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Cited by 144Genetic and Clinical Aspects of Sex Determination and Chromosomal AbnormalitiesSexual Differentiation and DisordersSperm and Testicular Function

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Four Core Genotypes mouse model: localization of the Sry transgene and bioassay for testicular hormone levels

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Itoh, Yuichiro, Mackie, Ryan, Kampf, Kathy, Domadia, Shelly, Brown, Judith D, O’Neill, Rachel, Arnold, Arthur P·2015·BMC Research Notes
DOI 10.1186/s13104-015-0986-2PMC PMC4354741
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Abstract

BACKGROUND: The "four core genotypes" (FCG) mouse model has emerged as a major model testing if sex differences in phenotypes are caused by sex chromosome complement (XX vs. XY) or gonadal hormones or both. The model involves deletion of the testis-determining gene Sry from the Y chromosome and insertion of an Sry transgene onto an autosome. It produces XX and XY mice with testes, and XX and XY mice with ovaries, so that XX and XY mice with the same type of gonad can be compared to assess phenotypic effects of sex chromosome complement in cells and tissues. FINDINGS: We used PCR to amplify the Sry transgene and adjacent genomic sequences, to resolve the location of the Sry transgene to chromosome 3 and confirmed this location by fluorescence in situ hybridization (FISH) of the Sry construct to metaphase chromosomes. Using quantitative PCR, we estimate that 12-14 copies of the transgene were inserted. The anogenital distance (AGD) of FCG pups at 27-29 days after birth was not different in XX vs. XY males, or XX vs. XY females, suggesting that differences between XX and XY mice with the same type of gonad are not caused by difference in prenatal androgen levels. CONCLUSION: The Sry transgene in FCG mice is present in multiple copies at one locus on chromosome 3, which does not interrupt known genes. XX and XY mice with the same type of gonad do not show evidence of different androgen levels prenatally.

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Figure 1 Location of the Sry transgene in four core genotypes mouse model. Samples were from XX and XY mice with testes (XXSry, XY− Sry) and XX and XY mice with ovaries (XX, XY−). Confirmation of transgene-genome boundary by PCR using transgene-specific primer d and Chr3 primer e (A) , with transgene-specific primer c and Chr3 primer b (B), and with transgene-specific primer c and Chr3 primer a (C). (D) The concatemer of Sry transgene is inserted into a repetitive sequence present genome-wide. (E) Sry transgene location on chromosome 3. (F) A visual estimate of the difference in copy number of Sry between wildtype and XY− Sry genomic DNA in agarose gel.
Figure 1 Location of the Sry transgene in four core genotypes mouse model. Samples were from XX and XY mice with testes (XXSry, XY− Sry) and XX and XY mice with ovaries (XX, XY−). Confirmation of transgene-genome boundary by PCR using transgene-specific primer d and Chr3 primer e (A) , with transgene-specific primer c and Chr3 primer b (B), and with transgene-specific primer c and Chr3 primer a (C). (D) The concatemer of Sry transgene is inserted into a repetitive sequence present genome-wide. (E) Sry transgene location on chromosome 3. (F) A visual estimate of the difference in copy number of Sry between wildtype and XY− Sry genomic DNA in agarose gel.CC BY · PubMed Central
Figure 2 FISH mapping of the Sry transgene to Chromosome 3. The Sry transgene signal (A, red) and chromosome Chromosome 3 paint (B, green) hybridize to the same metaphase chromosome. (C) Sry transgene hybridization in three additional metaphase cells demonstrating its location with respect to the p- and q-arm ends.
Figure 2 FISH mapping of the Sry transgene to Chromosome 3. The Sry transgene signal (A, red) and chromosome Chromosome 3 paint (B, green) hybridize to the same metaphase chromosome. (C) Sry transgene hybridization in three additional metaphase cells demonstrating its location with respect to the p- and q-arm ends.CC BY · PubMed Central
Figure 3 Anogenital distance measurement of FCG mice. Asterisk shows the significant effect of sex (two way ANOVA, p < 0.00001).
Figure 3 Anogenital distance measurement of FCG mice. Asterisk shows the significant effect of sex (two way ANOVA, p < 0.00001).CC BY · PubMed Central

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Metadata

Authors
Itoh, Y., Mackie, R., Kampf, K., Domadia, S., Brown, J. D., O’Neill, R., & Arnold, A. P.
Affiliations
  • University of California, Los Angeles (US)
  • University of Connecticut (US)
Corresponding author
Arnold, Arthur P
ORCID
Venue
BMC Research Notes
Publisher
Springer Science and Business Media LLC
Published
2015-03-07
Volume
8
Issue
1
Pages
69-69
ISSN
1756-0500 (ISSN-L)
DOI
10.1186/s13104-015-0986-2
PMC
PMC4354741
OpenAlex
W2000906752
Cited by
109
References
81
Citations by year
2026: 8 · 2025: 9 · 2024: 22 · 2023: 6 · 2022: 14 · 2021: 6 · 2020: 12 · 2019: 6 · 2018: 8 · 2017: 10 · 2016: 6 · 2015: 2
FWCI
5.96
Type
journal article
Language
en
Open access
Licence
Locations
  • BMC Research Notes (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Science Foundation
  • National Institutes of Health (DK 083561)
  • National Institute of Diabetes and Digestive and Kidney Diseases (R56DK083561, R01DK083561, DK083561)
  • National Institute of Neurological Disorders and Stroke (NS 043196, R01NS043196)
  • Eunice Kennedy Shriver National Institute of Child Health and Human Development (R01HD076125, HD076125)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic and Clinical Aspects of Sex Determination and Chromosomal Abnormalities
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Sexual Differentiation and Disorders
  • Health Sciences › Medicine › Reproductive Medicine › Sperm and Testicular Function
Keywords
testis-determining gene, sex chromosome complement, gonadal hormones, anogenital distance, transgene copy number, chromosome 3, sex differences in phenotype
MeSH terms
Androgens, Animals, Biological Assay, Female, Genotype, Male, Mice, Transgenic, Ovary, Phenotype, Sex Characteristics, Testis, X Chromosome, Y Chromosome, In Situ Hybridization, Fluorescence, Gene Dosage, Transgenes, Genes, sry, Mice

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