Sensory neuron regulation of gastrointestinal inflammation and bacterial host defence.

Sensory neurons in the gastrointestinal tract have multifaceted roles in maintaining homeostasis, detecting danger and initiating protective responses. The gastrointestinal tract is innervated by three types of sensory neurons: dorsal root ganglia, nodose/jugular ganglia and intrinsic primary afferent neurons. Here, we examine how these distinct sensory neurons and their signal transducers participate in regulating gastrointestinal inflammation and host defence. Sensory neurons are equipped with molecular sensors that enable neuronal detection of diverse environmental signals including thermal and mechanical stimuli, inflammatory mediators and tissue damage. Emerging evidence shows that sensory neurons participate in host-microbe interactions. Sensory neurons are able to detect pathogenic and commensal bacteria through specific metabolites, cell-wall components, and toxins. Here, we review recent work on the mechanisms of bacterial detection by distinct subtypes of gut-innervating sensory neurons. Upon activation, sensory neurons communicate to the immune system to modulate tissue inflammation through antidromic signalling and efferent neural circuits. We discuss how this neuro-immune regulation is orchestrated through transient receptor potential ion channels and sensory neuropeptides including substance P, calcitonin gene-related peptide, vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide. Recent studies also highlight a role for sensory neurons in regulating host defence against enteric bacterial pathogens including Salmonella typhimurium, Citrobacter rodentium and enterotoxigenic Escherichia coli. Understanding how sensory neurons respond to gastrointestinal flora and communicate with immune cells to regulate host defence enhances our knowledge of host physiology and may form the basis for new approaches to treat gastrointestinal diseases.

The use of air plasma in surface modification of peripheral nerve conduits.

Surface modification is a conventional approach in biomaterials development, but most of the modification processes are intricate and time inefficient. In this study, a convenient open air plasma treatment was employed to modify the surface of poly(d,l-lactide) (PLA). Chitosan and fibroblast growth factor 1 (FGF1) were sequentially grafted with the assistance of open air plasma treatment onto the PLA nerve conduits with designed micropores and surface microgrooves. Grafting of these components was verified by electron spectroscopy for chemical analysis. The modified nerve conduits showed enhanced ability in the repair of 10-mm sciatic nerve transection defects in rats. The sequential air plasma treatment can be a convenient way to introduce biocompatible (e.g., chitosan) and bioactive components (e.g., growth factors) onto the surface of biomaterials.

Multiple controlling mechanisms of FGF1 gene expression through multiple tissue-specific promoters.

We now know that fibroblast growth factor-1 (FGF1) transcription is controlled by at least four distinct promoters in a tissue-specific manner. Thus, promoter 1.A is active in the kidney, 1.B in the brain, and 1.C and 1.D in a variety of cultured cells including vascular smooth muscle cells. These promoters are separated from each other by up to 70 kbp. Multiple FGF1 transcripts arise from alternate promoter usage and alternative splicing of different 5'-untranslated exons. The 1.A and 1.B promoters are constitutively active in their respective cell types. In contrast, different biological response modifiers, including serum and transforming growth factor beta, can induce the 1.C and 1.D promoters. The 540-bp sequence upstream of the 1B transcription initiation site is sufficient to drive the expression of a heterologous luciferase reporter in cultured cells, and an 18-bp sequence within this region is important for the regulation of brain-specific gene expression. Furthermore, regulation occurs through the binding of the 18-bp sequence to a brain-specific 37-kDa protein and a ubiquitous basic helix-loop-helix protein, E2-2. We have produced transgenic mice bearing the brain-specific promoter of the human FGF1 gene joined to the SV40 immediate-early gene, which encodes the large T antigen. The resulting mice developed brain tumors that originated in the pontine gray, just rostral to the fourth ventricle. We have also identified a serum response element, comprising a CarG box and an Ets-binding site, in the 1.D promoter. Continued characterization of the mechanistic events that control the tissue-specific activation of FGF1 promoters will help us to understand the role of FGF1 in cancer, atherosclerosis, and neural development.

Thermodynamic characterization of the human acidic fibroblast growth factor: evidence for cold denaturation.

