Fluorescence In Situ Hybridization on Early Porcine Embryos

Foster, Helen A.; Sturmey, Roger G.; Stokes, Paula J.; Leese, Henry J.; Bridger, Joanna M.; Griffin, Darren K.

doi:10.1007/978-1-60761-789-1_33

Cited by 7 publications

(3 citation statements)

References 22 publications

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“…One milliliter of the supernatant was filtered through polycarbonate filters of 0.22 μm pore size (Isopore Membrane Filters, Merck Millipore Ltd). Bacteria remained on the filters were fixed, stored, and hybridized as described in [ 13 ] with slight modification. The dehydrated samples were double hybridized with two probes labeled with different fluorophores for 3 h at 46°C and washed for 15 min at 48°C.…”

Section: Methodsmentioning

confidence: 99%

Bacterial Community Associated with the Intestinal Tract of Chinese Mitten Crab (Eriocheir sinensis) Farmed in Lake Tai, China

Chen

Wang

et al. 2015

PLoS ONE

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Chinese mitten crab (CMC, Eriocheir sinensis) is an economically valuable species in South-East Asia that has been widely farmed in China. Characterization of the intestinal bacterial diversity of CMC will provide insights into the aquaculturing of CMCs. Based on the analysis of cloned 16S rRNA genes from culture-independent CMC gut bacteria, 124 out of 128 different clones reveal ＞95% nucleotide similarity to the species belonging to the four phyla of Tenericutes, Bacteroidetes, Firmicutes and Proteobacteria; one clone shows 91% sequence similarity to the member of TM7 (a candidate phylum without cultured representatives). Fluorescent in situ hybridization also reveals the abundance of Bacteroidetes in crab intestine. Electron micrographs show that spherical and filamentous bacteria are closely associated with the microvillus brush border of the midgut epithelium and are often inserted into the space between the microvilli using a stalk-like cell appendage. In contrast, the predominant rod-shaped bacteria in the hindgut are tightly attached to the epithelium surface by an unusual pili-like structure. Both 16S rRNA gene denaturing gel gradient electrophoresis and metagenome library indicate that the CMC Mollicutes group 2 appears to be present in both the midgut and hindgut with no significant difference in abundance. The CMC Mollicutes group 1, however, was found mostly in the midgut of CMCs. The CMC gut Mollicutes phylotypes appear to be most closely related to Mollicutes symbionts detected in the gut of isopods (Crustacea: Isopoda). Overall, the results suggest that CMCs harbor diverse, novel and specific gut bacteria, which are likely to live in close relationships with the CMC host.

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Section: Methodsmentioning

confidence: 99%

Bacterial Community Associated with the Intestinal Tract of Chinese Mitten Crab (Eriocheir sinensis) Farmed in Lake Tai, China

Chen

Wang

et al. 2015

PLoS ONE

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“…Another molecular cytogenetic technique, primed in situ DNA synthesis (PRINS), has been applied to pig's genome to visualize the interstitial telomeric signal in the genome. It is an attractive complement to FISH for detection of DNA repetitive sequences and unlike conventional FISH, it displays lower level of non-specific hybridization [15,21,24,25]. In the field of in vitro embryo production and other reproductive biotechnologies, cytogenetics in collaboration with other aspects of molecular biology are expected to play vital role in understanding the mechanisms underlying chromosome instability in embryos and the impact of the in vitro environment on embryo's chromosomes [26,27] Researchers have also been working to optimize the hybridization of molecular probes specific to the X chromosomes in mare.…”

Section: Cytogenetics and Domestic Animal Studiesmentioning

confidence: 99%

Application of Veterinary Cytogenetics in Domestic Animals: A Review

Yahaya

Salisi

Isa

et al. 2019

ARRB

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Cytogenetics is the study of chromosomes; their structure and properties, chromosome behavior during cell division, their influence on traits and factors which cause changes in chromosomes. Veterinary cytogenetics is the application of cytogenetics to clinical problems that occur in animal production. It has been applied to understand problems such as infertility and its types, embryonic and fetal death, abnormality in sexual and somatic development and hybrid sterility and also prenatal sex determination and other forms of chromosomal abnormalities. These are achieved through conventional and banded karyotyping techniques and molecular cytogenetic techniques. Although conventional techniques are still useful and very widely applied, the nature of cytogenetics has gradually changed as a result of advances achieved in the molecular cytogenetic techniques for example fluorescent in situ hybridization and array-based techniques. These changes are evident in both molecular diagnostics and basic research. The combination of conventional and molecular cytogenetics has given rise to high resolution techniques which have enabled the study of fundamental questions regarding biological processes. It enables the study of inherited syndromes, the mechanisms of tumorigenesis at molecular level, genome organization and the determination of chromosome homologies between species. It allows the ease with which animals are selected in breeding programs and other important aspects of animal production. In this paper we discussed a number of techniques employed in cytogenetics and their methodologies, and recommend where future focus should be for the benefits of animal production.

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“…Fluorescence in situ hybridization (FISH, Figure 8) is a well-established technique for chromosome analysis and aneuploidy screening (Foster et al, 2010;Lechniak et al, 2007). In contrast to previously described methods, FISH data can be obtained from interphase nuclei or at least in cases where not all chromosomes are separated during spreading.…”

Section: Fluorescence In Situ Hybridization (Fish)mentioning

confidence: 99%