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Gene: 1715	Name: Prss12	Family: Protease	Subfamily: Serine Protease	Accession: Y13192	GI: 2113885

Gene 1715
Summary of Phenotypic Analysis

Changes related to genotype:

Behavior: Homozygous mutant mice exhibited decreased total distance traveled as well as decreased time spent in the central region during open field testing.

There were no other significant differences detected in the homozygous mutant mice 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 differential cell counts
Clinical chemistry panels
Densitometry
Behavioral tests

Gene 1715
Behavior

Homozygous mutant mice exhibited significantly decreased total distance traveled during open field testing when compared with age- and gender-matched wild-type control mice.
Homozygous mutant mice exhibited significantly decreased time spent in the central region during open field testing 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, Startle response/PPI test, Tail flick test, Hot plate test, and Metrazol test.

Mouse ID numbers are as follows for the N1 generation:
10 homozygous mutant males (383690, 383692, 404363, 405889, 408927, 415711, 415712, 422093, 422094, 422096)
33 wild-type control males (382404, 382406, 382408, 382494, 401077, 402563, 404359, 405612, 405888, 405895, 406082, 407069, 408506, 408925, 409023, 411653, 412719, 412721, 413474, 414237, 414238, 414648, 414650, 415709, 417825, 417828, 418343, 418450, 418831, 418969, 419029, 419144, 421447)

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:
When compared to age- and gender-matched wild-type control mice, homozygous mutant mice exhibited significantly decreased total distance traveled during open field testing, indicating a possible hypoactivity phenotype.

When compared to age- and gender-matched wild-type control mice, homozygous mutant mice exhibited a significant decrease in time spent in the central region during open field testing, indicating a possible increased anxiety-like phenotype.

There were no other genotype-related differences noted between homozygous mutant and wild-type control mice for any other parameters evaluated during behavior testing.

Gene 1715
Expression Summary

Taqman Summary:
The highest levels of RNA transcripts are detectable in: lung, pancreas, gallbladder, adrenal gland, skeletal muscle, epididymis, seminal vesicle, prostate gland and uterus.

Lower levels of RNA transcripts are also detectable in: whole brain, cortex, subcortical region, brainstem, olfactory bulb, spinal cord, eye, Harderian glands, kidney, spleen, lymph nodes, skin, urinary bladder, pituitary gland, salivary gland, tongue, stomach, small intestine, testis, coagulating gland, ovary and white fat.

No RNA transcripts are detectable in: cerebellum, heart, liver, thymus, bone marrow, large intestine and cecum.

LacZ Summary:
LacZ expression was detected in several organs. Cerebral cortex showed striking staining in the wholemount brain and moderate to strong expression on coronal sections. Sections of brain also showed weak to moderate staining in amygdala and hippocampus, and in a few cells of the brainstem and gray matter of the spinal cord. Also of note, several nerves showed strong staining. In the kidney, strong expression was detected in some cells of the renal medulla.

LacZ expression was detected in: brain, spinal cord, sciatic nerve, lymph nodes, kidney, urinary bladder, skeletal muscle, skin, testis, ovary and cervix.

LacZ expression was not detected in: eyes, thymus, spleen, bone marrow, aorta, heart, lung, liver, pancreas, thyroid gland, parathyroid gland, larynx, pituitary gland, adrenal glands, prostate gland and uterus.

Gene 1715
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 (358303, 358304, 358305)
3 homozygous mutant males (370190, 370191, 370193)
2 wild-type control females (358301, 392285)
2 wild-type control males (358295, 358297)

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. 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 1715
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.

The following tissues were examined and considered to have no genotype-related abnormality: brain, pituitary gland, ears, nasal cavity, salivary glands, oral cavity, lymph nodes, aorta, lungs, liver, gallbladder, pancreas, spleen, kidneys, adrenal glands, urinary bladder, stomach, small and large intestines, larynx, esophagus, trachea, thyroid gland, thymus, 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 were 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, and/or lesions of a nonspecific etiology. They were not considered to be genotype related.

Gene 1715
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 (358303, 358304, 358305)
3 homozygous mutant males (370190, 370191, 370193)
2 wild-type control females (358301, 392285)
2 wild-type control males (358295, 358297)

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, 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 the 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 were 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, and/or lesions of a nonspecific etiology. They were not considered to be related to genotype.

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 1715
Clinical Chemistry

There were no significant differences 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 chemistry panel. The data are compiled from the F2N0 and F2N1 generations.

49 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (358303, 358304, 358305, 358306)
3 homozygous mutant males (370190, 370191, 370193)
2 wild-type control females (358301, 392262)
3 wild-type control males (358295, 358297, 358298)

Values for the various analytes evaluated were generally similar between homozygous mutant and wild-type control mice. Although variations in clinical chemistry values were present in some mice, they were not related to genotype and, thus, were not considered phenotypically relevant.

Gene 1715
Hematology

There were no significant differences 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. The data are compiled from the F2N0 and F2N1 generations.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (358303, 358304, 358305)
3 homozygous mutant males (370190, 370193, 383679)
2 wild-type control females (358301, 392285)
2 wild-type control males (358295, 358297)

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

Gene 1715
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 (358303, 358304, 358305)
3 homozygous mutant males (370190, 370191, 370193)
2 wild-type control females (358301, 392285)
2 wild-type control males (358295, 358297)

Mice were examined in detail as follows: 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 the appropriate gender.)

Individual homozygous mutant mice had only occasional minor differences in observed physical features compared to wild-type control mice. 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, and/or findings of a nonspecific etiology. However, none of these differences were regarded as biologically significant or genotype related.