Gene: 1231Name: ADAMTS4Family: ProteaseSubfamily: ADAMAccession: BC027773GI: 20381316

 

Gene 1231
Summary of Phenotypic Analysis

Behavior:  When compared to age- and gender-matched wild-type control mice, homozygous mutant mice fell from the rotarod at significantly slower speeds during rotarod testing, and homozygous mutant mice required a significantly lower dose of metrazol to reach the first seizure stage during metrazol 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
  • Fertility
  • Densitometry
  • Behavior Tests
  • Aging studies

Keywords

brain; nervous system; brain; rotarod test; neuroscience

brain; nervous system; brain; coordination; neuroscience

brain; nervous system; brain; metrazol test; neuroscience

brain; nervous system; brain; seizure; neuroscience


 

Gene 1231
Expression Summary

Taqman Summary:
The highest levels of RNA transcripts are detectable in whole brain, cortex, subcortical region, cerebellum, brainstem, olfactory bulb, spinal cord, ovary and uterus.

Moderate levels of RNA transcripts are detectable in eye, Harderian glands, lung, pancreas, kidney, spleen, thymus, lymph nodes, bone marrow, skin, gallbladder, urinary bladder, pituitary gland, adrenal gland, salivary gland, skeletal muscle, tongue, stomach, small intestine, cecum, colon, testis, epididymis, seminal vesicle, coagulating gland, prostate gland, and white fat.

Lower levels of RNA transcripts are also detectable in heart.

No RNA transcripts are detectable in liver.

LacZ Summary:
LacZ (beta-galactosidase) expression is detectable in brain, spinal cord, eyes, bone marrow, tongue, skin, ear, male and female reproductive systems.


Expression:
Brain
In the wholemount brain staining, strong to very strong LacZ expression is detected in the corpus callosum, anterior commissure, fornix, lateral and medial septal nuclei, cerebellar white matter and in the brainstem.  Weaker and more scattered LacZ expression is also found in the retrospenial cortex, thalamus, superior colliculus, and granular layer of the cerebrum.
In the coronal sections taken through the brain, strong to very strong LacZ expression is localized to the following regions of the cerebrum:  amygdala, anterior commisure, caudate-putamen, corpus callousum, cortex, dorsal penduncular cortex, fimbria, habenular nuclei, hypothalamus, lateral septal nucleus, medial septal nucleus, and thalamus.  Additional strong to very strong LacZ expression is also found the following areas of the cerebellum:  granular and purkinje cell layers.  Moderate LacZ expression is also detected in the hippocampus (CA field, pyramidal cell and molecular layers) and hypothalamus while very weak expression is detected in some occasional purkinje cells of the cerebellum.

Spinal cord
Very strong LacZ expression is observed in many cells of both the gray and white matter.

Eyes
Very strong LacZ expression is detected in the optic nerve.

Bone Marrow Smear
Very strong LacZ expression is detected in a few cells of the bone marrow.

Tongue
Strong LacZ expression is found in some cells in the connective tissue layer surrounding the minor salivary gland.

Skin
Strong LacZ expression is localized in some cells of the dermis.

Ear, external
Strong LacZ expression is detected in scattered cells of the dermis.

Male Reproductive Systems
Penis
Strong LacZ expression is found in scattered cells of the dermis.

Female Reproductive Systems
Ovary
Weak to very weak LacZ expression is localized to a few cells in the corpus luteum and the stroma.

Oviduct/Uterus
Very weak LacZ expression is detected in epithelial cells of the oviduct.  Strong LacZ expression is detected in cells in the endometrial stroma while weaker expression is found in the myometrial cells of the uterus.

Vagina/Cervix
Very strong LacZ expression is found in cells of the lamina propria in both the cervix and vagina.  Additional weak expression is observed in the smooth muscle layer of the cervix.

No Expression: 
LacZ expression is not detected in: sciatic nerve, Harderian glands, thymus, spleen, lymph nodes, heart, lung, liver, gallbladder, pancreas, kidney, urinary bladder, trachea, larynx, esophagus, thyroid gland, parathyroid gland, pituitary gland, adrenal glands, salivary glands, skeletal muscle.


 

Gene 1231
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 were as follows:
3 homozygous mutant females (207412, 207415, 207566)
3 homozygous mutant males (207408, 207410, 207421)
2 wild-type control females (207411, 207567)
2 wild-type control males (207560, 207563)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (237722, 257895)
2 homozygous mutant males (237715, 257217)

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 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, 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. Thymus to body weight ratios were slightly decreased in all homozygous mutant females when compared to age- and gender-matched wild-type control mice but remained within the historical reference range.


