The chicken oviduct and embryonic red blood cell transferrin receptors are distinct molecules.

We recently described an estrogen-inducible transferrin receptor from the chicken oviduct. We now report on the comparison of the oviduct transferrin receptor with the transferrin receptor obtained from chick embryo red blood cells. Western blot analysis reveals that rabbit polyclonal antibodies raised against one receptor do not cross react with the heterologous receptor. Furthermore, peptide map analyses of either affinity purified, native [125I]-labelled transferrin receptors (dimers) or dissociated, and repurified monomers obtained from oviducts and embryonic red blood cells yield distinct patterns. Therefore, the estrogen-modulated oviduct transferrin receptor appears to be structurally distinct from the iron-modulated red cell transferrin receptor.

Localization of immunoglobulins in the chicken oviduct.

Studies were made on the distribution of lymphoid tissues and immunoglobulin (Ig: IgA, IgG and IgM)-containing cells (cIg: cIgA, cIgG and cIgM) and the localization of immunoglobulins (Igs) in the oviducal walls of laying hens. Lymphocyte accumulations were occasionally observed, located mainly in the middle infundibulum and in the regions from the isthmus to the vagina. The number of cIgG significantly predominated over that of cIgA or cIgM in the mucosal connective tissue of the magnum and the isthmus. In contrast, in the regions other than the magnum and the isthmus, these three types of cIg were fewer in number. Igs were localized in some superficial epithelial cells (SECs) and glandular cells (GlCs) of the oviduct. Many IgG-containing SECs were found in the infundibulum, the isthmus, and the cranial and major uterus. IgA- or IgM-containing SECs were rare throughout the oviduct. Three types of Ig-containing GlCs were numerously found in the magnum, though lymphocyte accumulations were scarce there. In the isthmus, many IgG-containing GlCs were found, while IgA- or IgM-containing GlCs were rarely observed. Ig-containing GlCs in the magnum were considerably decreased in number after the egg passage. The results suggest that the maternal Igs are transferred to the egg mainly through GlCs in the magnum of the chicken oviduct, and that the oviducal lymphoid tissues have little relationship to the passive immunity.

Accumulation of zinc in egg yolk, ovarian follicles and organs after forced resting by high dietary zinc.

1. Eighteen Warren SSL hens of 71 weeks of age were forced-moulted by ad libitum feeding of a high-zinc diet (10,000 ppm zinc for 2 days followed by 5,000 ppm zinc-supplement diet for 4 days). From the start of the treatment, eggs were collected and 3 hens were slaughtered on days 0, 2, 3, 4, 5 and 6 of the study. 2. Zinc analyses were carried out on the different components of the eggs and on liver, pancreas, kidney, different yolky follicles of the ovary and various segments of the oviduct. 3. Seven-, six- and threefold increases in zinc concentration were found in pancreas, liver and kidney, respectively. 4. The shell gland and isthmus, but not the magnum, also showed slight but significant increases in Zn content. 5. Zinc accumulation was also high and almost identical in ovarian follicles F1 to F4 but slightly less in F5 and F6 follicles. 6. In the egg, a significant increase in zinc concentration was only observed in the yolk.

Distribution of estrogen and progesterone receptors and steroid-regulated gene products in the chick oviduct.

The distribution of estrogen and progesterone receptors (ER and PR, respectively) was studied immunohistochemically in the chick oviduct. Estrogen receptor immunoreactivity was found only in the nuclei of glandular epithelial cells. Progesterone receptor was found in the nuclei of glandular and luminal epithelia, stroma, smooth muscle cells and in the mesothelium. The dissimilar distribution of ER and PR suggests that either ER concentration in the luminal epithelium and smooth muscle is very low (below the sensitivity of ER immunostaining) or that estrogens control their PR synthesis indirectly via ER in glandular cells. A known estrogen-inducible protein, ovalbumin, was localized in the same glandular epithelial cells as ER. A progestin-inducible protein, avidin, was found in part of the luminal and glandular epithelium cells but not in other PR-positive cell types. This indicates the importance of cellular differentiation in the regulation of avidin synthesis. Estrogen and progesterone administration had effects also on ER and PR immunoreactivity. Estrogen and progesterone administrations for 24 h decreased markedly the immunoreactivity of their receptors. The decrease in receptor immunoreactivity is most likely due to a transient loss of immunoreactive receptor protein, since the antibodies (H222, PR6) react both with transformed (4 S) and non-transformed (8 S) receptor forms. At the subcellular level, PR was localized in the chromatin by immunoelectron microscopy. Progestin administration seemed to decrease PR immunoreactivity especially in the heterochromatin area, suggesting that conformational chromatin rearrangements occur during down-regulation of PR.

