In vivo versus in vitro oviductal glycoprotein (OGP) association with the zona pellucida (ZP) in the hamster and baboon.

The goal of this study was to determine if differences exist between in vivo vs. in vitro OGP association with the ZP and to quantitate those differences. Ovarian oocytes were harvested 12.5 or 27 hr post-hCG from hyperstimulated hamsters or baboons, respectively. Hamster and baboon ovarian oocytes were incubated in vitro in media +/- homologous OGP (100 or 200 microg/100 microl) or in some studies with 100 microl oviductal fluid for 3, 6, or 24 hr at 37 degrees C. Some of the baboon ovarian oocytes were transferred immediately after harvesting to the ampulla of both oviducts using a tom cat catheter and retrieved after a 3 hr in situ incubation. Hamster oviductal oocytes were collected 3, 6, and 24 hr following ovulation. After incubation or oocyte retrieval from the oviduct, cumulus cells were removed, oocytes were washed extensively and binding of OGP to the ZP was examined by immunofluorescence. Fluorescence intensity was quantified using densitometric scanning of photographic negatives with the background of each negative as an internal control. In all studies, OGP association with the ZP was significantly greater in vivo than in vitro (P < 0.05). In vitro OGP association with the ZP did not significantly increase with incubation time or OGP concentration; however, a small nonsignificant increase in OGP association with the ZP in the oviduct was detected over time. Differences did not appear to be due to depletion of OGP from the in vitro incubation media, since Western blot analysis of the media showed that OGP was still present. Although OGP concentration in vivo is unknown, Western blots showed similar intensity comparing 100 microg of OGP media and oviductal fluid. Immunolocalization of OGP using laser confocal microscopy showed regional differences in OGP binding. The outer half of the zona pellucida had significantly more OGP bound than the inner half on oviductal oocytes. No regional differences were detected for in vitro incubated oocytes. In conclusion, OGP association with the ZP is greater in vivo vs. in vitro, suggesting that one must be cautious in designing and evaluating in vitro studies of OGP function.

Characterization of antibodies generated against a conserved portion of oviductal glycoprotein (OGP) and endogenous hamster OGP and their ability to decrease sperm binding to the zona pellucida in vitro.

PROBLEM: The effect of antibodies generated against hamster oviductal glycoprotein (OGP) on sperm binding to the zona pellucida (ZP) was evaluated. METHOD OF STUDY: Antibodies against a 17-amino-acid sequence of the OGP core protein (amino acids 52-68) and the denatured hamster OGP protein were generated, characterized, and tested in an in vitro sperm binding assay. RESULTS: Sperm binding was significantly decreased (P < 0.05) when oviductal oocytes were incubated for 2 hr with 4 or 8 mg/ml of immune IgG of both antibodies when compared with normal rabbit IgG. A fluorescence assay showed binding of both antibodies to the endogenous OGP associated with the ZP of ovulated hamster oocytes. CONCLUSIONS: These results suggest that OGP may be a potential immunocontraceptive target because both antibodies significantly decreased sperm binding to the ZP of oviductal oocytes. Immunocontraception may be accomplished by attempting to generate active immunity to a recombinant OGP, to the region selected in this study (amino acids 52-68) or to some other region of the core protein.

Species-specific effect of oviductal glycoproteins on hamster sperm binding to hamster oocytes.

The secretory cells of the oviductal epithelium secrete a high-molecular-weight glycoprotein (OGP). OGPs from different mammalian species show similar immunological characteristics, their cDNAs show high homologies, and they associate with the zona pellucida of oviductal oocytes in vivo. The purpose of this study was to determine the effect of OGP obtained from different species on the binding of hamster sperm to hamster oocytes. Hamster oocytes were inseminated (30 min) in the presence or absence of homologous or heterologous OGPs, and sperm bound/oocyte were counted after removing loosely attached sperm. Ovarian oocytes had an average of 2.9 +/- 0.6 sperm bound/oocyte, whereas oviductal oocytes had 36.3 +/- 2.7. Hamster OGP (0.1 mg/ml) significantly increased sperm binding to ovarian oocytes twofold and had no effect on sperm bound/oviductal oocytes. Human OGP (0.5 mg/ml) significantly decreased sperm binding to ovarian oocytes (0.9 +/- 0.3 sperm bound/oocyte). This effect was dose dependent for oviductal oocytes and could be blocked by preincubating human OGP with a specific antibody to human OGP. The presence of baboon and cow OGP during the insemination of hamster oviductal oocytes also resulted in a significant decrease in sperm bound/oocyte, whereas the addition of hamster OGP to hamster oviductal oocytes had no effect. These results show that homologous OGP enhances sperm binding to the ZP, whereas heterologous OGP inhibits that effect. Thus, our results suggest that OGP plays a role in the species-specific characteristics of sperm/ZP interaction, and that one must use a homologous system (OGP and gametes from the same species) to study the biological effect of OGP.

