Journal: Science Advances

Article Title: Opposing insulin signals underlie the same developmental switch across hemipteran insects

doi: 10.1126/sciadv.aea4413

Figure Lengend Snippet: ( A ) Identification of ecdysteroids as a target of IIS by comparative transcriptomics. Fifth-instar Wt SW (short-winged–destined) and PaInR2 De4 (long-winged–destined) P. apterus nymphs, and fifth-instar Wt SW (short-winged–destined) and NlInR2 E4 (long-winged–destined) N. lugens nymphs were collected for RNA sequencing (RNA-seq). Three KEGG pathways are commonly regulated in the context of inactivation of IIS in the firebug (via PaInR2 De4 ) and activation of IIS in the planthopper (via NlInR2 E4 ). The ecdysteroid biosynthesis pathway is outlined on the right. ( B ) Heatmap of gene regulation by PaInR2 De4 . Three genes key to ecdysteroid biosynthesis are down-regulated in the context of IIS inactivation (via PaInR2 De4 ). ( C ) Heatmap of gene regulation by NlInR2 E4 . Three genes key to ecdysteroid biosynthesis are down-regulated in the context of IIS activation (via NlInR2 E4 ). ( D ) PaInR2 De4 firebugs decrease titers of the ecdysteroid makisterone A. Fifth-instar nymphs at 1, 3, 5, 7, and 9 days (d) after eclosion were collected for makisterone A determination. The duration of fifth-instar PaInR2 De4 and Wt SW nymphs is indicated at the bottom. ( E ) NlInR2 E4 planthoppers decrease titers of the ecdysteroid 20E. Fifth-instar nymphs at 2, 24, 48, and 72 hours (h) after eclosion were collected for 20E determination. The duration of fifth-instar PaInR2 De4 and Wt SW nymphs is indicated at the bottom. Bars in (D) and (E) represent means ± SEM derived from three independent biological replicates. * P < 0.05, ** P < 0.01, and *** P < 0.001 (two-tailed unpaired t test).

Article Snippet: Standard 20E and makisterone A were purchased from MedChemExpress (HY-N6979) and Macklin (M912023), respectively.

Techniques: Transcriptomics, RNA Sequencing, Activation Assay, Derivative Assay, Two Tailed Test

Journal: Science Advances

Article Title: Opposing insulin signals underlie the same developmental switch across hemipteran insects

doi: 10.1126/sciadv.aea4413

Figure Lengend Snippet: ( A ) Wing morphs of PaInR2 De4 after makisterone A microinjection. ** P < 0.01 and *** P < 0.001 (Pearson’s χ 2 test: χ 2 = 7.501 and df = 1 for 0.1 μg/μl and χ 2 = 63.188 and df = 1 for 0.5 μg/μl). ( B ) Phenotypes of PaInR2 De4 microinjected with DMSO (long-winged morphs). ( C ) Phenotypes of PaInR2 De4 microinjected with makisterone A (short-winged morphs). ( D ) Wing morphs of NlInR2 E4 after 20E microinjection. *** P < 0.001 (Pearson’s χ 2 test: χ 2 = 40.675 and df = 1 for 0.2 μg/μl, χ 2 = 32.125 and df = 1 for 1 μg/μl, and χ 2 = 32.178 and df = 1 for 2.5 μg/μl). ( E ) Phenotypes of NlInR2 E4 microinjected with DMSO (long-winged morphs). ( F ) Phenotypes of NlInR2 E4 microinjected with 20E (short-winged and intermediate size–winged morphs). Forewings and hindwings are indicated by arrows and arrowheads, respectively.

Article Snippet: Standard 20E and makisterone A were purchased from MedChemExpress (HY-N6979) and Macklin (M912023), respectively.

Techniques: Microinjection

Journal: Science Advances

Article Title: Opposing insulin signals underlie the same developmental switch across hemipteran insects

doi: 10.1126/sciadv.aea4413

Figure Lengend Snippet: The IIS pathway acts as both a positive and a negative regulator to control ecdysteroid titers in P. apterus and N. lugens via the InRs-Chico-PDK1-Tsc1/2-TORC1 and InRs-Chico-PDK1-Akt-FoxO cascades, respectively. In P. apterus , inactivation of the IIS pathway decreases ecdysteroid (makisterone A) biosynthesis via targeting Torso , Phantom , and Disembodied genes. In N. lugens , activation of the IIS pathway decreases ecdysteroid (20E) biosynthesis via targeting Neverland , Spook , and Cyp314A1 genes. Low ecdysteroid titers may guide the bipotential wing buds to develop into long wings. In contrast, high levels of ecdysteroid titers may guide wing buds to develop into short wings. Down and up arrows in colors indicate inactivation and activation, respectively.

Article Snippet: Standard 20E and makisterone A were purchased from MedChemExpress (HY-N6979) and Macklin (M912023), respectively.

