Gene 1115
Summary of Phenotypic Analysis

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

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. F2 homozygous mutant mice were produced by intercrossing F1 heterozygous males and females.

Wild-type control mice and homozygous mutant mice were evaluated by the following examinations or tests:

· Physical examination.
· Necropsy, including body length, body weight, and organ weight measurements.
· Histological examination of tissues and organs.
· Bone marrow section evaluation.
· Complete blood counts and differentials.
· Clinical chemistry panels.
· Fertility.
· Densitometry.

Gene 1115
Expression Summary

RT-PCR Summary:
RNA transcripts are detectable in all tissues analyzed: brain, cortex, subcortical region, cerebellum, brainstem, olfactory bulb, spinal cord, eye, Harderian gland, heart, lung, liver, pancreas, kidneys, spleen, thymus, lymph nodes, bone marrow, 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.

LacZ Summary:
LacZ (beta-galactosidase) expression is detectable in brain, spinal cord, heart, pancreas, trachea, larynx, esophagus, tongue, skin and male reproductive system.

Expression:
Brain
In wholemount staining weak lacZ expression is detectable in ventricles and brainstem. On coronal sections weak lacZ expression is detectable in the pyramidal cell layer of the hippocampus.

Spinal cord
Faint X-Gal signals are detectable in dorsal horns.

Heart
LacZ expression is detectable in epicardium and valves.
 
Pancreas
Few cells lining ducts show strong X-Gal staining.

Trachea
Weak lacZ expression is detectable in submucosal glands.

Larynx
LacZ expression is detectable in lamina propria.

Esophagus
Few cells in the surrounding muscle layer express lacZ strongly.

Tongue
LacZ expression is detectable in lamina propria and in the muscle layer.

Skin
LacZ expression is detectable in hair follicles and distinct cells below the epidermis.

Skin of the Ear
LacZ expression is detectable in epidermis and hair follicles.

Male Reproductive Systems
Testis
LacZ expression is detectable in interstitial and spermatogenic cells.

Penis
LacZ expression is detectable in the body, in hair follicles and os penis.

Coagulating Gland
Faint lacZ expression is detectable in secretory cells; strong expression is detectable in flattened basal cells.

Prostate Gland
LacZ expression is detectable in flattened basal cells.

No Expression: 
LacZ expression is not detected in: sciatic nerve, eyes, Harderian glands, thymus, spleen, lymph nodes, bone marrow, aorta, lung, liver, kidney, urinary bladder, thyroid gland, pituitary gland, adrenal glands, salivary glands, skeletal muscle and female reproductive systems.

Gene 1115
Necropsy

There were no significant differences detected in the homozygous mutant animals 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:
4 homozygous mutant females (154612, 154616, 169632, 186639)
3 homozygous mutant males (186641, 260627, 260632)
2 wild-type control females (154615, 186640)
2 wild-type control males (169627, 169630)

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. 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:
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 1115
Histopathology

There were no significant differences detected in the homozygous mutant animals 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:
4 homozygous mutant females (154612, 154616, 169632, 186639)
3 homozygous mutant males (186641, 260627, 260632)
2 wild-type control females (154615, 186640)
2 wild-type control males (169627, 169630)

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, 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 1115
Hematology

There were no significant differences detected in the homozygous mutant animals 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:
4 homozygous mutant females (154612, 154616, 169632, 186639)
3 homozygous mutant males (186641, 260627, 260632)
2 wild-type control females (154615, 186640)
2 wild-type control males (169627, 169630)

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

Gene 1115
Clinical Chemistry

There were no significant differences detected in the homozygous mutant animals 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:
4 homozygous mutant females (154612, 154616, 177922, 186639)
3 homozygous mutant males (186641, 260632, 285876)
3 wild-type control females (154615, 186638, 186640)
2 wild-type control males (169627, 169630)

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

Gene 1115
Densitometry

There were no significant differences detected in the homozygous mutant animals 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:
4 homozygous mutant females (154612, 154616, 169632, 186639)
3 homozygous mutant males (186641, 260627, 260632)
2 wild-type control females (154615, 186640)
2 wild-type control males (169627, 169630)

No Significant Abnormalities:
Evaluations of densitometric data included Bone Mineral Density (BMD presented as g/cm2), Bone Mineral Content (BMC in g), bone and tissue area, total tissue mass, and fat as a percent of body soft tissue (presented as fat %).  When compared with age- and gender-matched wild-type control mouse, three mutant homozygote female mice (No's.: 154616, 169632, 154612) all had slightly lower BMC, bone area, tissue area, % fat, and total tissue mass. No genotypically significant differences between mice were observed.

Gene 1115
Physical Examination

There were no significant differences detected in the homozygous mutant animals 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:
4 homozygous mutant females (154612, 154616, 169632, 186639)
3 homozygous mutant males (186641, 260627, 260632)
2 wild-type control females (154615, 186640)
2 wild-type control males (169627, 169630)

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. These findings are 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.

Gene 1115
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.  Three weeks later, the live pups were counted and weaned.

Mouse ID numbers are as follows:

3 homozygous mutant males (234828, 234829, 260658)

3 homozygous mutant females (246475, 248648, 260674)