Prenatal differentiation of bovine oviductal epithelium: an electron microscopic study

In this study, we investigated the ultrastructural changes during the prenatal differentiation of oviductal epithelium in 16 bovine embryos and fetuses from CRL of 18.0 cm to a CRL of 94.0 cm. Ciliated and secretory cells of bovine uterine tube, a derivative of the Müllerian duct, differentiate to distinct development stages in the prenatal period. The typical cellular pattern, which is generally characteristic for the adult bovine oviduct, is also obtained during fetal life. In the early stages (CRL 18.0/20.4 cm), the bovine oviductal epithelium appears mostly undifferentiated. The epithelial cells show only a few mitochondria, some cisternae of rough endoplasmic reticulum (rER) and a small Golgi-complex. Most of the cytoplasm is filled with a large amount of glycogen, which decreases during later development. Interspersed between the undifferentiated epithelial cells, a few cells undergoing ciliogenesis can be observed. Ciliogenesis increased significantly during the later prenatal developmental stages. At a CRL of 55.0 cm, ciliated cells appear fully differentiated with mature cells covering their luminal surface. Formation of cilia usually use the acentriolar pathway. Fibrous granules occurred initially in association with the Golgi-apparatus and r(ER) in the supranuclear cytoplasm. Fibrous granules later fuse with deuterosomes and give rise to procentrioles, which are translocated to the luminal plasma membrane. There they become arranged in a line just beneath the apical cell membrane and further differentiate to basal bodies from which the formation of cilia and striated rootlets take place. Clear signs of differentiation of secretory cells were first seen in our material in fetuses with a CRL of 51.0 cm and 64.0 cm. These cells contain a well developed rER and Golgi-apparatus with dilated cisterns. In the supranuclear cytoplasm, the number of secretory granules continuously increases during later development and the cells adapt to the morphology of mature secretory cells at the CRL 94.0 cm.     

Twin embryos in mares. II: Post fixation embryo reduction

Recent findings on the development and natural outcome of twins from Day 17 (immediately after fixation) to Day 40 are reviewed. Incidence of embryo reduction was increased significantly when the vesicles became fixed unilaterally, rather than bilaterally, and when the vesicles were unequal in diameter. Of 68 mares with twins on the day of fixation, post fixation embryo reduction occurred in 41 (60 per cent). The incidence of reduction was 41 of 48 (85 per cent) following unilateral fixation; reduction occurred in all of 22 mares with vesicles of dissimilar size (4 mm or more difference in diameter) and in 19 (73 per cent) of 26 mares with vesicles of similar size (0 to 3 mm difference). Embryo reductions were complete (vesicle no longer visible ultrasonically) by Day 20 (59 per cent of the reductions), during Days 21 and 30 (27 per cent), or Days 31 to 38 (14 per cent). In 80 per cent of the early reductions (by Day 20) the eliminated vesicle disappeared within one day. Reductions that occurred after Day 20 were preceded by a gradual decrease in size. As the number of days after Day 17 increased, the frequency of reduction decreased and the time required for completion of reduction increased. When the twins were dissimilar in diameter (4 mm or more), they were more likely to undergo reduction by Day 20. In summary, dissimilarity in diameter increased the likelihood of unilateral fixation, increased the incidence of reduction for unilaterally fixed vesicles, hastened the day of occurrence of reduction and shortened the interval from initiation to completion of reduction. The deprivation hypothesis proposes that embryo reduction occurs when a major portion of the three walled area of the yolk sac or the vascularised wall of the yolk sac or allantoic sac is in apposition with the wall of the adjacent vesicle rather than with the endometrium; the vesicle is deprived of adequate embryonal-maternal exchange and therefore regresses.     

On the development of the papillary body in the feline claw

The pre- and post-natal development of the feline claw was studied in 22 feline fetuses with a crown-rump length (CRL) ranging from 40 to 160 mm, in six kittens up to an age of 22 days, and in four adult cats. In fetuses up to a CRL of 75 mm, the characteristic shape of the feline claw was developed. Segment-specific dermal modifications in the various segments, especially the dorsal ridge, started to develop in fetuses with a CRL between 75 and 105 mm. Modifications of the papillary body in the different claw segments took place in the last third of prenatal development and were continued postnatally. At first, the basal lamina became wavy, followed by the formation of small dermal microridges, which would be enlarged to dermal ridges and lamellae. In the claw of adult cats, the papillary body was very small. The dermal tissue of the proximal part of the coronet formed low microridges with short papillae originating on and between these low ridges. In the wall segment, dermal microridges were formed which were arranged in a parallel fashion, and in the distal part of the wall, short dermal micropapillae arose on the crest of each microridge. In the sole segment, thin dermal lamellae were developed. The sequence of papillary body development and the varying conformations of the papillary body in the different segments of the feline claw are compared to those in the nail, the canine claw and hooves.     

