A new RT-PCR method for the identification of reoviruses in seawater samples

Muscillo, Michele; Rosa, Giuseppina La; Marianelli, Cinzia; Zaniratti, S.; Capobianchi, Maria Rosaria; Cantiani, L; Carducci, Annalaura

doi:10.1016/s0043-1354(00)00282-7

Cited by 38 publications

(32 citation statements)

References 21 publications

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“…2; data for reovirus types 2 and 3 are not shown). These detection sensitivities are comparable to the 7-to 70-PFU/reaction mixture RT-PCR detection sensitivities recently reported for reovirus types 1, 2, and 3 (18).…”

Section: Specificities Of the Primerssupporting

confidence: 85%

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Detection and Identification of Mammalian Reoviruses in Surface Water by Combined Cell Culture and Reverse Transcription-PCR

Spinner

Giovanni

2001

Appl Environ Microbiol

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Section: Specificities Of the Primerssupporting

confidence: 85%

“…However, the few ecological studies that have monitored the occurrence of reoviruses in water sources have found that they occur quite commonly (8,12,18,27). One study that examined secondary sewage treatment plant effluents showed that reoviruses were present in 84% of the samples and that enteroviruses were present in only 46% of the samples (8).…”

mentioning

confidence: 99%

Detection and Identification of Mammalian Reoviruses in Surface Water by Combined Cell Culture and Reverse Transcription-PCR

Spinner

Giovanni

2001

Appl Environ Microbiol

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“…PCR products were isolated from a low melting agarose gel and purified using the Wizard PCR Preps DNA purification System (Promega; Madison, WI). Their identity was confirmed to correspond to fragments from rat AQPs by automated sequencing using an ABI Prism310 sequencer (Perkin-Elmer) as described by Muscillo et al (38).…”

Section: Methodsmentioning

confidence: 99%

Regulation of the immunoexpression of aquaporin 9 by ovarian hormones in the rat oviductal epithelium

Brañes¹,

Morales²,

Ríos³

et al. 2005

American Journal of Physiology-Cell Physiology

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Brañ es, María C., Bernardo Morales, Mariana Ríos, and Manuel J. Villalón. Regulation of the immunoexpression of aquaporin 9 by ovarian hormones in the rat oviductal epithelium. Am J Physiol Cell Physiol 288: C1048 -C1057, 2005. First published January 12, 2005; doi:10.1152/ajpcell.00420.2003.-The volume of oviductal fluid fluctuates during the estrous cycle, suggesting that water availability is under hormonal control. It has been postulated that sexsteroid hormones may regulate aquaporin (AQP) channels involved in water movement across cell membranes. Using a functional assay (oocytes of Xenopus laevis), we demonstrated that the rat oviductal epithelium contains mRNAs coding for water channels, and we identified by RT-PCR the mRNAs for AQP5, -8, and -9, but not for AQP2 and -3. The immunoreactivity for AQP5, -8, and -9 was localized only in epithelial cells of the oviduct. The distribution of AQP5 and -8 was mainly cytoplasmic, whereas we confirmed, by confocal microscopy, that AQP9 localized to the apical plasma membrane. Staining of AQP5, -8, and -9 was lost after ovariectomy, and only AQP9 immunoreactivity was restored after estradiol and/or progesterone treatments. The recovery of AQP9 reactivity after ovariectomy correlated with increased mRNA and protein levels after treatment with estradiol alone or progesterone administration after estradiol priming. Interestingly, progesterone administration after progesterone priming also induced AQP9 expression but without a change in mRNA levels. Levels of AQP9 varied along the estrous cycle with their highest levels during proestrus and estrus. These results indicate that steroid hormones regulate AQP9 expression at the mRNA and protein level and that other ovarian signals are involved in the expression of AQP5 and -8. Thus hormonal regulation of the type and quantity of water channels in this epithelium might control water transport in the oviductal lumen. water channels; epithelial cells; estradiol; progesterone THE FLUID PRODUCED and secreted by epithelial cells of the oviduct provides a physiological medium for fertilization and facilitates ovum transport toward the uterus and early embryonic development. Furthermore, the quality and quantity of the oviductal fluid are modified in correlation with fluctuations of estrogen and progesterone (P 4 ) plasma levels during the estrous and menstrual cycle (30). The rate of fluid secretion increases during the estrus about 2-10 times, depending on the species, compared with the luteal phase or pregnancy (23, 30). However, the mechanism by which ovarian hormones regulate water availability from epithelial cells toward the oviductal lumen for mucus hydration is unknown.In many tissues, water channel proteins known as aquaporins (AQPs) have been implicated in transmembrane water transport (2). Eleven mammalian AQPs have been cloned, several of which have been related to physiological processes (2, 52). Moreover, specific mutations of AQPs are responsible of some human and rodent inherited diseases (27).AQPs have been doc...

