Gene: 1381Name: Slc6a9Family: TransporterSubfamily: GlycineAccession: NM_008135GI: 6680028

Gene 1381
Summary of Phenotypic Analysis

Changes related to genotype:

There were no other 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. Fewer than expected homozygous mutant mice were identified by PCR, when compared with observed numbers of wild-type and heterozygous mutant mice. The genotypic ratio suggested either a 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:

Gene 1381
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 response/PPI test, Tail flick test, Hot plate test, and Metrazol test.

Mouse ID numbers are as follows for the N1 generation:
11 heterozygous mutant males (281218, 281219, 281222, 298418, 298419, 308845, 308853, 312735, 312739, 312741, 336110)
15 wild-type control males (281217, 281221, 298415, 298416, 298417, 312244, 312737, 317140, 317191, 317615, 318647, 331543, 334641, 334863, 336109)

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.

Gene 1381
Expression Summary

RT-PCR Summary:
The highest levels of RNA transcripts are detectable in pancreas. 

Lower levels of RNA transcripts are also detectable in: brain, cortex, subcortical region, cerebellum, brainstem, olfactory bulb, spinal cord, eye, Harderian glands, heart, lung, liver, kidney, spleen, lymph nodes, bone marrow, skin, gallbladder, urinary bladder, 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.

No RNA transcripts are detectable in thymus and pituitary gland.

LacZ Summary:
LacZ expression was observed in most tissues examined.  Of note, expression was not detected in immune-related organs.  Most striking expression was observed in the Purkinje cell layer of cerebellum, brain stem, spinal cord, ciliary body and inner nuclear layer of the eye and blood vessels of many organs.

LacZ (beta-galactosidase) expression was detected in: brain, spinal cord, eyes, aorta, heart, lung, liver, pancreas, kidney, urinary bladder, thyroid gland, pituitary gland, adrenal glands, skeletal muscle, skin, testis, prostate gland, and uterus.

LacZ expression was not detected in: thymus, spleen, lymph nodes, bone marrow and ovary.


 

Gene 1381
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. The data were compiled from the N0F2 and N1F2 generations.

49 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (270396)
1 homozygous mutant male (270391)
3 heterozygous mutant females (229937, 229938, 229941)
2 heterozygous mutant males (229936, 229945)
3 wild-type control females (229939, 229940, 270397)
3 wild-type control males (229943, 229944, 270392)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (246194, 246201)
2 heterozygous mutant males (261824, 261825)
2 wild-type control females (261822, 261827)
2 wild-type control males (261823, 270387)

Bone Mineral Density (BMD in g/cm2 ), fat % (fat percent 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 ), and total tissue mass (g) generated by a PIXImus densitometer.

Densitometric Findings:

When compared to historical but not contemporaneous wild-type control mice, total tissue mass was increased in 300 day heterozygous females and males, and fat % was increased in 300 day heterozygous females. 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 differences, and/or differences of a nonspecific etiology. They were not considered to be genotype related.

Gene 1381
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. The data were compiled from the N0F2 and N1F2 generations.

20 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (247678)

49 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (270396)
1 homozygous mutant male (270391)
3 heterozygous mutant females (229937, 229938, 229941)
3 heterozygous mutant males (229935, 229936, 229945)
3 wild-type control females (229939, 229940, 270397)
3 wild-type control males (229943, 229944, 270392)

300 Day Cohort Mouse ID numbers are as follows:
1 heterozygous mutant female (246194)
1 heterozygous mutant male (261824)

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 1381
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. The data are compiled from the N0F2 and N1F2 generations.

20 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (247678)

49 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (270396)
1 homozygous mutant male (270391)
3 heterozygous mutant females (229937, 229938, 229941)
3 heterozygous mutant males (229935, 229936, 229945)
3 wild-type control females (229939, 229940, 270397)
3 wild-type control males (229943, 229944, 270392)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (246194, 246201)
2 heterozygous mutant males (261824, 261825)

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, a 300 day heterozygous male (261825) was reported to have a penis mass for which there was no histopathological correlate. 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:

The 49 day cohort homozygous mutant male had a decreased body weight when compared to age- and gender-matched wild-type control mice. When compared to age- and gender-matched wild-type control mice, body weight was increased in one female (229937) and two male (229935, 229936) 49 day heterozygous mutants, and two female and two male 300 day heterozygous mutants. These findings have not been reported as phenotypic changes, but have been presented 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. The variability between mice usually fell within our historical reference ranges and was not correlated with genotype.