Receptor interconversion model of hormone action. 1. Purification of a factor involved in conferring estradiol binding properties to the estrogen receptor.

We previously demonstrated that the chick oviduct estrogen receptor exists in three interconvertible forms. Two of these forms bind estradiol with high but distinct affinities. A third form exists as a non-estrogen binding recyclable form, Rnb, which upon treatment with ATP/Mg2+ is quantitatively converted to the lower affinity estradiol binding form. We now describe the isolation from chick oviduct cytosol of a factor involved in this conversion and its 1100-fold purification by ammonium sulfate fractionation, DEAE ion-exchange chromatography, and size-exclusion HPLC. The factor elutes from the size-exclusion column with an apparent molecular weight of 40,000. This highly purified factor potentiates estradiol binding in a dose-dependent manner in the presence of ATP/Mg2+. Its activity is destroyed by heating or by trypsin treatment but is relatively stable to freezing and thawing and is inert to treatment with reducing agents. ATP is an essential nucleotide substrate; GTP and cyclic nucleotides are inactive. Studies of cation dependence demonstrate that Mg2+ is also essential; Ca2+ alone is completely ineffective in catalyzing receptor potentiation and does not synergize with Mg2+. In the presence of excess ATP/Mg2+ and a fixed concentration of Fy, the Km for potentiation of estradiol binding is approximately 0.4 nM.

Origin of type I collagen localized within oviduct epithelium of quail hyperstimulated by progesterone.

Bird oviduct development is controlled by sex steroid hormones. Estrogens (E) induce cell proliferation, formation of tubular glands by epithelial cell evagination and cell differentiation. Progesterone (P) strongly increases secretory processes in E-treated quails, but inhibits cell proliferation and cell evagination. The balance between E and P is very critical for the development and morphogenesis of the oviduct. After six daily injections of low doses of E (10 micrograms day-1) and high doses of P (5 mg day-1) into ovariectomized quails, cell proliferation and secretory process are stimulated but cell evagination is totally inhibited and distribution of striated collagen is perturbed. Using antibodies against type I collagen the stroma, which is mainly composed of fibroblasts, is brightly stained, as are some regions within the epithelium. Electron microscopy shows that bundles of striated collagen fibrils appear in extracellular spaces between the lateral membranes of the epithelial cells or between the basal lamina and the epithelial basal membrane. After in situ hybridization using a 35S riboprobe specific for mRNA of the alpha 2 chain of type I collagen, mRNA was detected only in the fibroblasts of the stroma and not in epithelial cells. Furthermore electron microscope studies of collagen bundles in serial sections clearly show collagen fibrils passing through the basal lamina. It is assumed that the type I collagen between epithelial cells originates from mesenchymal cells. In the oviduct of immature birds or after physiological E + P stimulation, striated collagen is localized only in the stroma and never within the epithelium. These results indicate a modulation of extracellular matrix by sex steroid hormones in the quail oviduct.

Hormone-dependent regulation of chicken progesterone receptor deoxyribonucleic acid binding and phosphorylation.

To further understand the structure-function relationships of the chicken oviduct progesterone receptor, the effects of in vivo and in situ progesterone treatment were studied. Immunoprecipitated receptors isolated from oviduct slices incubated in the presence of H(3)32PO4 exhibited hormone-dependent phosphorylation. This was correlated with an increase in the apparent mol wt of receptors when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and increased DNA binding of total cytosolic receptors. Further, in vivo progesterone treatment resulted in dissociation of both the A and B receptor forms from nonhormone-binding proteins (such as heat shock protein-90) in vitro when analyzed by sucrose gradient ultracentrifugation. The 4S and 8S receptors were separated by phosphocellulose column chromatography, treated with ammonium sulfate to convert all receptors to DNA-binding forms, and analyzed for binding to DNA cellulose. The 4S receptor produced as a consequence of in vivo hormone treatment had a 3.35-fold higher affinity for DNA and bound to about a 3-fold greater extent than receptor that did not show altered interaction with other proteins. Thus, in vivo progesterone treatment results in increased receptor phosphorylation, altered interaction with heat shock protein-90, and increased DNA binding.

