Morphologic stages of the equine embryo proper on days 17 to 40 after ovulation.

OBJECTIVE: To describe the gross and histologic changes that develop in the equine embryo proper (ie, the portion of the embryo that becomes the fetus) from days 17 to 40 after ovulation and to compare the external features of equine embryos with those of porcine, ovine, and human embryos. SAMPLE POPULATION: 34 embryos collected from mixed-breed pony mares. PROCEDURE: External features for each embryo proper, including length, number of branchial arches, growth of appendages, face and head features, and body features, were examined, using a dissecting microscope, for embryos collected on days 17 to 40. Internal features were histologically examined by serially sectioning embryos collected on days 20 to 35. RESULTS: Number of embryos recovered for each day ranged from 1 to 5. The initial detection of features was not related closely to age; typically, the first attainment of a given body length or characteristic varied over a 3-day period among embryos. Similarly, the period during which individual characteristics for a given Carnegie stage were attained ranged from 3 to 6 days. Age at first appearance of a characteristic was greater for equine embryos than ages reported for ovine and porcine embryos but less than for human embryos. Indicators of age included number of pairs of branchial arches, all limb buds present, retinal pigmentation, and prominence of the pontine flexure. CONCLUSIONS: No embryologic structures or changes were found that could be considered unique to equine embryos on days 17 to 40 after ovulation.

Cytoplasmic abnormalities in cultured cerebellar neurons from the trisomy 16 mouse.

This study represents a first effort to characterize the growth and development of murine trisomy 16 neurons using single-cell neuron culture techniques. Murine trisomy 16 is a model for the human Down syndrome, or trisomy 21. Both show similar nervous system abnormalities including decreases in cerebellar size and in numbers of cerebellar neurons. Trisomy 16 cerebellar neurons cultured from 17-gestational day conceptuses grew less extensive neuritic arbors than normal neurons. Unlike controls, the individual neurites of the trisomic neurons were not clearly distinguishable as axons or dendrites over the 10 day period that they were observed. The trisomic neurons were characterized by diminished levels of microtubules, abnormally shaped mitochondria, and the presence of dense bundles of abnormal filaments that were not observed in any of the normal littermate neurons.

Examination of the eyelid closure defect in trisomy 16 mice.

A characteristic feature of trisomy 16 mouse conceptuses is a failure of their eyelids to close. This defect was investigated by examining ocular development in serially sectioned heads of trisomy 16 and normal littermate fetuses from 10 to 18 gestational days. Other heads were examined by using scanning electron microscopy. Between 10 and 15 days, trisomy 16 ocular structures were delayed, but there was no striking abnormal morphology. At 16 days, when the eyelids were closed and fused in normal mice, trisomic eyes had a large cell mass near the inner canthus that protruded between the open lids. The mass was covered by bulbar conjunctiva and cells of the mass were continuous with developing corneal tissue. The mass was not present in the eyes of normal mice on any gestational day and was not present in trisomic eyes at 17 and 18 days, when the lids began to show varying degrees of closure. Based on its positioning at the inner canthus, the mass may represent a transient hyperplasia of the developing semilunar fold which physically impedes lid closure in the trisomic conceptuses. Previously, the defect has been attributed to the trisomy 16 conceptus's overall pattern of growth retardation and delayed development. Masses such as those seen in the trisomic eyes have not been observed in other murine lid-gap defects that have been investigated. A second finding in this study is that trisomic eyes are positioned more superiorly in the head than normal eyes. This variation may be related to alterations in cranial base morphology that are associated with trisomy 16.

Altered placental morphology associated with murine trisomy 16 and murine trisomy 19.

The morphology of placentas from trisomy 16 and trisomy 19 mouse conceptuses aged 12 to 18 gestational days was studied at the light microscopic level. Comparisons were made with placentas from normal littermate animals. Trisomy 16 placentas showed marked changes from normal: 1) the junctional zone showed little indication of normal morphologic differentiation throughout gestation; 2) clusters of germinal trophoblast cells persisted in the labyrinth throughout gestation, whereas these cells disappeared by gestational day 16 in the normal littermate placentas; 3) the labyrinth was reduced in size in the trisomic placentas, and the differentiation of the interhemal membranes was delayed. The size of the labyrinths from trisomy 19 placentas appeared to be decreased, but otherwise the placentas appeared to have normal morphology. These observations and others from the literature show that placental development is affected by the presence of a trisomic genome, and that different trisomies influence the development of the placenta differently. For trisomy 16, we propose that the striking changes of the junctional zone may be associated with the trisomy 16-related gene dosage effect for alpha- and beta-interferon cell surface receptors. Because of the homology for this and other genes on mouse chromosome 16 with genes on human chromosome 21, findings related to the altered development of the trisomy 16 mouse may be relevant to understanding some of the phenotypic variations associated with human trisomy 21, the Down syndrome.

Chromosome 18 aneuploidy: anatomical variations observed in cases of full and mosaic trisomy 18 and a case of deletion of the short arm of chromosome 18.

