Gene 1242
Expression Summary

RT-PCR Summary:
RNA transcripts are detectable in: brain, cortex, subcortical region, cerebellum, brainstem, olfactory bulb, spinal cord, eye, Harderian gland, heart, lung, pancreas, kidney, spleen, thymus, lymph nodes, skin, gallbladder, urinary bladder, pituitary gland, adrenal gland, salivary gland, skeletal muscle, tongue, stomach, small intestine, large intestine, cecum, testis, epididymis, seminal vesicle, coagulating gland, prostate gland, ovary, uterus and white fat.

No RNA transcripts are detectable in liver and bone marrow.

LacZ Summary:
LacZ (beta-galactosidase) expression is detectable in brain, spinal cord, sciatic nerve, eyes, Harderian glands, thymus, spleen, lymph nodes, aorta, heart, lung, liver, gallbladder, pancreas, kidney, urinary bladder, trachea, larynx, esophagus, thyroid gland, adrenal glands, salivary glands, tongue, skeletal muscle, skin, male and female reproductive systems. In all tissues analyzed lacZ expression is detectable in smooth muscle cells of blood vessels.

Expression:
Brain
In wholemount staining strong lacZ expression is detectable in cortex, olfactory bulb, choroid plexus, hypothalamus, cerebellum and blood vessels. On coronal sections strong lacZ expression is detectable in cortex, caudate putamen, hippocampus, ventricles, amygdala and blood vessels.
In thalamus and meninges distinct nuclei express lacZ moderately to strongly. In cerebellum, lacZ expression is detectable in Purkinje cells. In brainstem, X-Gal signals are scattered. 

Spinal cord
LacZ expression is detectable in dorsal horns, gray matter and blood vessels.

Sciatic Nerve
Distinct nuclei express lacZ strongly.

Eyes
LacZ expression is detectable in the inner nuclear and ganglion cell layer of the retina and in blood vessels.

Harderian Glands
LacZ expression is detectable in blood vessels.

Thymus
LacZ expression is detectable in blood vessels.

Spleen
LacZ expression is detectable in trabeculae and capsule.

Lymph Nodes
LacZ expression is detectable in blood vessels.

Aorta
Strong lacZ expression is detectable in the aortic wall.

Heart
LacZ expression is detectable in blood vessels.

Lung
LacZ expression is detectable in smooth muscle cells of bronchioli and blood vessels.

Liver
LacZ expression is detectable in blood vessels.

Gallbladder
Strong lacZ expression is detectable in smooth muscle cells of the wall.

Pancreas
LacZ expression is detectable in blood vessels.

Kidney
Strong lacZ expression is detectable in blood vessels.

Urinary Bladder
LacZ expression is detectable in smooth muscle cells of muscularis and blood vessels.

Trachea
LacZ expression is detectable in the trachealis muscle and blood vessels.

Larynx
LacZ expression is detectable in blood vessels.

Esophagus
LacZ expression is detectable in lamina propria and blood vessels.

Thyroid Gland
LacZ expression is detectable in blood vessels.

Adrenal Glands
LacZ expression is detectable in medulla and blood vessels.

Salivary Glands
LacZ expression is detectable in smooth muscle cells of ducts and blood vessels.

Tongue
LacZ expression is detectable in lamina propria, nerves and blood vessels.

Skeletal Muscle
LacZ expression is detectable in blood vessels.

Skin
LacZ expression is detectable in dermis and blood vessels.

Skin of the Ear
LacZ expression is detectable in nerves and blood vessels.

Male Reproductive Systems
Testis
LacZ expression is detectable in blood vessels.

Penis
LacZ expression is detectable in skin, nerves and blood vessels.

Seminal Vesicles
Very strong lacZ expression is detectable in myocytes of the capsule.

Coagulating Gland
Very strong lacZ expression is detectable in myocytes of the capsule and in blood vessels.

Prostate and Ampullary Gland
Myocytes of the capsule express lacZ moderately to strongly. LacZ expression is detectable in blood vessels.

Female Reproductive Systems
Ovary
LacZ expression is detectable in smooth muscle cells of follicles and blood vessels.

Oviduct/Uterus
LacZ expression is detectable in smooth muscle cells of Fallopian tubules, myometrium and blood vessels.

Vagina/Cervix
LacZ expression is detectable in the smooth muscle cell layer and blood vessels.

