| Gene: 898 | Name: Grik5 | Family: Channel | Subfamily: Glutamate Receptor | Accession: D10011 | GI: 220415 |
|---|
Gene 898
Summary
of Phenotypic Analysis
Changes related to genotype:
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 homozygous mutant and heterozygous mutant
mice, were evaluated by the following examinations or tests:
Behavior Findings:
When compared to age- and gender-matched wild-type control mice, homozygous mutants were significantly different from wild-type control mice in the Tail flick test. Homozygous mutants displayed an increased response latency to an acute thermal stimulus. This may indicate an analgesic effect.
Homozygous mutants were significantly different from wild-types in the Startle/PPI test. Homozygous mutant mice displayed an increased startle response to auditory stimuli at the 100 and 120 dB level. This may indicate increased anxiety.
Homozygous mutants were significantly different from wild-types in the Metrazol test. Homozygous mutant mice required a lower dose than wild-types to induce a tonic/clonic seizure. This may indicate increased seizure susceptibility.
Gene 898
Behavior
Changes related to genotype:
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, Tail flick test, Startle/PPI, and Metrazol test.
Mouse
ID numbers are as follows:
8 homozygous
mutant males (143752, 142747, 142752, 155897, 164511, 189170, 203653, 222913)
10 wild-type control males (143756, 142743, 142744, 142755, 142756, 155895,
164508, 189162, 203652, 222916)
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:
When compared to age- and gender-matched wild-type control mice, homozygous
mutants were significantly different from wild-type control mice in the Tail
flick test. Homozygous mutants displayed an increased response latency to an
acute thermal stimulus. This may indicate an analgesic effect.
Homozygous
mutants were significantly different from wild-types in the Startle/PPI
test. Homozygous mutant mice displayed an increased startle response to
auditory stimuli at the 100 and 120 dB level. This may indicate increased
anxiety.
Homozygous
mutants were significantly different from wild-types in the Metrazol test.
Homozygous mutant mice required a lower dose than wild-types to induce a
tonic/clonic seizure. This may indicate increased seizure susceptibility.
There were no other genotype-related differences noted between homozygous mutant and wild-type control mice for any other parameters evaluated during behavior testing.
Gene 898
Fertility
Both homozygous mutant males and females were fertile. Their progeny were viable until weaning.
Three homozygous mutant mice of each gender were set up in a fertility mating one on one with each other at seven to ten weeks of age. The number of pups born from three litters was recorded. Three weeks later, the live pups were counted and weaned.
Mouse ID numbers are as follows:
3 homozygous mutant males (105902, 105924, 108002)
3 homozygous mutant females (105940, 100693, 100691)
Gene 898
Expression
Summary
RT-PCR Summary:
The highest levels of RNA transcripts are detectable in brain, cortex,
subcortical region, cerebellum, brainstem, olfactory bulb, spinal cord and eye.
Lower levels of RNA transcripts are also detectable in 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, spinal cord,
sciatic nerve, eye, Harderian glands, aorta, lung, gall bladder, kidney,
urinary bladder, trachea, parathyroid gland, pituitary gland, adrenal glands,
salivary glands, tongue, male and female reproductive systems. The most
striking expression is seen in nervous tissue and smooth muscle cells.
Expression:
Brain
In wholemount staining, all regions of the brain including the olfactory bulb
show strong lacZ expression. On frozen sections, strong lacZ expression is
detectable throughout forebrain, midbrain and brainstem with practically all
cells showing lacZ staining. In the cerebellum, strong lacZ expression is
detectable in the granular layer, white matter and peduncles. Basket cells of
the Purkinje cell layer show X-Gal signals. Many nuclei in the molecular stain
display X-Gal staining.
Spinal cord
Practically all cells express lacZ. Very strong signals are detectable in motor
neurons.
Sciatic Nerve
LacZ expression is detectable in the perineurium.
Eyes
LacZ expression is detectable in the retina, with striking signals in the inner
nuclear layer, ganglion layer and pigment layer. Further lacZ expression is
detectable in the uvea, sclera and optic nerve.
Harderiain Glands
Faint lacZ expression is detectable in blood vessels.
Aorta
Faint lacZ expression is detectable in the wall of the aorta.
Lung
LacZ expression is detectable in smooth muscle cells of blood vessels and
bronchioles.
Gallbladder
Scattered lacZ expression is detectable in the wall of the gallbladder.
Kidney
Strong lacZ expression is detectable in tubule cells of the cortex and blood
vessel walls. Faint X-Gal signals are detectable in glomeruli. In medulla
scattered X-Gal staining is observed.
Urinary Bladder
LacZ expression is detectable in smooth muscle cells of the muscularis, in
epithelial cells of the mucosa and in blood vessel walls.
Trachea
Faint lacZ expression is detectable in myocytes.
Parathyroid Gland
Practically all cells express lacZ.
Pituitary Gland
LacZ expression is detectable in pars distalis, pars intermedia and pars
nervosa.
Adrenal Glands
X-Gal staining is detectable in capsule and all cells of the medulla.
Salivary Glands
Ganglia in the salivary glands display strong lacZ expression.
Tongue
LacZ expression is detectable in blood vessels.
Male Reproductive Systems
Testis
Many spermatogenic cells in the seminiferous tubules express lacZ.
Penis
Weak lacZ expression is detectable in myocytes.
Seminal Vesicles
Myocytes in the capsule express lacZ strongly.
Coagulating Glands
Myocytes in the capsule express lacZ strongly.
Prostate and Ampullary Glands
Myocytes in the capsule express lacZ.
Female Reproductive Systems
Ovary
Few follicles show faint X-Gal staining.
