The location of eggs retained in the oviducts of mares

The oviducts of 24 mares were examined to determine the site of retention of unfertilized eggs. The ampullary-isthmic junction regions of 42 of the 48 oviducts were serially sectioned and examined histologically. The remaining parts of the oviducts were flushed and the flushings searched microscopically. Of 45 eggs located, 40 were in the sectioned segments of 24 oviducts and only 5 were in the flushings. All but one of the sectioned segments contained prominent masses of material obstructing the lumen, but these were apparently not the direct cause of egg retention since eggs were found on both sides of them.     

Oviduct development inDrosophila

Summary The lateral oviducts ofDrosophila virilis were investigated in normal unoperated adult females, after unilateral and bilateral ovariectomy, and following the transplantation of genital discs. Subsequent to unilateral ovariectomy at larval stages of development, mature adult females exhibited reproductive systems with a free lateral oviduct which appeared somewhat shorter and less distended than a lateral oviduct normally attached to an ovary. Histological examination revealed that such free lateral oviducts have undergone considerable growth and differentiation in the absence of direct ovarian attachment, but exhibited a smaller lumen and more highly folded epithelium. They may be distinguished from attached lateral oviducts by conformational differences and by a possibly lesser size of the epithelial cells. Free lateral oviducts observed among bilaterally ovariectomized and sham-ovariectomized specimens were indistinguishable from those obtained after unilateral ovariectomy. The results are at variance with the previously accepted conclusion that oviduct growth inDrosophila is dependent upon inductive influences emanating from the ovary and directing the outgrowth of the oviducts. Differences in the developmental performance of the oviducts as a function of age at the time of ovariectomy were not evident in the study which included larvae ranging from the second instar to late in the third instar. Transplants of larval female genital discs to other larvae revealed a lesser development of the lateral oviducts than that exhibited by a genital disc developingin situ. A range of oviducal growth which lacked any relation to ovarian influences or to other internal organs of the hosts was obtained. In general, decreased amounts of oviducal muscle were found associated with the transplants.Presented in partial fulfillment of the requirements for the degree of Doctor of Philosophy at the University of Virginia. This investigation was supported in part by: a National Science Foundation Co-operative Fellowship, by the United States Public Health Service Predoctoral Grant 1F1-GM 19,262-1, and by a Summer Research Grant, University of Missouri-St. Louis.     

Microvascular architecture of the pregnant rabbit oviduct

Methyl-methacrylate vascular corrosion casts of the oviducts were prepared in 7 rabbits which were 2-3 weeks pregnant. Scanning electron microscopy of the acrylic casts revealed little change in tubal microvascular connections when compared with control oviducts. Venous distension in the isthmic subserosal venous plexus, ampullary subserosal vasculature and in the fimbrial core was substantially greater than that observed in controls. These changes are interpreted as indicating a sensitivity of tubal microvasculature to the increased levels of circulating placental hormones in pregnancy. The implications of this interpretation in the role of tubal microvasculature at the time of ovulation are discussed.     

Infertility in mice infected genitally with a human strain of Chlamydia trachomatis

Progesterone-treated C3H and TO mice were inoculated genitally with a human C. trachomatis strain, serovar E, designated N.I. 1 or with 2SP control medium. Of the C3H mice serving as controls 93% had litters by the end of a 6-month period compared to 31% of mice infected with chlamydiae. This infertility could not be explained by tubal occlusion, since the oviducts appeared normal at autopsy. Some of the mice were induced to superovulate. Eggs were never recovered from the oviducts on the inoculated sides of infertile mice although they were sometimes found in the lumen of the uninoculated oviducts. In contrast, eggs were recovered routinely from both oviducts of control mice. In addition, eggs and/or their accompanying cumulus cells could be seen in the periovarial space of mice inoculated with chlamydiae, indicating a failure of the transportation of eggs to the oviduct. This could explain the high incidence of ectopic pregnancies in women after chlamydial infection. No adverse effect on fertility was seen in TO mice inoculated genitally with strain N.I.1. Of the mice given 2SP medium, 73% had litters, but 87% of the mice inoculated with chlamydiae were also fertile. There was, however, a significantly greater variation in the birth weights of mice born to infected TO mothers than those born to control mice. This difference in the susceptibility of mouse strains suggests that a genetic predisposition should also be considered for man.     