No Expression: 
LacZ expression is not detected in bone marrow and pituitary gland.

Gene 1242
Necropsy

There were no significant differences detected in the homozygous mutant mice when compared with age- and gender-matched wild-type control mice.

The following mice were necropsied. Body weight, body length, and organ weights were obtained, and gross pathological findings were recorded.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (151624, 151651, 151661)
3 homozygous mutant males (151634, 151635, 151638)
2 wild-type control females (151620, 151621)
2 wild-type control males (158858, 158859)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (181263, 181264)
2 homozygous mutant males (198175, 229166)
2 wild-type control females (181265, 181278)
2 wild-type control males (190674, 205615)

Mice were examined for the following observables: adrenal glands, body length, body weight, bone marrow, bone - cranium, bone - femur, bone - sternum, bone - stifle joint, bone - vertebral column, brain, cecum, colon, duodenum, epididymis - seminal vesicle, esophagus, eyes, feces, gallbladder, general appearance, Harderian glands, heart, heart weight, ileum, jejunum, kidney weight, kidneys, liver, liver weight, lungs, lymph nodes, mesentery, ovaries, pancreas, penis, salivary glands, sciatic nerve, scrotum, skeletal muscle, skin, skinned mouse, spleen, spleen weight, stomach, testes, testes - epididymis weight, thymus, thymus weight, tongue, trachea, urinary bladder, urine, uterus, and vagina. (Gender-specific observables apply to appropriate gender.)

Necropsy Findings:
There were no genotype-related or biologically significant differences noted between mutant and wild-type control mice for any of the parameters evaluated at necropsy. Incidental lesions may have been present in some tissues. One 300 day cohort homozygous mutant female (181263) was reported to have a pancreatic nodule that had/ did not have a histopathological correlate. These findings were considered to represent background lesions occasionally seen in this strain of mice, lesions due to spontaneous disease, age-related lesions, lesions due to procedural artifacts, and/or lesions of a nonspecific etiology. They were not considered to be genotype related.

Body and Organ Weight Findings:
One 300 day cohort homozygous mutant male (198175) had a low body weight and high multiple organ weight or organ weight to body weight ratios. We have not presented these findings as phenotypic changes but have presented them here for your consideration. Other differences in body length, body weight, organ weights, and/or organ weight to body weight ratios were present between individual mice. The variability between mice usually fell within our historical reference ranges and was not correlated with genotype.

Gene 1242
Histopathology

There were no significant differences detected in the homozygous mutant mice when compared with age- and gender-matched wild-type control mice.

Tissues from the following mice were evaluated histologically.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (151624, 151651, 151661)
3 homozygous mutant males (151634, 151635, 151638)
2 wild-type control females (151620, 151621)
2 wild-type control males (158858, 158859)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (181263, 181264)
2 homozygous mutant males (198175, 229166)
1 wild-type control female (181278)
1 wild-type control male (190674)

No Significant Abnormalities:
Tissues examined and considered to have no genotypically significant abnormality: brain, pituitary gland, ears, nasal cavity, salivary glands, oral cavity, lymph nodes, aorta, lungs, gallbladder, pancreas, spleen, kidneys, adrenal glands, urinary bladder, stomach, small and large intestines, larynx, esophagus, trachea, thyroid gland, thymus gland, tongue, skeletal muscle, sciatic nerve, mammary glands, vertebrae, spinal cord, bone (skull, sternum, femur, tibia, and stifle joint), reproductive tract (including gonads), eyes, Harderian glands, integumentary system (skin and either clitoral or preputial glands), and bone marrow.

Bone marrow was examined in sections of sternum, vertebrae, and/or femur and tibia. Marrow cellularity, myeloid:erythroid (M:E) ratio, myeloid and erythroid maturation sequences, and numbers of megakaryocytes were evaluated.

Incidental lesions may have been present in some tissues. These findings are considered to represent background lesions occasionally seen in this strain of mice, lesions due to spontaneous disease, age-related lesions, lesions due to procedural artifacts, and/or lesions of a nonspecific etiology. They are not considered to be genotype related.

Gene 1242
Hematology


There were no significant differences detected in the homozygous mutant mice when compared with age- and gender-matched wild-type control mice.