Oviduct/Uterus
Many myocytes of the myometrium display X-Gal staining. Very faint signals are
detectable in blood vessels.
Vagina/Cervix
LacZ expression is detectable in blood vessels, ganglia and myocytes.
No Expression:
LacZ expression is not detected in: heart thymus, spleen, lymph nodes, bone
marrow, liver, pancreas, larynx, esophagus, thyroid gland, skeletal muscle and
skin.
:
Gene 898
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:
2 homozygous mutant females (99075, 105945)
2 homozygous mutant males (105894, 105895)
2 wild-type control females (99084, 105944)
2 wild-type control males (105897, 105930)
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. They are not considered to be genotype related.
Gene
898
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:
3 homozygous mutant females (97286, 99072, 99074)
3 homozygous mutant males (99081, 99085, 99086)
1 heterozygous mutant female (97285)
1 heterozygous mutant male (97287)
2 wild-type control females (97282, 97283)
2 wild-type control males (99078, 99079)
300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (99075, 105945)
2 homozygous mutant males (105894, 105895)
2 wild-type control females (99084, 105944)
2 wild-type control males (105897, 105930)
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.
Certain histopathological lesions are present that occasionally occur
spontaneously in mice of this age group. In this target, lymphoid depletion
accompanied by moderately increased apoptosis of thymic lymphocytes is present
in one 49 day cohort homozygous male mouse (99085). We are not reporting this
finding as a phenotypic change, but we present it here for your consideration.
Incidental lesions may have been present in some tissues. For example, fat
necrosis in one 300 day cohort wild-type male (105930) correlated with the the
presence of a mesenteric mass. 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
898
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:
3 homozygous mutant females (97286, 99072, 99074)
3 homozygous mutant males (99081, 99085, 99086)
1 heterozygous mutant female (97285)
1 heterozygous mutant male (97287)
2 wild-type control females (97282, 97283)
2 wild-type control males (99078, 99079)
300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (99075, 105945)
2 homozygous mutant males (105894, 105895)
2 wild-type control females (99084, 105944)
2 wild-type control males (105897, 105930)
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 other 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 mesenteric mass in one 300 day cohort wild-type male
(105930) correlated with the presence of fat necrosis. 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:
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
898
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 (97286, 99072, 99074)
3 homozygous mutant males (99081, 99086, 109507)
1 heterozygous mutant female (109510)
1 heterozygous mutant male (97287)
2 wild-type control females (97282, 109511)
2 wild-type control males (99078, 99079)
90 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (99075, 105945, 111852)
3 homozygous mutant males (105894, 105896, 111774)
3 wild-type control females (99084, 111793, 111797)
5 wild-type control males (105897, 105930, 111775, 111826, 164505)
180 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (99075, 105945, 111852)
2 homozygous mutant males (105894, 105895)
1 wild-type control female (99084)
2 wild-type control males (105897, 105930)
300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (99075, 105945)
2 homozygous mutant males (105894, 105895)
2 wild-type control females (105944, 111796)
2 wild-type control males (105897, 105930)
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
898
Hematology
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.
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:
2 homozygous mutant females (97286, 194020)
4 homozygous mutant males (99081, 99085, 99086, 109508)
1 heterozygous mutant female (109512)
1 heterozygous mutant male (97287)
1 wild-type control female (97282)
2 wild-type control males (99078, 99079)
90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (99075, 105945, 111805, 111852)
4 homozygous mutant males (105894, 105895, 105896, 111774)
4 wild-type control females (99084, 105944, 111793, 111795)
5 wild-type control males (105897, 105930, 111775, 111826, 164505)
180 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (99075, 105945, 111852)
3 homozygous mutant males (105894, 105895, 111774)
4 wild-type control females (99084, 105944, 111793, 111795)
4 wild-type control males (105897, 105930, 111775, 111826)
300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (105908, 105945)
2 homozygous mutant males (105894, 105895)
2 wild-type control females (99084, 105944)
1 wild-type control male (105897)
Although minor variations of hematological values were present in some animals,
these changes were not consistent with genotype and, thus, were not considered
phenotypically relevant.
Gene
898
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:
3 homozygous mutant females (97286, 99072, 99074)
3 homozygous mutant males (99081, 99085, 99086)
1 heterozygous mutant female (97285)
1 heterozygous mutant male (97287)
2 wild-type control females (97282, 97283)
2 wild-type control males (99078, 99079)
300 Day Cohort Mouse ID numbers are as follows:
2 homozygous mutant females (99075, 105945)
2 homozygous mutant males (105894, 105895)
2 wild-type control females (99084, 105944)
2 wild-type control males (105897, 105930)
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 898
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 (99075, 105945, 111805, 111852)
4 homozygous mutant males (105894, 105895, 105896, 111774)
4 wild-type control females (99084, 105944, 111793, 111795)
4 wild-type control males (105897, 111775, 111826, 164505)
90 Day Cohort Mouse ID numbers are as follows:
4 homozygous mutant females (99075, 105945, 111805, 111852)
4 homozygous mutant males (105894, 105895, 105896, 111774)
4 wild-type control females (99084, 105944, 111793, 111795)
5 wild-type control males (105897, 105930, 111775, 111826, 164505)
180 Day Cohort Mouse ID numbers are as follows:
3 homozygous mutant females (99075, 105945, 111852)
3 homozygous mutant males (105894, 105895, 111774)
4 wild-type control females (99084, 105944, 111793, 111795)
4 wild-type control males (105897, 105930, 111775, 111826)
300 Day Cohort Mouse ID numbers are as follows:
1 homozygous mutant female (111852)
1 homozygous mutant male (111774)
2 wild-type control females (111793, 111795)
1 wild-type control male (111826)
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.