Characteristics of an oviductal glycoprotein and its potential role in fertility control.

At the time of ovulation the lining epithelium of the mammalian oviduct consists of columnar ciliated and secretory cells. These mature cells are dependent on ovarian steroids in carnivores. Oestradiol induces differentiation of these cells and maintains their mature functional state, and progesterone induces dedifferentiation. The secretory cells synthesize and secrete an oestrogen-dependent high molecular weight glycoprotein. The cDNAs encoding oviductal glycoproteins from several species have been sequenced and show high similarity. The human cDNA hybridized with a single message on northern blots of total oviduct RNA obtained from oestradiol-treated cats (about 2.3 kb) and dogs (about 2.1 kb). This glycoprotein is the major nonserum protein present in the oviductal lumen at the time of ovulation, fertilization and early embryonic development. The glycoproteins associate with the zona pellucida of oviductal eggs in all species studied to date. Recent studies suggest that the bovine glycoprotein facilitates sperm capacitation and significantly increases the ability of bovine spermatozoa to fertilize bovine oocytes in vitro, that the hamster glycoprotein increases the sperm penetration rate of the zona pellucida by three times and that the human glycoprotein increases sperm binding to the zona pellucida by three times. All of the evidence for a biological function for this glycoprotein is derived from studies performed in several different species at reproductive stages before fertilization. The biological actions of this glycoprotein suggest a potential role for the glycoprotein in fertility control. Specifically, purified or recombinant glycoprotein may improve success in IVF procedures by enhancing binding of spermatozoa to the zona pellucida and improving fertilization rates. The glycoprotein may also be a potential immunocontraceptive target since antibodies generated against the oviductal glycoprotein may prevent fertilization by preventing binding of spermatozoa to the zona pellucida.

The baboon oviduct: characteristics of an oestradiol-dependent oviduct-specific glycoprotein.

The baboon oviductal epithelium differentiates into a tall columnar epithelium consisting of ciliated and secretory cells during the follicular phase of the menstrual cycle in response to rising oestradiol levels. The apical tips of these secretory cells are filled with membrane-bound secretory granules. During the luteal phase when progesterone levels are elevated, the epithelium regresses and deciliation occurs. Analysis of secretory proteins obtained from explant culture media by SDS-PAGE followed by fluorography or Western blots has revealed that the baboon oviduct synthesizes and secretes a high molecular weight glycoprotein during the follicular phase of the cycle. Immunocytochemistry demonstrated that this oviductal glycoprotein is localized to the secretory granules of epithelial secretory cells, is oviduct specific, and that following secretion the oviductal glycoprotein binds to the zona pellucida and perivitelline space of ovulated oocytes and embryos within the oviduct. Similar proteins have been characterized in other mammalian species. cDNA data show that the complete coding sequence is 2228 bp for a protein of 623 amino acids. A Genbank search showed that baboon oviductal glycoprotein has high homology to other oviductal glycoprotein sequences at both the nucleotide and amino acid levels. Studies conducted to date probing the biological function of oviductal glycoprotein indicate that this protein plays a role in prefertilization reproductive events (sperm capacitation; sperm-zona binding; zona penetration). Additional experiments are needed to reveal a specific function and mechanism for this molecule.

Regional distribution and hormonal control of estrogen-dependent oviduct-specific glycoprotein messenger ribonucleic acid in the baboon (Papio anubis).

