Development of cardiac rhythms in altricial avian embryos

Mean heart rate (MHR) was determined during incubation and in hatchlings of 14 altricial avian species to investigate (1) if there is a common developmental pattern of heart rate in altricial embryos and (2) if heart rate changes during incubation are correlated with changes in embryonic growth rate. On the basis of normalized incubation MHR increased approximately linearly in 12 of 14 species from as early as 30-40% of incubation to that of pipped embryos. The MHR of hatchlings was equal to or higher than that of pipped embryos in seven species. Passerine embryos and hatchlings maintained higher MHR in comparison to parrots of similar egg mass, which may reflect phylogenetic differences in development. Embryonic MHR increased at a higher rate while embryonic growth rates were highest during the first 40% of incubation in tit, budgerigar and crow embryos than during subsequent development when relative growth rates decreased. MHR became independent of yolk-free wet mass at a smaller fraction of hatchling mass in budgerigar and crow than in the tit, suggesting that MHR is more likely to increase continuously after 40% of incubation in small altricial species than larger species.     

The sex-chromosome constitution and early postimplantation development of diandric triploid mouse embryos

Diandric triploid mouse embryos were produced by standard micromanipulatory techniques, using eggs isolated from female mice with a normal chromosome constitution that had been mated to homozygous Rb(1.3)1Bnr males (which carry a large metacentric "marker" chromosome, viz., a Robertsonian translocation involving chromosomes 1 and 3). The tripronucleate embryos were transferred to the oviducts of pseudopregnant mice, which were subsequently autopsied at about midday on the 10th day of gestation. Although a relatively small number of the isolated conceptuses consisted of morphologically abnormal egg-cylinder-like structures or empty gestational sacs, most were at clearly distinguishable embryonic stages, from the primitive streak stage to embryos with about 20 pairs of somites present. These embryos all appeared to be morphologically normal but were substantially smaller than normal (diploid) fertilized embryos analyzed at similar stages of development. A total of 63 diandric triploid conceptuses were recovered and analyzed cytogenetically. They were G-banded to determine their sex-chromosome constitution and confirm their diandric triploid status. No obvious difference was observed in the developmental potential of the 58,XXX class of diandric triploids, compared to that of the 58,XXY class. The ratio of 58,XXX to 58,XXY embryos was close to the expected ratio of 1:2, assuming that unfertilized eggs have an equal chance of becoming fertilized by an X- or a Y-bearing spermatozoon and that the additional (i.e., "donor") male pronucleus also has an equal chance of having either an X or a Y sex chromosome present. However, the development of the 58,XYY class appeared to be restricted, even at the stage of gestation analyzed, in that no embryos with this genetic constitution were observed that had progressed beyond the early somite stage. The present findings are discussed in relation to the cytogenetic findings in human triploid conceptuses, the majority of which are spontaneously aborted during the first half of pregnancy. In man, the 69,XYY class (equivalent to the 58,XYY class in our study) is only rarely encountered, and it has been assumed that these triploid embryos are probably lost at a very early stage of gestation.     

The cleavage rate of digynic triploid mouse embryos during the preimplantation period

Triploidy is a lethal condition in mammals, with most dying at some stage between implantation and term. In humans, however, a very small proportion of triploids are liveborn but display a wide range of congenital abnormalities. In particular, the placentas of human diandric triploid embryos consistently display “partial” hydatidiform molar degeneration, while those of digynic triploids generally do not show these histopathological features. In mice, the postimplantation development of diandric and digynic triploid embryos also differs. While both classes are capable of developing to the forelimb bud stage, no specific degenerative features of their placentas have been reported. Diandric triploid mouse embryos are morphologically normal while digynic triploid mouse embryos consistently display neural tube and occasionally cardiac abnormalities. Previously it was shown that the preimplantation development of micromanipulated diandric triploid mouse embryos was similar to developmentally matched diploid control embryos. In this study, the preimplantation development of micromanipulated digynic triploid mouse embryos is analysed and compared with that of diandric triploid mouse embryos in order to determine whether there is any difference in cleavage rate between these two classes of triploids. Standard micromanipulatory procedures were used to insert a female or a male pronucleus into a recipient diploid 1-cell stage embryo. The karyoplast was fused to the cytoplasm of the embryo by electrofusion. These tripronucleate 1-cell stage embryos were then transferred to pseudopregnant recipients and, at specific times after the HCG injection to induce ovulation, the embryos were recovered and total cell counts made. These results were plotted and regression lines drawn. An additional control group of embryos was subjected to similar micromanipulatory procedures to those used in the experimental study. These embryos had a single pronucleus removed and this was then reinserted into the perivitelline space. Diploidy was immediately restored by electrofusion. These embryos were transferred to recipients and at specific times after the HCG injection the embryos were recovered and total cell counts made. These results were also plotted and regression lines drawn. The results show that the cell doubling time of the digynic triploid embryos was 14.84 h (± 1.19). This was not significantly different from that of the diandric triploid embryos (13.55 h ± 0.86; P > 0.05) or of the manipulated diploid controls (12.12 h ± 0.79; P > 0.05). © 1993 Wiley-Liss, Inc.     