Blood samples from the following mice were evaluated by a complete blood count and differential cell count.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (151624, 151651, 158836)
3 homozygous mutant males (151638, 159742, 159762)
2 wild-type control females (151621, 151622)
2 wild-type control males (158858, 159743)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (181263, 181264, 181279, 181281)
4 homozygous mutant males (198175, 213653, 213654, 255859)
4 wild-type control females (181265, 181278, 190668, 190671)
4 wild-type control males (190676, 200473, 250448, 250452)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (181263, 181264, 181279, 181281)
2 homozygous mutant males (198175, 229166)
4 wild-type control females (181265, 181278, 190668, 190671)
4 wild-type control males (190674, 200473, 205605, 250452)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (181263, 181264)
2 homozygous mutant males (198175, 229166)
2 wild-type control females (181265, 181278)
2 wild-type control males (190674, 205615)

Although minor variations of hematological values were present in some mice, these changes were not consistent with genotype and, thus, were not considered phenotypically relevant.

Gene 1242
Clinical Chemistry


There were no significant differences detected in the homozygous mutant mice when compared with age- and gender-matched wild-type control mice.

Serum samples from the following mice were evaluated by a clinical biochemistry panel.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (151651, 151661, 158836)
3 homozygous mutant males (151638, 159742, 159762)
2 wild-type control females (151621, 151660)
2 wild-type control males (158859, 158860)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (181263, 181264, 181279, 181281)
4 homozygous mutant males (213653, 213654, 229166, 255860)
4 wild-type control females (181265, 181278, 190668, 195320)
5 wild-type control males (190674, 190676, 200473, 205605, 250448)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (181263, 181264, 181279, 181281)
2 homozygous mutant males (198175, 229166)
4 wild-type control females (181265, 181278, 190668, 190671)
4 wild-type control males (190674, 205605, 205615, 250448)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (181263, 181264)
2 homozygous mutant males (198175, 229166)
2 wild-type control females (181265, 181278)
2 wild-type control males (190674, 205615)

Values for the various analytes evaluated were generally similar between homozygous mutant and wild-type control mice. Variations in clinical chemistry values, if present, were not consistent with genotype and, thus, were not considered phenotypically relevant.

Gene 1242
Densitometry

There were no significant differences detected in the homozygous mutant mice when compared with age- and gender-matched wild-type control mice.

The following mice were evaluated by dual-energy x-ray absorptiometry.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (151624, 151651, 151661)
3 homozygous mutant males (151634, 151635, 151638)
2 wild-type control females (151620, 151621)
2 wild-type control males (158858, 158859)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (181263, 181264)
2 homozygous mutant males (198175, 229166)
2 wild-type control females (181265, 181278)
2 wild-type control males (190674, 205615)

Bone Mineral Density (BMD in g/cm² ), fat % (fat percentage expressed as a percentage of body soft tissue compartment), and R-value of soft tissue were calculated from Bone Mineral Content (BMC in g), bone and tissue areas (cm² ), and total tissue mass (g) generated by a PIXImus densitometer.

Densitometric Findings:

Incidental densitometric differences were present between some mice. For example, a mutant homozygous male mouse (151634) had slightly decreased BMC, bone area, tissue area, and total tissue mass.  Additionally, a 49 day cohort homozygous mutant male (151635) had slightly decreased tissue area and fat %, and a homozygous mutant female (151624) had slightly decreased bone area, tissue area, and total tissue mass. A 300 day cohort homozygous mutant male (198175) had decreased fat % and total tissue mass. These findings were considered to represent background differences occasionally seen in this strain of mice, differences due to spontaneous disease, age-related differences, and/or differences of a nonspecific etiology. They were not considered to be genotype related.

Gene 1242
Physical Examination

There were no significant differences detected in the homozygous mutant mice when compared with age- and gender-matched wild-type control mice.

The following mice were evaluated by physical examination.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (151624, 151651, 151661)
3 homozygous mutant males (151634, 151635, 151638)
2 wild-type control females (151620, 151621)
2 wild-type control males (158858, 158859)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (181263, 181264)
2 homozygous mutant males (198175, 229166)
2 wild-type control females (181265, 181278)
2 wild-type control males (190674, 205615)

Mice were examined for the following observables: anus, behavior, body shape, claws, coat - fur, coat color - back, coat color - belly, ear - left, ear - right, eye - left, eye - right, eye color - left, eye color - right, feces, forelimb - left, forelimb - right, forelimb number of amputated digits - left, forelimb number of amputated digits - right, forelimb number of digits - left, forelimb number of digits - right, general appearance, genitals - female, genitals - male, hair type, head shape, hindlimb - left, hindlimb - right, hindlimb number of amputated digits - left, hindlimb number of amputated digits - right, hindlimb number of digits - left, hindlimb number of digits - right, injuries, lesions, limb shape, locomotor, lumps - masses, mammary glands, mice in cage, respiration, skin appearance, snout, swelling - joints, tail, teeth color, teeth length, urine, and whiskers. (Gender-specific observables apply to appropriate gender.)

