BackgroundThe “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.FindingsWe 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.ConclusionThe 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.

Epigenetic gene silencing by histone modifications and DNA methylation is essential for cancer development. The molecular mechanism that promotes selective epigenetic changes during tumorigenesis is not understood. We report here that the PIAS1 SUMO ligase is involved in the progression of breast tumorigenesis. Elevated PIAS1 expression was observed in breast tumor samples. PIAS1 knockdown in breast cancer cells reduced the subpopulation of tumor-initiating cells, and inhibited breast tumor growth in vivo. PIAS1 acts by delineating histone modifications and DNA methylation to silence the expression of a subset of clinically relevant genes, including breast cancer DNA methylation signature genes such as cyclin D2 and estrogen receptor, and breast tumor suppressor WNT5A. Our studies identify a novel epigenetic mechanism that regulates breast tumorigenesis through selective gene silencing.

The selective and temporal DNA methylation plays an important role in the self-renewal and differentiation of hematopoietic stem cells (HSCs), but the molecular mechanism that controls the dynamics of DNA methylation is not understood. Here, we report that the PIAS1 epigenetic pathway plays an important role in regulating HSC self-renewal and differentiation. PIAS1 is required for maintaining the quiescence of dormant HSCs and the long-term repopulating capacity of HSC. Pias1 disruption caused the abnormal expression of lineage-associated genes. Bisulfite sequencing analysis revealed the premature promoter demethylation of Gata1, a key myeloerythroid transcription factor and a PIAS1-target gene, in Pias1 À/À HSCs. As a result, Pias1 disruption caused the inappropriate induction of Gata1 in HSCs and common lymphoid progenitors (CLPs). The expression of other myeloerythroid genes was also enhanced in CLPs and lineage-negative progenitors, with a concurrent repression of B cell-specific genes. Consistently, Pias1 disruption caused enhanced myeloerythroid, but reduced B lymphoid lineage differentiation. These results identify a novel role of PIAS1 in maintaining the quiescence of dormant HSCs and in the epigenetic repression of the myeloerythroid program.

An experiment lasting 42 d was performed in 4 consecutive stages on 6 healthy Friesian cows during mid-lactation. Mean values for the different components of mammary secretion during normal lactation were established. Milking was then suspended on all quarters from d 1-14. The mean values for lactate increased 20-to 30-fold over the mean value for normal lactation. Over the same period the leucocyte count (polymorphonuclear leucocytes and lymphocytes) also increased, whereas the mean values for glucose and the estimated redox potential decreased. From d 15-28 milking was resumed on one half of the udder and from d 29-42 milking was also resumed on the other udder-half. When milking was resumed the above-mentioned changes were reversed, taking 5-7 d to reach values obtained during normal lactation in the udder-half which had not been milked for 14 d, but at least 14 d in the udderhalf which had not been milked for 28 d. The changes in the levels of lactate, glucose, leucocyte count and estimated redox potential are discussed in relation to changes in the secretory activity of the mammary gland (lactose, /Mactoglobulin and epithelial cell count), permeability changes of the mammary epithelium (Na + , K + and serum albumin) and the immune defence mechanism in the udder (immunoglobulins). The results indicate that lactate is formed during anaerobic glycolysis by the leucocytes in the mammary secretion, most probably due to reduced blood flow to the udder and the accumulation of secretion in the gland and teat cisterns on cessation of milking.Most studies on function and control of the bovine mammary gland have dealt with the gross composition of milk or with changes in the levels of tissue metabolites and enzymes during the onset of lactation, normal lactation and involution when changes in mammary secretory activity are greatest (Peaker, 1975;Fleet et al. 1975; Schanbacher & Smith, 1975). However, in terms of the physiology of the bovine mammary gland, the diagnosis of sub-clinical mastitis and the production of high quality milk, it is important to elucidate the phenomenon of premature regression. Premature regression is the premature degeneration of the lactating udder epithelium

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