Chicken egg white cystatin. Molecular cloning, nucleotide sequence, and tissue distribution.

A lambda gt11 chicken oviduct cDNA library was screened with a mixed synthetic oligonucleotide corresponding to amino acid residues 81-90 of chicken egg white cystatin, a cysteine proteinase inhibitor. Two initial cDNA clones of 367 and 431 bases were isolated. Both clones contained coding sequences for cystatin from amino acid residue 82 to the carboxyl end plus 3'-untranslated region and a poly(A)+ tail. The two clones utilized different polyadenylation signals located 55 nucleotides apart. Further screening of the library yielded a full-length cystatin cDNA. Sequence analysis indicated that cystatin contains an NH2-terminal extension of 23 amino acids which is probably a signal sequence. The cystatin cDNA hybridized to an mRNA of approximately 0.95 kilobase and was present in varying amounts in all chicken tissues examined. The highest concentration was found in the lung. Gizzard, brain, and heart contained lesser amounts of cystatin mRNA but considerably higher than oviduct. Among a limited number of embryonic tissues examined, significantly higher levels of the mRNA were found in liver and heart tissues when compared with the corresponding adult tissues. These results suggested that the expression of the chicken cystatin gene is tissue-dependent and under developmental control.

Characterization and partial purification of an estrogen type II binding site in chick oviduct cytosol.

An estrogen binding site of moderate affinity (Kd approximately 10 nM) and high capacity (approximately 25-70 pmol/g of tissue) was measured in DES-stimulated chick oviduct cytosol. Saturation analysis by [3H]estradiol exchange demonstrated that this binding site displayed sigmoidal binding characteristics suggesting a cooperative binding mechanism. Competition analysis with a number of compounds demonstrated that the bioflavonoid luteolin was a better competitor for binding to type II sites in chick than either estradiol or DES. Steroid specificity was demonstrated by the inability of 17 alpha-estradiol, progesterone, testosterone, corticosterone, and the triphenylethylene antiestrogen nafoxidine (U-1100A) to compete for [3H]-17 beta-estradiol binding to chick oviduct cytosol preparations. In addition, the binding site appeared to be sensitive to sulfhydryl reducing reagents as evidenced by a 75% reduction in binding activity in the presence of dithiothreitol. Both prelabeling and postlabeling procedures used in conjunction with Sephacryl S-300 chromatography resulted in a single major peak of type II binding activity representing a molecular weight in the 40,000 range. Type II binding activity was recoverable after precipitation with ammonium sulfate, and this material was subjected to a variety of column chromatography procedures in order to achieve further purification of the type II site. Significant purification of the site was achieved with a bioflavonoid-Sepharose (quercetin-Sepharose) affinity matrix. The purified type II sites eluted from quercetin-Sepharose displayed the same sigmoidal binding curves characteristic of native cytosol.

Site-specific location of covalent DNA-polypeptide complexes in the chicken genome.

A nitrocellulose filter binding assay was applied to isolate and to analyze the fraction of chicken DNA fragments associated with residual nuclear polypeptides resistant to SDS/proteinase K treatment and phenol extraction. It is shown that the DNA-polypeptide complexes retained on nitrocellulose filters are located on a non-random sub-set of DNA sequences. (a) Southern analysis reveals that the fractions of DNA fragments from chicken erythrocytes and from hen oviduct cells associated with the resistant polypeptides have a lower sequence complexity than unfractionated DNA. Moreover, the retained DNA fractions from different cell types of the same species are highly homologous. (b) All DNA fragments of the transcriptionally active and inactive ovalbumin gene map in the DNA fraction passing the filters indicating that the tight DNA-polypeptide complexes are not remnants of transcription complexes. (c) By use of a genomic sub-set library prepared from DNA retained on filters, clones were isolated with sequences mapping specifically in the DNA fraction associated with the tight DNA-polypeptide complexes. The results are consistent with fixed covalent DNA-polypeptide complexes in the chicken genome whose location is essentially identical in different cell types of the same species and apparently determined by DNA signal-sequences.

Immunological detection of spectrin during differentiation and in mature ciliated cells from quail oviduct.