Cases of full and mosaic trisomy 18 and a body of an infant with the 18p-syndrome were dissected in detail to compare the anatomical variations associated with these 3 chromosome imbalances involving autosome 18. The types and numbers of morphologic variations present in both the full and mosaic trisomy 18 bodies were similar to the types and numbers of variations seen in all other cases of full trisomy 18 that have been studied by gross dissection. Apart from an atrial septal defect, the body of the infant with the 18p- imbalance showed only 2 striking defects: 1) deficiencies of the levator palpebrae superioris muscle of the upper eyelid, and 2) absence of the ligament of the head of the femur. The first variation provides a morphologic basis to explain the ptosis which is observed frequently in affected individuals. Absence of the ligament of the head of the femur may be a factor contributing to congenital dislocation of the hip, which is reported occasionally in affected individuals. In addition to providing more detailed information about the phenotype of individual aneuploidy syndromes, studies of cases of different imbalances of single autosomes may provide additional insights about the genotype/phenotype relationships of specific chromosome segments.

An anatomical study of human otocephaly.

This study describes the gross anatomic variations observed in a 32-week male fetus diagnosed as having otocephaly. Special attention was given to the muscular, peripheral nervous, and vascular systems of the entire body. External features included approximation of the ears on the front of the neck, underdevelopment of the lower jaw, and a small oral cavity. The mandible, maxillae, and zygomatic bones were smaller than normal and appeared shifted in a ventrocaudal direction. The middle ear ossicles were fused and abnormally positioned. The tongue was positioned abnormally and malformed. The muscles of mastication were fused in the midline and formed the floor of the oral cavity. The variations were similar to the spectrum of abnormalities reported in two cases in the literature. Because of this finding, it is possible that the causative events leading to these deviations were similar in the three cases. Possible mechanisms are considered which could lead to the observed malformations seen in these cases. There were also several muscle and nerve anomalies outside of the head region.

Analysis of gross anatomical variations in human triploidy.

Three cases of human triploidy were dissected in detail to determine whether or not there are any specific patterns of morphologic variation associated with this form of polyploidy. No specific constellations of anatomic variation were observed. However, each body did have a greater than normal number of variations among muscles that tend to be variable in the normal population. Two bodies showed a marked increase in the size of the thigh muscles, two bodies had eyeballs that were larger than normal, and two bodies had anomalous attachments of the extraocular muscles. The sample size is not large enough to draw any specific conclusions concerning the pathogenesis of these variations. However, the findings do suggest that careful examination of additional specimens will be of value in providing further information about the genotype/phenotype relationships in human triploidy.

Morphologic development of the fetal trisomy 19 mouse.

Trisomy 19 mice show a characteristic pattern of delayed morphologic development when compared with normal littermates during the second half of gestation. Examinations of the external phenotype and sectioned fetuses, and skeletal maturation at later days, suggest that trisomy 19 has little recognizable effect on morphogenesis prior to 10 days. Trisomic conceptuses collected at subsequent 24-hour intervals show a 1-day lag in weight gain and begin to show a progressive and uniform delay in differentiation so that 14- and 15-day animals resemble 13- and 14-day normal fetuses, respectively. There is no further recognizable increase in this delay through term. There was no striking change in numbers of viable trisomic conceptuses collected throughout the period from 10 days to birth. These observations suggest that most trisomies surviving at 10 days will develop to term, with the major effect of the aneuploid genome being to delay the otherwise normal-appearing growth and development of the affected conceptuses. In all cases, the litters were from crosses between mice doubly heterozygous for the (5.19) and (9.19) Robertsonian translocation chromosomes.

Erythropoiesis in the fetal trisomy 19 mouse. I. Characterization of erythrocyte populations in peripheral blood.

Changes in the type, size, and relative percentage of different erythrocyte populations in the peripheral blood of individual trisomy 19 and normal littermate mouse fetuses were studied from 12 gestational days to term. Large nucleated erythrocytes of yolk-sac origin comprise the first population of cells and are gradually diluted out of the circulation by nonnucleated erythrocytes of hepatic origin. This transition occurs between 12 and 16 gestational days. The rate of decline of the nucleated erythrocytes in the trisomic animals lagged by approximately 1 day behind the normal littermates, so that they did not completely disappear from the peripheral circulation until day 17. A slight decrease in size of the nonnucleated erythrocytes which occurs with increasing gestational age was also delayed by approximately 1 day in the trisomic fetuses. These observations are consistent with an hypothesis that one effect of the murine trisomy 19 genome is to retard by 1 day the growth and development of the affected animal.

Growth characteristics of the murine trisomy 19 thymus.