Ultrastructural Studies of Chicken Embryo Chorioallantoic Membrane during Incubation

Structural changes taking place in the chorioallantoic membrane (CAM) of embryonated hen's eggs during incubation were followed up by histological, histochemical and electron microscopic methods. The allantoic sac surrounds the amnionic sac and also the yolk sac by the 7th to 11th day, respectively, and forms together with the chorion the CAM. The mesenchymal layer placing between the chorion of ectodermal and the allantoic layer of entodermal origin develops from the somatic and splanchnic-pleure of the parietal mesoderm. The CAM is composed of three different layers, i.e. chorionic epithelium, mesenchyme and allantoic epithelium. The chorion comprises 2 layers of cubical epithelial cells. In between the 2 layers capillaries and directly under the shell membrane vascular sinuses can be found. Blood circulating in the sinuses is separated from the air in the pores of the shell membrane by the adacent epithelial cell-projections of 0,2 μm thickness, the basal membrane (0.1 μm) and the sinus endothelial layer of 0.2 μm thickness. The mesenchymal cells are of star-like form. The allantioc epithelium is built of one layer of fusiform cells with oval-shaped or rod-like nuclei.     

Early organogenesis of the caprine stomach

Early organogenesis of the caprine stomach was studied in a series of 11 embryos ranging from 6.5 mm neck-rump length (NRL) to 13.3 mm crown-rump length (CRL). In embryos with 6.5-6.7 mm NRL, a part of the primordial proper esophagus extended to the dorsal side of the primordial stomach. The primordial proper esophagus and its extension were lined with a simple epithelium and stained dark brown with Con A III, while the primordial stomach was weakly stained. In embryo with 7.3 mm NRL, the esophageal extension was separated from the proper esophagus by constriction, and became a primordial forestomach situated in an area outside the omental sac. In embryos with 8.3 mm NRL-10.7 mm CRL, primordial forestomach and primordial stomach were united and formed a spindle shaped primordial ruminant stomach with foregut rotation. The primordial ruminant stomach was similar to the primordial simple stomach except that it was more flattened laterally with the convex at the area of 'lesser curvature'. Primordial rumen, omasum and abomasum appeared from the spindle shaped primordial ruminant stomach in an embryo with 12.9 mm CRL. In an embryo with 13.3 mm CRL, primordial reticulum originated from an area between the primordial rumen and omasum.     

The origin of bacteria recovered from the peritoneum and yolk sac of healthy chickens

Subclinical peritoneal and yolk sac infections were demonstrated in about 38 and 23 per cent, respectively, of 121 conventional chicks examined during the first 5 days of life; the incidence varied markedly from hatch to hatch. Organisms were demonstrated in the peritoneum and yolk sac of a proportion of gnotobiotic chicks following per os administration of pure cultures of bacteria isolated either from the peritoneum or yolk sac, or from the intestinal tract of conventional chickens. It is concluded that this subclinical infection arises as a result of the translocation across the gut wall of bacteria present in the lumen of the intestine.     

Early development and cell differentiation of the parasympathetic vagus-glossopharyngeal nucleus in cattle. Light and electron microscopic studies]

The early development, differentiation of the cell and cell migration of the nucleus parasympathicus nervi vagi et glossopharyngei were examined by light microscope in 32 bovine embryos with crown-rump-lengths (CRL) ranging from 1 cm to 53 cm. During this period the nucleus is being enlarged 6 to 7 times and the size of the cell increases to 35-40 microns. The ultrastructure during the differentiation of the cell is shown electron microscopically in embryos with CRL of 2.5 cm and 3.6 cm. Several layers of matrix cells arise from the neuroepithelium of the neural tube by mitosis. They migrate in the shape of dark nucleated cells into the parasympathetic cell column. With advancing age of the embryos the number of cells with light nucleus increases. They represent the presumptive neurons. In embryos of 2.5 cm CRL their cytoplasm surrounds the nucleus on three sides in the shape of a narrow rim while on the fourth side it is enlarged into an outgrowing process. In this process a smaller number of organelles and their preliminary stages appears. Their number is significantly increased in embryos of 3.6 cm CRL and they can be seen throughout the growing process. In the following stages of maturity cytological development proceeds. In embryos of 53 cm CRL topographical and cytological data are comparable to those in adult animals.     

