Localization of cells from the winter flounder gill expressing a skin type antifreeze protein gene

2002 ◽  
Vol 80 (1) ◽  
pp. 110-119 ◽  
Author(s):  
Harry M Murray ◽  
Choy L Hew ◽  
Ken R Kao ◽  
Garth L Fletcher
Keyword(s):  
In Situ Hybridization ◽  
Antifreeze Protein ◽  
Tissue Expression ◽  
Cross Reactivity ◽  
Winter Flounder ◽  
Skin Type ◽  
General Distribution ◽  
Pavement Cells ◽  
Peripheral Tissues

In situ hybridization on whole mounts and paraffin-sectioned winter flounder (Pleuronectes americanus) gill, using riboprobes specific to a skin type antifreeze protein (AFP) gene, showed a mRNA distribution associated with cells throughout the filament and the lamellae. Immunohistochemistry using antibodies for a skin-type AFP identified cells corresponding to those detected using in situ hybridization. Parallel experiments with antibodies for chloride-cell markers showed that these cells were not involved in antifreeze-protein expression. Similarly, goblet cells did not show cross-reactivity with the AFP antibodies. This general distribution suggested that pavement cells were likely involved. Reverse transcription polymerase chain reaction using gill cDNA templates and subsequent sequencing of the products confirmed the presence of skin type AFP transcripts in this tissue. Expression of a AFP in this area may act as a first line of defence against ice-crystal migration into peripheral tissues.

A low molecular weight peptide from snow mold with epitopic homology to the winter flounder antifreeze protein

1994 ◽  
Vol 72 (3-4) ◽  
pp. 152-156 ◽  
Author(s):  
W. Jay Newsted ◽  
Sandra Polvi ◽  
Bob Papish ◽  
Ed Kendall ◽  
Mohammed Saleem ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Antifreeze Protein ◽  
Cross Reactivity ◽  
Winter Flounder ◽  
Low Molecular Weight ◽  
Culture Temperature ◽  
Snow Mold ◽  
Typhula Incarnata ◽  
Magnetic Resonance Microimaging ◽  
Nuclear Magnetic Resonance Microimaging

Evidence for a small size protein (ca. 3500 kDa) exhibiting epitopic homology to the Atlantic winter flounder antifreeze protein (AFP) is found in the snow molds Coprinus psychromorbidus, Myriosclerotinia borealis, and Typhula incarnata. The protein shows strong cross-reactivity with antisera specific for the flounder AFP. Preliminary studies suggest that the protein is synthesized in response to lowering the culture temperature, and that it is membrane associated and, therefore, may function in an analogous capacity to the fish AFP. Also, the protein is shown to have antifreeze properties as determined by nuclear magnetic resonance microimaging experiments.Key words: antifreeze proteins, low temperature stress, snow mold, winter flounder.

scholarly journals Tissue Expression of the S100 Protein Family-related MRP8 Gene in Human Chorionic Villi by in situ Hybridization Techniques

1999 ◽  
Vol 76 (2-3) ◽  
pp. 123-129 ◽  
Author(s):  
Noriaki SATO ◽  
Kazuhiro ISONO ◽  
Isamu ISHIWATA ◽  
Mitsuo NAKAI ◽  
Koji KAMI
Keyword(s):  
In Situ Hybridization ◽  
S100 Protein ◽  
Tissue Expression ◽  
Protein Family ◽  
Chorionic Villi

scholarly journals Diurnal Rhythmicity of the Clock Genes Per1 and Per2 in the Rat Ovary

Endocrinology ◽  
2006 ◽  
Vol 147 (8) ◽  
pp. 3769-3776 ◽  
Author(s):  
Jan Fahrenkrug ◽  
Birgitte Georg ◽  
Jens Hannibal ◽  
Peter Hindersson ◽  
Søren Gräs
Keyword(s):  
In Situ Hybridization ◽  
Circadian Clock ◽  
Estrous Cycle ◽  
Clock Genes ◽  
Continuous Darkness ◽  
Peripheral Tissues ◽  
Hybridization Histochemistry ◽  
In Situ Hybridization Histochemistry ◽  
Rat Ovary

Circadian rhythms are generated by endogenous clocks in the central brain oscillator, the suprachiasmatic nucleus, and peripheral tissues. The molecular basis for the circadian clock consists of a number of genes and proteins that form transcriptional/translational feedback loops. In the mammalian gonads, clock genes have been reported in the testes, but the expression pattern is developmental rather than circadian. Here we investigated the daily expression of the two core clock genes, Per1 and Per2, in the rat ovary using real-time RT-PCR, in situ hybridization histochemistry, and immunohistochemistry. Both Per1 and Per2 mRNA displayed a statistically significant rhythmic oscillation in the ovary with a period of 24 h in: 1) a group of rats during proestrus and estrus under 12-h light,12-h dark cycles; 2) a second group of rats representing a mixture of all 4 d of the estrous cycle under 12-h light,12-h dark conditions; and 3) a third group of rats representing a mixture of all 4 d of estrous cycle during continuous darkness. Per1 mRNA was low at Zeitgeber time 0–2 and peaked at Zeitgeber time 12–14, whereas Per2 mRNA was delayed by approximately 4 h relative to Per1. By in situ hybridization histochemistry, Per mRNAs were localized to steroidogenic cells in preantral, antral, and preovulatory follicles; corpora lutea; and interstitial glandular tissue. With newly developed antisera, we substantiated the expression of Per1 and Per2 in these cells by single/double immunohistochemistry. Furthermore, we visualized the temporal intracellular movements of PER1 and PER2 proteins. These findings suggest the existence of an ovarian circadian clock, which may play a role both locally and in the hypothalamo-pituitary-ovarian axis.