Growth restriction of cranial sutures in the fetal lamb causes deformational changes, not craniosynostosis

Newborns with in utero cranial vault molding can present with severe forms of plagiocephaly. Intrauterine constraint has been proposed as one cause for craniosynostosis. The purpose of this experiment was to investigate whether rigid plate fixation across a fetal cranial suture, representing a severe form of growth restriction in utero, would lead to cranial suture fusion in a fetal lamb model. Six fetal lambs at 85 to 95 days gestation (term = 145 days) underwent laparotomy, hysterotomy, fetal coronal scalp incision, and miniplate screw fixation across the right coronal suture in utero. Two unoperated twins and four unoperated age-matched lambs were used as controls (n = 12). Animals were killed at both 4 and 8 weeks postoperatively. Fetal head analysis consisted of gross examination, photography, basilar and lateral radiographs, and three-dimensional computed tomographic scans. Cranial suture analysis consisted of imaging by computed tomographic scan (axial and sagittal cuts) and histology of experimentally plated coronal sutures, contralateral nonplated coronal sutures and twin control coronal sutures. Gross examination, radiographs, and three-dimensional computed tomographic analysis of heads with cranial suture plating showed ipsilateral forehead flattening, contralateral forehead bossing, superiorly displaced ipsilateral orbital rim, anterolateral projection of ipsilateral malar eminence, and anterior position of the ipsilateral ear point compared with the contralateral side of the same animal and normal controls. There was no change in nasal root, chin point, or predentition occlusal plane. Although analysis of the plated coronal sutures by computed tomographic scans showed diminished width or even stenosis, the histology revealed narrowed but patent experimental coronal sutures at 4 and 8 weeks. Contralateral, nonplated coronal sutures were not only patent, but widened compared with normal control sutures. This finding may have represented compensatory changes in the contralateral coronal suture caused by growth restriction at the plated suture. These data demonstrate that intrauterine growth restriction across a cranial suture caused by compression plate fixation resulted in deformational skull changes, not craniosynostosis. In addition, these data strongly support a role for in utero positional molding secondary to growth restriction in the maternal pelvis as a cause for nonsynostotic plagiocephaly seen in newborns.     

Basal cells in the mouse olfactory epithelium after axotomy: immunohistochemical and electron-microscopic studies

Summary The olfactory epithelium of mice after axotomy was investigated to clarify the stem cells of olfactory cells by double immunostaining using antikeratin (MA903) and anti-bromodeoxyuridine (BrdU) antibodies and by conventional electron microscopy. When a single dose of BrdU was given to mice 9 days after axotomy, immunostaining for BrdU was found in the globose basal cells which were negative for MA903, but not in the basal cells proper which were positive for MA903. The BrdU-immunoreactive cells increased 3-to 6-fold over the number of these cells in the controls, indicating active cell proliferation. At other postoperative days (4 and 14 days), fewer BrdU-immunoreactive cells were found. Furthermore, three pulses of BrdU resulted in numerous BrdU-immunolabelings in the globose basal cells and a few in the basal cells proper. There was no detectable difference in the number of labeled basal cells proper in operated and unoperated mice. In the electron micrographs 9 days after axotomy, the basal cells proper, flat-shaped in unoperated mice, appeared cylindrical or pyramidal in shape and the globose basal cells often lay between the basal cells proper. In unoperated controls, the globose basal cells were located above the flat-shaped basal cells proper. The results suggest that the stem cells of the olfactory cells are globose basal cells and not basal cells proper, and that the shape of basal cells proper changes in relation to the active proliferation of stem cells.     

