Genetic background influences the capacity for medial edge epithelium disintegration and phenotype of cleft palate in TGFβ3 knockout mice

The present study was undertaken to determine whether the severity of renal failure or hypertension in homozygous cyclooxygenase (COX)-2-deficient (COX-2−/−) mice affected by genetic background or gender. COX-2 deletion was introduced into three congenic genetic backgrounds, 129/Sv (129/COX-2−/−), C57/BL6 (C57/COX-2−/−), and BALB/c (BALB/COX-2−/−), by backcrossing the original mixed-background knockout mice with the respective inbred strains for 9 or 10 generations. Evaluation of the severity of hypertension and renal failure was performed in knockout and wild-type mice at the age of 2.5–3.5 mo. Blood pressure measured by tail-cuff plethysmography was significantly elevated in the male 129/COX-2−/− mice (165.8 ± 9.2 vs. 116 ± 5.1 mmHg, P < 0.05), and to a much lesser extent in the female 129/COX-2−/− mice (127.4 ± 3.3 vs. 102.4 ± 3.3), whereas it was unchanged in the C57- or BALB/COX-2−/− mice regardless of gender. Urinary excretion of albumin, determined by EIA, was remarkably increased in the 129/COX-2−/− (16.4 ± 4.1 vs. 0.16 ± 0.043 mg albumin/mg creatinine, P < 0.001), and to a lesser extent in the male C57/COX-2−/− mice (0.595 ± 0.416 vs. 0.068 ± 0.019). Albumin excretion was not elevated in the male BALB/COX-2−/− or in female COX-2−/− mice on any of the three genetic backgrounds. Histological analysis showed abundant protein casts, dilated tubules, and infiltration of inflammatory cells in the male 129/COX-2−/− mice, but not in COX-2−/− mice in other strains or gender. However, the presence of small glomeruli in the nephrogenic zone was observed in all strains of COX-2 knockout mice, regardless of genetic background and gender. Therefore, we conclude that the severity of hypertension and renal failure in COX-2-deficient mice is influenced by genetic background and gender, whereas the incomplete maturation of outer cortical nephrons appears to be independent of genetic background effects.

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Kiss1 −/− Mice Exhibit More Variable Hypogonadism than Gpr54−/− Mice

Endocrinology ◽

10.1210/en.2007-0078 ◽

2007 ◽

Vol 148 (10) ◽

pp. 4927-4936 ◽

Cited By ~ 318

Author(s):

Risto Lapatto ◽

J. Carl Pallais ◽

Dongsheng Zhang ◽

Yee-Ming Chan ◽

Amy Mahan ◽

...

Keyword(s):

Sexual Maturation ◽

Knockout Mice ◽

Genetic Background ◽

Phenotypic Variability ◽

Hypogonadotropic Hypogonadism ◽

Vaginal Opening ◽

Wild Type ◽

Males And Females ◽

G Protein Coupled ◽

Estrous Cycling

The G protein-coupled receptor Gpr54 and its ligand metastin (derived from the Kiss1 gene product kisspeptin) are key gatekeepers of sexual maturation. Gpr54 knockout mice demonstrate hypogonadotropic hypogonadism, but until recently, the phenotype of Kiss1 knockout mice was unknown. This report describes the reproductive phenotypes of mice carrying targeted deletions of Kiss1 or Gpr54 on the same genetic background. Both Kiss1 and Gpr54 knockout mice are viable but infertile and have abnormal sexual maturation; the majority of males lack preputial separation, and females have delayed vaginal opening and absence of estrous cycling. Kiss1 and Gpr54 knockout males have significantly smaller testes compared with controls. Gpr54 knockout females have smaller ovaries and uteri than wild-type females. However, Kiss1 knockout females demonstrate two distinct phenotypes: half have markedly reduced gonadal weights similar to those of Gpr54 knockout mice, whereas half exhibit persistent vaginal cornification and have gonadal weights comparable with those of wild-type females. FSH levels in both Kiss1 and Gpr54 knockout males and females are significantly lower than in controls. When injected with mouse metastin 43–52, a Gpr54 agonist, Gpr54 knockout mice fail to increase gonadotropins, whereas Kiss1 knockout mice respond with increased gonadotropin levels. In summary, both Kiss1 and Gpr54 knockout mice have abnormal sexual maturation consistent with hypogonadotropic hypogonadism, although Kiss1 knockout mice appear to be less severely affected than their receptor counterparts. Kiss1 knockout females demonstrate a bimodal phenotypic variability, with some animals having higher gonadal weight, larger vaginal opening, and persistent vaginal cornification.

