Gene: 108Name: Arts1-pendingFamily: ProteaseSubfamily: AminopeptidaseAccession: AB047552GI: 9886746

Gene 108
Summary of Phenotypic Analysis

There were no 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. 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.

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.
Densitometry.
Behavior Analysis.
Fertility tests.
Aging studies.

Gene 108
Behavior

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 for the N1 generation:
11 homozygous mutant males (173879, 173892, 173893, 173895, 173897, 179835, 182052, 206469, 206470, 209777, 209781)
10 wild-type control males (173880, 173896, 179831, 179834, 182048, 182049, 206468, 209775, 209782, 209786)

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 108
Fertility

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

Two 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:

2 homozygous mutant males (180886, 193956)

2 homozygous mutant females (179854,193960)

 

 

Gene 108
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, kidney, 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, spleen, liver, kidney, trachea, larynx, thyroid gland, pituitary gland, adrenal glands, skin, male and female reproductive systems.

Expression:
Brain
In wholemount staining weak lacZ expression is detectable in ventricles. On coronal sections no X-Gal signals are detectable.

Spleen
Weak lacZ expression is detectable in blood vessels and few cells.

Liver
Many hepatocytes express lacZ at various intensities from faint to strong.

Kidney
Scattered lacZ expression is detectable in cortex and medulla. In blood vessels, endothelial cells and smooth muscle cells express lacZ weakly.

Trachea
Few cells in the mucosal epithelium express lacZ weakly.

Larynx
Few cells in the laryngeal epithelium and in submucosal glands express lacZ weakly.

Thyroid Gland
Faint X-Gal signals are detectable in blood vessels.

Pituitary Gland
Weak lacZ expression is detectable in pars distalis.

Adrenal Glands
Weak lacZ expression is detectable in the medulla.

Skin of the Ear
Faint X-Gal signals are present in cartilage.

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

Female Reproductive Systems
Oviduct/Uterus
LacZ expression is detectable in Fallopian tubules.

No Expression: 
LacZ expression is not detected in: brain, spinal cord, sciatic nerve, eyes, Harderian glands, thymus, lymph nodes, bone marrow, aorta, heart, lung, gallblader, pancreas, urinary bladder, esophagus, parathyroid gland, salivary glands, tongue and skeletal muscle.

 

Gene 108
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 were as follows:
3 homozygous mutant females (158433, 158437, 180887)
3 homozygous mutant males (161530, 161534, 161535)
2 wild-type control females (158438, 158449)
2 wild-type control males (161536, 168349)

300 Day Cohort Mouse ID numbers were as follows:
2 homozygous mutant females (179842, 191759)
2 homozygous mutant males (217112, 217114)
2 wild-type control females (179840, 191758)
2 wild-type control males (217113, 281699)

Bone Mineral Density (BMD in g/cm
2 ), 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:

When compared with age- and gender-matched wild-type control mice, two 49 day homozygous mutant females (158437, 158433) had increased total tissue mass, and one of these (158437) had increased fat %.  Additionally, two 49 day homozygous mutant males (161535, 161530) had increased fat %.   However, in the 300 day cohort one female (179842) and one male (217112) homozygous mutants had slightly decreased fat %. We have not reported these findings as phenotypic changes but we have presented them here for your consideration.

Incidental densitometric differences also 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 108
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 were as follows:
3 homozygous mutant females (158433, 158437, 180887)
3 homozygous mutant males (161530, 161534, 161535)
2 wild-type control females (158438, 158449)
2 wild-type control males (161536, 168349)

300 Day Cohort Mouse ID numbers were as follows:
2 homozygous mutant females (179842, 191759)
2 homozygous mutant males (217112, 217114)

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.

Increased extramedullary hematopoiesis of the spleen in one female (179842) and one male (217114) 300 day homozygous mutants correlated with enlarged spleen size, increased spleen weight, and increased spleen weight/body weight ratio. We have not reported these findings as phenotypic changes but we have presented them here for your consideration.