EFFECTS OF SINUS NERVE STIMULATION ON CAROTID BODY GLOMUS CELLS

The sinus nerve or sympathetic trunk was stimulated unilaterally in one group of adult cats or Syrian hamsters while in another group the sinus nerve or sympathetic trunk was cut unilaterally and the animals were given reserpine. In a third group, atropine was administered prior to sinus nerve stimulation. All tissues were processed for the detection of primary monoamines. The carotid bodies on the operated sides were compared with those on the unoperated sides of the same animal in order to determine if amine depletion occurred following the experimental procedures. After sinus nerve stimulation alone, the density of the granules in the glomus cells was decreased, but changes were not noted in the granules following sympathetic nerve stimulation. Sinus nerve stimulation after atropine administration resulted in no change in granule density. Sinus nerve transection followed by reserpine treatment resulted in a greater decrease in granule density on the unoperated than on the operated side. Transection of the sympathetic components to the carotid body followed by reserpine injections resulted in a decrease in granule density in the glomus cells on both the operated and unoperated sides. These results suggest that the sinus nerve must be intact for reserpine to exert an effect and that the sinus nerve may contain efferent fibers which modulate amine secretion.     

STRUCTURE OF THE ROYAL ANN CHERRY CUTICLE AND ITS SIGNIFICANCE TO CUTICULAR PENETRATION

The structure of the Royal Ann cherry cuticle (Prunus avium L.) was determined and interpreted in the light of its possible significance to cuticular penetration by a SO₂-calcium bisulfite brine. The morphology of the cuticle was determined by standard histological and histochemical techniques. The surface structure of the cuticle was found to have a smooth to granular sheet or layer of surface wax, which when removed revealed a porous sponge-like surface. The cuticular surface showed intermittent birefringence, which increased as the fruit matured. Ectodesmata were found to occur over anticlinal walls and in guard cells on both sides of the fruit, with more on the side opposite the suture. Both sides had stomata with more occurring on the suture side. Secondary bleaching was found to alter the structure and permeability of the cuticle.     

Choline Acetyltransferase Activity in the Rat Brain Cortex Homogenate, Synaptosomes, and Capillaries After Lesioning the Nucleus Basalis Magnocellularis

Stereotaxic lesions of the nucleus basalis magnocellularis were made unilaterally in male Wistar rats with either kainic or ibotenic acid, using the contralateral side as control. Differences in behavior, body weight, and survival were observed between the kainic and ibotenic acid-treated rats. One week after surgery, the rats were sacrificed and the effect of the lesions on choline acetyltransferase activity was measured in brain cortex homogenate, synaptosomes, and capillaries. In kainic acid-lesioned rats, choline acetyltransferase activity decreased in homogenate and synaptosomes of the ipsilateral side with respect to that of the contralateral side; but the ibotenic acid lesion, which also reduced the ipsilateral choline acetyltransferase activity in homogenate, showed a rather different effect on the enzymatic activity of the synaptosomes. There were also differences between the effect of kainic and ibotenic acid lesions on choline acetyltransferase activity in the capillaries of the ipsilateral side with respect to that of the contralateral one. However, capillary choline acetyltransferase activity of the treated rats was in both sides three times higher than that of unoperated rats.     

The effect of rigid fixation on growth of the neurocranium

The effects on skull growth of plating the coronal suture and frontal bone were studied in New Zealand White rabbits. Three-dimensional coordinate landmarks were digitized and analyzed to determine the differences in form between operated and unoperated animals using Euclidian distance matrix analysis. This method compares sets of interlandmark distances in three dimensions and was used to demonstrate changes induced by plating. We interpret these changes in morphology to be the result of differences in growth between the operated and unoperated groups. Periosteal elevation alone (n = 6) resulted in a minimal local growth increase. Coronal suture plating (n = 8) resulted in local growth restriction with contralateral and adjacent size increases. Frontal bone plating (n = 6) without crossing a suture line also resulted in local growth restriction and adjacent bone size increases. The timing of intervention in relation to the completion of bone growth may explain the magnitude of clinically apparent effects. Changes in bones adjacent to those directly manipulated may be an attempt to maintain a normal skull volume.     

Morphological observations on sperm–egg interactions during in vivo fertilization in the dasyurid marsupial Sminthopsis crassicaudata

The aim of the present study was to determine the morphological changes that take place in the male and female gametes during in vivo fertilization in the Australian marsupial, the fat-tailed dunnart, Sminthopsis crassicaudata. Plastic sections were cut of sperm and eggs recovered from the oviducts of recently mated individuals, and light microscopy of thick, and transmission EM of thin, sections was carried out. It was found that, before penetration of the zona, the spermatozoon came to lie along the outer surface with its rostral tip forming a depression in the zona substance. During penetration, zona material was packed tightly around the spermatozoon, and no large hole was formed. A spermatozoon within the perivitelline space had made contact with the oolemma by way of its apical tip. In a spermatozoon partly incorporated into the ooplasm, fusion appeared to have taken place between its plasma membrane and that of the oolemma. Mucoid coat material became deposited outside the zona at this time; its existence and/or the release of cortical granule content probably prevented polyspermy. Once inside the egg cytoplasm, the sperm head sometimes travelled a considerable distance before chromatin decondensation occurred. In addition, it appeared to rotate somewhat on its axis at this time. Finally, some membranous structures were found around two condensed sperm heads in the ooplasm, which may have been part of the pronuclear envelope. Thus this study on in vivo fertilization in the dunnart documents, for the first time, some aspects of fertilization in an Australian marsupial as seen with the transmission electron microscope; it indicates a few differences from those previously found for the American opossum.     

Effects of orbitofrontal lesions on social behavior in a confined group of stump-tail macaques (Macaca speciosa)

We grouped 14 stump-tail macques, five males and nine females, and observed social behavior before subjecting four of them to resection of orbitofrontal cortex. Four monkeys also received control lesions of superior temporal cortex and the remaining animals served as unoperated controls. Observations of social behavior continued after surgery on the following behavior: Joining, grooming, self-grooming, threat, aggression, and presenting (total hours of observation equaled 154). Monkeys with orbitofrontal lesions showed decreases in threat and aggression but only one such monkey fell in dominance. Control (operated and unoperated) monkeys displayed little change in these behaviors. Monkeys with orbitofrontal lesions also increased joining and self-grooming but showed a decrease in grooming of others. Presenting did not change. The role of orbitofrontal cortex in modulating different aspects of social interaction is emphasized by these results. However, in primates this area of the frontal lobes appears to have its major influence in the emotional loading of such complex behaviors.     

A new interpretation of the female genitalia in Macrocyclops albidus (Copepoda, Cyclopidae)

The female genital structures of Macrocyclops albidus (Cyclopidae, Eucyclopinae) were studied using light and electron microscopy. The results confirm that the exterior genital area shows only a copulatory pore, located anteromedially on the ventral face of the genital double-somite, and paired gonopores (not directly visible), situated laterally under the P6 plates. An internal seminal receptacle, composed of several parts, is connected to the gonopores by ventro-lateral cuticular extensions or seminal ducts. The lateral site of communication shows a complex set of connections between the seminal receptacle and the oviducts (via the egg-laying ducts). The structure until now designated as ‘transverse ducts’, visible by transparency on the ventral face, is in fact constituted of internal cuticular thickenings resulting of the fusion of the 6th thoracic somite and the 1st abdominal somite forming the genital double-somite and appearing externally as a part of the suture line; the term ‘suture cord’ is proposed to designate it. The functioning of the system is explained.     

Sperm binding properties and secretory activity of the bovine oviduct immediately before and after ovulation

