title: The Ins2Akita mouse as a model of early retinal complications in diabetes.
abstract: OBJECTIVE
  This study tested the Ins2(Akita) mouse as an animal model of retinal complications in diabetes. The Ins2(Akita) mutation results in a single amino acid substitution in the insulin 2 gene that causes misfolding of the insulin protein. The mutation arose and is maintained on the C57BL/6J background. Male mice heterozygous for this mutation have progressive loss of beta-cell function, decreased pancreatic beta-cell density, and significant hyperglycemia, as early as 4 weeks of age.
  
  
  METHODS
  Heterozygous Ins2(Akita) mice were bred to C57BL/6J mice, and male offspring were monitored for hyperglycemia, beginning at 4.5 weeks of age. After 4 to 36 weeks of hyperglycemia, the retinas were analyzed for vascular permeability, vascular lesions, leukostasis, morphologic changes of micro- and macroglia, apoptosis, retinal degeneration, and insulin receptor kinase activity.
  
  
  RESULTS
  The mean blood glucose of Ins2(Akita) mice was significantly elevated, whereas the body weight at death was reduced compared with that of control animals. Compared with sibling control mice, the Ins2(Akita) mice had increased retinal vascular permeability after 12 weeks of hyperglycemia (P < 0.005), a modest increase in acellular capillaries after 36 weeks of hyperglycemia (P < 0.0008), and alterations in the morphology of astrocytes and microglia, but no changes in expression of Muller cell glial fibrillary acidic protein. Increased apoptosis was identified by immunoreactivity for active caspase-3 after 4 weeks of hyperglycemia (P < 0.01). After 22 weeks of hyperglycemia, there was a 16.7% central and 27% peripheral reduction in the thickness of the inner plexiform layer, a 15.6% peripheral reduction in the thickness of the inner nuclear layer (P < 0.001), and a 23.4% reduction in the number of cell bodies in the retinal ganglion cell layer (P < 0.005). In vitro insulin receptor kinase activity was reduced (P < 0.05) after 12 weeks of hyperglycemia.
  
  
  CONCLUSIONS
  The retinas of heterozygous male Ins2(Akita) mice exhibit vascular, neural, and glial abnormalities generally consistent with clinical observations and other animal models of diabetes. In light of the relatively early, spontaneous onset of the disease and the popularity of the C57BL/6J inbred strain as a background for the generation and study of other genetic alterations, combining the Ins2(Akita) mutation with other engineered mutations will be of great use for studying the molecular basis of retinal complications of diabetes.
authors.affiliation: Penn State Retina Research Group, The Ulerich Ophthalmology Research Laboratory, Penn State College of Medicine, Hershey, 17033, USA. abarber@psu.edu
authors.forename: Alistair J
authors.initials: AJ
authors.lastname: Barber
authors.forename: David A
authors.initials: DA
authors.lastname: Antonetti
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Kern
authors.forename: Chad E N
authors.initials: CE
authors.lastname: Reiter
authors.forename: Rohit S
authors.initials: RS
authors.lastname: Soans
authors.forename: J Kyle
authors.initials: JK
authors.lastname: Krady
authors.forename: Steven W
authors.initials: SW
authors.lastname: Levison
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Gardner
authors.forename: Sarah K
authors.initials: SK
authors.lastname: Bronson
mesh_terms: D000818:Animals; D017209:Apoptosis; D001253:Astrocytes / Q000378:metabolism / Q000473:pathology; D001786:Blood Glucose / Q000378:metabolism; D001835:Body Weight; D002199:Capillary Permeability; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D003930:Diabetic Retinopathy / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology*; D004195:Disease Models, Animal; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006943:Hyperglycemia / Q000473:pathology; D007328:Insulin / Q000235:genetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D009457:Neuroglia / Q000378:metabolism / Q000473:pathology; D011972:Receptor, Insulin / Q000378:metabolism; D012162:Retinal Degeneration; D012171:Retinal Vessels / Q000473:pathology
chemical_list: D001786:Blood Glucose; D005904:Glial Fibrillary Acidic Protein; D007328:Insulin; D011972:Receptor, Insulin; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases