Sulfated glycoprotein-1 (SGP-1) expression in ovine endometrium during the oestrous cycle and early pregnancy

1995 ◽  
Vol 7 (5) ◽  
pp. 1053 ◽  
Author(s):  
TE Spencer ◽  
GH Graf ◽  
FW Bazer
Keyword(s):  
Early Pregnancy ◽  
Blot Hybridization ◽  
Immunohistochemical Localization ◽  
Oestrous Cycle ◽  
Northern Hybridization ◽  
Mrna Levels ◽  
Slot Blot ◽  
Slot Blot Hybridization ◽  
Endometrial Epithelium ◽  
Sulfated Glycoprotein

This study determined effects of day of oestrous cycle and early pregnancy on sulfated glycoprotein-1 (SGP-1) expression in ovine endometrium. A 364-bp clone of the ovine SGP-1 mRNA was amplified from reverse transcribed Day-15 cyclic endometrial mRNA using the polymerase chain reaction (PCR) and primers specific for the rat SGP-1 mRNA sequence. Nucleotide sequence of the ovine SGP-1 cDNA shared significant identity with rat SGP-1 and human prosaposin. Ewes (n = 40) were hysterectomized on either Day 1, 6, 11, 13 or 15 of the oestrous cycle or on Day 11, 13, 15, 17 or 25 of early pregnancy. Total cellular RNA was isolated from endometrium and subjected to Northern and slot blot hybridization analyses using an antisense cRNA probe transcribed from the ovine SGP-1 cDNA clone. A single 2.6-kb mRNA transcript was detected by Northern hybridization analyses. Slot blot hybridization analyses indicated that steady-state levels of endometrial SGP-1 mRNA varied during the oestrous cycle (cubic, P < 0.02) and increased between Day 11 and Day 25 of early pregnancy (linear, P < 0.01). On Days 11, 13 and 15, endometrial SGP-1 mRNA levels were greater in pregnant ewes than in cyclic ewes (day x pregnancy status, P < 0.01). Immunohistochemical localization of SGP-1 in uterine tissues with rabbit anti-rat SGP-1 antibody revealed intense immunoreactivity associated primarily with the endometrial epithelium. These results indicate that the ovine endometrium expresses SGP-1, a prosaposin, and that SGP-1 expression varies during the oestrous cycle and is enhanced by the conceptus. The presence of SGP-1 in the endometrium suggests intracellular and extracellular roles for this protein in glycosphingolipid metabolism or transport in the uterine environment.

scholarly journals Diurnal rhythm of rat liver cytosolic 3-hydroxy-3-methylglutaryl-CoA synthase

1991 ◽  
Vol 280 (1) ◽  
pp. 61-64 ◽  
Author(s):  
T Royo ◽  
J Ayté ◽  
F Albericio ◽  
E Giralt ◽  
D Haro ◽  
...  
Keyword(s):  
Rat Liver ◽  
Diurnal Rhythm ◽  
Translational Regulation ◽  
Blot Hybridization ◽  
Normal Diet ◽  
Mrna Levels ◽  
Slot Blot ◽  
Dark Cycle ◽  
Slot Blot Hybridization ◽  
Sds Page

Rat liver cytosolic 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase exhibits a diurnal rhythm of enzyme activity which coincides with the diurnal rhythm of HMG-CoA synthase protein. The peaks of activity and protein (determined by SDS/PAGE and immunoblotting) both occur at D10 (the tenth hour of the daily 12 h dark cycle). The peak of mRNA levels (measured by slot-blot hybridization of liver RNA) is slightly advanced with respect to that of protein, by about 4 h, and shows a maximum at D6. Cytosolic HMG-CoA synthase activity and protein in rats fed on a normal diet were approx. 2-fold higher during the peak at D10 than in the nadir at D2. HMG-CoA synthase mRNA levels were approx. 4-fold higher during the peak at D6 than in the nadir at D2. These results point to a transcriptional and translational regulation of the cytosolic HMG-CoA synthase.

Comparison between the polymerase chain reaction and slot blot hybridization for the detection of HPV sequences in cervical scrapes

Cytopathology ◽  
1990 ◽  
Vol 1 (1) ◽  
pp. 19-23 ◽  
Author(s):  
P. WARD ◽  
G. N. PARRY ◽  
R. YULE ◽  
D. V. COLEMAN ◽  
A. D. B. MALCOLM
Keyword(s):  
Polymerase Chain Reaction ◽  
Blot Hybridization ◽  
Slot Blot ◽  
Chain Reaction ◽  
Slot Blot Hybridization ◽  
Polymerase Chain

Using genomic slot blot hybridization to assess intergeneric Saccharum x Erianthus hybrids (Andropogoneae – Saccharinae)

Genome ◽  
1997 ◽  
Vol 40 (4) ◽  
pp. 428-432 ◽  
Author(s):  
P. Besse ◽  
C. L. McIntyre ◽  
D. M. Burner ◽  
C. G. de Almeida
Keyword(s):  
Genomic Dna ◽  
Blot Hybridization ◽  
Female Parent ◽  
Saccharum Officinarum ◽  
Rapid Screening ◽  
Hybridization Technique ◽  
Slot Blot ◽  
Saccharum Species ◽  
Slot Blot Hybridization