Our objective in this study was to complete the sequence of the baboon oviductal glycoprotein, examine the hormonal regulation of the oviductal glycoprotein mRNA, and determine whether there was a regional variation within the oviduct in the level of oviductal glycoprotein mRNA expression. Finally, because of the structural similarity of the amino terminal end of the oviductal glycoprotein to chitinases, we sought to determine whether the oviductal glycoprotein functions as a glycosyl hydrolase. The total transcript length of the baboon oviductal glycoprotein was determined to be 2228 nucleotides in length plus a poly(A) tail. The largest open reading frame was 623 amino acids, which would produce a protein of 69.3 kDa. The first 420 amino acids were highly homologous to the amino acid sequence of other oviductal glycoproteins, but the remainder of the sequence differed considerably from that of all other species except the human. Although the N-terminal region exhibited sequence similarity to chitinases, the oviductal glycoprotein did not exhibit any activity towards typical chitinase substrates. The oviductal glycoprotein mRNA levels were elevated to approximately the same extent in the fimbria, ampulla, and isthmus of the oviduct after estradiol treatment and in the late follicular stage of the menstrual cycle. The oviductal glycoprotein mRNA levels were lower in the early follicular stage and early luteal stage and were not detectable in the late luteal stage or in progesterone-treated baboons. These results indicate that the oviductal glycoprotein mRNA is induced by estradiol and is present at the highest levels at the time of fertilization. Although there is structural homology with chitinases, no such glycosyl hydrolase activity could be detected. However, the common structure of the N-terminal region of the oviductal glycoproteins implies that it has the same, as yet unknown, function in all species.

Ontogenesis of IGF regulation of longitudinal bone growth in rat metatarsal rudiments cultured in serum-free medium.

The aim of this study was to compare the effect of two cytokines, IGF-I and IGF-II on skeletal development in the rat. The three medial metatarsal rudiments were dissected out from fetuses at days 19, 20 or 21 of gestation and from newborns at days 1, 3, 6 and 9 after birth, then grown in serum-free MEM medium at 37 degrees C and 5% CO2 in air. From day 19 of gestation to the end of experiment, longitudinal bone growth (mm) was significantly increased by IGF-I (2.975 +/- 0.050) and IGF-II (2.530 +/- 0.062), compared to controls (2.188 +/- 0.060). In the same way, the width (mm) at the last experimental day was 0.360 +/- 0.010 in IGF-I- and 0.327 +/- 0.008 in IGF-II-treated bones, respectively (vs 0.313 +/- 0.012 in controls). Mineralization was also stimulated under both growth factors (length of the calcified diaphysis (mm): 0.691 +/- 0.019 in IGF-I- and 0.446 +/- 0.017 in IGF-II-treated bones; vs 0.383 +/- 0.024 in controls). IGF-I and IGF-II (but to a lesser extent) stimulation was due to an increased DNA synthesis (3H-thymidine uptake) as well as protein anabolism (incorporated proline). In addition, cartilage activity (35S captation) and mineralization (45Ca fixed) were involved in the action of these cytokines. An age dependency of bone response to IGFs was pointed out, the effect being higher during the fetal period than after birth. In conclusion, our results raise the possibility that IGF-II, as well as IGF-I, is involved in the control of osteogenesis.

Association of oviduct-specific glycoproteins with human and baboon (Papio anubis) ovarian oocytes and enhancement of human sperm binding to human hemizonae following in vitro incubation.

The objectives of this study were 1) to determine whether or not human and baboon oviduct-specific glycoproteins (human OGP, baboon OGP) would associate with ovarian oocytes during in vitro incubation in a manner similar to that detected in vivo for oviductal oocytes and 2) to determine whether the association of OGP with ovarian oocytes influenced sperm binding. In vitro association of OGP with ovarian oocytes was assessed by indirect immunofluorescence assay using a polyclonal antibody prepared against human or baboon OGP. Human and baboon ovarian oocytes incubated in culture media containing OGP showed association of OGP with the zona pellucida (ZP) as detected by bright fluorescence. A similar pattern of fluorescence was observed in baboon oviductal oocytes (positive control). No fluorescence of the ZP was detected from ovarian oocytes incubated with culture medium alone. The pattern of fluorescence for ovarian oocytes incubated with OGP and serum albumin, the major oviductal fluid protein, was similar to that for oocytes incubated with OGP alone. A modified hemizona assay was used to assess whether association of human OGP with human ovarian oocytes influenced sperm binding. The number of sperm bound to hemizonae in the presence of human OGP was significantly greater (p < 0.01) than the number bound to hemizonae in the control culture medium. Addition of antibodies specific for human OGP to the incubation medium 1 h prior to addition of gametes blocked the enhancement of sperm binding seen in the presence of human OGP alone. Finally, human hemizona assays conducted in the presence of baboon OGP resulted in a significant decrease (p < 0.05) in the number of sperm bound per zona compared with that in culture medium alone despite high homology between human and baboon OGP. These results 1) suggest that human OGP associates with ovulated oocytes in vivo; 2) support the hypothesis that association of OGP with the ZP may play a role in fertilization, possibly through enhancing the binding of sperm to the ZP within the oviduct; and 3) suggest that a homologous system (i.e., gametes and oviductal glycoprotein from the same species) is necessary for study of the function of oviductal glycoproteins.