A protein that was immunologically related to the erythrocyte and brain alpha-240-subunit and to the brain beta-235-subunit of spectrin was characterized by immunoblotting and was detected by immunofluorescence in the apical part of ciliated cells from quail oviduct. After immunogold-labeling electron immunocytochemistry, spectrin was detected mainly in a fibrillar meshwork located between the proximal parts of the basal bodies. It was also observed to be in contact with the basal foot of basal bodies, but was not found to be associated with the apical plasma membrane. Cilia and microvilli were unlabeled. In contrast, spectrin was detected in close contact with the lateral plasma membrane of mature ciliated cells as well as in stem epithelial cells in unstimulated oviduct. During ciliogenesis induced by estrogen, spectrin gradually appeared at the apex of the cells as the apical cytoskeleton differentiated.

Two progesterone receptors in the oviduct of the freshwater turtle Chrysemys picta: possible homology to mammalian and avian progesterone receptor systems.

Two progesterone receptors in the oviduct of the freshwater turtle Chrysemys picta: possible homology to mammalian and avian receptor systems. Here we report the characterization of two specific progesterone receptors in nuclear extracts of the turtle oviduct. The receptors differ in dissociation constants (2.8 nM vs 27 nM) which can be separated on DEAE-Sepharose, the former eluting at 0.08 M KCl and the latter at 0.20 M KCl. [3H]R5020 photoaffinity labeling SDS-PAGE revealed that the 2.8 nM moiety migrates with an apparent molecular weight of 80 +/- 5 kDa and the 27 nM moiety migrates with an apparent molecular weight of 120 +/- 5 kDa. These receptors are termed PR-A and PR-B due to their molecular mass and elution profiles. DNA-cellulose chromatographic studies show that both bind DNA-cellulose with the PR-A eluting at 0.09 M NaCl and PR-B eluting between 0.20-0.21 M NaCl. In reproductively inactive turtles (from the months of January and February) estradiol is undetectable, and PR-B is absent as determined by Scatchard analysis, [3H]R5020 photoaffinity labeling electrophoretic studies and DEAE-Sepharose and DNA-cellulose chromatography. In these animals PR-B can be replenished by estrogen treatment, suggesting a physiological role for both PR-A and PR-B and dependence of PR-B on estradiol.

New freeze-dry and vapor fixation method for immunohistochemistry of soluble proteins: subcellular location of the progesterone receptor.

We describe a new application of freeze-drying and vapor fixation for immunohistochemical location of soluble proteins. The method avoids the liquid phase, which eliminates the possible diffusion of soluble proteins. Two vapor fixatives, paraformaldehyde and p-benzoquinone, were tested and p-benzoquinone was found to preserve antigenicity of progesterone receptor (PR) and ovalbumin better than paraformaldehyde. The method proved to be highly sensitive, since higher concentrations of antigen were found in some tissues and some tissues found to be antigen negative by earlier liquid fixation methods proved to contain antigen. The location of PR as a highly soluble protein was studied. With the present method, both unoccupied and occupied PR were located in the nuclei, a similar finding as with the earlier liquid fixation method. The results further support the concept that PR is an intranuclear protein independent of its ligand occupation. PR was detected in a few cells inside the follicles of the bursa of Fabricius and in the smooth muscle cell nuclei of the small intestine, observations not previously made owing to the insensitivity of the earlier methods.

Rapid inhibition of c-myc gene expression by a glucocorticoid in the avian oviduct.

The glucocorticoid dexamethasone (DEX) causes a rapid, reversible reduction in c-myc mRNA level in the oviducts of estrogen-treated, immature chickens. The c-myc mRNA level begins to decrease by 5 min after injection of 0.5 mg DEX, reaches a minimum of 10% of the control value by 30 min, and returns to 30-40% of the control value by 4 h post injection. This rapid effect of DEX on the c-myc mRNA level occurs in both diethylstilbestrol-stimulated and diethylstilbestrol-withdrawn oviducts. The effect is dose dependent, with reduction of the c-myc mRNA measured with as little as 10 micrograms DEX injection (0.03 micrograms/g BW). The effect of the steroid is gene specific with H2B histone mRNA displaying a significantly reduced response. The effect is also tissue specific with liver displaying an increase of 170% of control values in c-myc mRNA level by 30 min after injection of 0.5 mg DEX. The reduction of avian oviduct c-myc mRNA levels by DEX may play a role in glucocorticoid inhibition of cell proliferation in this tissue. The rapidity of the steroid effects on c-myc expression makes it likely that the steroid-induced reduction of c-myc mRNA levels represents a direct primary action of the steroid-receptor complex on the c-myc gene expression.