Incidental lesions may have been present in some tissues. For example, there was atypical hyperplasia of the thymus gland in one female (191759) and one male (217112) 300 day homozygous mutants. 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 108
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 (158433, 158437, 180887)
3 homozygous mutant males (161530, 161534, 161535)
2 wild-type control females (158438, 158449)
2 wild-type control males (161536, 168349)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (179842, 191759)
2 homozygous mutant males (217112, 217114)
2 wild-type control females (179840, 191758)

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:
Kidney size was enlarged in two 300 day homozygous mutant males. The spleen was reported to be enlarged in two female and one male (217114) 300 day homozygous mutant mice. These findings correlated with increased extramedullary hematopoiesis of the spleen in one female (179842) and one male (217114) 300 day homozygous mutants. We have not reported these findings as phenotypic changes but we have presented them here for your consideration.

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 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:
Kidney weight was increased in two 300 day homozygous mutant males. Spleen weight and spleen weight/body weight ratio were increased in one female (179842) and one male (217114) 300 day homozygous mutant mice. These findings correlated with increased extramedullary hematopoiesis of the spleen in one female (179842) and one male (217114) 300 day homozygous mutants. We have not reported these findings as phenotypic changes but we 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 108
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. The data are compiled from the N1F2 and N2F2 generations.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (158433, 180887, 192011)
3 homozygous mutant males (161530, 161534, 161535)
2 wild-type control females (158438, 158449)
2 wild-type control males (161536, 168349)

90 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (173890, 173899, 191759)
4 homozygous mutant males (217112, 217114, 281700, 323899)
4 wild-type control females (173889, 173902, 179840, 191758)
3 wild-type control males (217113, 323897, 323898)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (173890, 173899, 191759, 193939)
4 homozygous mutant males (217112, 217114, 281700, 323899)
4 wild-type control females (173891, 173902, 191758, 193941)
4 wild-type control males (217113, 281699, 323897, 323898)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (179842, 191759)
2 homozygous mutant males (217112, 217114)
2 wild-type control females (179840, 191758)
2 wild-type control males (217113, 281699)

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 108
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.The data are compiled from the N1F2 and N2F2 generations.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (180887, 192011, 192029)
3 homozygous mutant males (161530, 161534, 161535)
2 wild-type control females (158438, 158449)
2 wild-type control males (161536, 184317)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (173890, 173899, 179842, 191759)
4 homozygous mutant males (217112, 217114, 281700, 323899)
4 wild-type control females (173889, 173902, 179840, 191758)
4 wild-type control males (217113, 281699, 323897, 323898)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (173890, 173899, 179842, 191759)
4 homozygous mutant males (217112, 217114, 281700, 323899)
4 wild-type control females (173889, 173902, 179840, 191758)
4 wild-type control males (217113, 281699, 323897, 323898)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (179842, 191759)
2 homozygous mutant males (217112, 217114)
2 wild-type control females (179840, 191758)
2 wild-type control males (217113, 281699)

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 108
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 were as follows:
3 homozygous mutant females (158433, 158437, 180887)
3 homozygous mutant males (161530, 161534, 161535)
2 wild-type control females (158438, 158449)
2 wild-type control males (161536, 168349)

300 Day Cohort Mouse ID numbers were as follows:
2 homozygous mutant females (179842, 191759)
2 homozygous mutant males (217112, 217114)
2 wild-type control females (179840, 191758)
2 wild-type control males (217113, 281699)

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 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 was regarded as biologically significant or genotype related.

Gene 108
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 (173890, 173899, 179842, 191759)
3 homozygous mutant males (217112, 217114, 281700)
4 wild-type control females (173889, 173902, 179840, 191758)
2 wild-type control males (217113, 281699)

90 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (173890, 173899, 179842)
4 homozygous mutant males (217112, 217114, 281700, 323899)
3 wild-type control females (173889, 173902, 179840)
4 wild-type control males (217113, 281699, 323897, 323898)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (173890, 173899, 179842, 191759)
1 homozygous mutant male (323899)
4 wild-type control females (173889, 173902, 179840, 191758)
2 wild-type control males (323897, 323898)

300 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (173890, 173899, 179842)
2 homozygous mutant males (281700, 323899)
3 wild-type control females (173891, 173902, 179840)
4 wild-type control males (217113, 281699, 323897, 323898)


Body Weight and Length Findings:

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.