 

Gene 1231
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 (207412, 207415, 207566)
3 homozygous mutant males (207408, 207410, 207421)
2 wild-type control females (207411, 207567)
2 wild-type control males (207560, 207563)

300 Day Cohort Mouse ID numbers were as follows:
1 homozygous mutant female (237722)
1 homozygous mutant male (257217)

No Significant Abnormalities:

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, 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 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 1231
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.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (207568, 211155, 215450)
3 homozygous mutant males (207408, 207429, 207562)
2 wild-type control females (207411, 215443)
2 wild-type control males (207563, 207564)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (237722, 247030, 257893, 257895)
4 homozygous mutant males (237715, 257217, 257220, 257221)
4 wild-type control females (237717, 237727, 237728, 244830)
4 wild-type control males (235525, 235526, 235527, 237713)

180 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (237722, 257893, 257895)
4 wild-type control females (237717, 237727, 237728, 244830)
4 wild-type control males (235525, 235526, 235527, 237713)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (237722, 257895)
2 homozygous mutant males (237715, 257217)
2 wild-type control females (237717, 237727)
2 wild-type control males (235525, 235526)

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 1231
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.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (207415, 207566, 207568)
3 homozygous mutant males (207408, 207410, 207429)
2 wild-type control females (207411, 216744)
2 wild-type control males (207560, 207564)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (237722, 247030, 257893, 257895)
4 homozygous mutant males (237715, 248351, 257217, 257221)
4 wild-type control females (237717, 237727, 237728, 244830)
4 wild-type control males (235525, 235526, 235527, 247024)

180 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (237722, 267701)
1 homozygous mutant male (237715)
4 wild-type control females (237717, 237727, 237728, 244830)
4 wild-type control males (235525, 235526, 235527, 237713)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (237722, 257895)
2 homozygous mutant males (237715, 257217)
1 wild-type control female (237727)
2 wild-type control males (235525, 235526)

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 1231
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 (207412, 207415, 207566)
3 homozygous mutant males (207408, 207410, 207421)
2 wild-type control females (207411, 207567)
2 wild-type control males (207560, 207563)

300 Day Cohort Mouse ID numbers were as follows:
2 homozygous mutant females (237722, 257895)
2 homozygous mutant males (237715, 257217)
2 wild-type control females (237717, 237727)
2 wild-type control males (235525, 235526)

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

Densitometric Findings:

Incidental densitometric differences may have been present between some mice. For example, fat % was high in one 300 day homozygous mutant female (237722). These findings were considered to represent background differences occasionally seen in this strain of mice, differences due to spontaneous disease, age-related changes, differences due to procedural artifacts, and/or differences of a nonspecific etiology. They were not considered to be genotype related.


 

Gene 1231
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 (207412, 207415, 207566)
3 homozygous mutant males (207408, 207410, 207421)
2 wild-type control females (207411, 207567)
2 wild-type control males (207560, 207563)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (237722, 257895)
2 homozygous mutant males (237715, 257217)
2 wild-type control females (237717, 237727)
2 wild-type control males (235525, 235526)

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 was regarded as biologically significant or genotype related.


 

Gene 1231
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:
3 homozygous mutant females (237722, 247030, 257893)
4 homozygous mutant males (237715, 257217, 257220, 257221)
7 wild-type control females (237717, 237727, 237728, 244830, 261026, 261027, 261028)
4 wild-type control males (235525, 235526, 235527, 237713)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (237722, 247030, 257893, 257895)
4 homozygous mutant males (237715, 257217, 257220, 257221)
4 wild-type control females (237717, 237727, 237728, 244830)
4 wild-type control males (235525, 235526, 235527, 237713)

180 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (237722)
1 homozygous mutant male (237715)
4 wild-type control females (237717, 237727, 237728, 244830)
4 wild-type control males (235525, 235526, 235527, 237713)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (257895, 267701)
2 homozygous mutant males (257220, 257221)
3 wild-type control females (237717, 237727, 237728)
4 wild-type control males (235525, 235526, 235527, 237713)

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.


 

Gene 1231
Behavior

  • Homozygous mutant mice displayed significantly decreased motor performance on the rotarod test compared to age- and gender-matched wild-type control mice.
  • Homozygous mutant mice required a significantly lower dose of metrazol to reach the first seizure stage 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:
11 homozygous mutant males (237707, 255278, 257210, 257211, 257886, 278641, 278859, 294420, 296669, 296670, 296672)
12 wild-type control males (237706, 255280, 255281, 257209, 257890, 257891, 257892, 278640, 278858, 293162, 297840, 297847)

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 fell from the rotarod at significantly slower speeds, indicating a possible decreased motor performance phenotype.

When compared to age- and gender-matched wild-type control mice, homozygous mutant mice required a significantly lower dose of metrazol to reach the first seizure stage during metrazol testing, indicating a possible higher propensity towards seizure 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 1231
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 (248351, 255269, 255274)

3 homozygous mutant females (248353, 255275, 255283)