Gene: 824Name: Avpr1bFamily: GPCRSubfamily: VasopressinAccession: NM_011924GI: 6753151

Gene 824
Summary of Phenotypic Analysis

Changes related to genotype:

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:

When compared to age- and gender-matched wild-type control mice, homozygous mutant mice exhibited a shorter latency to reach the stages of tonic extension and death during metrazol testing, suggesting a heightened propensity towards seizure in the knockout mice. 

Gene 824
Behavior


Changes related to genotype:

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:

10 homozygous mutant males (187659, 189400, 189403, 191486, 191487, 210987, 217717, 217721, 218412, 218480)
12 wild-type control males (187656, 187657, 189402, 191484, 191485, 191488, 191489, 191490, 210984, 210985, 217719, 217720)

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 a shorter latency to reach the stages of tonic extension and death during metrazol testing, suggesting a heightened propensity towards seizure in the knockout mice. 

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 824
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 nine 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 (210967, 212444, 242752)

3 homozygous mutant females (210968, 218488, 242760)

 

 

Gene 824
Expression Summary

Taqman Summary:
High 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, pituitary gland and testis.

Expression:
Brain
In the wholemount staining lacZ expression is detectable in a small layer surrounding the corpus callosum. On coronal sections lacZ expression is not detectable.

Pituitary Gland
Scattered lacZ expression is detectable in pars distalis.

Male Reproductive Systems
Testis
Spermatogenic cells of the seminiferous tubules express lacZ.

No Expression:
LacZ expression is not detected in: spinal cord, sciatic nerve, eye, Harderian glands, thymus, spleen, lymph nodes, bone marrow, aorta, heart, lung, liver, gallbladder, pancreas, kidney, urinary bladder, trachea, larynx, esophagus, thyroid gland, parathyroid gland, adrenal glands, salivary glands, tongue, skeletal muscle, skin and female reproductive systems.

Gene 824
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. The data are compiled from the N0F2 and N1F2 generations.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (144385, 144386, 150836)
3 homozygous mutant males (144390, 152857, 152858)
2 wild-type control females (144383, 146949)
2 wild-type control males (144387, 144389)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (174344, 183219)
2 homozygous mutant males (200177, 226036)
2 wild-type control females (174348, 180365)
2 wild-type control males (183208, 183210)

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:

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 824
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 data are compiled from the N0F2 and N1F2 generations.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (144385, 144386, 150836)
3 homozygous mutant males (144390, 152857, 152858)
2 wild-type control females (144383, 146949)
2 wild-type control males (144387, 144389)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (174344, 183219)
1 homozygous mutant male (200177)

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 824
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. The data are compiled from the N0F2 and N1F2 generations.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (144385, 144386, 150836)
3 homozygous mutant males (144390, 152857, 152858)
2 wild-type control females (144383, 146949)
2 wild-type control males (144387, 144389)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (174344, 183219)
2 homozygous mutant males (200177, 226036)
2 wild-type control females (174348, 180365)
2 wild-type control males (183208, 183210)

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 824
Clinical Chemistry


There were no significant differences detected 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 biochemistry panel.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (144385, 144386, 152862)
3 homozygous mutant males (144390, 152857, 152858)
2 wild-type control females (144383, 146949)
2 wild-type control males (144387, 144389)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (174344, 183219, 189398, 191482)
4 homozygous mutant males (200177, 226036, 232584, 242750)
4 wild-type control females (174348, 180365, 180369, 183217)
4 wild-type control males (183208, 183210, 195048, 195050)

180 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (174344, 183219, 189398)
4 homozygous mutant males (200177, 226036, 232584, 242750)
4 wild-type control females (174348, 180365, 180369, 183217)
3 wild-type control males (183208, 183210, 195050)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (174344, 183219)
2 homozygous mutant males (200177, 226036)
2 wild-type control females (174348, 180365)
2 wild-type control males (183208, 183210)

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 824
Hematology


There were no significant differences detected 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 (144385, 144386, 152862)
3 homozygous mutant males (144390, 152857, 152858)
2 wild-type control females (144383, 146949)
2 wild-type control males (144387, 144389)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (174344, 183219, 189398, 189399)
4 homozygous mutant males (200177, 226036, 232584, 242750)
4 wild-type control females (174348, 180365, 180369, 183217)
3 wild-type control males (183208, 195048, 195050)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (174344, 183219, 189398, 189399)
4 homozygous mutant males (200177, 226036, 232584, 242750)
4 wild-type control females (174348, 180365, 180369, 183217)
3 wild-type control males (183208, 183210, 195050)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (174344, 183219)
2 homozygous mutant males (200177, 226036)
2 wild-type control females (174348, 180365)
2 wild-type control males (183208, 183210)

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 824
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. The data are compiled from the N0F2 and N1F2 generations.

49 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (144385, 144386, 150836)
3 homozygous mutant males (144390, 152857, 152858)
2 wild-type control females (144383, 146949)
2 wild-type control males (144387, 144389)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (174344, 183219)
2 homozygous mutant males (200177, 226036)
2 wild-type control females (174348, 180365)
2 wild-type control males (183208, 183210)

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.

Gene 824
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. The data are compiled from the N0F2 and N1F2 generations.

49 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (174344, 183219, 189398, 189399)
4 homozygous mutant males (200177, 226036, 232584, 242750)
4 wild-type control females (174348, 180365, 180369, 183217)
4 wild-type control males (183208, 183210, 195048, 195050)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (174344, 183219, 189398, 189399)
3 homozygous mutant males (200177, 226036, 242750)
3 wild-type control females (174348, 180365, 183217)
4 wild-type control males (183208, 183210, 195048, 195050)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (174344, 183219, 189398, 189399)
3 homozygous mutant males (200177, 232584, 242750)
3 wild-type control females (180365, 180369, 183217)
4 wild-type control males (183208, 183210, 195048, 195050)

300 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (174344, 183219, 189398, 189399)
1 homozygous mutant male (242750)
3 wild-type control females (174348, 180369, 183217)
2 wild-type control males (183208, 183210)

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.