A human oviduct-specific glycoprotein: synthesis, secretion, and localization during the menstrual cycle.

The major objective of this study was to examine the hormonal regulation of a human oviduct-specific glycoprotein (huOGP) throughout the menstrual cycle and in all regions of the human oviduct. Regulation of synthesis and secretion was examined at both the protein (Western immunoblots and immunocytochemistry) and mRNA (Northern and slot blots) levels and correlated with changes in the morphological features of the oviductal epithelial cells throughout the cycle. Immunoblot analysis of oviductal fluid and explant culture media from all regions of the oviduct demonstrated that huOGP is primarily found during the follicular stage of the cycle and is not present in serum, follicular fluid, or uterine endometrium. Moreover, two-dimensional (2-D) immunoblots showed that all major isoelectric variants of huOGP observed on 2-D fluorographs are immunologically related. Light microscopic immunocytochemistry localized huOGP to oviductal secretory cells in both ampulla and isthmic regions, with the most intense immunoperoxidase staining seen in midcycle samples. Using an indirect immunogold technique at the electron microscopic level, huOGP was specifically localized to secretory granules of the ampullary and isthmic nonciliated epithelial cells. The ultrastructural characteristics of these secretory cells during the mid to late follicular phase of the cycle suggested elevated protein synthetic activity. In addition, mRNA expression for huOGP was elevated in all regions of the oviduct in midcycle specimens. Collectively, these data indicate that huOGP is a major tissue-specific, stage-specific secretory product of the human oviduct during the periovulatory stage of the cycle and support the hypothesis that huOGP synthesis and secretion may be regulated by fluctuations in the levels of estrogen and progesterone.

Metabolic end products inhibit sarcoplasmic reticulum Ca2+ release and [3H]ryanodine binding.

Sarcoplasmic reticulum (SR) Ca2+ release channel function is modified by ligands (Mg2+, Ca2+, ATP, and H+) that are generated during a bout of exercise. We have examined the effects of changing intracellular metabolites on Ca2+ release, [3H]ryanodine binding, and single-Ca2+ release channel activity of SR isolated from white rabbit skeletal muscle. Increasing Mg2+ (from 0 to 4 mM) and decreasing pH (7.1-6.5) inhibited SR Ca2+ release and [3H]-ryanodine binding. In addition, increasing lactate concentrations from 2 to 20 mM inhibited [3H]ryanodine binding to SR vesicles, inhibited SR Ca2+ release, and decreased the single-channel open probability. These findings suggest that intracellular modifications that disrupt excitation-contraction coupling and decrease Ca2+ transients will promote a decline in tension development and contribute to muscle fatigue. In addition, we show that hydrogen peroxide induces Ca2+ release and increases [3H]ryanodine binding to its receptor, suggesting that reactive oxygen species produced during exercise may compromise muscle function by altering the normal gating of the SR Ca2+ release channel.

In vitro incubation of golden (Syrian) hamster ovarian oocytes and human sperm with a human oviduct specific glycoprotein.

The objective of this study was to determine if human oviduct specific glycoprotein (huOGP) would associate with hamster ovarian oocytes and human sperm during in vitro incubation. The huOGP used in these studies was partially purified from human hydrosalpinx fluid. Hamster ovarian oocytes and human sperm samples were incubated in culture medium with and without huOGP. Association of huOGP was assessed by indirect immunofluorescence assay using a polyclonal antibody prepared against huOGP. Intense fluorescence of the zona pellucida, and bright but uneven fluorescence of the perivitelline space, were observed in hamster ovarian oocytes following incubation in the presence of huOGP. A similar but more uniform pattern of fluorescence was observed when hamster oviductal oocytes (positive controls) were incubated in culture medium alone. Fluorescence was absent when oocytes were assayed with preimmune serum. The association of huOGP with the zona pellucida and perivitelline space appeared to be specific since thyroglobulin, a large molecular weight glycoprotein, and human serum albumin, the major protein in oviduct fluid, did not associate with the hamster oocytes nor inhibit huOGP association when included in the culture medium. Fluorescence was absent when human sperm incubated with huOGP were assayed with antiserum to huOGP. However, human sperm fluoresced when incubated with a uterine glycoprotein, CUPED, which had previously been shown to bind to cat sperm during in vitro incubation. Sperm also fluoresced brightly when human sperm antibody was used as a positive control. Solubilization of sperm membrane proteins postincubation and analysis of these proteins by 1-D SDS-PAGE followed by immunoblotting also failed to show an association of huOGP with human sperm. Electron microscopy of sperm both pre- and postsolubilization confirmed that the sperm membranes were removed by this process. In conclusion, the association of huOGP with hamster oocytes in vitro suggests that huOGP may associate with human oocytes in vivo, whereas that may not be true for human sperm in vivo. The association of huOGP with oocytes may serve to facilitate the process of fertilization and early embryonic development within the oviduct.