Individual homozygous mutant mice had only occasional minor differences in observed physical features compared to wild-type control mice. For example, two 49 day cohort homozygous mutant male mice (151634, 151635) had a rough fur coat. These findings were considered to represent individual variability, background features occasionally seen in this strain of mice, findings due to spontaneous disease, age-related findings, procedural artifacts, and/or findings of a nonspecific etiology. However, none of these differences was regarded as biologically significant or genotype related.

There were no other genotype-related or biologically significant differences noted between mutant and wild-type control mice for any of the parameters evaluated at physical examination.

Gene 1242
Aging Metrics

There were no significant differences detected in the homozygous mutant mice when compared with age- and gender-matched wild-type control mice.

Body weights and body lengths were measured for mice at 49, 90, 180, and 300 days of age.

49 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (181263, 181264, 181279, 181281)
4 homozygous mutant males (198175, 213653, 213654, 229166)
4 wild-type control females (178471, 178472, 181265, 181278)
6 wild-type control males (190674, 190676, 200473, 205617, 250448, 250452)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (181263, 181264, 181279, 181281)
6 homozygous mutant males (198175, 213653, 213654, 229166, 255859, 255860)
6 wild-type control females (178471, 178472, 181265, 181278, 190668, 190671)
6 wild-type control males (190674, 190676, 200473, 205617, 250448, 250452)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (181263, 181264, 181279, 181281)
4 homozygous mutant males (198175, 229166, 255859, 255860)
4 wild-type control females (181265, 181278, 190668, 190671)
6 wild-type control males (190674, 200473, 205605, 205615, 250448, 250452)

300 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (181263, 181264, 181279, 188429)
1 homozygous mutant male (198175)
4 wild-type control females (181265, 181278, 190668, 190671)
3 wild-type control males (200473, 205605, 205615)

Body Weight and Length Findings:

One 300 day cohort homozygous  mutant male (198175) had a low body weight and a low body weight/ body length ratio. We have not reported these findings as phenotypic changes but have presented them here for your consideration.

Other differences in body length and body weight were present between individual mice. The variability between mice usually fell within our historical reference ranges and was not correlated with genotype.

Gene 1242
Behavior

Changes related to genotype:

• There were no significant differences detected in the homozygous mutant animals when compared with age- and gender-matched wild-type control mice.

Homozygous mutant and wild-type control mice were evaluated for phenotypic changes by testing on seven behavioral tasks: Open field test, Tail suspension test, Rotarod test, Hot plate test, Startle/PPI, Tail flick test, and Metrazol test.

Mouse ID numbers are as follows:

11 homozygous mutant males (188559, 188560, 195315, 195316, 195297, 195299, 198194, 208303, 221864, 221890, 221891)
12 wild-type control males (188573, 188575, 195312, 195300, 198193, 208299, 221853, 221854, 221873, 221874, 221875, 221897)

ES cells derived from the 129/OlaHsd mouse substrain were used to generate chimeric mice.  F1 mice were generated by breeding with C57BL/6 females. The resultant F1N0 heterozygotes were backcrossed to C57BL/6 mice to generate F1N1 heterozygotes.  F2N1 homozygous mutant mice were produced by intercrossing F1N1 heterozygous males and females.

Behavior Findings:
There were no genotype-related or biologically significant differences noted between homozygous mutant and wild-type control mice for any of the parameters evaluated during behavior testing.

Gene 1242
Fertility

Both homozygous mutant males and females were fertile.  Their progeny were viable until weaning.

Three homozygous mutant mice of each gender were set up in a fertility mating one on one with each other at seven to ten weeks of age.  The number of pups born from three litters was recorded.  One mating pair(188537, 188548) had no pups born. Three weeks later, the live pups were counted and weaned.

Mouse ID numbers are as follows:

3 homozygous mutant males (188553, 188554, 188537)

3 homozygous mutant females (188585, 188568, 188548)