The thermodynamic parameters characterizing the conformational stability of the human acidic fibroblast growth factor (hFGF-1) have been determined by isothermal urea denaturation and thermal denaturation at fixed concentrations of urea using fluorescence and far-UV CD circular dichroism (CD) spectroscopy. The equilibrium unfolding transitions at pH 7.0 are adequately described by a two-state (native <--> unfolded state) mechanism. The stability of the protein is pH-dependent, and the protein unfolds completely below pH 3.0 (at 25 degrees C). hFGF-1 is shown to undergo a two-state transition only in a narrow pH range (pH 7.0-8.0). Under acidic (pH <6.0) and basic (pH >8.0) conditions, hFGF-1 is found to unfold noncooperatively, involving the accumulation of intermediates. The average temperature of maximum stability is determined to be 295.2 K. The heat capacity change (DeltaC(p)()) for the unfolding of hFGF-1 is estimated to be 2.1 +/- 0.5 kcal.mol(-1).K(-1). Temperature denaturation experiments in the absence and presence of urea show that hFGF-1 has a tendency to undergo cold denaturation. Two-dimensional (1)H-(15)N HSQC spectra of hFGF-1 acquired at subzero temperatures clearly show that hFGF-1 unfolds under low-temperature conditions. The significance of the noncooperative unfolding under acidic conditions and the cold denaturation process observed in hFGF-1 are discussed in detail.

15N NMR relaxation studies of free and ligand-bound human acidic fibroblast growth factor.

15N NMR relaxation data have been used to characterize the backbone dynamics of the human acidic fibroblast growth factor (hFGF-1) in its free and sucrose octasulfate (SOS)-bound states. (15)N longitudinal (R(1)), transverse (R(2)) relaxation rates and (1H)-(15)N steady-state nuclear Overhauser effects were obtained at 500 and 600 MHz (at 25 degrees C) for all resolved backbone amide groups using (1)H- detected two-dimensional NMR experiments. Relaxation data were fit to the extended model free dynamics for each NH group. The overall correlation time (tau(m)) for the free and SOS-bound forms were estimated to be 10.4 +/- 1.07 and 11.1 +/- 1.35 ns, respectively. Titration experiments with SOS reveals that the ligand binds specifically to the C-terminal domain of the protein in a 1:1 ratio. Binding of SOS to hFGF-1 is found to induce a subtle conformational change in the protein. Significant conformational exchange (R(ex)) is observed for several residues in the free form of the protein. However, in the SOS-bound form only three residues exhibit significant R(ex) values, suggesting that the dynamics on the micro- to millisecond time scale in the free form is coupled to the cis-trans-proline isomerization. hFGF-1 is a rigid molecule with an average generalized parameter (S(2)) value of 0.89 +/- 0.03. Upon binding to SOS, there is a marked decrease in the overall flexibility (S(2) = 0.94 +/- 0.02) of the hFGF-1 molecule. However, the segment comprising residues 103-111 shows increased flexibility in the presence of SOS. Significant correlation is found between residues that show high flexibility and the putative receptor binding sites on the protein.

The small GTPases Ras, Rac, and Cdc42 transcriptionally regulate expression of human fibroblast growth factor 1.

Four distinct promoters (1A, 1B, 1C, and 1D) of fibroblast growth factor 1 (FGF1), spaced up to 70 kilobase pairs apart, direct the expression of alternatively spliced transcript variants (FGF1.A, -1. B, -1.C, and -1.D) that encode FGF1. These FGF1 transcripts can be detected in cultured cells as well as in normal and diseased tissues. These transcripts are differentially regulated in a cell-specific manner. To further delineate the biological function of multiple promoter usage by a single gene, we investigated the transcriptional regulation of these promoters by defined signaling pathways associated with cell proliferation and cell survival. Here we show a specific association of two of the FGF1 promoters, 1C and 1D, with signaling cascades of the Ras superfamily of GTPases. A serum-response element, comprised of the Ets and CArG motifs, present in promoter 1D was shown to be the target of distinct signaling cascades; the Ets motif target of Ras, Rac1, and Cdc42 regulation; and the CArG motif target of de novo protein synthesis-independent cascade. Ras and Rac1 also activated the FGF2 promoter. Further, the transcription factor Ets2 synergistically activated FGF1 gene, but not FGF2, in a Ras- and Rac1-dependent signaling pathway. In support of these conclusions high levels of intracellular FGF1 were detected in cells undergoing cytokinesis. Altogether, our results suggest that FGF1 may play a fundamental role in cell division, spreading, and migration, in addition to cell proliferation.