The use of genomic slot blot hybridization enabled the differentiation of hybrids from selfs in Saccharum × Erianthus intergeneric crosses in which Saccharum was used as the female parent. Based on the genomic in situ hybridization technique, slot blots of DNA from the parents and the progeny were blocked with the Saccharum parent DNA and hybridized with the labelled male Erianthus genomic DNA. This technique allowed a rapid screening for hybrids and was sensitive enough to detect a 1/20 dilution of Erianthus in Saccharum DNA, which should enable the detection of most partial hybrids. The genomic slot blot hybridization technique was shown to be potentially useful for assessing crosses involving Saccharum species with either Old World Erianthus section Ripidium or North American Erianthus (= Saccharum) species. The effectiveness of the technique was assessed on 144 progeny of a Saccharum officinarum × Erianthus arundinaceus cross, revealing that 43% of the progeny were selfs. The importance of this test as a tool to support intergeneric breeding programs is discussed.Key words: slot blot, Erianthus, genomic DNA, Saccharum, sugarcane.

scholarly journals Using a CCD Camera Imaging System as a Recording Device to Quantify Human DNA by Slot Blot Hybridization

BioTechniques ◽  
2001 ◽  
Vol 30 (3) ◽  
pp. 680-685 ◽  
Author(s):  
Bruce Budowle ◽  
William R. Hudlow ◽  
Steven B. Lee ◽  
Leonard Klevan
Keyword(s):  
Imaging System ◽  
Blot Hybridization ◽  
Ccd Camera ◽  
Recording Device ◽  
Slot Blot ◽  
Camera Imaging ◽  
Human Dna ◽  
Slot Blot Hybridization

Analysis of RNA by Northern and Slot Blot Hybridization

1997 ◽  
Vol 37 (1) ◽  
Author(s):  
Terry Brown ◽  
Karol Mackey
Keyword(s):  
Blot Hybridization ◽  
Slot Blot ◽  
Slot Blot Hybridization

scholarly journals The expression of the IGF system in the bovine uterus throughout the oestrous cycle and early pregnancy

2000 ◽  
Vol 165 (2) ◽  
pp. 231-243 ◽  
Author(s):  
RS Robinson ◽  
GE Mann ◽  
TS Gadd ◽  
GE Lamming ◽  
DC Wathes
Keyword(s):  
Cross Sections ◽  
Early Pregnancy ◽  
In Situ Hybridisation ◽  
Endometrial Biopsy ◽  
Oestrous Cycle ◽  
Luminal Epithelium ◽  
Mrna Levels ◽  
Igf Binding Proteins ◽  
Igf System ◽  
Igf I

The IGF system is expressed in the uterus during the oestrous cycle and early pregnancy and is likely to play an important role in regulating the development of the embryo and uterus. The IGF peptides (IGF-I and -II) mediate their effects through the type 1 IGF receptor (IGF-1R), while the IGF-binding proteins (IGFBP-1 to -6) modulate their interaction with the receptor. In this study, the expression of the IGF system in the bovine uterus was determined throughout the oestrous cycle and on day 16 of pregnancy. Endometrial biopsy samples were collected from four cows over three cycles such that there were samples for every 2 days from day 0 (oestrus) to day 14 and then every day until day 21. To assess the effect of pregnancy, uterine horn cross-sections were collected on day 16 from 15 pregnant (PREG), five inseminated non-pregnant (INP) and nine uninseminated cyclic controls (CONT). The expression of mRNA for the IGFs, IGF-1R and IGFBP-1 to -5 was determined by in situ hybridisation and the results were quantified by measuring the optical density units from autoradiographs. The main region of IGF-I mRNA expression was the sub-epithelial stroma underlying the luminal epithelium. The expression of IGF-I mRNA was highest at oestrus and lowest during the early and late luteal phases. On day 16, IGF-I mRNA levels were low in all groups, with pregnancy having no effect on the IGF-I mRNA concentrations. The strongest expression of IGF-II mRNA was in the caruncular stroma, with pregnancy having no significant effect in this region. IGF-1R mRNA was also present in the caruncles and was strongly expressed in all epithelial cells both throughout the oestrous cycle and during early pregnancy. The expression of IGFBP-1 mRNA was confined to the luminal epithelium, with the strongest expression seen on day 14 of the cycle. On day 16 the expression of IGFBP-1 mRNA was higher in the PREG group compared with the CONT group. The expression of IGFBP-2 mRNA was localised to the sub-epithelial stroma with more INP than PREG cows showing detectable levels of IGFBP-2. The strongest expression of IGFBP-3 mRNA was in the caruncular stroma; expression in the endometrial stroma was similarly decreased during early pregnancy. IGFBP-5 mRNA was mainly expressed in the inner ring of myometrium and was not affected by pregnancy on day 16. In conclusion, these results show that many components of the uterine IGF system are differentially regulated during the oestrous cycle and early pregnancy and suggest that modulation of the IGF system may influence uterine activity during this period.

Identification of rare Candida species and other yeasts by polymerase chain reaction and slot blot hybridization

2000 ◽  
Vol 38 (4) ◽  
pp. 207-212 ◽  
Author(s):  
Juergen Loeffler ◽  
Holger Hebart ◽  
Stella Magga ◽  
Diethard Schmidt ◽  
Lena Klingspor ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Candida Species ◽  
Blot Hybridization ◽  
Slot Blot ◽  
Chain Reaction ◽  
Slot Blot Hybridization ◽  
Polymerase Chain
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