representative pictures of RMTg-projecting TVA expressing cells and first-order rabies infected cells in the LHb. (E) Representative pictures of second-order rabies infected cells in the VP, LPO, rEPN, LH, and VTA after Cav2-Cre injection into RMTg, indicating neurons disynaptically connected to RMTg.

2019

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Figure 1. VTAGad67+ neurons project to multiple areas in the brain. ; (a) Schematic of AAV injection to express Cre-inducible hrGFP in Gad67-Cre mice. The dotted brain map area is shown to the right. White rectangular area is shown below. (b) Immunohistochemical studies showing expression of hrGFP in Gad67+ neurons (arrowhead), but not in the nearby DA (arrow) neurons. (c) Pie chart showing the percent of hrGFP expression in DA and non-DA neurons in the VTA (n = 4 mice). (d and e) Expression of hrGFP in VTAGad67+ neurons and some of their projected brain areas are shown. (f) Table showing the comparative scoring of hrGFP signals across different brain areas. Abbreviations- AcbC: Nucleus accumbens core, AcbSh: Nucleus accumbens shell, BLA: Basolateral amygdala, CeA: Central nucleus of the amygdala, CM: Central medial thalamic nucleus, CnF: Cuneiform nucleus, CPu: Caudate putamen (striatum), DEn: Dorsal endopiriform nucleus, DR: Dorsal raphe nucleus, IMD: Intermediodorsal thalamic nucleus, InG: Intermediate gray layer of the superior colliculus, LH: Lateral hypothalamus, LPO: Lateral preoptic area, MM: Medial mammillary nucleus, Pa4: Paratrochlear nucleus, PAG: Periaqueductal gray, PF: Parafascicular thalamic nucleus, PnO: Pontine reticular nucleus, SubB: Subbrachial nucleus, VLL: Ventral nucleus of the lateral lemniscus, VP: Ventral pallidum, Xi: Xiphoid thalamic nucleus and xscp: Decussation of the superior cerebellar peduncles. DAPI (4′,6-diamidino-2-phenylindole) staining was used to label nuclear DNA and also to assist understanding of the anatomical position of VTAGad67+ neuronal projections.

2019

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Figure 2—figure supplement 2. Growth rate determination of lpo mutants across an expanded set of pH conditions. ; Mean mass doublings per hour and transformed percent parental growth for ∆lpoA (EAM657) and ∆lpoB (EAM659) mutants compared to the parental strain (MG1655) from pH 4.8–8.4. Significance was determined by an unpaired t-test corrected for multiple comparisons using the Holm-Sidak method. Error bars represent SD from six independent biological replicates. Asterisks denote significance as follows: *, p

2019

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Supplementary file 1. Phylogenetic trees of the markers investigated. ; (A) Simplified Maximum Likelihood (ML) tree for Myosin Regulatory Light Chain (full tree in panel M), rooted with Calmodulin, which shares an EF-hand calcium-binding domain with MRLC. (B) ML tree for FoxF, rooted with FoxQ1, the probable closest relative of the FoxF family (Shimeld et al., 2010). (C) MrBayes tree for bilaterian ZASP/LBD3, rooted with the cnidarian ortholog (Steinmetz et al., 2012). (D) ML tree for bilaterian Myosin Heavy Chain, rooted at the (pre-bilaterian) duplication between smooth and striated MHC (Steinmetz et al., 2012). (E) MrBayes tree for Mef2, rooted by the first splice isoform of the cnidarian ortholog (Genikhovich and Technau, 2011). (F) MrBayes tree for Titin, rooted at the protostome/deuterostome bifurcation (Titin is a bilaterian novelty). (G) MrBayes tree for Troponin T, rooted at the protostome/deuterostome bifurcation (Troponin T is a bilaterian novelty). (H) MrBayes tree for Troponin I, rooted by the Calponin/Transgelin family, which shares an EF-hand calcium-binding domain with Troponin I. (I) MrBayes tree for MyoD, rooted at the protostome/deuterostome bifurcation (MyoD is a bilaterian novelty). (J) MrBayes tree for Myocardin, rooted at the protostome/deuterostome bifurcation (the Drosophila myocardin ortholog is established [Han et al., 2004]). (K) Complete MRLC tree. Species names abbreviations: Pdu: Platynereis dumerilii; Xenla: Xenopus laevis; Mus: Mus musculus; Hsa: Homo sapiens; Dre: Danio rerio; Gga: Gallus gallus; Dme: Drosophila melanogaster; Cte: Capitella teleta; Patvu: Patella vulgata; Brafl: Branchiostoma floridae; Nve or Nemv: Nematostella vectensis; Acdi: Acropora digitifera; Expal: Exaiptasia pallida; Rat: Rattus norvegicus; Sko: Saccoglossus kowalevskii; Limu or Lpo: Limulus polyphemus; Trib or Trca: Tribolium castaneum; Daph: Daphnia pulex; Prcau: Priapulus caudatus; Cgi or Cgig: Crassostrea gigas; Ling or Linan: Lingula anatina; Hdiv: Haliotis diversicolor; Apcal or Aca: Aplysia californica; Spu: Strongylocentrotus purpuratus; Poli or Polis: Polistes dominula; Cin or Cint: Ciona intestinalis; Hro: Helobdella robusta; Bos: Bos taurus; Capsa: Capsaspora owczarzaki; Thtr: Thecamonas trahens; Lpo:; Bga: Biomphalaria glabrata; Cel: Caenorhabditis elegans; Tt: Terebratalia transversa; Octo: Octopus vulgaris; Sma: Schmidtea mediterranea; Bbe: Branchiostoma belcheri, Batden: Batrachochytrium dendrobaditis; Monve: Mortierella verticillata; Alloma: Allomyces macrogynus; Salpun: Spizellomyces punctatus; Mucor: Mucor racemosus; Lichco: Lichtheimia corymbifera; Ephmu: Ephydatia muelleri; Sycon: Sycon ciliatum; Amqu: Amphimedon queenslandica; Osc: Oscarella lobularis; Metse: Metridium senile; Pfu: Pinctada fucata; Rypa: Riftia pachyptila; Plma: Placopecten magellanicus; Air: Argopecten irradians; Scolop: Scolopendra gigantea; Artfra: Artemia franciscana; Bmor: Bombyx mori; Loa: Loa loa; Necator-am: Necator americanus; Trichi: Trichinella spiralis; Asc: Ascaris lumbricoides; Wuch: Wucheria bancrofti; Ancy: Ancylostoma duodenale; Callorinc: Callorhinchus milii; Dana: Danaus plexippus; Anop: Anopheles gambiae; Asty: Astyanax mexicanus; Oreo: Oreochromis niloticus; Icta: Ictalurus punctatus.

2016

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Detailansicht für Treffer 8 öffnen

Figure 1. Experimental and analysis procedures for cell-type-specific circuit tracing. ; (A) RV-mediated transsynaptic retrograde tracing of BF inputs. Upper panel, viral vectors and injection procedure. Lower panel, fluorescence images of BF in the region of the NDB (red box in coronal diagram) in ChAT-, VGLUT2-, PV-, and SOM-Cre mice. Scale bar, 200 µm. Inset, enlarged view of the region in white box showing starter cells (yellow, expressing both eGFP and tdTomato, indicated by white arrowheads). Scale bar, 50 µm. NDB, diagonal band nucleus; SIB, substantia innominata, basal part; MCPO, magnocellular preoptic nucleus; VP, ventral pallidum; LPO, lateral preoptic area. (B) Viral vector and injection procedure for tracing BF axonal projections. (C) Flow chart showing the main steps in data generation and processing.

2016

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