Changes in expression and localization of GPRC5B and RARalpha in the placenta and yolk sac during middle to late gestation in mice

The mRNA expression of GPRC5B, an orphan G protein-coupled receptor, is induced by retinoic acid (RA). Because RA plays critical roles in embryonic development, reproductive functions, metabolism and homeostasis, GPRC5B is also considered crucial in these physiological events. We investigated the changes in expression of GPRC5B and RA receptor (RAR) alpha mRNAs and immunohistochemical localization of their proteins in the murine placenta and yolk sac at 13.5, 15.5 and 17.5 days post coitus. Stable levels of GPRC5B and RARalpha mRNAs were detected in the placenta and yolk sac. In the placenta, GPRC5B was present in maternal and fetal vascular endothelial cells, stromal cells, fibroblast-like cells and glycogen cells. A strong reaction to RARalpha was detected in maternal and fetal vascular endothelial cells and stromal cells. The levels of GPRC5B and RARalpha proteins in maternal and fetal vascular endothelial cells decreased with gestation. In the yolk sac, GPRC5B and RARalpha proteins were detected in vascular endothelial cells, but their levels did not change during the gestation period. These findings indicate that GPRC5B is involved in RA-dependent morphogenesis/angiogenesis and regulation of extracellular matrix synthesis in the murine placenta and yolk sac.     

Yolk sac, body dimensions and hatchling quality of ducklings, chicks and poults

1. One-day-old chicks of Pekin duck, turkey, layer fowl and broiler fowl were examined for bacteria in the yolk sac and yolk fluid. 2. Whole hatchling, yolk-free hatchling and yolk sac weights were recorded for all species along with crown-rump length and beak-tip to toe-tip length. 3. Bacteriology revealed positive results for the whole yolk sacs of 43 to 64% of the birds in the sample of ducklings, poults and layer chicks. Broiler chicks had a 6.6% incidence of bacteria isolated from the whole yolk sac. By contrast, there were very few positive results from swabs of yolk fluid for any of the bird types. 4. The presence of bacteria in the yolk sac of hatchlings suggests that there is colonisation, rather than infection, of the yolk sac membrane during the hatching period or the first few hours post-hatching. Isolation of bacteria from the yolk sacs of young chicks might no longer be considered as solely indicative of yolk sac infection but further research is required to confirm this result. 5. Contrary to what is being suggested in commercial practice relationships between linear dimensions and hatchling weight suggest that measurement of chick length is at best a very crude measure of chick quality.     

On the Development of the Feline Skeleton

Light microscopic observations revealed that in camel foetuses of 25 mm crown-to-rump length (CRL) the primordial tubular system of the prospective lung was formed of several tubules lined by undifferentiated columnar epithelium. Intra-epithelial neuroendocrine cells were the first elements to be differentiated in the lining epithelium of the primordial tubular system of the prospective lung as early as 25 mm CRL. On reaching 50–67 mm CRL, the primordial tubular system started to differentiate into two systems of primordial tubules, the prospective bronchial or light tubules and the future respiratory or dark tubules. The lining epithelium of the prospective bronchial tubules revealed a clear evidence of ciliogenesis as early as 80 mm CRL. From 800 mm CRL onwards, the bronchial epithelium demonstrated ciliated and non-ciliated secretory cells. The non-ciliated secretory cells of the bronchial epithelium of fetal camel lung showed moderate reaction to AB/PAS technique, for the first time, in fetuses reaching 600 mm CRL.     

Yolk sac utilization in ostrich (Struthio camelus) chicks

The mass of residual yolk sac expressed as a percentage of initial mass of the egg from which the chick hatched decreased sharply in the first 2 days post-hatching. A gradual reduction occurred between 3 and 10 days after which a sharp decline was noted between 11 and 13 days post-hatching. The highest number of chicks with unabsorbed yolk sac was noted on day 5 post-hatching followed by days 6 and 7. Chick mortality followed the same pattern. The dynamics, causes and clinical consequences of yolk sac utilization are discussed.     