Techniques: Control, Activation Assay

Journal: Science Advances

Article Title: Opposing insulin signals underlie the same developmental switch across hemipteran insects

doi: 10.1126/sciadv.aea4413

Figure Lengend Snippet: ( A ) Identification of ecdysteroids as a target of IIS by comparative transcriptomics. Fifth-instar Wt SW (short-winged–destined) and PaInR2 De4 (long-winged–destined) P. apterus nymphs, and fifth-instar Wt SW (short-winged–destined) and NlInR2 E4 (long-winged–destined) N. lugens nymphs were collected for RNA sequencing (RNA-seq). Three KEGG pathways are commonly regulated in the context of inactivation of IIS in the firebug (via PaInR2 De4 ) and activation of IIS in the planthopper (via NlInR2 E4 ). The ecdysteroid biosynthesis pathway is outlined on the right. ( B ) Heatmap of gene regulation by PaInR2 De4 . Three genes key to ecdysteroid biosynthesis are down-regulated in the context of IIS inactivation (via PaInR2 De4 ). ( C ) Heatmap of gene regulation by NlInR2 E4 . Three genes key to ecdysteroid biosynthesis are down-regulated in the context of IIS activation (via NlInR2 E4 ). ( D ) PaInR2 De4 firebugs decrease titers of the ecdysteroid makisterone A. Fifth-instar nymphs at 1, 3, 5, 7, and 9 days (d) after eclosion were collected for makisterone A determination. The duration of fifth-instar PaInR2 De4 and Wt SW nymphs is indicated at the bottom. ( E ) NlInR2 E4 planthoppers decrease titers of the ecdysteroid 20E. Fifth-instar nymphs at 2, 24, 48, and 72 hours (h) after eclosion were collected for 20E determination. The duration of fifth-instar PaInR2 De4 and Wt SW nymphs is indicated at the bottom. Bars in (D) and (E) represent means ± SEM derived from three independent biological replicates. * P < 0.05, ** P < 0.01, and *** P < 0.001 (two-tailed unpaired t test).

Article Snippet: Standard 20E and makisterone A were purchased from MedChemExpress (HY-N6979) and Macklin (M912023), respectively.

Techniques: Transcriptomics, RNA Sequencing, Activation Assay, Derivative Assay, Two Tailed Test

Journal: Science Advances

Article Title: Opposing insulin signals underlie the same developmental switch across hemipteran insects

doi: 10.1126/sciadv.aea4413

Figure Lengend Snippet: ( A ) Wing morphs of PaInR2 De4 after makisterone A microinjection. ** P < 0.01 and *** P < 0.001 (Pearson’s χ 2 test: χ 2 = 7.501 and df = 1 for 0.1 μg/μl and χ 2 = 63.188 and df = 1 for 0.5 μg/μl). ( B ) Phenotypes of PaInR2 De4 microinjected with DMSO (long-winged morphs). ( C ) Phenotypes of PaInR2 De4 microinjected with makisterone A (short-winged morphs). ( D ) Wing morphs of NlInR2 E4 after 20E microinjection. *** P < 0.001 (Pearson’s χ 2 test: χ 2 = 40.675 and df = 1 for 0.2 μg/μl, χ 2 = 32.125 and df = 1 for 1 μg/μl, and χ 2 = 32.178 and df = 1 for 2.5 μg/μl). ( E ) Phenotypes of NlInR2 E4 microinjected with DMSO (long-winged morphs). ( F ) Phenotypes of NlInR2 E4 microinjected with 20E (short-winged and intermediate size–winged morphs). Forewings and hindwings are indicated by arrows and arrowheads, respectively.

Article Snippet: Standard 20E and makisterone A were purchased from MedChemExpress (HY-N6979) and Macklin (M912023), respectively.

Techniques: Microinjection

Journal: Science Advances

Article Title: Opposing insulin signals underlie the same developmental switch across hemipteran insects

doi: 10.1126/sciadv.aea4413

Figure Lengend Snippet: The IIS pathway acts as both a positive and a negative regulator to control ecdysteroid titers in P. apterus and N. lugens via the InRs-Chico-PDK1-Tsc1/2-TORC1 and InRs-Chico-PDK1-Akt-FoxO cascades, respectively. In P. apterus , inactivation of the IIS pathway decreases ecdysteroid (makisterone A) biosynthesis via targeting Torso , Phantom , and Disembodied genes. In N. lugens , activation of the IIS pathway decreases ecdysteroid (20E) biosynthesis via targeting Neverland , Spook , and Cyp314A1 genes. Low ecdysteroid titers may guide the bipotential wing buds to develop into long wings. In contrast, high levels of ecdysteroid titers may guide wing buds to develop into short wings. Down and up arrows in colors indicate inactivation and activation, respectively.

Article Snippet: Standard 20E and makisterone A were purchased from MedChemExpress (HY-N6979) and Macklin (M912023), respectively.

Techniques: Control, Activation Assay