Gene: 669Name: Fzd6Family: GPCRSubfamily: WNTAccession: U43319GI: 1151255

Gene 669
Summary of Phenotypic Analysis

There were no significant differences detected in the homozygous or heterozygous 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. 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, as well as homozygous mutant and heterozygous mutant mice, were evaluated by the following examinations or tests:

Gene 669
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 (168552, 316000, 336804)

3 homozygous mutant females (166309, 315994, 352116)

 

 

Gene 669
Expression Summary

RT-PCR Summary:
Low levels of RNA transcripts are detectable in all tissues analyzed: brain, cortex, subcortical region, cerebellum, brainstem, olfactory bulb, spinal cord, eye, Harderian glands, 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, ovaries, uterus and white fat.

LacZ Summary:
LacZ (beta-galactosidase) expression is detectable in brain, heart, pituitary gland, tongue, skin, male and female reproductive system. The most striking expression is seen in endothelial cells of blood vessels.

Expression:
Brain
In wholemount staining, strong lacZ expression is detectable in the thalamus and in the vasculature. Faint signals are detectable in cerebellum and brainstem.
In frozen sections strong signals are detectable in the thalamus and third ventricle.

Heart
Strong X-Gal signals are detectable in the endothelium layer of blood vessels.

Trachea
Strong X-Gal signals are detectable in endothelial cells of surrounding blood vessels.

Pituitary Gland
Scattered lacZ expression is detectable in pars distalis and pars nervosa of the pituitary gland.

Tongue
Many epithelial cells of the epithelium, and endothelial cells of blood vessels stain strongly. Few epithelial cells of the mucous glands stain strongly.

Skin
Hair follicles show variable degrees of X-Gal staining.

Skin of the Ear
Scattered X-Gal staining is detectable in the epithelial cell layer and in hair follicles.

Male Reproductive Systems
Penis
Strong lacZ expression is detectable in endothelial cells of blood vessels, in epithelial cells.
Scattered X-Gal staining is detectable in hair follicles.

Prostate Gland
Scattered, strong X-Gal staining is detectable in epithelial cells.

Female Reproductive Systems
Ovary
Strong lacZ expression is seen in blood vessels

Oviduct/Uterus
Many epithelial cells of the Fallopian tubes stain.

No Expression:
LacZ expression is not detected in: spinal cord, sciatic nerve, eye, Harderian glands, thymus, spleen, lymph nodes, bone marrow, aorta, lung, liver, gall bladder, pancreas, kidney, urinary bladder, esophagus, larynx, salivary glands, thyroid gland, parathyroid gland, adrenal glands, skeletal muscle,

Gene 669
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:
1 homozygous mutant female (111231)
2 homozygous mutant males (111236, 117662)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (114182, 134051)
2 homozygous mutant males (140195, 145236)
2 wild-type control females (114181, 114184)
1 wild-type control male (140206)

Bone Mineral Density (BMD in g/cm2 ), 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. These findings are 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 are not considered to be genotype related.

Gene 669
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 (106616, 106631, 111231)
3 homozygous mutant males (106623, 111236, 117662)
1 heterozygous mutant female (106630)
1 heterozygous mutant male (106637)
2 wild-type control females (106633, 106634)
3 wild-type control males (106635, 106636, 111234)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (114182, 134051)
2 homozygous mutant males (140195, 145236)
2 wild-type control females (114181, 114184)
1 wild-type control male (140206)

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

Gene 669
Necropsy

There were no significant differences detected in the homozygous or heterozygous 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 changes were recorded.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (106616, 106631, 111231)
3 homozygous mutant males (106623, 111236, 117662)
1 heterozygous mutant female (106630)
1 heterozygous mutant male (106637)
2 wild-type control females (106633, 106634)
2 wild-type control males (106635, 106636)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (114182, 134051)
2 homozygous mutant males (140195, 145236)
2 wild-type control females (114181, 114184)
1 wild-type control male (140206)

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 nonspecific etiology. They were not considered to be genotype related.

