Endothelial NOS knockout mice and methods of use

A method of screening compounds for anti-hypertensive activity, comprising:

• (a) providing a transgenic mouse comprising a disrupted eNOS gene, wherein said transgenic mouse is homozygous for said disrupted eNOS gene, and wherein said transgenic mouse exhibits hypertension;
• (b) administering a test compound to said transgenic mouse, wherein said test compound is not a NOS inhibitor;
• (c) determining the effect of said test compound on the blood pressure of said transgenic mouse;
• and
• (d) correlating a decrease in the blood pressure of said transgenic mouse with an anti-hypertensive effect of said test compound.

A method of screening compounds for wound healing activity, comprising:

• (a) providing a transgenic mouse comprising a disrupted eNOS gene, wherein said transgenic mouse is homozygous for said disrupted eNOS gene, and wherein said transgenic mouse exhibits wound healing abnormalities;
• (b) administering a test compound to said transgenic mouse, wherein said test compound is not a NOS inhibitor;
• (c) determining the effect of said test compound on the wound healing properties of said mouse;
• and
• (d) correlating the effect of said test compound on the wound healing properties of said transgenic mouse with a wound healing effect of said test compound.

A method of screening compounds for potential utility to treat cerebral ischemia or stroke, comprising:

• (a) providing a transgenic mouse comprising a disrupted eNOS gene, wherein said transgenic mouse is homozygous for said disrupted eNOS gene, and wherein said transgenic mouse develops significantly larger infarct volume following induction of focal cerebral ischemia by occlusion of middle cerebral artery, than a mouse that does not have a disrupted eNOS gene;
• (b) administering a test compound to said transgenic mouse, wherein said test compound is not a NOS inhibitor;
• (c) determining the effect of said test compound on infarct size following induction of focal cerebral ischemia in the brain of said transgenic mouse;
• and
• (d) correlating a decrease in infarct size with a potential therapeutic utility to treat cerebral ischemia or stroke.

A method of screening compounds for potential utility to treat atherosclerosis, comprising:

• (a) providing a transgenic mouse comprising a disrupted eNOS gene, wherein said transgenic mouse is homozygous for said disrupted eNOS gene, and wherein, in the cuff model of vessel injury, said transgenic mouse exhibits neointimal formation significantly greater than that in a mouse that does not have a disrupted eNOS gene, or wherein, in the filament model of vessel injury, said transgenic mouse exhibits significantly slower endothelial resurfacing or injured blood vessels than a mouse that does not have a disrupted eNOS gene;
• (b) administering a test compound to said transgenic mouse, wherein said test compound is not a NOS inhibitor;
• (c) determining the effect of said test compound on atherosclerosis in said transgenic mouse;
• and
• (d) correlating the effect of said test compound on atherosclerosis in said transgenic mouse with a potential utility to treat atherosclerosis.

A method of making a transgenic mouse having a disrupted eNOS gene, comprising:

• (a) providing a murine embryonic stem cell comprising an intact eNOS gene;
• (b) providing a targeting vector capable of disrupting said eNOS gene upon homologous recombination;
• (c) introducing said targeting vector into said murine embryonic stem cell under conditions where said targeting vector will undergo homologous recombination with the eNOS gene of said murine embryonic stem cell to produce a disrupted gene;
• (d) introducing said murine embryonic stem cell into a blastocyst;
• (e) implanting said blastocyst into a pseudopregnant female mouse;
• and
• (f) delivering a first transgenic mouse comprising a disrupted eNOS gene from said pseudopregnant female
• (g) repeating steps (a) through (f) to obtain a second transgenic mouse comprising a disrupted eNOS gene;
• and
• (h) breeding said first transgenic mouse comprising a disrupted eNOS gene to said second transgenic mouse comprising a disrupted eNOS gene to obtain one or more mice homozygous for a disrupted eNOS gene.