Gene: 1841Name: Slc39a1Family: TransporterSubfamily: IronAccession: AF215637GI: 8895486

Gene 1841
Summary of Phenotypic Analysis

Changes related to genotype:

         Behavior:  Heterozygous mutant mice exhibited a significant decrease in thermal response latency during hot plate testing 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, whereas wild-type and heterozygous mutant mice were present. Partially resobed homozygous mutant embryos were detected by PCR at E8.5, but no homozygous embryo was found at later stages.

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 heterozygous mutant mice were evaluated by the following examinations or tests:

Gene 1841
Behavior

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:
10 heterozygous mutant males (299190, 299192, 306801, 306805, 306815, 314368, 316959, 316963, 320310, 323727)
7 wild-type control males (299189, 299191, 306791, 306804, 306812, 316962, 320309)

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:
When compared to age- and gender-matched wild-type control mice, heterozygous mutant mice exhibited a significantly decreased latency to lick a hindpaw or jump during hot plate testing, indicating a possible decreased pain threshold phenotype.

There were no other genotype-related differences noted between heterozygous mutant and wild-type control mice for any other parameters evaluated during behavior testing.

Gene 1841
Expression Summary

Taqman Summary:
Moderate levels of RNA transcripts are detectable in liver, kidney, spleen, thymus, bone marrow, skin, gallbladder, urinary bladder, adrenal gland, salivary gland, stomach, small intestine, colon, ovary and uterus.         

No RNA transcripts are detectable in whole brain, cortex, subcortical region, cerebellum, brainstem, olfactory bulb, spinal cord, eye, Harderian glands, heart, lung, pancreas, lymph nodes, pituitary gland, skeletal muscle, tongue, cecum, testis, epididymis, seminal vesicle, coagulating gland, prostate gland and white fat.

LacZ Summary:
LacZ expression was detected in several of the organs examined.  Most striking expression was detected in red pulp of the spleen, hepatocytes of the liver and in the kidney, which showed strong staining in tubules and very strong staining in papilla.  Staining was observed in different tissue and cell types, including epithelium, smooth and cardiac muscle, fibroblasts, adipose tissue, lymphatic tissue and blood vessels.

LacZ expression was detected in:  brain, spinal cord, eyes, thymus, spleen, bone marrow, heart, lung, liver, pancreas, kidney, urinary bladder, thyroid gland, trachea, pituitary gland, adrenal glands, skeletal muscle, skin, ovary, uterus and cervix.

LacZ expression was not detected in:  sciatic nerve, lymph nodes, aorta, testis and prostate gland.

Gene 1841
Densitometry


There were no significant differences detected in the 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:
3 heterozygous mutant females (273271, 273273, 273276)
3 heterozygous mutant males (273274, 273275, 273277)
2 wild-type control females (273272, 295607)
3 wild-type control males (262987, 295617, 295618)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (273278, 273280)
2 heterozygous mutant males (295605, 295614)
2 wild-type control females (295632, 299193)
2 wild-type control males (295603, 295604)

Bone Mineral Density (BMD in g/cm
2 ), fat % (fat percent 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 1841
Histopathology

There were no significant differences detected in the heterozygous mutant mice 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 heterozygous mutant females (273271, 273273, 273276)
3 heterozygous mutant males (273274, 273275, 273277)
2 wild-type control females (273272, 295607)
3 wild-type control males (262987, 295617, 295618)

300 Day Cohort Mouse ID numbers are as follows:
1 heterozygous mutant female (273278)
2 heterozygous mutant males (295605, 295614)

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 1841
Necropsy 

There were no significant differences detected in the 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.

49 Day Cohort Mouse ID numbers are as follows:
3 heterozygous mutant females (273271, 273273, 273276)
3 heterozygous mutant males (273274, 273275, 273277)
2 wild-type control females (273272, 295607)
3 wild-type control males (262987, 295617, 295618)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (273278, 273280)
2 heterozygous mutant males (295605, 295614)

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 1841
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. The data are compiled from the N0F2 and N1F2 generations.

