Paracrine cell signaling is believed to be important for ovarian follicle development, and a role for some members of the fibroblast growth factor (FGF) family has been suggested. In the present study, we tested the hypothesis that FGF-8 and its cognate receptors (FGFR3c and FGFR4) are expressed in bovine antral follicles. RT-PCR was used to analyze bovine Fgf8, Fgfr3c and Fgfr4 mRNA levels in oocytes, and granulosa and theca cells. Fgf8 expression was detected in oocytes and in granulosa and theca cells; this expression pattern differs from that reported in rodents. Granulosa and theca cells, but not oocytes, expressed Fgfr3c, and expression in granulosa cells increased significantly with follicle estradiol content, a major indicator of follicle health. Fgfr4 expression was restricted to theca cells in the follicle, and decreased significantly with increasing follicle size. To investigate the potential regulation of Fgfr3c expression in the bovine granulosa, cells were cultured in serum-free medium with FSH or IGF-I; gene expression was upregulated by FSH but not by IGF-I. The FSH-responsive and developmentally regulated patterns of Fgfr3c mRNA expression suggest that this receptor is a potential mediator of paracrine signaling to granulosa cells during antral follicle growth in cattle. IntroductionAntral ovarian follicle growth in monovular species is regulated by a number of factors, the most well known of which are the gonadotropins. Follicles are considered to be follicle-stimulating hormone (FSH)-dependent until dominance occurs, after which they become luteinizing hormone-dependent (reviewed by Fortune et al. 2001, Ginther et al. 2001. It has also become clear that growth factors are key stimulatory/regulatory molecules. Several lines of evidence point to a critical role for members of the transforming growth factor-b (TGF-b) superfamily, especially growth/differentiation factor 9 and bone morphogenetic protein 15 (reviewed by Gilchrist et al. 2004, Juengel et al. 2004, Shimasaki et al. 2004.The fibroblast growth factor (FGF) family is emerging as a group of factors that are potentially important for follicle growth. For example, FGF-7 is expressed in theca cells, its receptor is expressed in granulosa cells (Parrott & Skinner 1998, Berisha et al. 2004, and FGF-7 stimulated bovine granulosa cell proliferation and inhibited steroidogenesis (Parrott & Skinner 1998). Another potentially interesting member of this family is FGF-8. Widely expressed in fetal tissues, this factor is predominantly expressed in the gonads of adult rodents and ruminants (MacArthur et al. 1995a, Buratini et al. 2005. Within the ovary, Fgf8 gene expression occurs only in the oocyte in adult mice (Valve et al. 1997), which suggests a potential role in signaling of follicular cells by the oocyte.There are five known FGF receptor (FGFR) genes (Kim et al. 2001, Sleeman et al. 2001, of which FGF-8 preferentially activates FGFR4 and the 'c' splice form of FGFR3 (Ornitz et al. 1996). mRNAs encoding Fgfr4 or Fgfr3c were not consiste...

Fibroblast growth factor 17 (FGF17) is a member of the FGF8 subfamily that appears to be relevant to folliculogenesis and oogenesis, as the prototype member FGF8 is an oocyte-derived protein that signals to cumulus cells. FGF8 has structural and receptor-binding similarities to FGF17, whose expression in the ovary has not been reported. In this study, we demonstrate localization of FGF17 protein to the oocyte of preantral follicles, and to the oocyte and granulosa cells of antral follicles. Real-time PCR demonstrated the presence of mRNA in oocytes and, to a lesser extent, in granulosa and theca cells. FGF17 mRNA abundance was low in granulosa and theca cells from healthy follicles and increased significantly in atretic follicles. Addition of FSH or IGF-I to granulosa cells in vitro decreased FGF17 mRNA abundance, and treatment with FGF17 inhibited estradiol and progesterone secretion from granulosa cells in relation to control cultures without these additives. We conclude that FGF17 is a potential mediator of granulosa cell differentiation.

This study compared the oral hygiene and oral microbiota in children and young people with neurological impairment and oropharyngeal dysphagia with and without gastrostomy. Forty children and young people participated in this study: 19 females and 21 males, aged 2 to 22 years (mean age 8.6 years). Participants were divided into two groups: group I (GI = 20) with gastrostomy and group II (GII = 20) without gastrostomy (with oral feeding). Oral hygiene was assessed using the Simplified Oral Hygiene Index (SOHI). Analysis of two bacteria, Streptococcus mutans and Streptococcus sobrinus, was performed by collecting saliva using an oral swab, then mRNA expression was evaluated using the polymerase chain reaction (PCR) technique. The oral hygiene index had a general median of 2.2, and the two groups were statistically different (Group I: median 2.9 and Group II: median 2.0) (p = 0.01751). Bacterial analysis indicated 13 individuals with S. mutans and none with S. sobrinus. Of the 13 individuals with S. mutans, 6 were from Group I and 7 from Group II. Those with gastrostomy had worse oral hygiene, and both groups harbored the bacterium S. mutans.

