Dynamic glucoregulation and mammalian-like responses to metabolic and developmental disruption in zebrafish.
Zebrafish embryos are emerging as models of glucose metabolism. However, patterns of endogenous glucose levels, and the role of the islet in glucoregulation, are unknown. We measured absolute glucose levels in zebrafish and mouse embryos, and demonstrate similar, dynamic glucose fluctuations in both species. Further, we show that chemical and genetic perturbations elicit mammalian-like glycemic responses in zebrafish embryos. We show that glucose is undetectable in early zebrafish and mouse embryos, but increases in parallel with pancreatic islet formation in both species. In zebrafish, increasing glucose is associated with activation of gluconeogenic phosphoenolpyruvate carboxykinase1 (pck1) transcription. Non-hepatic Pck1 protein is expressed in mouse embryos. We show using RNA in situ hybridization, that zebrafish pck1 mRNA is similarly expressed in multiple cell types prior to hepatogenesis. Further, we demonstrate that the Pck1 inhibitor 3-mercaptopicolinic acid suppresses normal glucose accumulation in early zebrafish embryos. This shows that pre- and extra-hepatic pck1 is functional, and provides glucose locally to rapidly developing tissues. To determine if the primary islet is glucoregulatory in early fish embryos, we injected pdx1-specific morpholinos into transgenic embryos expressing GFP in beta cells. Most morphant islets were hypomorphic, not a genetic, but embryos still exhibited persistent hyperglycemia. We conclude from these data that the early zebrafish islet is functional, and regulates endogenous glucose. In summary, we identify mechanisms of glucoregulation in zebrafish embryos that are conserved with embryonic and adult mammals. These observations justify use of this model in mechanistic studies of human metabolic disease.
University of Massachusetts Medical School, Diabetes Center of Excellence, 373 Plantation Street, Suite 218, Worcester, MA 01605, USA.
Agata
A
Jurczyk
Nicole
N
Roy
Rabia
R
Bajwa
Philipp
P
Gut
Kathryn
K
Lipson
Chaoxing
C
Yang
Laurence
L
Covassin
Waldemar J
WJ
Racki
Aldo A
AA
Rossini
Nancy
N
Phillips
Didier Y R
DY
Stainier
Dale L
DL
Greiner
Michael A
MA
Brehm
Rita
R
Bortell
Philip
P
diIorio
D000818:Animals; D030801:Animals, Genetically Modified; D004625:Embryo, Nonmammalian / Q000378:metabolism*; D047108:Embryonic Development / Q000187:drug effects; D005947:Glucose / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000032:analysis; D017403:In Situ Hybridization; D050417:Insulin-Secreting Cells / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D010802:Phylogeny; D010848:Picolinic Acids / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D015027:Zebrafish / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
D010848:Picolinic Acids; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; C008363:3-mercaptopicolinic acid; D010729:Phosphoenolpyruvate Carboxykinase (GTP); D005947:Glucose