Hormonal control of the cervix in pregnant gilts. IV. Relaxin promotes changes in the histological characteristics of the cervix that are associated with cervical softening during late pregnancy in gilts.

We recently described relaxin-induced changes that occur in both the physical properties and the biochemical composition of the uterine and vaginal portions of the cervix during the last third of gestation in the gilt. This study employed morphometric analysis to examine both the changes that occur in the histological characteristics of the uterine and vaginal portions of the cervix between days 80 and 110 of pregnancy in intact gilts and the effects of relaxin on these changes in ovariectomized gilts given progesterone to maintain pregnancy. There were four treatment groups: intact day 80 control gilts, sham-ovariectomized day 110 control gilts, ovariectomized progesterone-treated day 110 gilts, and ovariectomized progesterone- plus relaxin-treated day 110 gilts. The histological characteristics of the uterine portion of cervices obtained from intact controls on day 110 differed markedly from those of intact controls on day 80; there was a reduction in the density and organization of collagen fiber bundles, a reduction in the density of smooth muscle fiber bundles, and an increase in amorphous ground substance. The histological characteristics of cervices removed on day 110 from ovariectomized gilts given progesterone only did not differ from those of day 80 controls. After replacement therapy with progesterone plus relaxin, however, the histological characteristics of the cervix on day 110 did not differ from those in intact day 110 controls. Additionally, this study demonstrates that the relaxin-dependent changes are more dramatic in the uterine than in the vaginal portion of the cervix. These findings are consistent with and extend our earlier findings that relaxin-dependent changes in the physical properties and biochemical composition of the uterine portion of the cervix far exceed those in the vaginal portion of the cervix during late pregnancy in the gilt. We conclude that the relaxin-dependent changes in the histological characteristics of the cervix described in this report may contribute at least in part to the cervical softening that occurs during late pregnancy in gilts.

Immunological characterization and immunocytochemical localization of an oviduct-specific glycoprotein in the human.

The objective of the current study was to generate a polyclonal antibody toward a previously described 110- to 130-kilodalton (kDa) human oviductal glycoprotein and to use the antibody to detect the protein in tissue sections, tissue culture media, and oviductal flushings. The polyclonal antibody was generated in male rabbits against the 110- to 130-kDa glycoprotein partially purified from hydrosalpinx fluid. Segments of human oviducts were either cut into 2- to 3-mm pieces and cultured for 24 h, or fixed and embedded in Araldite for light and electron microscopic immunocytochemistry. The protein was only present in midcycle oviductal flushings and was most evident in culture medium samples obtained at midcycle when analyzed on Western blots. No cross-reactivity was observed with proteins in human serum or human endometrial and cervical explant culture media. Immunoperoxidase staining was observed in the apical granules of the secretory cells lining the oviductal lumen. No staining was noted in other parts of the oviduct or in sections of human endometrium and cervix. Indirect immunogold localization demonstrated specific clustering of gold particles over the apical granules of the secretory cells. In summary, a polyclonal antibody to a 110- to 130-kDa human oviductal glycoprotein was successfully generated. This protein is found in the secretory cells and is released into the oviductal lumen. The synthesis of this protein appears to require elevated levels of estrogen and may play a role in early reproductive events occurring within the oviduct.

The physiological effects of relaxin during pregnancy: studies in rats and pigs.

