Category Archives: Myometrium

Adenylyl Cyclase Messenger Ribonucleic Acid: DISCUSSION(4)

The rat uterus is composed of two separate smooth muscle layers: an outer longitudinal and an inner circular layer. Responses to a variety of pharmacological agents differ between the two layers. The longitudinal layer demonstrates greater sensitivity to isoproterenol, which … Continue reading

Adenylyl Cyclase Messenger Ribonucleic Acid: DISCUSSION(3)

Adenylyl cyclase types II-VI were detected in endothelial cells from a wide variety of peripheral tissues. In the current study, the very sensitive technique of RT-PCR demonstrated expression of multiple adenylyl cyclase isoforms in uterine smooth muscle cells, consistent with … Continue reading

Adenylyl Cyclase Messenger Ribonucleic Acid: DISCUSSION(2)

The pattern of expression of mRNA for all adenylyl cyclase isoforms in myometrium has not been previously described. One study, using the relatively insensitive technique of Northern blotting, identified mRNA for adenylyl cyclase isoforms II and IV in uteri from … Continue reading

Adenylyl Cyclase Messenger Ribonucleic Acid: DISCUSSION(1)

Results of this study show that late-gestation rat myometrium expresses mRNA for multiple isoforms of ad-enylyl cyclase. Messenger RNA for each of the isoforms (II-IX) was expressed both in fresh uterine muscle and in cultured, immortalized rat myometrial cells, while … Continue reading

Adenylyl Cyclase Messenger Ribonucleic Acid: RESULTS(3)

To confirm that these adenylyl cyclase isoforms were expressed in a pure population of myometrial cells, RT-PCR was performed using RNA from a cell line derived from rat myometrial cells. These cells, which were transformed by the E7 opening reading … Continue reading

Adenylyl Cyclase Messenger Ribonucleic Acid: RESULTS(2)

Two PCR products were obtained with type IV primers (Fig. 3) corresponding to the published sequence for rat type IV and a newly identified splice variant of type IV containing 24 nucleotides with two possible splice sites. It is possible … Continue reading

Adenylyl Cyclase Messenger Ribonucleic Acid: RESULTS(1)

We found expression of eight of nine subtypes of ad-enylyl cyclase in freshly isolated rat myometrial cells. Isoforms II-IX were identified by appropriately sized PCR products after RT-PCR of rat total myometrial RNA (Figs. 1 and 2). These products were … Continue reading

Adenylyl Cyclase Messenger Ribonucleic Acid: MATERIALS AND METHODS(9)

Type VII primers were sense 5′-CCAGTTATTTAGAGAGA-GACCTG and antisense 5′ -CTTGCTCATCAGGGCCA-TGCTAA, which correspond to bases 3098-3657 of the murine type VII adenylyl cyclase sequence. Type VIII primers were sense 5′ -GGACAGCAGCTGGAGTACA-CAGC and antisense 5′ -CCTGATCCTTCAGGATGAGA-TAG, which correspond to bases 3513-4196 of … Continue reading

Adenylyl Cyclase Messenger Ribonucleic Acid: MATERIALS AND METHODS(8)

The components listed above were mixed and incubated at 42°C for 2 min before the addition of 200 units of Superscript I reverse transcriptase (Gibco BRL) and further incubated for 50 min at 42°C. Parallel reactions were performed as outlined … Continue reading

Adenylyl Cyclase Messenger Ribonucleic Acid: MATERIALS AND METHODS(7)

Total RNA was first treated with DNase I to remove residual genomic DNA that might yield false-positive results during RT-PCR amplification of mRNA. Four micrograms of total RNA was incubated with 4 units of amplification-grade ribonuclease (RNase)-free DNase I (Gibco … Continue reading

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