Homoeobox gene Hox-1.5 expression in mouse embryos: earliest detection by in situ hybridization is during gastrulation

Development ◽  
1987 ◽  
Vol 101 (1) ◽  
pp. 51-60 ◽  
Author(s):  
S.J. Gaunt

We showed earlier (Gaunt, Miller, Powell & Duboule, 1986) that the mouse homoeobox gene Hox-1.5 is expressed in posterior ectoderm and mesoderm of 7 1/2- and 7 3/4-day embryos, and in the 9 1/2-day nervous system posterior to a discrete boundary within the hindbrain. In further in situ hybridization experiments, it is now shown that restriction of Hox-1.5 expression to the posterior regions of the embryo can be detected at stages of development between 7 1/2 and 9 1/2 days. During this period, the intensity of transcription in presomitic and somitic mesoderm declines relative to that in the overlying neural ectoderm, and the transcription boundary within the presumptive hindbrain region sharpens. Hox-1.5 expression posterior to the hindbrain boundary is detected in the 10 1/2- and 12 1/2-day embryo, but this is no longer found in newborn mice. Embryos of ages 3 1/2, 6 1/2 and 7 1/4 days showed no evidence of Hox-1.5 transcripts. It is concluded that embryos undergoing gastrulation (at 7 1/2 days) are the earliest stage at which Hox-1.5 transcripts can be detected by the in situ hybridization technique. In discussion, it is shown how this lies within the period of development during which tissues become determined along the anteroposterior axis of the mouse. Since there may be anterior-to-posterior variation in the time of determination along the body axis, it is suggested that homoeobox genes expressed more posteriorly, such as Hox-3 (Awgulewitsch et al. 1986), might start expression at times later in development.

2003 ◽  
Vol 77 (7) ◽  
pp. 4383-4388 ◽  
Author(s):  
Brian P. Schlitt ◽  
Matthew Felrice ◽  
Mary Lou Jelachich ◽  
Howard L. Lipton

ABSTRACT Theiler's murine encephalomyelitis virus (TMEV) persists in the mouse central nervous system principally in macrophages, and infected macrophages in culture undergo apoptosis. We have detected abundant apoptotic cells in perivascular cuffs and inflammatory, demyelinating lesions of SJL mice chronically infected with TMEV. T cells comprised 74% of apoptotic cells, while 8% were macrophages, 0.6% were astrocytes, and ∼17% remained unidentified. In situ hybridization revealed viral RNA in ∼1% of apoptotic cells.


Foods ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1502
Author(s):  
Jorge García-Hernández ◽  
Manuel Hernández ◽  
Yolanda Moreno

Vibrio parahaemolyticus is a human food-borne pathogen with the ability to enter the food chain. It is able to acquire a viable, non-cultivable state (VBNC), which is not detected by traditional methods. The combination of the direct viable count method and a fluorescent in situ hybridization technique (DVC-FISH) makes it possible to detect microorganisms that can present VBNC forms in complex samples The optimization of the in vitro DVC-FISH technique for V. parahaemolyticus was carried out. The selected antibiotic was ciprofloxacin at a concentration of 0.75 μg/mL with an incubation time in DVC broth of 5 h. The DVC-FISH technique and the traditional plate culture were applied to detect and quantify the viable cells of the affected pathogen in artificially contaminated food matrices at different temperatures. The results obtained showed that low temperatures produced an important logarithmic decrease of V. parahaemolyticus, while at 22 °C, it proliferated rapidly. The DVC-FISH technique proved to be a useful tool for the detection and quantification of V. parahaemolyticus in the two seafood matrices of oysters and mussels. This is the first study in which this technique has been developed to detect viable cells for this microorganism.


1986 ◽  
pp. 181-191 ◽  
Author(s):  
J. Stephen Fink ◽  
Marc R. Montminy ◽  
Toshihiko Tsukada ◽  
Heinz Hoefler ◽  
Linda A. Specht ◽  
...  

1999 ◽  
Vol 39 (7) ◽  
pp. 41-47 ◽  
Author(s):  
Satoshi Okabe ◽  
Hisashi Satoh ◽  
Tsukasa Itoh ◽  
Yoshimasa Watanabe

The vertical distribution of sulfate-reducing bacteria (SRB) in microaerophilic wastewater biofilms grown on fully submerged rotating disk reactors (RDR) was determined by the conventional culture-dependent MPN method and in situ hybridization of fluorescently-labelled 16S rRNA-targeted oligonucleotide probes for SRB in parallel. Chemical concentration profiles within the biofilm were also measured using microelectrodes for O2, S2-, NO3- and pH. In situ hybridization revealed that the SRB probe-stained cells were distributed throughout the biofilm even in the oxic surface zone in all states from single scattered cells to clustered cells. The higher fluorescence intensity and abundance of SRB probe-stained cells were found in the middle part of the biofilm. This result corresponded well with O2 and H2S concentration profiles measured by microelectrodes, showing sulfate reduction was restricted to a narrow anaerobic zone located about 500 μm below the biofilm surface. Results of the MPN and potential sulfate reducing activity (culture-dependent approaches) indicated a similar distribution of cultivable SRB in the biofilm. The majority of the general SRB probe-stained cells were hybridized with SRB 660 probe, suggesting that one important member of the SRB in the wastewater biofilm could be the genus Desulfobulbus. An addition of nitrate forced the sulfate reduction zone deeper in the biofilm and reduced the specific sulfate reduction rate as well. The sulfate reduction zone was consequently separated from O2 and NO3- respiration zones. Anaerobic H2S oxidation with NO3- was also induced by addition of nitrate to the medium.


Author(s):  
Elena Perea-Atienza ◽  
Brenda Gavilán ◽  
Simon G. Sprecher ◽  
Pedro Martinez

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