Oestrogen and progesterone receptors and their relationship to histological grades of epithelial tumours of the magnum region of the oviduct of the domestic fowl.

Tumours of the proprial tubular glands of the magnum region of the oviduct of the domestic fowl (Gallus domesticus) were found to contain receptors for oestrogen and progesterone. The distribution of the receptors in the cytosol and nucleus of the tumour cells was similar to that of the normal magnum under oestrogen stimulation. Receptor content showed a positive correlation with increasing clinical stage and histological grade, i.e., malignancy and undifferentiation of the tumours. These tumours are unusual for hormonally responsive tumours in that their receptor mechanism remains intact with loss of differentiation.

Compared intracellular localization of the glucocorticosteroid and progesterone receptors: an immunocytochemical study.

The intracellular distribution of the glucocorticosteroid and progesterone receptors (GR and PR, respectively) was studied immunohistochemically. In control adrenalectomized (Adx) rat liver, immunostaining of paraffin sections revealed GR in cell nuclei, with a wide range of intensity between individuals. Following dexamethasone (Dex) treatment, the nuclear staining was uniformly high in all animals; the cytoplasmic staining was always weak and remained unchanged after Dex treatment. In frozen sections, the GR immunoreactivity in cell nuclei was weak in the absence and very strong in the presence of Dex, while no GR-specific cytoplasmic staining was observed. In frozen sections fixed in vapor of formaldehyde to avoid any artifactual redistribution of the receptor, some GR immunostaining was observed in the cytoplasm and the nucleus. In contrast, in paraffin as well as in frozen sections of chick oviduct, fixed by immersion or in vapor, PR was exclusively nuclear, including in the absence of progesterone, and the intensity of immunostaining was not modified by progesterone treatment. In order to verify if loss of nuclear receptors during tissue preparation could explain the differences in nuclear immunostaining observed between hormone-free and hormone-occupied GR, and between GR and PR, frozen sections of Adx rat liver and chick oviduct were preincubated at 4 degrees C in buffer solutions before the fixation procedure. It was found that hormone-free GR diffused out of the nucleus faster than hormone-occupied GR nuclei, and that nuclear GR diffused faster than nuclear PR. Based on these results, we propose that, during the fixation procedure, the fraction of nuclear GR which diffuses out of the nucleus is much smaller in the presence than in the absence of Dex. This lesser loss of nuclear GR after Dex treatment results in an increase of immunostaining after hormonal administration, which might have been erroneously interpreted as a sign of translocation from cytoplasm to nucleus. That the nuclear PR detection is not modified by progesterone treatment may be explained by its reduced diffusibility as compared to nuclear GR. This hypothesis does not rule out the existence of some cytoplasmic GR, whose significance remains unclear, but it offers a unified mechanism of action for all steroid hormone receptors. In the case of glucocorticosteroids, as already proposed for estradiol and progesterone, no step of cytoplasm to nucleus translocation would be required for hormone action, and transformation-activation would occur in the nucleus, resulting in tighter binding of the hormone receptor complexes.(ABSTRACT TRUNCATED AT 250 WORDS)

[Dynamics of amino acid and protein metabolism in laying hens after the administration of 15N-labeled wheat protein. 10. 15N-labeling of crude protein and amino acids of the oviducts]. / Dynamik des Aminosäuren- und Proteinstoffwechsels der Legehennen nach Applikation von 15N-markiertem Weizenprotein. 10. Mitteilung. 15N-Markierung des Rohproteins und der Aminosäuren des Oviduktes.