49 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (273271, 273273, 273276, 295631)
3 heterozygous mutant males (273274, 273275, 273277)
2 wild-type control females (273272, 303176)
3 wild-type control males (262987, 295617, 295618)

90 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (273278, 273279, 273280, 273281)
2 wild-type control females (306817, 306820)

180 Day Cohort Mouse ID numbers are as follows:
3 heterozygous mutant females (273278, 273280, 273281)
3 heterozygous mutant males (295605, 295614, 295616)
4 wild-type control females (295632, 299193, 306817, 306820)
4 wild-type control males (295603, 295604, 295606, 295629)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (273278, 273280)
2 heterozygous mutant males (295605, 295614)
2 wild-type control females (295632, 299193)
2 wild-type control males (295603, 295604)

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 1841
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. The data are compiled from the N0F2 and N1F2 generations.

49 Day Cohort Mouse ID numbers are as follows:
3 heterozygous mutant females (273273, 273276, 306821)
3 heterozygous mutant males (273274, 273275, 273277)
2 wild-type control females (273272, 295607)
3 wild-type control males (262987, 295617, 295618)

90 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (273278, 273279, 273280, 273281)
4 heterozygous mutant males (295605, 295614, 295615, 295616)
1 wild-type control female (295632)
4 wild-type control males (295603, 295604, 295606, 295629)

180 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (273278, 273279, 273280, 273281)
4 heterozygous mutant males (295605, 295614, 295615, 295616)
4 wild-type control females (295632, 299193, 306817, 306820)
4 wild-type control males (295603, 295604, 295606, 295629)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (273278, 273280)
2 heterozygous mutant males (295605, 295614)
2 wild-type control females (295632, 299193)
2 wild-type control males (295603, 295604)

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 1841
Physical Examination

There were no significant differences detected in the heterozygous mutant mice 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 heterozygous mutant females (273271, 273273, 273276)
3 heterozygous mutant males (273274, 273275, 273277)
2 wild-type control females (273272, 295607)
3 wild-type control males (262987, 295617, 295618)

300 Day Cohort Mouse ID numbers are as follows:
2 heterozygous mutant females (273278, 273280)
2 heterozygous mutant males (295605, 295614)
2 wild-type control females (295632, 299193)
2 wild-type control males (295603, 295604)

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 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 were regarded as biologically significant or genotype related.

Gene 1841
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 (273278, 273279, 273280, 273281)
4 heterozygous mutant males (295605, 295614, 295615, 295616)
4 wild-type control females (295632, 299193, 306817, 306820)
4 wild-type control males (295603, 295604, 295606, 295629)

90 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (273278, 273279, 273280, 273281)
4 heterozygous mutant males (295605, 295614, 295615, 295616)
1 wild-type control female (295632)
4 wild-type control males (295603, 295604, 295606, 295629)

180 Day Cohort Mouse ID numbers are as follows:
4 heterozygous mutant females (273278, 273279, 273280, 273281)
4 heterozygous mutant males (295605, 295614, 295615, 295616)
4 wild-type control females (295632, 299193, 306817, 306820)
4 wild-type control males (295603, 295604, 295606, 295629)

300 Day Cohort Mouse ID numbers are as follows:
1 heterozygous mutant female (273281)
1 heterozygous mutant male (295616)
3 wild-type control females (295632, 299193, 306817)
4 wild-type control males (295603, 295604, 295606, 295629)

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.

Gene  1841
Summary of Embryonic Development

Homozygous mutant embryos die at ~ E8.5

Embryos were isolated at 8 .5 to 12.5 days post coitum. Homozygous offspring were detected by PCR at E8.5, but not at later stages.  All homozygous embryos detected were partially resorbed.These preliminary data suggests that death is around day 8.5 of embryonic development.  

Embryos were isolated at E8.5 to E12.5
Four litters were examined comprising of 36 embryos, resorptions and partial resorptions, of which 27 were successfully genotyped.

Litter

Embryonic stage

+/+

+/-

-/-

complete resorption or unknown

1

8.5

4

2

3

0

2

8.5

0

4

3

2

3

10.5

1

6

0

3

4

12.5

2

2

0

4