Paracrine cell signaling is thought to be important for ovarian follicle development, and a role for some members of the fibroblast growth factor (FGF) family have been suggested. In the present study, we tested the hypothesis that FGF-8 and its cognate receptors (FGFR-3c and FGFR-4) are expressed in bovine preantral follicles. Reverse transcription-polymerase chain reaction was used to amplify bovine FGF-8, FGFR-3c, and FGFR-4 from preantral follicle samples and a variety of fetal and adult tissues. All three genes were widely expressed in fetal tissues, with a restricted expression pattern in adult tissues. FGF-8 and FGFR-3c were expressed in secondary follicles in 70% of fetuses examined, whereas FGFR-4 expression was significantly less frequent (20%). FGFR-3c expression frequency was significantly lower in primordial compared to secondary follicles, and FGF-8 expression showed a similar trend. FGFR-4 was only observed when all follicle classes of an individual were expressing both FGF-8 and FGFR-3c. We conclude that FGF-8 and its receptors are expressed in preantral follicles in a developmentally regulated manner.

The rat tail artery has been used for the study of vasoconstriction mediated by ␣ 1A -adrenoceptors (ARs). However, rings from proximal segments of the tail artery (within the initial 4 cm, PRTA) were at least 3-fold more sensitive to methoxamine and phenylephrine (n ϭ 6 -12; p Ͻ 0.05) than rings from distal parts (between the sixth and 10th cm, DRTA). Interestingly, the imidazolines N- [5-(4,5-dihydro-1H-imidazol-2-yl)-2-hydroxy-5,6,7,8-tetrahydronaphthalen-1-yl]methanesulfonamide hydrobromide (A-61603) and oxymetazoline, which activate selectively ␣ 1A -ARs, were equipotent in PRTA and DRTA (n ϭ 4 -12), whereas buspirone, which activates selectively ␣ 1D -AR, was Ϸ70-fold more potent in PRTA than in DRTA (n ϭ 8; p Ͻ 0.05).dihydrochloride (BMY-7378) was Ϸ70-fold more potent against the contractions induced by phenylephrine in PRTA (pK B of Ϸ8.45; n ϭ 6) than in DRTA (pK B of Ϸ6.58; n ϭ 6), although the antagonism was complex in PRTA. 5-Methylurapidil, a selective ␣ 1A -antagonist, was equipotent in PRTA and DRTA (pK B of Ϸ8.4), but the Schild slope in DRTA was 0.73 Ϯ 0.05 (n ϭ 5). The noncompetitive ␣ 1B -antagonist conotoxin -TIA reduced the maximal contraction induced by phenylephrine in DRTA, but not in PRTA. These results indicate a predominant role for ␣ 1A -ARs in the contractions of both PRTA and DRTA but with significant coparticipations of ␣ 1D -ARs in PRTA and ␣ 1B -ARs in DRTA. Semiquantitative reverse transcription-polymerase chain reaction revealed that mRNA encoding ␣ 1A -and ␣ 1B -ARs are similarly distributed in PRTA and DRTA, whereas mRNA for ␣ 1D -ARs is twice more abundant in PRTA. Therefore, ␣ 1 -ARs subtypes are differentially distributed along the tail artery. It is important to consider the segment from which the tissue preparation is taken to avoid misinterpretations on receptor mechanisms and drug selectivities.Three ␣ 1 -adrenoceptor (AR) subtypes (␣ 1A -, ␣ 1B -, and ␣ 1D -ARs) are involved in the contractions of vascular smooth muscle in response to AR agonists. However, depending on the vessel, the participation of one particular subtype may predominate over the other two ␣ 1 -ARs. The use of subtype selective ␣ 1 -AR agonists and antagonists has allowed the identification of a predominant role for ␣ 1A -ARs in the contractions of the rat tail artery in response to norepinephrine (Gisbert et al., 2003), phenylephrine (Piascik et al., 1995(Piascik et al., , 1997, methoxamine (Villalobos-Molina and Ibarra, 1996), NS-49 (Murata et al., 1999), and A-61603 (Lachnit et al., 1997;Chang et al., 2000;Jahnichen et al., 2004), suggesting that this artery is an interesting model for the study of the mechanisms involved in the vasoconstriction mediated by ␣ 1A -ARs and for the characterization of the properties of selective ligands.Interestingly, studies evaluating the expression of mRNA encoding ␣ 1 -ARs reveal that the ␣ 1A -AR is probably not the unique subtype expressed in the rat tail artery, since mRNA encoding ␣ 1B -and ␣ 1D -AR has been detected in this vessel (Piascik et al., 19...