Gene: 527Name: Lats2Family: KinaseSubfamily: Serine/Threonine Kinas...Accession: AB023958GI: 7212785

Gene 527
Summary of Phenotypic Analysis

 Changes related to genotype:

There were no other significant differences detected in the 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 suggested an embryonic 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 heterozygous mutant mice were evaluated by the following examinations or tests:

Gene 527
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 (381266, 381267, 381272, 381278, 381280, 381281, 381282, 381283, 381284, 381285)
12 wild-type control males (374383, 380467, 381268, 381273, 381279, 382327, 382351, 382404, 382406, 382408, 382494, 384440)

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 significant decrease in prepulse inhibition, indicating a stimulus processing deficit similar to that observed in some human schizophrenic patients.

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 527
Expression Summary

Taqman Summary:
RNA transcripts are detectable in all tissues analyzed.

The highest levels of RNA transcripts are detectable in testis.

Moderate levels of RNA transcripts are detectable in: eye, lung, kidney, lymph nodes, skin, gallbladder, urinary bladder, adrenal gland, prostate gland, ovary and white fat.     

Lower levels of RNA transcripts are also detectable in: whole brain, cortex, subcortical region, cerebellum, brainstem, olfactory bulb, spinal cord, Harderian glands, heart, liver, pancreas, spleen, thymus, bone marrow, pituitary gland, salivary gland, skeletal muscle, tongue, stomach, small intestine, large intestine, cecum, epididymis, seminal vesicle, coagulating gland and uterus.

LacZ Summary:
LacZ expression was detected in all organs examined.  Striking expression was detected in cerebral ventricles, purkinje cells in the cerebellum, myocardium in the heart and smooth muscle of the urinary bladder.  Staining was observed in different tissue and cell types, including skeletal, cardiac and smooth muscle, neurons, epithelium, adipose tissue, cartilage, lymphatic tissue and blood vessels.

LacZ expression was detected in:  brain, spinal cord, sciatic nerve, eyes, thymus, spleen, lymph nodes, bone marrow, aorta, heart, lung, liver, pancreas, kidney, urinary bladder, thyroid gland, larynx, pituitary gland, skeletal muscle, skin, testis, prostate gland, ovary, uterus and cervix.

Gene 527
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 (350889, 350890, 350891)
3 heterozygous mutant males (350592, 350593, 350594)
1 wild-type control female (359784)
2 wild-type control males (350591, 350595)

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.

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 527
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 (350889, 350890, 350891)
3 heterozygous mutant males (350592, 350593, 350594)
2 wild-type control females (355721, 359784)
2 wild-type control males (350591, 350595)

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, adrenal glands, urinary bladder, stomach, small and large intestines, larynx, esophagus, trachea, thyroid gland, thymus, 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 527
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 (350889, 350890, 350891)
3 heterozygous mutant males (350592, 350593, 350594)
2 wild-type control females (355721, 359784)
2 wild-type control males (350591, 350595)

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:

At 49 days, low thymus weights and thymus weight to body weight ratios were observed in the heterozygous male mice and this finding did not have any histological correlate. 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. The variability between mice usually fell within our historical reference ranges and was not correlated with genotype.

Gene 527

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:
3 heterozygous mutant females (350889, 350890, 350891)
3 heterozygous mutant males (350592, 350593, 350594)
2 wild-type control females (355721, 359784)
2 wild-type control males (350591, 350595)

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 527

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 (350889, 350890, 350891)
3 heterozygous mutant males (350592, 350593, 350594)
2 wild-type control females (355721, 359784)
2 wild-type control males (350591, 359777)

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 527
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 (350889, 350890, 350891)
3 heterozygous mutant males (350592, 350593, 350594)
2 wild-type control females (355721, 359784)
2 wild-type control males (350591, 350595)

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 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 527
Summary of Embryonic Development

Homozygous mutant embryos die at ~ E12.5

Embryos were isolated at 8 .5 to 10.5 days post coitum. Homozygous offspring were detected by PCR at E9.5 and E10.5.   Grossly normal homozygous embryos were detected at E9.5 and E10.5. No homozygous embryo was detected at later stages

Embryos were isolated at E8.5 to E10.5
Four litters were examined comprising of 28 embryos, resorptions and partial resorptions, of which 28 were successfully genotyped.

Litter

Embryonic stage

+/+

+/-

-/-

complete resorption or unknown

1

8.5

1

5

0

0

2

9.5

2

4

1

0

3

10.5

0

4

2

0

4

12.5

5

4

0

0