Expression and activity of human Na+/I- symporter in human glioma cells by adenovirus-mediated gene delivery.

Radioiodide concentrating activity in the thyroid, mediated by human Na+/I- symporter (hNIS), provides a mechanism for effective radioiodide treatment for patients who have invasive, recurrent, and metastatic thyroid cancers after total thyroidectomy. In an attempt to develop hNIS gene transfer for radioiodide therapy for patients with brain tumors, we have constructed recombinant adenoviruses, rAd-CMV-hNIS9 and rAd-CMV-FLhNIS, to express exogenous hNIS in U1240 and U1240Tag human glioma cells. U1240Tag differs from U1240 glioma cells in that it expresses the SV40 large T antigen oncoprotein. In both U1240 and U1240Tag cells, radioiodide uptake (RAIU) activity in the cells infected with rAd-CMV-hNIS9 or rAd-CMV-FLhNIS increases as the adenoviral MOI increases. The protein expression profile of hNIS in infected cells is generally in agreement with their RAIU activity profile. Although the expressed hNIS9 protein appeared to have a shorter half-life than FLhNIS, hNIS9 expression could be maintained by multiple infections in these cells. In addition, we show that hNIS can be expressed and function in a xenografted human glioma by intratumoral injection of rAd-CMV-hNIS9.

Tumorigenesis in transgenic mice in which the SV40 T antigen is driven by the brain-specific FGF1 promoter.

Gene expression can be manipulated by the introduction of a hybrid gene formed by linking a highly tissue-specific regulatory element to a gene whose expression might be expected to alter cellular function. Previously, we have shown that the human FGF1 gene contains four distinct tissue-specific promoters. In an effort to perturb the programming of proliferation and differentiation in a subset of neural cells, we have produced transgenic mice bearing the brain-specific promoter of the human FGF1 gene joined to the SV40 immediate early gene, which encodes the large T antigen. The resulting mice, and offspring from four individual lines, developed brain tumors that originated in the pontine gray, just rostral to the fourth ventricle. Tumors were moderately vascularized, as demonstrated by staining with both hematoxylin and eosin and antibodies to three different endothelial cell markers, but vessels were histologically normal. Scattered tumor foci were present as early as postnatal day 26; and affected animals died between 5 - 8 months of age. In mature animals, tumors lacked terminal differentiation markers for astrocytes (glial fibrillary acidic protein) or neurons (synaptophysin and neuron-specific enolase). However, they expressed high levels of proliferating cell nuclear antigen and vimentin, markers for proliferating cells. This immunophenotype is consistent with the tumor being at an early stage of differentiation. Therefore, these mice may provide a valuable tool for the study of tumorigenesis, replenishment and differentiation of neural stem cells.

Site-directed mutagenesis and molecular modeling identify a crucial amino acid in specifying the heparin affinity of FGF-1.

Heparin potentiates the mitogenic activity of FGF-1 by increasing the affinity for its receptor and by extending its biological half-life. During the course of labeling human FGF-1 with Na(125)I and chloramine T, it was observed that the protein lost its ability to bind to heparin. In contrast, bovine FGF-1 retained its heparin affinity even after iodination. To localize the region responsible for the lost heparin affinity, chimeric FGF-1 proteins were constructed from human and bovine FGF-1 expression constructs and tested for their heparin affinity after iodination. The results showed that the C-terminal region of human FGF-1 was responsible for the loss of heparin affinity. This region harbors a single tyrosine residue in human FGF-1 in contrast to a phenylalanine at this position in bovine FGF-1. Mutating this tyrosine residue in the human FGF-1 sequence to phenylalanine did not restore the heparin affinity of the iodinated protein. Likewise, changing the phenylalanine to tyrosine in the bovine FGF-1 did not reduce the ability of the iodinated protein to bind to heparin. In contrast, a mutant human FGF-1 that has cysteine-131 replaced with serine (C131S) was able to bind to heparin even after iodination while bovine FGF-1 (S131C) lost its binding affinity to heparin upon iodination. In addition, the human FGF-1 C131S mutant showed a decrease in homodimer formation when exposed to CuCl(2). Molecular modeling showed that the heparin-binding domain of FGF-1 includes cysteine-131 and that cysteine-131, upon oxidation to cysteic acid during the iodination procedures, would interact with lysine-126 and lysine-132. This interaction alters the conformation of the basic residues such that they no longer bind to heparin.

