Gene: 1520Name: Adcy3Family: CyclaseSubfamily: AdenylateAccession: AF458089GI: 18092642

Gene 1520
Summary of Phenotypic Analysis

Changes related to genotype:

·         Lethality: Homozygous mice frequently do not survive to weaning age. 

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

Weaned progeny from the heterozygous matings were genotyped. No homozygous mutant mice were identified by PCR, whereas wild-type and heterozygous mutant mice were present. The genotypic ratio suggests either an embryonic, perinatal or juvenile lethal phenotype.

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 mutant mice were produced by intercrossing F1 heterozygous males and females.

Wild-type control mice, and homozygous and heterozygous mutant mice, were evaluated by the following examinations or tests:

 

Keywords

developmental lethality

perinatal lethal

 


 

Gene 1520
Expression Summary

RT-PCR Summary:
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, Harderian glands, aorta, heart, lung, kidney, urinary bladder, salivary glands, tongue, external ear, male and female reproductive systems.  In many of the tissues with detectable expression, lacZ expression is detectable in blood vessels.

Expression:
Brain
In wholemount staining very weak lacZ expression is detectable in a few regions of the cerebrum, the fourth ventricle and the more caudal regions of the cerebellum.

Harderian Glands
Moderate lacZ expression is detectable in some smooth muscle cells of blood vessels.

Aorta
Strong lacZ expression is detectable in some cells of the periaortic adipose tissue.

Heart
Strong lacZ expression is detectable in some cells of the pericardial adipose tissue.

Lung
Moderate lacZ expression is detectable in some smooth muscle cells of blood vessels.

Kidney
Moderate lacZ expression is detectable in some smooth muscle cells of blood vessels.

Urinary Bladder
Moderate lacZ expression is detectable in some smooth muscle cells of blood vessels and in a few cells of the perinephric adipose tissue.

Salivary Glands
Moderate lacZ expression is detectable in some smooth muscle cells of blood vessels.

Tongue
Moderate lacZ expression is detectable in some smooth muscle cells of blood vessels.

Ear, external
Moderate lacZ expression is detectable in some epidermal cells and in a few chondrocytes.

Male Reproductive Systems
Testis
Moderate lacZ expression is detectable in some smooth muscle cells of blood vessels.  Weak lacZ expression is detectable in a few cells of the interstitium.

Penis
Strong lacZ expression is detectable in some smooth muscle cells of blood vessels. Weak lacZ expression is detectable in some epithelial cells of the skin and hair follicles.

Coagulating Gland
Moderate lacZ expression is detectable in some smooth muscle cells of blood vessels.

Prostate and Ampullary Gland
Moderate lacZ expression is detectable in some smooth muscle cells of blood vessels.

Female Reproductive Systems
Vagina/Cervix
Moderate lacZ expression is detectable in some smooth muscle cells of blood vessels.


No Expression: 
LacZ expression is not detected in: spinal cord, sciatic nerve, eyes, thymus, spleen, lymph nodes, bone marrow, liver, gallbladder, pancreas, trachea, larynx, esophagus, thyroid gland, pituitary gland, adrenal glands, and skeletal muscle.

 


 

Gene 1520
Necropsy 

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

20 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (240182)
2 wild-type control males (240179, 276366)

49 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (228353, 228355, 228356, 228357)
3 heterozygous mutant males (228350, 228352, 235699)
2 wild-type control females (221121, 221123)
2 wild-type control males (228351, 228354)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (254859, 256704)
2 heterozygous mutant males (256721, 256726)
1 wild-type control male (256727)

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 heterozygous mutant female (256704) increased subcutaneous fat correlated with high body weight at necropsy and high fat % at densitometer examination. 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:

Body weight and several organ weights, but not organ weight/body weight ratios, were high in a 300 day heterozygous mutant female (256704). The first finding correlated with high body weight in another 300 day heterozygous mutant female at aging metrics examination. We have not reported these findings as phenotypic changes but we have presented them here for your consideration.

Other differences in body length, body weight, organ weights, and/or organ weight to body weight ratios were present between individual mice. For example, The variability between mice usually fell within our historical reference ranges and was not correlated with genotype.


 

Gene 1520
Histopathology

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

Tissues from the following mice were evaluated histologically.

20 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (240182)
2 wild-type control males (240179, 276366)

49 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (228353, 228355, 228356, 228357)
3 heterozygous mutant males (228350, 228352, 235699)
2 wild-type control females (221121, 221123)
2 wild-type control males (228351, 228354)

300 Day Cohort Mouse ID numbers are as follows:
1 heterozygous mutant female (254859)
1 heterozygous mutant male (256726)

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 1520
Hematology

There were no significant differences in the heterozygous 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 heterozygous mutant females (228355, 228356, 228357)
4 heterozygous mutant males (228350, 228352, 235698, 235699)
2 wild-type control females (221121, 270277)
3 wild-type control males (228351, 228354, 231223)