The possibility that differences in hormonal regimes between the two oviducts in the cow around ovulation affects secretory activity of the oviduct epithelial cells and/or sperm-oviduct binding was studied. Oviducts were collected immediately after slaughter at 6 hr before to 5 hr after timed ovulation of 14 normally cyclic cows that had been inseminated (n = 6) or not (n = 8) and material obtained from the same cows was processed in three ways. First, in vivo, after artificial insemination of the cows, low numbers of sperm cells (approx. 15 per oviduct) were found within the entire oviducts as observed by scanning electron microscopy (SEM). Almost all sperm were located in the isthmus and then only on ciliated cells and showed without exception fully matured, intact morphology. Secretory activity of noninseminated oviduct epithelia was induced after ovulation which was most predominant in the pockets of the ipsi-lateral ampulla compared to the contra-lateral ampulla (P < 0.01). Second, ex vivo, explants dissected from oviducts of the noniseminated cows were incubated with sperm. In all cases, the sperm bound to the explants in a similar pattern as observed in vivo and this binding was strictly fucose-dependent. The main difference with in vivo experiments was the high numbers of sperm bound at any site of the oviduct ( approximately 3,000 cells per mm(2)) indicating the high sperm binding capacity of the oviduct epithelia. Ovulation induced a striking drop in sperm binding capacity in the oviducts and was most pronounced in the isthmus ( approximately 1,300 cells per mm(2); P < 0.001) and to a lesser extent in the ampulla ( approximately 2,000 cells per mm(2), P < 0.01). Third, in vitro, pieces of tissue dissected from oviducts of the noninseminated cows were cultured to mono-layers. Culturing epithelial cells resulted in loss of their normal morphological appearance. In all cases, the sperm binding capacity in monolayers was very low (<50 cells per mm(2)) when compared to corresponding explants (P < 0.0001). Sperm binding to monolayers originating from the isthmus (<25 cells per mm(2)) was lower than in those from the ampulla (40-50 cells per mm(2); P < 0.01) and remained similar after ovulation. In all three approaches, no significant differences were found in sperm-oviduct binding characteristics and sperm-distribution in the ipsi- versus contra-lateral oviducts. This indicates, that systemic endocrine changes around ovulation rather than specific oviduct changes at the ipsi-lateral oviduct induce secretion in oviduct epithelial cells, and thus induce sperm release.     

Cell of the mucous membrane of the female genital tract in culture: A comparative study with regard to the histogenesis of endometriosis

Summary Cellular elements from the mucous membrane of the uterus and oviducts and from peritoneal washings were cultured. The in vitro behavior of these cells was compared to elucidate the histogenesis of endometriosis and the role of various diagnostic procedures. In 65% of the cultured matrial obtained by uterine-tubal flushing, proliferating cells of the uterine-tubal mucous membrane were present. Their morphology and behavior corresponded to those of cultured cells obtained by separate washing of the uterine cavity and the tubes, respectively, curetted material, and biopsies of endometriosis lesions. Epithelial and stromal cells were identified using phase contrast microscopy, electron microscopy, and immunohistochemical methods. These cell types did not occur in peritoneal washings before the flushing of uterus and tubes. It was therefore assumed that they were detached and transported to the pelvic cavity during the above-mentioned procedures. In view of their intensive proliferation they may form the basis in the development of nodules of endometriosis. This would support the implantation theory concerning the pathogenesis of endometriosis. Interactions between epithelial and mesothelial cells point to the possible role of the latter in encapsulating the endometrial elements.     

Comparison of residual osseous mass between vascularized and nonvascularized onlay bone transfers

Composite flaps containing vascularized frontal bone were transferred on muscle pedicles in immature rabbits. Vascular continuity was maintained on one side and interrupted on the other. Bone weights at 16 weeks following transfer were compared with those of unoperated controls. The conventional bone graft demonstrated significant reduction in osseous mass. The vascularized bone maintained its mass compared with unoperated controls. Vascularized bone transfer appears to be the preferred surgical technique whenever possible.     

Fiber type composition of unoperated rat soleus and extensor digitorum longus muscles after unilateral isotransplantation of a foreign muscle in long-term experiments

We examined the effects of the unilateral heterochronous isotransplantation on the fiber type composition and myosin heavy chain (MyHC) isoform content of unoperated slow soleus and fast extensor digitorum longus muscles of female inbred Lewis strain rats. Comparison was made between "control" unoperated muscles of experimental rats (after intramuscular transplantation surgery) with the corresponding muscles of completely naive (unoperated) rats of three age groups (5-, 8- and 14-month-old). This was done in order to ascertain whether these muscles can be used as reliable controls to the transplanted and host muscles for our ongoing grafting experiments. The fiber type composition was determined by assessing the histochemical reaction for myofibrillar adenosine triphosphatase, the MyHC isoform content was determined immunocytochemically using monoclonal antibodies specific to different MyHC isoforms and by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Our experiments show that the heterochronous intramuscular isotransplantation procedure had no significant effect on the fiber type composition and MyHC isoform content of the "control" unoperated muscles of the experimental rats when compared to the corresponding muscles of the naive animals. Furthermore, the duration and type of isotransplantation did not also lead to differences among corresponding "control" muscles of experimental animals. We conclude that the unoperated muscles of the experimental rats can be used as controls in our current transplantation project dealing with long-term grafting experiments.     