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Pathogenesis of cleft palate in TGF-beta3 knockout mice

Development ◽

10.1242/dev.126.17.3869 ◽

1999 ◽

Vol 126 (17) ◽

pp. 3869-3879 ◽

Cited By ~ 3

Author(s):

Y. Taya ◽

S. O'Kane ◽

M.W. Ferguson

Keyword(s):

Organ Culture ◽

Cleft Palate ◽

Transforming Growth Factor ◽

Outer Cell ◽

Complete Disappearance ◽

Surface Receptors ◽

Palatal Fusion ◽

Medial Edge

We previously reported that mutation of the transforming growth factor-beta3 (TGF-beta3) gene caused cleft palate in homozygous null (−/−) mice. TGF-beta3 is normally expressed in the medial edge epithelial (MEE) cells of the palatal shelf. In the present study, we investigated the mechanisms by which TGF-beta3 deletions caused cleft palate in 129 × CF-1 mice. For organ culture, palatal shelves were dissected from embryonic day 13.5 (E13.5) mouse embryos. Palatal shelves were placed singly or in pairs on Millipore filters and cultured in DMEM/F12 medium. Shelves were placed in homologous (+/+ vs +/+, −/− vs −/−, +/− vs +/−) or heterologous (+/+ vs −/−, +/− vs −/−, +/+ vs +/−) paired combinations and examined by macroscopy and histology. Pairs of −/− and −/− shelves failed to fuse over 72 hours of culture whereas pairs of +/+ (wild-type) and +/+ or +/− (heterozygote) and +/−, as well as +/+ and −/− shelves, fused within the first 48 hour period. Histological examination of the fused +/+ and +/+ shelves showed complete disappearance of the midline epithelial seam whereas −/− and +/+ shelves still had some seam remnants. In order to investigate the ability of TGF-beta family members to rescue the fusion between −/− and −/− palatal shelves in vitro, either recombinant human (rh) TGF-beta1, porcine (p) TGF-beta2, rh TGF-beta3, rh activin, or p inhibin was added to the medium in different concentrations at specific times and for various periods during the culture. In untreated organ culture −/− palate pairs completely failed to fuse, treatment with TGF-beta3 induced complete palatal fusion, TGF-beta1 or TGF-beta2 near normal fusion, but activin and inhibin had no effect. We investigated ultrastructural features of the surface of the MEE cells using SEM to compare TGF-beta3-null embryos (E 12. 5-E 16.5) with +/+ and +/− embryos in vivo and in vitro. Up to E13.5 and after E15.5, structures resembling short rods were observed in both +/+ and −/− embryos. Just before fusion, at E14.5, a lot of filopodia-like structures appeared on the surface of the MEE cells in +/+ embryos, however, none were observed in −/− embryos, either in vivo or in vitro. With TEM these filopodia are coated with material resembling proteoglycan. Interestingly, addition of TGF-beta3 to the culture medium which caused fusion between the −/− palatal shelves also induced the appearance of these filopodia on their MEE surfaces. TGF-beta1 and TGF-beta2 also induced filopodia on the −/− MEE but to a lesser extent than TGF-beta3 and additionally induced lamellipodia on their cell surfaces. These results suggest that TGF-beta3 may regulate palatal fusion by inducing filopodia on the outer cell membrane of the palatal medial edge epithelia prior to shelf contact. Exogenous recombinant TGF-beta3 can rescue fusion in −/− palatal shelves by inducing such filopodia, illustrating that the effects of TGF-beta3 are transduced by cell surface receptors which raises interesting potential therapeutic strategies to prevent and treat embryonic cleft palate.

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Genetic background influences tumour phenotype in heterozygous Men1 knockout mice

Endocrine Abstracts ◽

10.1530/endoabs.31.oc5.1 ◽

2013 ◽

pp. 1-1

Author(s):

Kate E Lines ◽

Mahsa Javid ◽

Anita A C Reed ◽

Sian E Piret ◽

Gerard V Walls ◽

...