90 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (254859, 256704, 256705, 256708)
4 heterozygous mutant males (256721, 256726, 256766, 264369)
4 wild-type control females (254829, 254861, 256703, 256706)
4 wild-type control males (256727, 266378, 266639, 266641)

180 Day Cohort Mouse ID numbers are as follows:
3 heterozygous mutant females (254859, 256704, 256708)
2 heterozygous mutant males (256721, 256766)
4 wild-type control females (254829, 254861, 256703, 256706)
2 wild-type control males (256727, 266378)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (254859, 256704)
2 heterozygous mutant males (256721, 256726)
2 wild-type control females (254829, 254861)
1 wild-type control male (256727)

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 1520
Clinical Chemistry

There were no significant differences in the heterozygous 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:
4 heterozygous mutant females (228355, 228356, 231222, 231224)
3 heterozygous mutant males (228350, 228352, 235699)
2 wild-type control females (221121, 231217)
3 wild-type control males (228351, 228354, 231216)

90 Day Cohort Mouse ID numbers are as follows:
3 heterozygous mutant females (254859, 256704, 256705)
4 heterozygous mutant males (256721, 256726, 256766, 264369)
4 wild-type control females (254829, 254861, 256703, 256706)
4 wild-type control males (256727, 266378, 266639, 266641)

180 Day Cohort Mouse ID numbers are as follows:
1 heterozygous mutant male (264369)
2 wild-type control males (266639, 266641)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (254859, 256704)
2 heterozygous mutant males (256721, 256726)
2 wild-type control females (254829, 254861)
1 wild-type control male (256727)

Values for the various analytes evaluated were generally similar between 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 1520
Densitometry

There were no significant differences detected in the homozygous or heterozygous 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:
4 heterozygous mutant females (228353, 228355, 228356, 228357)
3 heterozygous mutant males (228350, 228352, 235699)
2 wild-type control females (221121, 221123)
2 wild-type control males (228351, 228354)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (254859, 256704)
2 heterozygous mutant males (256721, 256726)
2 wild-type control females (254829, 254861)
1 wild-type control male (256727)

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:

In a 300 day heterozygous mutant female (256704) high fat % and total tissue mass correlated with increased subcutaneous fat and high body weight at necropsy examination, and with high body weight in another 300 day heterozygous mutant female at aging metrics examination. We have not reported these findings as phenotypic changes but we have presented them here for your consideration.

Other incidental densitometric differences may have been 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 changes, differences due to procedural artifacts, and/or differences of a nonspecific etiology. They were not considered to be genotype related.


 

Gene 1520
Physical Examination

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

The following mice were evaluated by physical examination.

20 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (240182)
2 wild-type control males (240179, 276366)

49 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (228353, 228355, 228356, 228357)
3 heterozygous mutant males (228350, 228352, 235699)
2 wild-type control females (221121, 221123)
2 wild-type control males (228351, 228354)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (254859, 256704)
2 heterozygous mutant males (256721, 256726)
2 wild-type control females (254829, 254861)
1 wild-type control male (256727)

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 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 1520
Aging Metrics

There were no significant differences detected in the heterozygous 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 heterozygous mutant females (254859, 256704, 256705, 256708)
4 heterozygous mutant males (256721, 256726, 256766, 264369)
4 wild-type control females (254829, 254861, 256703, 256706)
4 wild-type control males (256727, 266378, 266639, 266641)

90 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (254859, 256704, 256705, 256708)
4 heterozygous mutant males (256721, 256726, 256766, 264369)
4 wild-type control females (254829, 254861, 256703, 256706)
4 wild-type control males (256727, 266378, 266639, 266641)

300 Day Cohort Mouse ID numbers are as follows:
1 heterozygous mutant female (256708)
2 heterozygous mutant males (256766, 264369)
4 wild-type control females (254829, 254861, 256703, 256706)
2 wild-type control males (266639, 266641)

Body Weight and Length Findings:

Body weight and/or body weight/body length ratio were high compared to historical but not contemporaneous wild-type control mice in three 90 day heterozygous mutant females (256704, 256705, 256708), one 90 day heterozygous mutant male (256766), one 300 day heterozygous mutant female (256708) and one 300 day heterozygous mutant male (256766). These findings correlated with increased subcutaneous fat and high body weight at necropsy examination and high fat % and total tissue mass at densitometer examination in another 300 day heterozygous mutant female. We have not reported these findings as phenotypic changes but we have presented them here for your consideration.

Other 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 1520
Behavior


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

Heterozygous 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 test, Tail flick test, Hot plate test, and Metrazol test.

Mouse ID numbers are as follows for the N1 generation:
13 heterozygous mutant males (254833, 254834, 254835, 254855, 254858, 264863, 264864, 264865, 266589, 266590, 276334, 276343, 276349)
8 wild-type control males (254854, 254856, 254857, 264862, 266587, 266588, 276346, 276348)

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 heterozygous mutant mice were produced by intercrossing F1N1 heterozygous males and females.

Behavior Findings:
There were no genotype-related or biologically significant differences noted between heterozygous mutant and wild-type control mice for any of the parameters evaluated during behavior testing.