Limits...
Novel Genes Critical for Hypoxic Preconditioning in Zebrafish Are Regulators of Insulin and Glucose Metabolism.

Manchenkov T, Pasillas MP, Haddad GG, Imam FB - G3 (Bethesda) (2015)

Bottom Line: Severe hypoxia is a common cause of major brain, heart, and kidney injury in adults, children, and newborns.However, mild hypoxia can be protective against later, more severe hypoxia exposure via "hypoxic preconditioning," a phenomenon that is not yet fully understood.Using a functional genomic approach, we used this zebrafish model to identify and validate five novel hypoxia-protective genes, including irs2, crtc3, and camk2g2, which have been previously implicated in metabolic regulation.

View Article: PubMed Central - PubMed

Affiliation: Division of Neonatology, University of California San Diego School of Medicine, La Jolla, California 92093.

No MeSH data available.


Related in: MedlinePlus

Multiple acute hypoxia response genes are required for the protective effect of hypoxic preconditioning. Hypoxia survival graphs with fractional lethality at 24 hr are shown on the y-axis. (A) h-MO knockdown of candidate hypoxia-protective genes is shown in comparison to ctl-MO in untreated controls (injection alone), prolonged sH(1.5:38), and hPC + prolonged sH embryos. Impaired hPC is demonstrated at P < 0.0001 in the following morphants: camk2g2 (P = 6.7 × 10−4); btr01 (P = 7.3 × 10−4); ncam2 (P = 1.7 × 10−6); crtc3 (P = 6.2 × 10−7); and irs2 (P= 2.6 × 10−7); but not in ttll11. (B) h-MO knockdown inhibits hPC and is rescued by mRNA coinjection (btr01, P = 0.0038, 0.015; crct3, P = 1.0 × 10−6, 7.2 × 10−4; irs2, P = 0.033, 0.049). (C) Hypoxia response gene overexpression via mRNA injection is sufficient to confer partial protection from prolonged hypoxia [btr01, P = 0.027; irs2, P = 0.050; sH(1.5d:38h) protocol]; n = 2 to 8 biological replicates with SEM for (A–C).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4478541&req=5

fig5: Multiple acute hypoxia response genes are required for the protective effect of hypoxic preconditioning. Hypoxia survival graphs with fractional lethality at 24 hr are shown on the y-axis. (A) h-MO knockdown of candidate hypoxia-protective genes is shown in comparison to ctl-MO in untreated controls (injection alone), prolonged sH(1.5:38), and hPC + prolonged sH embryos. Impaired hPC is demonstrated at P < 0.0001 in the following morphants: camk2g2 (P = 6.7 × 10−4); btr01 (P = 7.3 × 10−4); ncam2 (P = 1.7 × 10−6); crtc3 (P = 6.2 × 10−7); and irs2 (P= 2.6 × 10−7); but not in ttll11. (B) h-MO knockdown inhibits hPC and is rescued by mRNA coinjection (btr01, P = 0.0038, 0.015; crct3, P = 1.0 × 10−6, 7.2 × 10−4; irs2, P = 0.033, 0.049). (C) Hypoxia response gene overexpression via mRNA injection is sufficient to confer partial protection from prolonged hypoxia [btr01, P = 0.027; irs2, P = 0.050; sH(1.5d:38h) protocol]; n = 2 to 8 biological replicates with SEM for (A–C).

Mentions: Hypoxia-activated genes identified by microarrays and validated by qPCR and/or in situ hybridization were then tested using the optimized preconditioning protocol sH(1d:5h + 1.5d:38h), depicted in Figure 1. Of the six morphants without developmental defects tested, five showed significant impairment of hPC in comparison to control MO–injected embryos: irs2, crct3, btr01, camk2g2, and ncam2 (Figure 5A). For btr01, crct3, and irs2, we additionally demonstrated that the loss of protective hPC in morphants could be restored by full-length mRNA rescue (Figure 5B).


