Studies on Expression of the c-ski Gene in Equine (Thoroughbred) Tissues.

Abstract

A partial cDNA fragment of equine (Thoroughbred) , obtained on day 180 of pregnancy, were analyzed by means of reverse transcriptase-mediated PCR (RT-PCR). In all the tissues examined, the expression of c-ski genes was confirmed. Northern blot analysis was carried out to assess the relative expression levels of c-ski genes in the male fetal equine tissues.In all tissues examined, two forms of the transcripts were detected at molecular sizes of 7.8 kb and 6 kb, respectively. In the cerebellum and the lung, the relative expression levels were high. The expression levels in skeletal muscles were moderate compared to the other tissues. The small population size of satellite cells may explain the relatively low expression levels of c-ski genes in the well developed skeletal muscles.

Expression of Ski in the Granulosa Cells of Atretic Follicles in the Rat Ovary

“…As shown in Fig. 1, the c-ski gene was expressed in all the tissues examined, which was in agreement with the previous findings made in mice by Lyons et al [8] and in equine by Yamanouchi et al [13]. The ovary was one of the tissues that intensively expressed the c-ski gene.…”

“…Ski has been demonstrated in a variety of tissues [8,13] as in the present study and is suggested to play multiple roles in a variety of cell types [27][28][29]. For example, Ski is expressed in proliferating myoblast [6] and uterine epithelial cells [7].…”

“…The primer set used for amplification of the partial rat c-ski cDNA fragment consisted of the forward primer 5'-ACC ATC TCG TGC TTC GTG GTG GGA-3' and reverse primer 5'-CTC CTT GCC CGT GTA ATC CTG GCT-3' and was designed on the basis of the DNA sequences of the mouse and human c-ski genes published in the literature [12][13][14]. Rat-specific glyceraldehyde-3-phosphate dehydrogenase (GAPDH) primers were designed using the Primer3 software (available at http:// fokker.wi.mit.edu/primer3/) and the forward primer was 5'-CAT TGT TGC CAT CAA CGA CCC CTT-3' and the reverse primer was 5'-ACT CAG CAC CAG CAT CAC CCC ATT-3'.…”

Endometrial expression of cellular protooncogene c‐ski and its regulation by estradiol‐17β

“…The primer set used for amplification of the partial rat c‐ ski cDNA fragment consists of the forward primer, 5′‐ACC ATC TCG TGC TTC GTG GTG GGA‐3′ and the reverse primer, 5′‐CTC CTT GCC CGT GTA ATC CTG GCT‐3′, and was designed on the basis of the DNA sequences of the mouse [10] and human [11] c‐ ski gene published in the literature [12]. The predicted PCR amplified sequence was set within exon 1 of the c‐ ski gene because of its high homology with the mouse [10] and human [11] c‐ ski , and a 561 bp product was expected to be generated [12]. Fifty microliters of the reaction mixture contained 2 μl of reverse transcription reaction as above, 60 mM Tris‐HCl (pH 8.5), 15 mM (NH 4 ) 2 SO 4 , 2 mM MgCl 2 , 200 μM each of dATP, dTTP, dGTP and dCTP, 10 pmol of each primer, and 1.25 U Taq polymerase (rTaq polymerase, Takara, Japan).…”