Over a period of 4 days 12 colostomized laying hens daily received 36 g coarse wheat meal containing 14.37 atom-% 15N excess (15N') together with a conventional ration. After the homogenisation of each oviduct N and 15N' were determined. After the precipitation with TCA the 15N' of the amino acids was analysed in both the precipitate and the supernatant. In addition, the free amino acids and the peptides were determined in the TCA soluble fraction. The atom-% 15N' in the total N and in the non-basic amino acid N showed a parallel decrease; it diminished from 1.75 atom-% 15N' to 0.64. Of the three basic amino acids, lysine shows the lowest labelling at all four measuring points. The quotas of non-basic amino acid 14N and 15N' in the total 14N and 15N' of the oviduct are the same and amount to 53%. In contrast to this, the quota of the 14N of the basic amino acids in the total 14N of the oviduct only amounts to 21.6% and that of 15N' only to 15.4%. The average atom-% 15N' of the free amino acids 12 h after the last 15N application is 1.54 and is considerably above that of the peptides with 1.15 atom-% 15N'. 36 h after the last 15N application the ascertained value of 1.25 is identical in both fractions. The labelling of the free amino acids decreases more quickly than that of the peptides the more time has passed after the last 15N application.

Alteration of the spermatozoal glycocalyx and its effect on duration of fertility in the fowl (Gallus domesticus).

The hypothesis that sialic acid has a role in spermatozoal sequestration within the hen's oviduct was tested by treating spermatozoa with Clostridium perfringens neuraminidase. Spermatozoal content of sialic acid ranged from 94 to 135 micrograms per 10(9) spermatozoa (n = 12 roosters). Spermatozoa contained 80% of total seminal sialic acid (coefficient of variation = 4.6%). Spermatozoal sialic acid content was reduced by 18% when 10(9) spermatozoa were incubated at pH 6.5 with 10 IU neuraminidase activity (Type V, Sigma Chemical Co.). Such treatment had no effect on spermatozoal viability as evidenced by ethidium bromide uptake. However, treatment of spermatozoa with neuraminidase prior to intravaginal insemination reduced fertility by 24 percentage units (p less than 0.001). In contrast, when similarly treated spermatozoa were deposited in the magnum via laparatomy, fertility was not affected (p greater than 0.05). The preceding work was done with neuraminidase prepared by salt fractionation (Type V, Sigma Chemical Co.). Type V neuraminidase was absorbed to diethylaminoethyl-Sephacel and then eluted with a stepwise KCl gradient. Treatment of spermatozoa with this preparation of neuraminidase (10 IU/10(9) spermatozoa) prior to intravaginal insemination reduced fertility by 19 percentage units (p less than 0.001). Decreased fertility could not be attributed to contamination of neuraminidase preparation with proteolytic activity. We conclude that spermatozoal sialic acid has a role in spermatozoal sequestration within the hen's utero-vaginal glands.

Species crossreactivity of human progesterone receptor monoclonal antibodies: Western blot analysis.

The crossreactivity of monoclonal antibodies (hPRa 1, 2, 3 and 6) generated against human progesterone receptor was examined in six mammalian and an avian species using the techniques of sodium-dodecylsulfate polyacrylamide gel electrophoresis and Western blot analysis. Immunoreactive bands were detected on protein blots of receptor-containing preparations from human endometrial carcinoma grown in nude mice, human T47D breast cancer cells, rabbit, cow and mouse uteri, and chick oviduct. No receptor-associated, immunoreactive bands were detected in rat, guinea pig or hamster uteri. The number and molecular weights of the receptor subunits detected varied between species, and only human progesterone receptor displayed electrophoretic microheterogeneity in its high molecular weight subunit. These data demonstrate that the human progesterone receptor antibodies recognize epitopes not common to all species.

Autoradiographic localization of beta-adrenergic receptors in rat oviduct.

Catecholamines are known to have an inhibitory effect on oviductal smooth musculature by changing the adrenergic receptors activity. In order to further investigate the role of epinephrine and/or norepinephrine in oviduct, the distribution of beta-adrenergic receptors has been studied in the rat oviduct, using in vitro autoradiography and [125I]cyanopindolol (CYP), as radioligand. The specificity of the labelling and the characterization of receptor subtypes in different cell types was achieved by displacement of radioligand with increasing concentrations of zinterol, a beta-adrenergic agonist with preferential affinity for the beta 2-adrenoreceptor subtype and practolol, a beta-adrenergic antagonist that binds preferentially to beta 1-subtype. Quantitative estimation of ligand binding was achieved by densitometry. It was shown that the vast majority of beta-receptors were of the beta 2-subtype and were found in smooth muscle layers as well as in the epithelium. The latter localization suggests a role for epinephrine and/or norepinephrine on the oviductal epithelium.