Morphological Studies on the Development of the Bronchial Epithelium of the Fetal Camel Lung

Light microscopic observations revealed that in camel foetuses of 25 mm crown‐to‐rump length (CRL) the primordial tubular system of the prospective lung was formed of several tubules lined by undifferentiated columnar epithelium. Intra‐epithelial neuroendocrine cells were the first elements to be differentiated in the lining epithelium of the primordial tubular system of the prospective lung as early as 25 mm CRL. On reaching 50–67 mm CRL, the primordial tubular system started to differentiate into two systems of primordial tubules, the prospective bronchial or light tubules and the future respiratory or dark tubules. The lining epithelium of the prospective bronchial tubules revealed a clear evidence of ciliogenesis as early as 80 mm CRL. From 800 mm CRL onwards, the bronchial epithelium demonstrated ciliated and non‐ciliated secretory cells. The non‐ciliated secretory cells of the bronchial epithelium of fetal camel lung showed moderate reaction to AB/PAS technique, for the first time, in fetuses reaching 600 mm CRL.     

Labelling of rat endothelial cells with antibodies to vWF, RECA-1, PECAM-1, ICAM-1, OX-43 and ZO-1

Labelling with endothelium specific monoclonal antibodies, von Willebrand Factor (vWF), rat endothelial cell antigen-1 (RECA-1), platelet-endothelial cell adhesion molecule-1 (PECAM-1), intercellular adhesion molecule-1 (ICAM-1), OX-43 and zonula occludentes-1 (ZO-1), was investigated in cryostat sections of vessels from rats of different ages using a confocal microscope. The results showed that labelling of the vWF was positive in endothelial cells from adult, fetal and different ages of embryonic rat. Labelling with RECA-1 was weakly positive in adult rat aorta and lung endothelial cells but not in embryonic yolk sac endothelial cells. Labelling using PECAM-1, ICAM-1 and OX-43 was negative in both adult and embryonic endothelial cells. ZO-1 showed positive but very weak reactivity in embryonic yolk sac endothelial cells. The expression of vWF on vessels from adult and 19.5-day fetal tissues was strongly positive. However, the expression of vWF in embryonic endothelial cells was dependent on the gestational age. While the 11.5-day yolk sac vessels stained weakly, staining gradually increased in 13.5-, 15.5- and 17.5-day-old yolk sac vessels. The results suggest that vWF is a reliable endothelial cell marker in rat vascular endothelial cells, including both fetal and embryonic stages.     

Morphological aspects of prenatal haematopoietic development in the cat

The appearance of haematopoietic cells and the development of haematopoiesis in certain embryonic organs of the cat were studied using light microscopic (cytological smears and paraffin embedded tissues) and transmission electron microscopic methods. Primitive erythropoiesis occurred predominantly in the yolk sac whereas definitive erythropoiesis occurred in the yolk sac, liver, bone marrow and, to a lesser extent, in the spleen. In the yolk sac, erythropoiesis was predominantly intravascular whereas in the liver and bone marrow it was usually extravascular. Granulopoiesis occurred mainly in the liver and bone marrow. In the liver it was predominantly extravascular and occurred around vessels, bile ducts and in perisinusoidal spaces. Megakaryocytopoiesis occurred in the yolk sac, liver, bone marrow and spleen. The megakaryocytic line of cells were similar to those occurring in adult cats except that in the yolk sac unusual small, mononuclear to binuclear thrombocytogenic megakaryocytes were present also. The relative contributions of the embryonic organs of the cat to haematopoiesis were similar to those described for man and certain other mammals but the results for the sites of development and the appearance of early forms in the erythrocytic, granulocytic and megakaryocytic lines varied at times from those reported for man and other mammals.     

幼龄鸡卵黄囊中^3H—VA的吸收代谢及其利用的研究

选用出壳后１２时龄的ＡＡ肉雏１４５只，采用放射性同位素＾Ｈ示踪方法，分别进行了饲养试验，屠补试验和代射试验研究表明，在全价日粮和无ＶＡ纯日粮条件下，雏鸡卵黄囊中的内源性３＾Ｈ－ＶＡ在体内发挥持续时间分别为２１天和１４天。卵黄囊（内源）３＾Ｈ－ＶＡ在鸡肠道中的吸收代谢，与饲料（外源）中营养物质相比，有着独自的特点。卵黄囊中内源３＾Ｈ－ＶＡ在雏鸡肠料中外源ＶＡ之间存在有动态交换关系，内源３＾Ｈ－ＶＡ主     