Keyword(s):

Knockout Mice ◽

Genetic Background

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Genetic Background Influences Cataractogenesis, but Not Lens Growth Deficiency, in Cx50-Knockout Mice

Investigative Opthalmology & Visual Science ◽

10.1167/iovs.02-1311 ◽

2003 ◽

Vol 44 (6) ◽

pp. 2669 ◽

Cited By ~ 22

Author(s):

Dwan A. Gerido ◽

Caterina Sellitto ◽

Leping Li ◽

Thomas W. White

Keyword(s):

Knockout Mice ◽

Genetic Background ◽

Growth Deficiency ◽

Lens Growth

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Cleft Lip and/or Palate with Monogenic Autosomal Recessive Transmission in Pyrenees Shepherd Dogs

The Cleft Palate-Craniofacial Journal ◽

10.1597/06-229.1 ◽

2009 ◽

Vol 46 (1) ◽

pp. 81-88 ◽

Cited By ~ 19

Author(s):

Cordula Kemp ◽

Holger Thiele ◽

Anja Dankof ◽

Gül Schmidt ◽

Carola Lauster ◽

...

Keyword(s):

Linkage Analysis ◽

Cleft Palate ◽

Candidate Genes ◽

Autosomal Recessive ◽

Cleft Lip ◽

Pedigree Analysis ◽

Recessive Trait ◽

Medial Edge ◽

Mesenchymal Transformation ◽

Cleft Formation

Objective: To document the genetic background of Pyrenees shepherd dogs as it relates to the incidence of cleft lip and/or cleft palate, to describe the phenotype, and to determine possible candidate genes. Design: Pedigree analysis was performed and blood samples were taken from five affected pups, their siblings, and parents. Seven candidate genes were selected and linkage analysis was performed. Further methods used included sequencing and histology. Results: In 37 litters consisting of 163 pups, we found 47 affected pups in a total population of 2104. The male:female ratio was 1:0.96. Affected pups showed isolated cleft lip and/or cleft palate; no attendant disorders have been reported. Despite a high degree of relationship, two affected pups displayed a cleft palate (– H S H –) and a cleft lip with or without cleft palate (L A —) cleft formation. Histology of affected pups showed that the medial edge epithelium remained intact and did not undergo an epithelial-mesenchymal transformation. There was no evidence for linkage between the trait and TGFb3 or Msx1. Subsequent sequencing excluded the coding sequence of Fst as well. Conclusion: Pedigree analysis showed that cleft palate is not genetically distinct from cleft lip with or without cleft palate but is inherited in this breed as a monogenic autosomal recessive trait. Linkage analysis and sequencing excluded TGFb3, Msx1, and Fst as candidate genes. Histology of affected pups showed that the medial edge epithelium is still intact.

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Cleft Palate as Distinguishing Feature in a Patient with GABRB3 Epileptic Encephalopathy

Neuropediatrics ◽

10.1055/s-0039-1693143 ◽

2019 ◽

Vol 50 (06) ◽

pp. 378-381

Author(s):

Daniel Bamborschke ◽

Matthias Pergande ◽

Hülya Sevcan Daimagüler ◽

Elisabeth Mangold ◽

Jörg Dötsch ◽

...

Keyword(s):

Cleft Palate ◽

Gabaa Receptor ◽

Knockout Mice ◽

De Novo ◽

Index Patient ◽

Epileptic Encephalopathy ◽

De Novo Mutation ◽

Neonatal Onset ◽

Whole Exome ◽

Patients With Epilepsy

Mutations in GABAA-receptor subunit genes are associated with a heterogeneous spectrum of epilepsies. Patients with epilepsy caused by mutations in a specific GABAA-receptor (GABRA3) occasionally present with orofacial dysmorphism (e.g., cleft palates). While cleft palates have been described in Gabrb3 knockout mice and in humans with GABRB3 variants without epilepsy, the specific combination of epilepsy and cleft palate in humans with GABRB3 mutations has not yet been reported.We describe a patient with epileptic encephalopathy (EE) who presented with therapy-refractory neonatal-onset myoclonic seizures and severe developmental delay. Electroencephalogram showed burst suppression pattern at neonatal age and hypsarrhythmia at infantile age. Initial magnetic resonance imaging was unremarkable. As he additionally presented with a cleft palate, we were curious whether cleft palate and EE had the same genetic origin. Whole exome sequencing of the index patient revealed a novel pathogenic heterozygous de novo mutation in GABRB3 (c.899T > C; p.I300T). In consistency with Gabrb3 knockout mice data, this is the first report of cleft palate in a patient with GABRB3 associated EE.We suggest to add cleft palate to the phenotypic GABRB3 spectrum and to screen for mutations in GABAA-receptors in patients with EE and orofacial dysmorphism.

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