arl 67156 trisodium salt  (Tocris)

Journal: Experimental & Molecular Medicine

Article Title: Adenosine transmission from hypothalamic tanycytes to AGRP/NPY neurons regulates energy homeostasis

doi: 10.1038/s12276-025-01449-6

Figure Lengend Snippet: a A schematic diagram of ATP release from tanycytes and conversion of nucleotides by ectonucleotidases. ATP is released through Cx43 from tanycytes and is converted to ADP and AMP by ENTPD. AMP is converted to adenosine by NT5E. Adenosine is transported by ENT1 and is converted to inosine by ADA. b Relative expression levels of ectonucleotidases families and nucleoside transporters in A2/29 cells ( n = 6). c Relative intracellular and extracellular adenosine levels in si-scram and si- Tspo A2/29 cells ( n = 9 per group). d , f Relative intracellular and extracellular ATP levels in si-scram and si- Tspo A2/29 cells in the presence or absence of ENTPD inhibitor ( ARL67156 , 0.1 mM) ( d ) or NT5E inhibitor (APCP, 0.1 mM) ( f ) for 3 h ( n = 9 per group). e, g – i Relative intracellular and extracellular adenosine levels in si-scram and si- Tspo A2/29 cells in the presence or absence of ARL67156 ( e ) APCP ( g ) ADA inhibitor (EHNA, 0.01 mM) ( h ) or ENT1 inhibitor (NBMPR, 0.01 mM) ( i ) for 3 h ( n = 9 per group). Data are mean ± s.e.m. or boxes indicating the interquartile range with whiskers, and dotted lines indicate that significance was calculated separately for each. Significance was determined by one-way ANOVA with Dunnett’s multiple-comparisons test (** P < 0.01, **** P < 0.0001) in b two-tailed unpaired Student’s t -test (*** P < 0.001) in c or otherwise by two-way ANOVA with Šidák’s multiple-comparisons test (** P < 0.01, *** P < 0.001, **** P < 0.0001). n.s., not significant.

Article Snippet: Tan Tspo WT and Tan Tspo cKO mice were implanted with a unilateral cannula (PlasticsOne) at the 3V (1.3 mm posterior, 0.0 mm medial, and 5.3 mm ventral to bregma) to inject ARL67156 (0.1 nmol; Tocris, 1283) or vehicle or with a bilateral cannula (PlasticsOne) at the ARC (1.3 mm posterior, ±0.5 mm lateral and 5.6 mm ventral to bregma) to inject DPCPX (0.4 nmol; Tocris, 0439) or vehicle.

Techniques: Expressing, Two Tailed Test

Journal: Experimental & Molecular Medicine

Article Title: Adenosine transmission from hypothalamic tanycytes to AGRP/NPY neurons regulates energy homeostasis

doi: 10.1038/s12276-025-01449-6

Figure Lengend Snippet: a , d , g , j Schematic illustrations of ARL67156 ( a ) APCP ( d ) EHNA ( g ) and NBMPR ( j ) treatments in the indirect coculture system. b , e , h , k Relative extracellular ATP or adenosine levels in si-scram or si- Tspo A2/29 cells indirectly cocultured with N41 cells in the presence or absence of ARL67156 ( b ) APCP ( e ) EHNA ( h ) or NBMPR ( k ) under 0.06 mM oleic acid (OL) treatment for 3 h ( n = 9 per group). c , f , i , l Relative mRNA levels of Agrp and Npy in N41 cells cocultured with si-scram or si- Tspo A2/29 cells in the presence or absence of ARL67156 ( c ), APCP ( f ) EHNA ( i ) or NBMPR ( l ) under OL treatment for 3 h ( n = 9 per group). Data are mean ± s.e.m. or boxes indicating the interquartile range with whiskers, and dotted linesindicate that significance was calculated separately for each. Significance was determined by two-way ANOVA with Šidák’s multiple-comparisons test (* P < 0.5, ** P < 0.01, *** P < 0.001, **** P < 0.0001; n.s., not significant).

Article Snippet: Tan Tspo WT and Tan Tspo cKO mice were implanted with a unilateral cannula (PlasticsOne) at the 3V (1.3 mm posterior, 0.0 mm medial, and 5.3 mm ventral to bregma) to inject ARL67156 (0.1 nmol; Tocris, 1283) or vehicle or with a bilateral cannula (PlasticsOne) at the ARC (1.3 mm posterior, ±0.5 mm lateral and 5.6 mm ventral to bregma) to inject DPCPX (0.4 nmol; Tocris, 0439) or vehicle.

Techniques:

Journal: Experimental & Molecular Medicine

Article Title: Adenosine transmission from hypothalamic tanycytes to AGRP/NPY neurons regulates energy homeostasis

doi: 10.1038/s12276-025-01449-6

Figure Lengend Snippet: a The schematic strategy and schedule of ARL67156 or Veh ICV injection into the 3V through an implanted cannula in Tan Tspo WT or Tan Tspo cKO mice. b , c Food intake ( b ) and body weight change ( c ) of Tan Tspo WT and Tan Tspo cKO mice for 24 h after ARL67156 (0.1 nmol) or Veh ICV injection and HFD feeding ( n = 6 per group). d Relative Agrp and Npy expression levels in Tan Tspo WT and Tan Tspo cKO mice 4 h after ARL67156 or Veh ICV injection and HFD feeding ( n = 6 per group). e Relative expression levels of adenosine receptors in the mouse hypothalamus ( n = 6). f Representative image of A1R (red) expressed on NPY neurons (green) in the hypothalamus and colocalization coefficients of A1R with NPY neurons ( n = 10). g Representative image of A2bR (red) expressed on NPY neurons (green) in the hypothalamus and colocalization coefficients of A2bR with NPY neurons ( n = 10). h The schematic strategy and schedule of A1R inhibitor (DPCPX) or Veh ICV injection into the ARC through an implanted cannula in Tan Tspo WT or Tan Tspo cKO mice. i , j Food intake ( i ) and body weight change ( j ) of Tan Tspo WT and Tan Tspo cKO mice for 24 h after DPCPX (0.4 nmol) or Veh ICV injection and HFD feeding ( n = 6 per group). k Relative Agrp and Npy expression levels in Tan Tspo WT and Tan Tspo cKO mice 4 h after DPCPX or Veh ICV injection and HFD feeding ( n = 6 per group). Scale bars, 20 μm ( f and g ). Data are mean ± s.e.m. or boxes indicating the interquartile range with whiskers, and dotted lines indicate that significance was calculated separately for each. Significance was determined by one-way ANOVA with Dunnett’s multiple-comparisons test (**** P < 0.0001; n.d., not detected) in e or otherwise by two-way ANOVA with Šidák’s multiple-comparisons test (* P < 0.5, ** P < 0.01, *** P < 0.001, **** P < 0.0001; n.s., not significant).

Article Snippet: Tan Tspo WT and Tan Tspo cKO mice were implanted with a unilateral cannula (PlasticsOne) at the 3V (1.3 mm posterior, 0.0 mm medial, and 5.3 mm ventral to bregma) to inject ARL67156 (0.1 nmol; Tocris, 1283) or vehicle or with a bilateral cannula (PlasticsOne) at the ARC (1.3 mm posterior, ±0.5 mm lateral and 5.6 mm ventral to bregma) to inject DPCPX (0.4 nmol; Tocris, 0439) or vehicle.

Techniques: Injection, Expressing