Characterization of the entire transcription unit of the mouse fibroblast growth factor 1 (FGF-1) gene. Tissue-specific expression of the FGF-1.A mRNA.

Fibroblast growth factor 1 (FGF-1, also known as acidic FGF) is a mitogen for a variety of mesoderm- and neuroectoderm-derived cells, as well as an angiogenic factor in vivo. It has been implicated in angiogenic diseases including atherosclerosis, cancer and inflammatory diseases. In the present study, the entire transcriptional unit of the mouse FGF-1 gene, including four promoters, is characterized. By nucleotide sequence and RNase protection analyses, we have determined that its 3'-end resides 3.2 kilobase pairs downstream from the stop codon. We have previously cloned and characterized the mouse homologue of the human 1B promoter, as well as a novel upstream untranslated exon. In order to elucidate the regulatory mechanism of FGF-1 gene expression, the mouse promoter containing TATA and CAAT consensus sequences (FGF-1. A) was isolated from a P1 library and characterized. We further determined that the mouse heart is the most abundant source for the FGF-1.A mRNA. Finally, via both RNase protection analysis and 5'-rapid amplification of cDNA ends, we determined the transcription start site of the FGF-1.A mRNA.

Isolation of yeast artificial chromosomes containing the entire transcriptional unit of the human FGF1 gene: a 720-kb contig spanning human chromosome 5q31.3-->q32.

The q31-q33 region of chromosome 5 includes a number of genes encoding growth factors, growth factor receptors, and hormone/neurotransmitter receptors. The human fibroblast growth factor 1 locus (FGF1) resides in this region of chromosome 5, which is frequently lost in myelodysplastic syndromes and acute myeloid leukemia patients. Other disease loci, including the loci for limb-girdle muscular dystrophy and an autosomal dominant deafness, have been mapped on this region, but their genes have not been isolated. It was shown that the critical region lost in two patients with the 5q- syndrome resides between FGF1 and IL12B. We previously reported the construction of a yeast artificial chromosome (YAC) contig spanning 330 kb around the FGF1 gene. Here we report the isolation of additional YAC clones that extend 290 kb from the previous contig. Sequence-tagged sites developed from the outermost YAC ends were utilized in the contig cloning of two P1 clones P1Y2 and P1Y8. Together, these YAC and P1 clones span 720 kb around the FGF1 locus. With the use of fluorescence in situ hybridization, a physical map has been constructed of these P1 and GRL (glucocorticoid receptor locus) probes on metaphase and interphase chromosomes. On the basis of our work and the known orientation of GRL transcription, the determined order of these loci on chromosome 5q31.3-q32 is centromere-P1Y8-3'[FGF1]5'-P1Y2-5'[GRL]3'-telome re. Knowing the transcriptional orientation of the FGF1 gene relative to the centromere will now facilitate the directional cloning of clinically important genes that may reside in this region.

Regulation of a promoter of the fibroblast growth factor 1 gene in prostate and breast cancer cells.

FGF-1 mRNA is expressed in the prostate cancer cell lines LNCaP and PC-3 and in the breast carcinoma cell line MDA-MB-231. Levels of FGF-1 mRNA have been shown to be up-regulated by serum, phorbol esters, and combinations of growth factors. It was shown that the major FGF-1 mRNA species expressed following serum stimulation in MDA-MB-231 cells is FGF-1.C. To better understand the potential role of FGF-1 in human prostate and breast cancer, we began an analysis of the cis- and trans-acting elements of one of its promoters required for the serum, PMA, and androgen regulation in breast and prostate cancer cell lines. We show that FGF-1.C steady-state mRNA levels are increased following serum or PMA stimulation of PC-3 cells. Further, we determine the FGF-1.C transcription start site in PC-3 cells. By sequence analysis, we show that consensus AP1, AP2, and Sp1 sites and ARE- and CRE-near consensus elements are present in the immediate 5' region of the FGF-1.C transcription start site. Gel-shift assays show that oligonucleotides containing FGF-1.C AP1, AP2, or Spl sequences form specific DNA-protein complexes with nuclear extracts from PC-3 cells. To determine if these or other cis-acting sequences are responsible for the serum, androgen, or growth factor regulation of FGF-1 expression, fragments of the FGF-1.C promoter region were cloned upstream of the luciferase reporter gene. We show that FGF-1 synergizes with androgen to enhance FGF-1.C transcription in LNCaP cells. We further show that the DNA fragment containing sequence up to 1614 nucleotides upstream of the FGF-1.C transcription start site is sufficient for stimulating promoter activity following serum treatment of MDA-MB-231 cells. Thus, FGF-1.C promoter contains sequences that are important for androgen or serum stimulation in prostate and breast cancer cells.