Correlation of infertility with altered tubal morphology and function in mice with salpingitis induced by a human genital-tract isolate of Chlamydia trachomatis

Progesterone-treated C3H mice were inoculated into the uterus or ovarian bursa with a human genital tract isolate of C. trachomatis (serovar E), or with control medium alone. The mice were then observed at various times up to 260 days after inoculation. Before being killed the mice were given pituitary gonadotrophins to induce ovulation. Eggs were sought in the oviducts and ciliary activity in the fimbrial and ampullary sections of the oviducts was determined by light microscopy, before detailed examination by scanning electron microscopy. Eggs were visible in all control oviducts and both mucosal ultrastructure and ciliary activity appeared normal. By contrast, eggs were not recovered from the inoculated oviducts of mice infected intrabursally, nor was ciliary activity observed up to 28 days after inoculation. After this, ciliary activity reappeared but eggs were still not transported to the oviduct. Ultrastructural studies suggested that severe mucus congestion accompanied by tubal oedema and loss of ciliated epithelia play a major role in the aetiology of chlamydial-induced tubal damage. Infertility following chlamydial salpingitis could be associated with failure of egg transportation to the oviduct. Egg transport was still impaired even when luminal ciliary activity, ultrastructural integrity and patency had recovered. Our results suggest that chlamydial salpingitis in this mouse model closely resembles the human disease in its pathology and consequences for fertility, making the model particularly relevant for research on chlamydial vaccine development.     

Induced inflammatory process in Peripatus acacioi Marcus et Marcus (Onychophora)

The inflammatory response induced by the implant of a suture thread in Peripatus acacioi muscle was characterized under light and transmission electron microscopy (TEM). After 24 and 48 h granulocytes were observed migrating through the connective tissue toward the suture thread. These cells contain cytoplasmic eosinophilic granules as well as free granules near to the thread. There were few spherule cells with eccentric smooth kidney-shaped acidophilic nuclei and basophilic granules. Cells with intermediary characteristics as well as cells with a central basophilic nucleus with scarce acidophilic cytoplasm devoid of granules were also found. Under TEM, the granulocytic coelomocytes show small and homogeneous electron dense granules, while the spherule cells possess spherules that can be heterogeneous, granular, or with myelin figures. An acute induced inflammatory process is described for the first time in Onychophora and contributes to the scarce available literature on the function of the coelomocytes within this group.     

Effects of unilateral ovariectomization on the numbers of sperm in the oviducts, uterus and cervix of rabbits

The experiment was designed to determine the effect of removing one of the rabbits' ovaries on the numbers of sperm in the oviducts, uterine horns and cervices. The possible relationship between the anatomical asymmetry of the two sides of the reproductive tract and the distribution of sperm within different segments of the tract was also studied. Fifteen female rabbits were used; five were kept intact as the control group, five were right ovariectomized and five were left ovariectomized. All rabbits were injected with 50 IU HCG to induce ovulation and then inseminated with 60x10(6) sperm in 0.25 ml semen. Does were inseminated one month after unilateral ovariectomy. Animals were sacrificed 10 hrs later and sperm was recovered from the right and left oviducts, uterine horns and cervices. Unilateral ovariectomy significantly reduced the total numbers of sperm recovered as compared with intact does. The total numbers of sperm recovered from each of the two sides of the tract were not affected by the site of the removed ovary. Sperm numbers were high in the cervices of all groups and then decreased gradually in the upper segments of the tract. Sperm numbers that reached the oviduct of the intact rabbits were greater than those of both unilateral ovariectomized groups of rabbits. Differences between the length of the left and right cervices and uterine horns were not significant, but the right oviduct was significantly longer than the left oviduct.