Gene 1381
Clinical Chemistry

There were no significant differences in the homozygous or 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. The data are compiled from the N0F2 and N1F2 generations.

20 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (247678)

49 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (270396)
1 homozygous mutant male (270391)
3 heterozygous mutant females (229937, 229938, 229941)
4 heterozygous mutant males (229935, 229936, 229945, 248369)
3 wild-type control females (229939, 229942, 270397)
3 wild-type control males (229943, 229944, 270392)

90 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (246194, 246201, 246202, 261826)
4 heterozygous mutant males (261824, 261825, 261829, 261831)
3 wild-type control females (261822, 261827, 270398)
3 wild-type control males (261823, 270387, 281206)

180 Day Cohort Mouse ID numbers are as follows:
1 heterozygous mutant female (261826)
4 heterozygous mutant males (261824, 261825, 261829, 261831)
3 wild-type control females (261822, 261827, 270398)
4 wild-type control males (261823, 270387, 281206, 292994)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (246194, 246201)
2 heterozygous mutant males (261824, 261825)
2 wild-type control females (261822, 261827)
2 wild-type control males (261823, 270387)

Certain differences, including elevated  high density lipoprotein and cholesterol levels, were present that occasionally occur spontaneously in mice of this sex and age group.  In this target, such differences were present in homozygous females and males at 90, 180, and 300 days. We have not reported these findings as phenotypic changes, but we have presented them here for your consideration.

Other 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 1381

Hematology

There were no significant differences in the homozygous or 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. The data are compiled from the N0F2 and N1F2 generations.

49 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (270396)
1 homozygous mutant male (270391)
3 heterozygous mutant females (229937, 229938, 229941)
3 heterozygous mutant males (229936, 248369, 248370)
3 wild-type control females (229942, 246973, 270397)
3 wild-type control males (229944, 246981, 270392)

90 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (246194, 246201, 246202, 261830)
4 heterozygous mutant males (261824, 261825, 261829, 261831)
3 wild-type control females (261822, 261827, 270398)
4 wild-type control males (261823, 270387, 281206, 292994)

180 Day Cohort Mouse ID numbers are as follows:
1 heterozygous mutant female (261826)
4 heterozygous mutant males (261824, 261825, 261829, 261831)
3 wild-type control females (261822, 261827, 298423)
3 wild-type control males (261823, 281206, 292994)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (246194, 246201)
2 heterozygous mutant males (261824, 261825)
1 wild-type control female (261827)
2 wild-type control males (261823, 270387)

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 1381
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. The data were compiled from the N0F2 and N1F2 generations.

20 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (247678)

49 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (270396)
1 homozygous mutant male (270391)
3 heterozygous mutant females (229937, 229938, 229941)
3 heterozygous mutant males (229935, 229936, 229945)
3 wild-type control females (229939, 229940, 270397)
3 wild-type control males (229943, 229944, 270392)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (246194, 246201)
2 heterozygous mutant males (261824, 261825)
2 wild-type control females (261822, 261827)
2 wild-type control males (261823, 270387)

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 or 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 1381
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 (246194, 246201, 246202, 261828)
4 heterozygous mutant males (261824, 261825, 261829, 261831)
4 wild-type control females (261822, 261827, 270398, 298423)
4 wild-type control males (261823, 270387, 281206, 292994)

90 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (246194, 246201, 246202, 261828)
4 heterozygous mutant males (261824, 261825, 261829, 261831)
3 wild-type control females (261822, 261827, 270398)
4 wild-type control males (261823, 270387, 281206, 292994)

180 Day Cohort Mouse ID numbers are as follows:
1 wild-type control female (298423)
2 wild-type control males (281206, 292994)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (246202, 261826)
2 heterozygous mutant males (261829, 261831)
4 wild-type control females (261822, 261827, 270398, 298423)
3 wild-type control males (261823, 270387, 292994)

Body Weight and Length Findings:

When compared to age- and gender-matched wild-type control mice, body weight was increased in one 49 day male (261825), three 90 day female (261824, 261825, 261829), two 300 day female (246202, 261826) and one 300 day male (261829) heterozygous mutants. 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.