Gene: 609Name: BdkrbFamily: GPCRSubfamily: BradykininAccession: U47281GI: 1255048

Gene 609
Summary of Phenotypic Analysis

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

Gene 609
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, and Metrazol test.

Mouse ID numbers are as follows for the N1 generation:
8 homozygous mutant males (116282, 116291, 116293, 116294, 127344, 134116, 138362, 138363)
7 wild-type control males (114265, 114266, 116286, 116295, 127357, 127359, 138360)

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 609

Fertility


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

Two to three homozygous mutant mice of each gender were set up in a fertility mating at seven to eight weeks of age. The number of pups born from one to three litters were recorded. Three weeks later, the live pups were counted and weaned.

Mouse ID numbers are as follows:

3 homozygous mutant males (112827, 116315, 140512)
3 homozygous mutant females (105462, 116289, 134098)

Gene 609
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 609
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.

300 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (105394)
2 homozygous mutant males (105443, 105456)
2 wild-type control females (105408, 105410)
2 wild-type control males (105441, 105457)


No Significant Abnormalities:

Evaluations of densitometric data included Bone Mineral Density (BMD presented as g/cm2), Bone Mineral Content (BMC in g), bone and tissue area, total tissue mass, and fat as a percent of body soft tissue (presented as fat %). 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 differences, differences due to procedural artifacts, and/or differences of a nonspecific etiology. For example, fat % was decreased in a homozygous female (105394) with a malignant lymphoma that was considered a spontaneous disease. They are not considered to be genotype related.

Gene 609
Histopathology

There were no significant differences detected in the homozygous mutant and heterozygous 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:
6 homozygous mutant females (101723, 101724, 105409, 111295, 112807, 112823)
6 homozygous mutant males (105432, 105436, 105447, 111297, 112811, 112824)
1 heterozygous mutant female (100182)
1 heterozygous mutant male (100179)
4 wild-type control females (100183, 101722, 112810, 112819)
4 wild-type control males (100180, 100181, 112826, 116283)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (105394, 105398)
2 homozygous mutant males (105443, 105456)
2 wild-type control females (105408, 105410)
2 wild-type control males (105441, 105457)

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 may have been present in some tissues. For example, there is a small lymphocytic lymphoma in one 300 day cohort homozygous female (105394). These findings are 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 nonspecific etiology. They are not considered to be genotype related.

Gene 609
Necropsy

There were no significant differences detected in the homozygous mutant and 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:
6 homozygous mutant females (101723, 101724, 105409, 111295, 112807, 112823)
6 homozygous mutant males (105432, 105436, 105447, 111297, 112811, 112824)
1 heterozygous mutant female (100182)
1 heterozygous mutant male (100179)
4 wild-type control females (100183, 101722, 112810, 112819)
4 wild-type control males (100180, 100181, 112826, 116283)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (105394, 105398)
2 homozygous mutant males (105443, 105456)
2 wild-type control females (105408, 105410)
2 wild-type control males (105441, 105457)

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. For example, in a 300 day cohort homozygous female (105394) found to have malignant lymphoma, there were enlarged lymph nodes, an enlarged mesentery, an enlarged spleen, and an abnormal vertebral column. 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 nonspecific etiology. They were not considered to be genotype related.

Body and Organ Weight Findings:

Three 49 day cohort homozygous mice had increased spleen and/or liver weights; however, there were no gross pathologic or histopathologic findings present for these tissues. It is not clear whether these changes are genotype-related. They are presented here for your consideration. The spleen weight and spleen weight to body weight ratio was increased in one 300 day cohort homozygous female (105394) found to have a malignant lymphoma that was considered to be an incidental finding.

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 609
Clinical Chemistry

There were no significant differences detected in the homozygous mutant and 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:
3 homozygous mutant females (101723, 101724, 111295)
5 homozygous mutant males (105432, 105436, 105447, 111297, 116314)
1 heterozygous mutant female (100182)
1 heterozygous mutant male (100179)
2 wild-type control females (100183, 105423)
3 wild-type control males (100181, 112826, 116283)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (105394, 105398, 116278, 116280)
4 homozygous mutant males (105438, 105440, 105443, 105456)
4 wild-type control females (105408, 105410, 105424, 105426)
4 wild-type control males (105441, 105457, 105458, 105459)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (105394, 105398, 116278, 116280)
4 homozygous mutant males (105438, 105440, 105443, 105456)
4 wild-type control females (105408, 105410, 105424, 105426)
4 wild-type control males (105441, 105457, 105458, 105459)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (105394, 105398)
2 homozygous mutant males (105443, 105456)
2 wild-type control females (105408, 105410)
2 wild-type control males (105441, 105457)

Values for the various analytes evaluated were generally similar between homozygous mutant, heterozygous 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 609
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 (112807, 112823, 116278)
4 homozygous mutant males (105432, 105436, 112824, 116314)
2 wild-type control females (100183, 101722)
3 wild-type control males (100180, 112826, 116283)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (105394, 105398, 116278, 116280)
4 homozygous mutant males (105440, 105443, 105456, 116315)
4 wild-type control females (105408, 105410, 105424, 105426)
4 wild-type control males (105441, 105457, 105458, 118581)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (105394, 105398, 116278, 116280)
4 homozygous mutant males (105438, 105440, 105443, 105456)
4 wild-type control females (105408, 105410, 105424, 105426)
3 wild-type control males (105441, 105458, 105459)

300 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (105394, 121375, 123408, 123414)
2 homozygous mutant males (105456, 152285)
2 wild-type control females (105408, 105461)
2 wild-type control males (105441, 105457)

In a 300 day cohort homozygous female (105394) with a small lymphocytic lymphoma that is considered to be incidental, there is a moderate lymphocytosis. Although minor variations of hematological values were present in other animals, these changes were not consistent with genotype and, thus, were not considered phenotypically relevant.

Gene 609
Physical Examination

There were no significant differences detected in the homozygous mutant 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:
6 homozygous mutant females (101723, 101724, 105409, 111295, 112807, 112823)
6 homozygous mutant males (105432, 105436, 105447, 111297, 112811, 112824)
1 heterozygous mutant female (100182)
2 heterozygous mutant males (100179, 111296)
4 wild-type control females (100183, 101722, 112810, 112819)
5 wild-type control males (100180, 100181, 112817, 112826, 116283)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (105394, 105398)
2 homozygous mutant males (105443, 105456)
2 wild-type control females (105408, 105410)
2 wild-type control males (105441, 105457)

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

Gene 609
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 mice at 49, 90, 180, and 300 days of age.

49 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (105394, 105398, 116278, 116280)
4 homozygous mutant males (105440, 105443, 105455, 105456)
4 wild-type control females (105408, 105410, 105424, 105426)
4 wild-type control males (105441, 105457, 105458, 105459)

90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (105394, 105398, 116278, 116280)
4 homozygous mutant males (105440, 105443, 105455, 105456)
4 wild-type control females (105408, 105410, 105424, 105426)
4 wild-type control males (105441, 105457, 105458, 105459)

180 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (105394, 105398, 116278, 116280)
4 homozygous mutant males (105438, 105440, 105443, 105456)
4 wild-type control females (105408, 105410, 105424, 105426)
4 wild-type control males (105441, 105457, 105458, 105459)

300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (116278, 116280)
3 homozygous mutant males (105438, 105440, 105443)
4 wild-type control females (105408, 105410, 105424, 105426)
3 wild-type control males (105441, 105458, 105459)

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.