A splice variant of E2-2 basic helix-loop-helix protein represses the brain-specific fibroblast growth factor 1 promoter through the binding to an imperfect E-box.

We previously demonstrated that a cis-element (-489 to -467) in the brain-specific fibroblast growth factor (FGF)-1 promoter (FGF-1.B) binds multiple nuclear factors, and this binding enhances transcriptional activity of this promoter. Here we report the isolation of three cDNA clones, VL1, VL2 and VL3, from a human brain stem cDNA expression library using four tandem repeats of the 26-base pair sequence (-492 to -467) as the probe. These cDNA clones represent the variant of bHLH protein E2-2/SEF2-1 in having 12 additional nucleotides encoding the amino acids RSRS. The glutathione S-transferase (GST) fusion proteins of VLl, VL2, and VL3 immunologically react with anti-E2-2 antibody and anti-GST-VL2 antibody. Electrophoretic mobility shift assay and methylation interference assay revealed that the GST fusion proteins specifically bind to an imperfect E-box sequence (GACCTG) present in the 26-base pair sequence. Transient expression of the full-length E2-2 without RSRS in U1240MG glioblastoma cells resulted in repression of FGF-1.B promoter activity. We further showed a significant repression of promoter activity (>40 fold) by E2-2 (lacking the amino acid sequence RSRS) when the E47 reporter construct, containing a hexameric E-box site, was used. In contrast, the E2-2 variant containing the RSRS sequence has no significant effect on either the FGF-1 promoter or E47 promoter. These results suggest that the relative abundance of the two splice variants of E2-2 in brain could be an important determinant for the expression of FGF-1.

Fibroblast variants nonresponsive to fibroblast growth factor 1 are defective in its nuclear translocation.

Fibroblast growth factors (FGF) elicit biological effects by binding to high affinity cell-surface receptors and activation of receptor tyrosine kinase. We previously reported that two NIH/3T3 derivatives, NR31 and NR33 (NR cells), express high levels of full-length FGF-1 and exhibit a complete spectrum of transformed phenotype. In the present study, we report that NR cells respond to the mitogenic stimulation of truncated FGF-1 but not to the full-length FGF-1. Incubation of the NR cells with either form of FGF-1 resulted in its binding to cell-surface FGF receptors, activation of mitogen-activated protein (MAP) kinase, and induction of c-fos and c-myc. These data demonstrate that the FGF receptor-mediated, MAP kinase-dependent signaling pathway is not defective in the NR cells. Our data further suggest that the activation of MAP kinase in response to full-length FGF-1 is not sufficient for mitogenesis. Subcellular distribution of exogenously added FGF-1 demonstrated that full-length FGF-1 fails to translocate to the nuclei of NR31 cells. Although the full-length FGF-1 was detected in the nuclear fractions of both NIH/3T3 and NR33 cells, its half-life is much shortened in NR33 than in NIH/3T3 cells. These observations suggest that non-responsiveness of the two NR cell lines may be due to defectiveness at different steps of nuclear translocation mechanism of FGF-1.

Novel receptor protein tyrosine phosphatase (RPTPrho) and acidic fibroblast growth factor (FGF-1) transcripts delineate a rostrocaudal boundary in the granule cell layer of the murine cerebellar cortex.

We have identified a novel receptor-like protein tyrosine phosphatase (RPTPrho) transcript whose expression in the cerebellar cortex is restricted to the granule cell layer of lobules 1-6. Acidic fibroblast growth factor (FGF-1) mRNA follows a similar cerebellar expression pattern. Together, the two markers define a sharp boundary in lobule 6, slightly caudal to the primary fissure. Anterior and posterior compartments became discernible only during postnatal weeks two and six, for RPTPrho and FGF-1, respectively. A rostrocaudal boundary in lobule 6 of the murine cerebellar cortex has also been identified morphologically by the effects of the meander tail mutation. The position of the RPTPrho and FGF-1 boundary on the rostrocaudal axis of the cerebellar cortex was close to, but not coincident with, the caudal extent of the disorganized anterior lobe of meander tail and the rostral extent of Otx-2 expression. The restricted pattern of FGF-1 and RPTPrho implies that these molecules may have specific signaling roles in the tyrosine phosphorylation/dephosphorylation pathway in the anterior compartment of the adult cerebellar cortex.