Development of the glandular epithelium of the bovine parotid gland during ontogenesis

The development of the parotid gland was examined in 36 bovine embryos and foetuses with a crown-rump-length (CRL) from 28 up to 1000 mm by light, transmission electron microscopical and actin-immunohistochemical methods. The anlage of the parotid gland in an embryo with 28 mm CRL can be found at the lateral angle of the primitive oral cavity as a local thickening of the epithelium. During the second month, the differentiation of primary ducts and endbuds starts and a lumen develops in the primary ducts. At the end of the second month a lumen appears in the terminal endbuds. In the immature endpiece cells first secretory granules can be seen from a CRL of 240 mm. In the third month differentiation between intra- and inter-lobular ducts is possible. Immature myoepithelial cells present as a basal layer of flattened cells between the epithelial cells and the basement membrane at the end of the second month. During further development they increase in number, become more flattened and form long cellular processes. At the end of the fourth month isolated actin filament bundles are formed, which were also detected by an antibody against smooth muscle actin. The actin filaments condense continuously until they fill the cell processes completely at the end of foetal development.     

Effect of aspiration pressure during oocyte harvesting on oocyte recovery and in vitro development of ovine oocytes.

Oocytes from abattoir-sourced ovine ovaries were aspirated from 2- to 4-mm follicles using 25, 50 or 100 mmHg pressure and an aspiration pump, or a needle (20-G) and syringe (2.5 ml) and subjected to in vitro maturation, fertilization and culture to determine the effect of aspiration pressure on the number and quality of oocytes recovered, and early embryonic development. Oocyte recovery rate was similar between groups (range: 57.1-73.1%; p > 0.05). The number and percentage of grade I and II oocytes recovered was reduced for 100 mm (24.5 +/- 3.6 and 31.1 +/- 6.1%) compared with 25 mm (51.4 +/- 7.0 and 60.2 +/- 7.8%) and 50 mm pressure (40.8 +/- 5.6 and 50.3 +/- 4.4%) and a syringe (40.3 +/- 12.0 and 45.2 +/- 2.1%; p < 0.05). Oocyte cleavage was similar for all groups at 24 (range: 30.9-49.6%) and 48 h post-insemination (49.7-65.5%), but blastocyst formation (% cleaved oocytes) was lower for oocytes aspirated with 25 mm (37.8%) than 50 (69.2%) or 100 mm (67.2%) pressure, and a syringe (72.0%; p < 0.05). Embryo production efficiency (% of oocytes cultured developing to the blastocyst stage) was higher for oocytes aspirated with 50 mm (45.4%) and 100 mm pressure (43.8%) and a syringe (45.0%) than 25 mm pressure (18.8%; p < 0.05). These results demonstrate that the aspiration of ovine oocytes with an aspiration pressure of 50 mm, or aspiration with a needle and syringe are equally efficacious for the in vitro production of embryos.     

氨基酸和牛磺酸对绵羊体外受精胚胎体外培养的影响

本文在KSOM培养液中添加牛磺酸和NEAA、EAA ,研究氨基酸对绵羊胚胎体外培养的影响。研究表明 :①牛磺酸、NEAA可以显著提高胚胎的桑椹胚和囊胚发育率 ;②EAA能促进胚胎由囊胚向孵化囊胚发育 ;③NEAA和EAA可促进胚胎的囊胚发育和孵化囊胚的发育。     

家蚕马氏管组织构造的研究

通过对家蚕 (Bombyxmori) 5龄第 5日幼虫马氏管外部形态和内部组织结构的观察 ,发现不同部位马氏管的结构差异很大 :膀胱呈囊状体开口于小肠两侧 ,肌肉层厚、细胞层较薄 ,内腔很大 ;下行管靠膀胱处圆形 ,较膨大 ,其余大部分外型呈扁平形且表面光滑 ,细胞层较薄 ,内缘有微绒毛状突起、管腔较大 ;上行管前部管体稍扁平 ,结构与下行管相似 ,中后部管体圆形、细胞层肥厚、内缘呈蜂窝状、管腔狭小 ;隐肾管的外列马氏管细胞层薄、管腔较大 ,而内列马氏管细胞层肥厚、管腔很小。马氏管结构的差异与各部位的功能有密切关系。