Development of the trisomy 19 mouse thymus was studied by (1) evaluation of cell and nuclear counts and volume distributions and (2) examination of the in vivo cell cycle characteristics using flow cytometry. Direct preparations of thymuses from affected animals and their normal littermates at 17, 18, 19 gestational days, and at term were used. Cell numbers in aliquots from the trisomic thymus suspensions were always significantly less than those in the normal animals. There were no differences in cellular or nuclear volumes between the two groups. However, the populations of cells in the trisomic suspensions showed a lag in the normally occurring transition from a larger to a smaller size. DNA distribution histograms from these suspensions were generated using a TPS-1 Cell Sorter, and percentages of cells in different phases of the cell cycle were estimated using computer analysis. The resulting graphs and numerical data from the trisomic mice, when compared with the normals, showed consistent increases in the relative numbers of cells in the S and G2 + M phases of the cell cycle. Because growth retardation is a major feature of trisomy 19, as well as in other murine trisomies, these results suggest that cell cycle alterations may, in part, contribute to the reduced cell number, delayed development, and smaller size of the trisomic animals.

Anatomical analysis of the developmental effects of aneuploidy in man: the Down syndrome.

Detailed anatomical dissections of five Down syndrome (DS) bodies revealed a unique and consistent "internal phenotype" composed of: 1) variations in muscles, 2) peripheral artery variations, and 3) the presence of dilatations and nerve rootlets associated with the spinal accessory and first cervical nerves. Most of these variations may occur occasionally in the normal population, but their more frequent occurrence in the five DS cases provides further support for Shapiro's (1975) hypothesis that one effect of the extra autosome is to increase the instability of normal developmental processes. Three of the variations: 1) the presence of an extra facial muscle, 2) multiple vertebral arteries, and 3) the presence of dilatations and nerve rootlets associated with the spinal accessory and 1st cervical nerves, may be the result of a failure of regression of otherwise transient embryonic structures. A variation of the midfacial muscles of expression, explained as an arrest in development, has not been described previously and, to date, remains a manifestation unique to the trisomy 21 phenotype. Aside from their use in developmental speculation, the sum of these variations may be useful for corroborating diagnoses of suspected post-mortem cases of the DS without cytogenetic confirmation.

Anatomical analysis of the developmental effects of aneuploidy in man--the 18-trisomy syndrome: II. Anomalies of the upper and lower limbs.

We report the anatomical variations of the limbs in eight infants with the trisomy-18 syndrome that were dissected and studied in detail. In each case, the upper limbs showed defects which further define the specific influence of this aneuploidy on the development of its preaxial (radial) component, and the tendency towards reduction defects. Abnormalities included muscle variations concentrated along the radial margin of the forearm and hand, the absence of the definitive musculocutaneous nerve in all of the limbs, and reductions of the radial artery in four of the bodies. Pathogenetic mechanisms explaining the observed defects are discussed, and include: 1) a defect in peripheral nerve development; or 2) tissue necrosis. The characteristic flexion deformities of the fingers seem to be due to a displacement of the tendons of extensors digitorum and digiti minimi. The lower limbs did not show a consistent pattern of defects, except for the absence of some muscles (psoas minor, the tendon of flexor digitorum brevis to digit V), and the presence of several supernumerary muscles. These variations are discussed as possible nonspecific effects of 18-trisomy on development. The additional anatomical data from this and the first paper in this series [Bersu and Ramirez-Castro, 1977] provide a more detailed picture of the trisomy-18 phenotype which may be useful in corroborating an unconfirmed clinical diagnosis of the syndrome.

Anatomical analysis of the developmental effects of aneuploidy in man--the 18-trisomy syndrome: I. Anomalies of the head and neck.

This paper describes the anatomical variations observed in the head and neck in eight infants with the 18-trisomy syndrome that were dissected and studied in detail. Of the usual muscles of facial expression, occipitofrontalis and the auricular and nasal muscles were hypolastic in all eight bodies and each subject showed extensive fusion of the muscles around the corner of the mouth. In each body there was a supernumerary muscle band that extended from the region near the corner of the mouth to the occipital attachment of trapezius. The otomandibular region in each body showed a variable spectrum of muscular, skeletal, arterial, and salivary gland variations bilaterally. Three of the bodies had infrahyoid muscle variations. The sum of these observations provides a more complete delineation of the variations that occur in the 18-trisomy syndrome. Tentative pathogenetic hypotheses for several of the defects are discussed, based on observations from human descriptive embryology. Poswillo's teratological model [1973] implicating hemorrhaging as a causal mechanism in human first and second branchial arch malformations is suggested as a possible mechanism to explain the bilateral otomandibular defects.

Studies of malformation syndromes of man XXXXIA: anatomical studies in the Hanhart syndrome--a pathogenetic hypothesis.

Two infants with the Hanhart syndrome, i.e. micrognathia, microglossia, terminal deficiency of all limbs and imperforate anus in one, were dissected and studied in detail. The interrelationships of the muscular and skeletal defects suggested that they were the result of incomplete rather than abnormal morphogenesis. We speculate that the oral and limb abnormalities resulted from deficient mesodermal proliferation caused by disturbances in the ectodermal-mesodermal interactions beginning about the 4th week of development. The imperforate anus may also relate to the proposed defect.