Cloning and interspecies comparisons of three newt (Notophthalmus viridescens) fibroblast growth factor receptor sequences.

We report the nucleotide sequences of two fibroblast growth factor receptor (FGFR) cDNAs, FGFR1 and FGFR3, from the newt species Notophthalmus viridescens. These two cDNA sequences and a previously published newt FGFR cDNA, FGFR2, were used to derive the amino acid sequences which were then compared with their homologues from other species. This comparison shows that the intracellular tyrosine kinase domain is highly conserved across the species examined with the second half of the domain slightly more conserved than the first half. The 3' portion of the carboxyl terminal tail is not very highly conserved. The comparison of the extracellular portion of FGFR2 shows a high degree of conservation among the Ig-like domains and a low degree of conservation in the region that links the third Ig-like domain with the transmembrane domain.

Differential regulation of human fibroblast growth factor 1 transcripts provides a distinct mechanism of cell-specific growth factor expression.

Four variants of fibroblast growth factor 1 (FGF-1) mRNA, FGF-1.A, -1.B, -1.C, and -1.D, originate from four discrete promoters of the gene. Each promoter is coupled with its 5'-untranslated exon. These four promoters are separated by as much as 70 kbp in the FGF-1 locus. The present study indicates that expression of these transcripts in different cell types is regulated by distinct mechanisms. FGF-1.C mRNA requires de novo protein synthesis and de novo transcription for expression and processing, and this mRNA increases acutely in response to TGF-beta and serum stimulation. The serum-induced FGF-1.C correlates with a marked increase in protein level. In addition, FGF-1.C mRNA also increases significantly (more than 100-fold) in response to phorbol 12-myristate 13-acetate. FGF-1.D mRNA is uniquely superinduced by serum in the presence of cycloheximide and displays delayed early kinetics, suggesting that this mRNA does not require de novo protein synthesis for expression. In sharp contrast to the FGF-1.C and -1.D mRNAs, FGF-1.B mRNA levels do not increase in response to serum or phorbol 12-myristate 13-acetate and are in fact slightly down-regulated. Furthermore, FGF-1.B mRNA is stable and appears to have a long half-life (> 12 h). Thus, the unique cell-specific regulation of these FGF-1 transcripts and subsequent protein synthesis indicate that each transcript may have a distinct role in development, normal cellular processes, and, upon aberrant regulation, disease. In support of these conclusions, multiple FGF-1 transcripts in normal, fetal, and diseased tissues, containing mixed cell types, were detected. Our results suggest that FGF-1 transcripts FGF-1.C and -1.D arising from promoters 1C and 1D are specific and are potential markers for proliferation of certain cells, whereas transcripts FGF-1.A and -1.B arising from promoters 1A and 1B are specific for maintenance and survival of cells, particularly cardiac and neuronal cells. Together, these data provide evidence for a biological function for multiple promoter usage of a single gene. The discrete mechanisms for expression of the FGF-1 gene further underscore the biological significance of this growth factor.

Activation of fibroblast growth factor 8 gene expression in human embryonal carcinoma cells.

To study the role of fibroblast growth factor 8 (FGF-8) in human development and malignancies, we have isolated and characterized its gene. The gene spans 6.0 kbp and is comprised of five exons. Using reverse transcription-polymerase chain reaction, we were able to show that FGF-8 is expressed in two of the seven human mammary carcinoma cell lines tested and in only one of nine breast tumors. In contrast, both of the two normal breast tissues tested express FGF-8. FGF-8 was previously shown to be present in adult testis and ovary. Surprisingly, only one of the seven testis carcinomas and one of 12 ovary carcinomas express FGF-8, whereas all three kidney carcinomas tested express FGF-8. We further showed that fetal brain and lung express FGF-8, whereas fetal intestine and liver do not. Finally, we showed that a teratocarcinoma cell line, Tera-2, can be induced to express FGF-8 mRNA by fetal bovine serum.