Novel Genes Critical for Hypoxic Preconditioning in Zebrafish Are Regulators of Insulin and Glucose Metabolism.

Manchenkov T, Pasillas MP, Haddad GG, Imam FB - G3 (Bethesda) (2015)

Multiple acute hypoxia response genes are required for the protective effect of hypoxic preconditioning. Hypoxia survival graphs with fractional lethality at 24 hr are shown on the y-axis. (A) h-MO knockdown of candidate hypoxia-protective genes is shown in comparison to ctl-MO in untreated controls (injection alone), prolonged sH(1.5:38), and hPC + prolonged sH embryos. Impaired hPC is demonstrated at P < 0.0001 in the following morphants: camk2g2 (P = 6.7 × 10−4); btr01 (P = 7.3 × 10−4); ncam2 (P = 1.7 × 10−6); crtc3 (P = 6.2 × 10−7); and irs2 (P= 2.6 × 10−7); but not in ttll11. (B) h-MO knockdown inhibits hPC and is rescued by mRNA coinjection (btr01, P = 0.0038, 0.015; crct3, P = 1.0 × 10−6, 7.2 × 10−4; irs2, P = 0.033, 0.049). (C) Hypoxia response gene overexpression via mRNA injection is sufficient to confer partial protection from prolonged hypoxia [btr01, P = 0.027; irs2, P = 0.050; sH(1.5d:38h) protocol]; n = 2 to 8 biological replicates with SEM for (A–C).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4478541&req=5

fig5: Multiple acute hypoxia response genes are required for the protective effect of hypoxic preconditioning. Hypoxia survival graphs with fractional lethality at 24 hr are shown on the y-axis. (A) h-MO knockdown of candidate hypoxia-protective genes is shown in comparison to ctl-MO in untreated controls (injection alone), prolonged sH(1.5:38), and hPC + prolonged sH embryos. Impaired hPC is demonstrated at P < 0.0001 in the following morphants: camk2g2 (P = 6.7 × 10−4); btr01 (P = 7.3 × 10−4); ncam2 (P = 1.7 × 10−6); crtc3 (P = 6.2 × 10−7); and irs2 (P= 2.6 × 10−7); but not in ttll11. (B) h-MO knockdown inhibits hPC and is rescued by mRNA coinjection (btr01, P = 0.0038, 0.015; crct3, P = 1.0 × 10−6, 7.2 × 10−4; irs2, P = 0.033, 0.049). (C) Hypoxia response gene overexpression via mRNA injection is sufficient to confer partial protection from prolonged hypoxia [btr01, P = 0.027; irs2, P = 0.050; sH(1.5d:38h) protocol]; n = 2 to 8 biological replicates with SEM for (A–C).
Mentions: Hypoxia-activated genes identified by microarrays and validated by qPCR and/or in situ hybridization were then tested using the optimized preconditioning protocol sH(1d:5h + 1.5d:38h), depicted in Figure 1. Of the six morphants without developmental defects tested, five showed significant impairment of hPC in comparison to control MO–injected embryos: irs2, crct3, btr01, camk2g2, and ncam2 (Figure 5A). For btr01, crct3, and irs2, we additionally demonstrated that the loss of protective hPC in morphants could be restored by full-length mRNA rescue (Figure 5B).

Bottom Line: Severe hypoxia is a common cause of major brain, heart, and kidney injury in adults, children, and newborns.However, mild hypoxia can be protective against later, more severe hypoxia exposure via "hypoxic preconditioning," a phenomenon that is not yet fully understood.Using a functional genomic approach, we used this zebrafish model to identify and validate five novel hypoxia-protective genes, including irs2, crtc3, and camk2g2, which have been previously implicated in metabolic regulation.

View Article: PubMed Central - PubMed

Affiliation: Division of Neonatology, University of California San Diego School of Medicine, La Jolla, California 92093.

No MeSH data available.


Related in: MedlinePlus