scholarly journals Identification of avian alpha-melanocyte-stimulating hormone in the eye: temporal and spatial regulation of expression in the developing chicken

2001 ◽  
Vol 168 (3) ◽  
pp. 527-537 ◽  
Author(s):  
K Teshigawara ◽  
S Takahashi ◽  
T Boswell ◽  
Q Li ◽  
S Tanaka ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Neural Retina ◽  
Melanocortin Receptor ◽  
Receptor Subtypes ◽  
Peripheral Tissues ◽  
Rpe Cells ◽  
Melanocyte Stimulating Hormone ◽  
Cone Cells ◽  
Stimulating Hormone

The presence and possible physiological roles of alpha-melanocyte-stimulating hormone (alpha-MSH) in the peripheral tissues of birds have not been established. By a combination of RT-PCR, immunocytochemistry and in situ hybridization, we have examined alpha-MSH expression in the eye of the chicken during development. In the 1-day-old chick, alpha-MSH was expressed in the retinal pigment epithelial (RPE) cells, and also at a lower level in the cone cells. The melanocortin receptor subtypes, CMC1, CMC4 and CMC5, were expressed in the layers of the choroid and the neural retina, but not in the RPE cells. It is probable that the RPE cells secrete alpha-MSH to exert paracrine effects on the choroid and neural retina. During embryonic development, alpha-MSH immunoreactivity in the RPE cells was initially detected at embryonic day 10, and increased in intensity as development proceeded. No cone cells were stained with anti-alpha-MSH antiserum in any of the embryonic stages tested. The immunoreactivities for two prohormone convertases, PC1 and PC2, were co-localized to the RPE cells with a pattern of staining similar to that of alpha-MSH. Despite containing alpha-MSH immunoreactivity, the RPE cells in 1-day-old chicks expressed no immunoreactivity for the endoproteases. Furthermore, in a 3-day-old chick, pro-opiomelanocortin mRNA was detectable by in situ hybridization only in the photoreceptor layer and not in the RPE cells. These results suggest that the RPE cells and the cone cells are intraocular sources of alpha-MSH in the embryonic and postnatal life of the chicken respectively. Embryonic expression of alpha-MSH in the RPE cells implies a possible role for the peptide in ocular development.

scholarly journals Human eosinophils express transforming growth factor alpha.

1990 ◽  
Vol 172 (3) ◽  
pp. 673-681 ◽  
Author(s):  
D T Wong ◽  
P F Weller ◽  
S J Galli ◽  
A Elovic ◽  
T H Rand ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Growth Factor ◽  
Transforming Growth Factor ◽  
Biological Effects ◽  
Hypereosinophilic Syndrome ◽  
Enzyme Analysis ◽  
Transforming Growth Factor Alpha ◽  
Peripheral Tissues ◽  
Factor Alpha

Transforming growth factor alpha (TGF-alpha) is a pleuripotential cytokine with diverse biological effects, including the ability to influence the proliferation of normal cells or neoplastic epithelial cells. Eosinophils are a subset of granulocytes that normally enter the peripheral tissues, particularly those beneath gastrointestinal, respiratory, and urogenital epithelium, where they reside in close proximity to the epithelial elements. In this study, we demonstrate that the great majority of eosinophils infiltrating the interstitial tissues adjacent to two colonic adenocarcinomas and two oral squamous cell carcinomas labeled specifically by in situ hybridization with a 35S-riboprobe for human TGF-alpha (hTGF-alpha). No other identifiable leukocytes in these lesions contained detectable hTGF-alpha mRNA. We also examined leukocytes purified from a patient with the idiopathic hypereosinophilic syndrome. 80% of these eosinophils, but none of the patient's neutrophils or mononuclear cells, were positive for hTGF-alpha mRNA by in situ hybridization, and 55% of these eosinophils were positive by immunohistochemistry with a monoclonal antibody directed against the COOH terminus of the mature hTGF-alpha peptide. Finally, the identification of the purified eosinophil-associated transcript as hTGF-alpha was confirmed by polymerase chain reaction product restriction enzyme analysis followed by Southern blot hybridization. In contrast to eosinophils from the patient with hypereosinophilic syndrome, the peripheral blood eosinophils from only two of seven normal donors had detectable TGF-alpha mRNA and none of these eosinophils contained immunohistochemically detectable TGF-alpha product. Taken together, these findings establish that human eosinophils can express TGF-alpha, but suggest that the expression of TGF-alpha by eosinophils may be under microenvironmental regulation. Demonstration of TGF-alpha production by tissue-infiltrating eosinophils and the eosinophils in the hypereosinophilic syndrome identifies a novel mechanism by which eosinophils might contribute to physiological, immunological, and pathological responses.

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