Body and Organ Weight Findings:

One 300 day cohort homozygous mutant female (134051) weighed less than both age- and gender-matched control mice and the historical reference range. This finding is not being presented as a phenotype but is included for your consideration. The 300 day cohort wild-type females were large with increased body weight. 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 669
Clinical Chemistry

There were no significant differences detected in the homozygous or heterozygous 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:
5 homozygous mutant females (106631, 117644, 117654, 123886, 138142)
2 homozygous mutant males (111236, 117662)
1 heterozygous mutant female (111233)
1 heterozygous mutant male (106637)
2 wild-type control females (106633, 106634)
2 wild-type control males (106635, 106636)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (114182, 134058, 138153, 140201)
2 homozygous mutant males (140195, 145236)
5 wild-type control females (114181, 114184, 134052, 134055, 138156)
2 wild-type control males (140206, 145235)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (114182, 134051, 140201, 154285)
2 homozygous mutant males (140195, 145236)
5 wild-type control females (114181, 114184, 134052, 134055, 147865)
2 wild-type control males (140206, 145235)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (114182, 134051)
2 homozygous mutant males (140195, 145236)
2 wild-type control females (114181, 114184)
1 wild-type control male (140206)

Values for the various analytes evaluated were generally similar between homozygous or heterozygous 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 669
Hematology

There were no significant differences detected in the homozygous or heterozygous 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:
5 homozygous mutant females (106631, 117644, 117654, 119999, 138142)
2 homozygous mutant males (111236, 117662)
1 heterozygous mutant female (111233)
1 heterozygous mutant male (106637)
2 wild-type control females (106633, 106634)
2 wild-type control males (106635, 106636)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (114182, 134051, 134058, 154285)
2 homozygous mutant males (140195, 145236)
4 wild-type control females (114181, 134052, 134055, 138156)
2 wild-type control males (140206, 145235)

180 Day Cohort Mouse ID numbers are as follows:
5 homozygous mutant females (114182, 117659, 134051, 138153, 140201)
2 homozygous mutant males (140195, 145236)
4 wild-type control females (114184, 134052, 134055, 138156)
2 wild-type control males (140206, 145235)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (114182, 134051)
2 homozygous mutant males (140195, 145236)
2 wild-type control females (114181, 147865)
1 wild-type control male (140206)

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 669
Physical Examination

There were no significant differences detected in the homozygous and heterozygous 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 (106616, 106631, 111231)
3 homozygous mutant males (106623, 111236, 117662)
1 heterozygous mutant female (106630)
1 heterozygous mutant male (106637)
2 wild-type control females (106633, 106634)
2 wild-type control males (106635, 106636)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (114182, 134051)
2 homozygous mutant males (140195, 145236)
2 wild-type control females (114181, 114184)
1 wild-type control male (140206)

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 and heterozygous 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, and/or findings of a nonspecific etiology. However, none of these differences was regarded as biologically significant or genotype related.

Gene 669
Aging Metrics

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

Body weights and body lengths were measured for the following micee.

49 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (114182, 134051, 134058, 138153)
2 homozygous mutant males (140195, 145236)
9 wild-type control females (106615, 106617, 106618, 106619, 114181, 114184, 134052, 134055, 138156)
2 wild-type control males (140206, 145235)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (114182, 134051, 134058, 138153)
2 homozygous mutant males (140195, 145236)
5 wild-type control females (114181, 114184, 134052, 134055, 138156)
2 wild-type control males (140206, 145235)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (114182, 134051, 138153, 140201)
2 homozygous mutant males (140195, 145236)
5 wild-type control females (114181, 114184, 134052, 134055, 138156)
2 wild-type control males (140206, 145235)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (138153, 140201)
3 wild-type control females (134052, 134055, 138156)
1 wild-type control male (140206)

Body Weight and Length Findings:

When compared to age- and gender-matched wild-type control mice, two 300 day cohort homozygous females (134051, 138153) had a decreased body weight. However, only one (138153) weighs less than historical control mice. Therefore, this finding is not being reported as a phenotypic change but is included for your consideration. 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.