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“…The thermal cycling conditions included an initial activation step at 95 8C for 25 min, followed by 40 cycles of denaturizing and annealing-amplification (95 8C for 15 s, 60 8C for 15 s and 72 8C for 30 s) and finally one cycle of melting (95-60 8C). To verify specificity of the product, amplified products were subject to melting curve analysis as well as electrophoresis, and product sequencing was performed to confirm identity as described by Muscillo et al (2001). The expression of transcripts was determined using a method previously reported (Livak & Schmittgen 2001, Parada-Bustamante et al 2010.…”

Section: Real-time Pcrmentioning

confidence: 99%

Tumour necrosis factor-α is the signal induced by mating to shutdown a 2-methoxyestradiol nongenomic action necessary to accelerate oviductal egg transport in the rat

et al. 2013

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Mating shut down a 2-methoxyestradiol (2ME) nongenomic action necessary to accelerate egg transport in the rat oviduct. Herein, we investigated whether tumour necrosis factor-α (TNF-α) participates in this mating effect. In unmated and mated rats, we determined the concentration of TNF-α in the oviductal fluid and the level of the mRNA for Tnf-a (Tnf) and their receptors Tnfrsf1a and Tnfrsf1b in the oviduct tissues. The distribution of the TNFRSF1A and TNFRSF1B proteins in the oviduct of unmated and mated was also assessed. Finally, we examined whether 2ME accelerates oviductal egg transport in unmated rats that were previously treated with a rat recombinant TNF-α alone or concomitant with a selective inhibitor of the NF-κB activity. Mating increased TNF-α in the oviductal fluid, but Tnf transcript was not detected in the oviduct. The mRNA for TNF-α receptors as well as their distribution was not affected by mating, although they were mainly localized in the endosalpinx. Administration of TNF-α into the oviduct of unmated rats prevented the effect of 2ME on egg transport. However, the NF-κB activity inhibitor did not revert this effect of TNF-α. These results indicate that mating increased TNF-α in the oviductal fluid, although this not associated with changes in the expression and localization of TNF-α receptors in the oviductal cells. Furthermore, TNF-α mimicked the effect of mating on the 2ME-induced egg transport acceleration, independently of the activation of NF-κB in the oviduct. We concluded that TNF-α is the signal induced by mating to shut down a 2ME nongenomic action in the rat oviduct.

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A new RT-PCR method for the identification of reoviruses in seawater samples

Cited by 38 publications

References 21 publications

Detection and Identification of Mammalian Reoviruses in Surface Water by Combined Cell Culture and Reverse Transcription-PCR

Detection and Identification of Mammalian Reoviruses in Surface Water by Combined Cell Culture and Reverse Transcription-PCR

Regulation of the immunoexpression of aquaporin 9 by ovarian hormones in the rat oviductal epithelium

Tumour necrosis factor-α is the signal induced by mating to shutdown a 2-methoxyestradiol nongenomic action necessary to accelerate oviductal egg transport in the rat

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