For 50 years after its discovery in 1926, there was a general lack of interest in relaxin among both reproductive biologists and clinicians. A key reason for this lack of interest was the lack of information concerning relaxin's physiological importance during pregnancy in any species. Research conducted since the early 1980s has established that the hormone relaxin is essential during pregnancy in at least two species--rats and pigs. Two vital roles for relaxin during pregnancy have been identified. Relaxin promotes growth and softening of the uterine cervix and thereby enables rapid and safe delivery in both rats and pigs. Relaxin also promotes growth and development of the mammary apparatus in both species. Interestingly, the major effects of relaxin on mammary growth and development are targeted on the nipple in the rat, whereas they are targeted on the glandular parenchyma in the pig. Relaxin-dependent growth of the nipple in rats is required for normal lactational performance. Although likely, it remains to be established that relaxin's profound effects upon mammary gland development in pigs are required for normal lactational performance. The fact that relaxin has effects upon cervical and mammary gland development during pregnancy in both rats and pigs encourages the view that relaxin may have similar effects during pregnancy in other species. Nevertheless, one must keep in mind that there is great diversity in the physiology of relaxin among species (reviewed by Sherwood 1988). This diversity includes not only relaxin's source, regulation of synthesis and secretion, and secretory profiles during pregnancy, but also its biological effects. It seems essentially certain that relaxin's effects during pregnancy differ among species. For example, transformation of the pubic joint cartilage to a flexible and elastic interpubic ligament occurs during pregnancy in several species, including guinea pigs, mice, and bats. This pelvic adaptation, which is nearly certainly relaxin dependent, does not occur in species such as rats and sheep. It is possible that relaxin may have little or no physiological significance during pregnancy in some species. Although considerable progress has been made toward an understanding of the physiological role(s) of relaxin in pregnant rats and pigs, many fundamental questions remain unanswered.(ABSTRACT TRUNCATED AT 400 WORDS)

Hormonal control of the cervix in pregnant gilts. III. Relaxin's influence on cervical biochemical properties in ovariectomized hormone-treated pregnant gilts.

The effects of relaxin on the biochemical properties of both the uterine and vaginal portions of the cervix were examined between days 80-110 of pregnancy in ovariectomized gilts given progesterone to maintain pregnancy. In the cervix of control gilts and those ovariectomized and given progesterone plus relaxin, wet and dry weights, water content, and the glycosaminoglycan/collagen ratio increased between days 80-110 of gestation. Collagen concentrations based on wet or dry weight and glycosaminoglycan concentrations based on wet weight decreased during this period. After ovariectomy, there were no changes in these cervical connective tissue components when relaxin was not given. The glycosaminoglycans hyaluronic acid, heparan sulfate, and dermatan sulfate were found in the cervices of all treatment groups, with dermatan sulfate predominating. The ratio of the individual glycosaminoglycans did not change during pregnancy or with treatment. The major dermatan sulfate proteoglycan from the pig cervix was isolated and found to be similar in size, immunoreactivity, amino acid composition, and amino acid sequence to the major dermatan sulfate proteoglycans isolated from the cervices of other mammals. It is concluded that the relaxin-induced changes in the connective tissue composition of the cervix may contribute at least in part to increased extensibility and growth of the cervix during the last third of gestation in pigs.

Effect of relaxin on mammary development in ovariectomized pregnant gilts.

The role of relaxin in mammary development was studied between days 80-110 of pregnancy in ovariectomized gilts given progesterone to maintain pregnancy. To obtain an objective measurement of lobulo-alveolar (parenchymal) composition, mammary glands were cut in cross-section through the teat, and the area of parenchymal tissue on the exposed face of the gland was determined. Ovariectomy on day 80 or 100 followed by progesterone replacement therapy resulted in a dramatic reduction in the rate of growth of mammary parenchymal cross-section area on days 100 and 110 of gestation, respectively, compared to that in controls. In contrast, progesterone plus relaxin therapy, with highly purified porcine relaxin, restored the mammary parenchymal cross-section area to control values in ovariectomized gilts. Morphometric analysis of mammary tissue on day 110 of pregnancy indicated that both the absence of relaxin after ovariectomy and replacement therapy with porcine relaxin in ovariectomized gilts had little if any effect on the percentages of the lumen, stroma, or epithelial that comprised the mammary parenchyma. It is concluded that relaxin has a stimulatory effect on the growth of mammary parenchymal tissue during late gestation in the pig.

The copper hypothesis of schizophrenia: a review.

The hypothesis that excess tissue copper can cause schizophrenia is a relatively old theory that has never been compellingly demonstrated nor convincingly refuted. This article traces the development and abandonment of the copper hypothesis and examines the evidence for and against the etiological involvement of copper in schizophrenia. A plausible mechanism by which copper excesses could result in schizophrenia is presented and evaluated, and various attempts to reconcile the contradictory data are considered.