Gene 315
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/Sv-+P+Mgf-SLJ/J 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, as well as heterozygous 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
·
Aging Studies
· Fertility tests
Gene 315
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 (223780, 292809, 407713)
3 homozygous mutant females (264106, 267682, 408769)
Gene 315
Expression
Summary
RT-PCR Summary:
RNA transcripts are detectable in all tissues examined: brain, cortex, subcortical
region, cerebellum, brainstem, olfactory bulb, eye, heart, lung, liver,
pancreas, kidneys, spleen, thymus, lymph nodes, bone marrow, skin, gall
bladder, 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
and uterus.
Gene 315
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 (215361, 215369, 225022)
2 homozygous mutant males (215359, 216688)
2 wild-type control females (215371, 215374)
2 wild-type control males (215377, 215380)
300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (223789, 225037)
2 homozygous mutant males (223771, 223772)
2 wild-type control females (216691, 216693)
2 wild-type control males (216689, 225016)
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 315
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:
3 homozygous mutant females (215361, 215369, 225022)
3 homozygous mutant males (215354, 215359, 216688)
2 wild-type control females (215371, 215374)
2 wild-type control males (215377, 215380)
300 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (225037)
1 homozygous mutant male (223771)
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, liver, 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 315
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.
Mouse ID numbers are as follows:
49
Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (215361, 215369, 225022)
3 homozygous mutant males (215354, 215359, 216688)
2 wild-type control females (215371, 215374)
2 wild-type control males (215377, 215380)
300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (223789, 225037)
2 homozygous mutant males (223771, 223772)
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 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
315
Clinical
Chemistry
There
were no significant differences 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 chemistry panel.
49
Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (223786, 225022, 225024)
4 homozygous mutant males (215359, 216688, 216690, 223777)
2 wild-type control females (215371, 215374)
2 wild-type control males (215357, 215377)
90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (223789, 225037, 238744, 238745)
4 homozygous mutant males (223771, 223772, 223773, 257778)
3 wild-type control females (216691, 216693, 232542)
4 wild-type control males (216689, 225016, 238739, 238740)
180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (223789, 225037, 238744, 238745)
3 homozygous mutant males (223771, 223772, 223773)
4 wild-type control females (216691, 216693, 232541, 232542)
4 wild-type control males (216689, 225016, 238739, 238740)
300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (223789, 225037)
2 homozygous mutant males (223771, 223772)
2 wild-type control females (216691, 216693)
2 wild-type control males (216689, 225016)
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 315
Hematology
There were no significant differences 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:
3 homozygous mutant females (223776, 223778, 225023)
3 homozygous mutant males (215359, 216688, 216690)
2 wild-type control females (215371, 223785)
2 wild-type control males (215357, 215377)
90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (223789, 225037, 238744, 238745)
4 homozygous mutant males (223771, 223772, 223773, 257778)
4 wild-type control females (216691, 216693, 232541, 232542)
4 wild-type control males (216689, 225016, 238739, 238740)
180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (223789, 225037, 238744, 238745)
3 homozygous mutant males (223771, 223772, 223773)
4 wild-type control females (216691, 216693, 232541, 232542)
4 wild-type control males (216689, 225016, 238739, 238740)
300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (223789, 225037)
2 homozygous mutant males (223771, 223772)
2 wild-type control females (216691, 216693)
2 wild-type control males (216689, 225016)
Although minor variations of hematological values were present in some animals,
these changes were not related to genotype and, thus, were not considered
phenotypically relevant.
Gene 315
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:
3 homozygous mutant females (215361, 215369, 225022)
3 homozygous mutant males (215354, 215359, 216688)
2 wild-type control females (215371, 215374)
2 wild-type control males (215377, 215380)
300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (223789, 225037)
2 homozygous mutant males (223771, 223772)
2 wild-type control females (216691, 216693)
2 wild-type control males (216689, 225016)
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,
feces color, feces exam, 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 exam, mice in cage, respiration, skin appearance, snout,
swelling - joints, tail, teeth color, teeth length, urine, urine color, urine
exam 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 nonspecific etiology. However, none of
these differences was regarded as biologically significant or genotype related.
Gene 315
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 (223789, 225037, 238744, 238745)
4 homozygous mutant males (223771, 223772, 223773, 257778)
4 wild-type control females (216691, 216693, 232541, 232542)
4 wild-type control males (216689, 225016, 238739, 238740)
90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (223789, 225037, 238744, 238745)
4 homozygous mutant males (223771, 223772, 223773, 257778)
2 wild-type control females (216691, 216693)
4 wild-type control males (216689, 225016, 238739, 238740)
180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (223789, 225037, 238744, 238745)
3 homozygous mutant males (223771, 223772, 223773)
2 wild-type control females (232541, 232542)
3 wild-type control males (225016, 238739, 238740)
300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (238744, 238745)
1 homozygous mutant male (223773)
2 wild-type control females (216691, 216693)
4 wild-type control males (216689, 225016, 238739, 238740)
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.