Chromosome abnormalities in chicken (Gallus domesticus) embryos: Types,frequencies and phenotypic effects

Cytological screening of 4182 chick embryos from 10 strains and 5 strain crosses was performed to determine the types and frequencies of chromosome abnormalities. Gross phenotypic effects, such as growth retardation and malformation, were noted. Clues to the etiology of such chromosome aberrations were also sought. The following euploid series was observed: Haploid mosaics (A-Z/2A-ZZ, A-Z/2A-ZZ/3A-ZZZ, A-Z/A-W/2A-ZW/2A-ZZ, A-Z/A-W/2A-ZW/ 3A-Z?), diploid (2A-ZZ and 2A-ZW), triploid (3A-ZWW, 3A-ZZW, 3A-ZZZ, 3A-ZZZW) and tetraploid (4A-ZZWW and 4A-ZZZZ). Aneuploidy was observed as follows: Trisomy for chromosome numbers 1, 2, 3, 4 and double trisomy 2/5. Trisomy-4 with deletion of 50% of the long arm of one member of the trisomic triplet was observed. A 3A-ZWW embryo was found with two cell populations: one, disomic for chromosome 2 and 6; the other, tetrasomic for 2 and 6. Of the 4182 embryos sampled 1.4% were haploids, 97.5% diploids, 0.8% triploids, 0.1% tetraploids and 0.2% trisomics. On the average 10.8% of the early dead embryos were euploid (excluding diploid) or aneuploid. However, the range for euploidy and aneuploidy among strains was 2.3–23.7% of early deads. Haploid embryos were consistently underdeveloped at 4 days of incubation (D.I.), and died by 5–7 D.I. About 90% of (36) triploid embryos died at or before 4 D.I. The remaining 10% (normal embryos) died prior to hatching. Trisomic embryos were dead or underdeveloped at 4 D.I. Tetraploidy appeared to be lethal at a very early stage. The various strains examined had different overall rates of chromosome aberrations (0.4–8.9%), and also showed different varieties of such aberrations. The modes and possible causes of meiotic, mitotic and fertilization errors are considered. Genetic control of chromosome abnormalities, particularly haploidy, is postulated.     

Osmotic regulation in late embryos and prelarvae of the American lobster Homarus americanus H. Milne-Edwards, 1837 (Crustacea,Decapoda)

Osmotic regulation of embryos, hatching prelarvae, and free prelarvae was studied in Homarus americanus H. Milne-Edwards, 1837. Adaptation time in dilute media was much longer in embryos than in hatchlings and prelarvae. Regulation in dilute media was hyperosmotic in embryos and hyperosmoconforming in hatchlings and prelarvae. The change in type of osmoregulation that occurred during the hatch is due to the rupture of the relatively impermeable outer egg membrane.     

Effect of hatching time on time to first feed intake,organ development,enzymatic activity and growth in broiler chicks hatched on-farm

The conventional commercial hatcheries used today do not allow the newly hatched chicks to consume feed or water. Combined with natural variation in hatching time, this can lead to early hatched chicks being feed-deprived for up to 72 h before being unloaded at the rearing site. This study investigated the effects of hatching time on time to first feed intake and development of organs, digestive enzymes and productivity in terms of growth and feed conversion ratio in chicks hatched on-farm. Chicks were divided into three hatching groups (early, mid-term and late), and assessed over a full production cycle of 34 days. The results revealed that chicks remain inactive for a considerable amount of time before engaging in eating-related activities. Eating activity of 5% (i.e. when 5% of birds in each hatching group were eating or standing close to the feeder) was recorded at an average biological age (BA) of 25.4 h and a proportion of 50% birds with full crop was reached at an average BA of 30.6 h. Considering that the hatching window was 35 h in this study, the average chick probably did not benefit from access to feed and water immediately post-hatch in this case. At hatch, mid-term hatchlings had a heavier small intestine (30.1 g/kg bw) than both early (26.4 g/kg bw) and late (26.0 g/kg bw) hatchlings. Relative length of the small intestine was shorter in late hatchlings (735 cm/kg bw) than in mid-term (849 cm/kg bw) and early (831 cm/kg bw) hatchlings. However, the relative weight of the bursa fabricii was greater in mid-term (1.30 g/kg bw) than in early hatchlings (1.01 g/kg bw). At hatch, late hatchlings were heavier than early and mid-term hatchlings (P < 0.05), but by 3 days of age early hatchlings were heavier than mid-term and late hatchlings (P < 0.01). The only effect persisting throughout the study was a difference in the relative weight of the small intestine, where late hatchlings had heavier intestines than early hatchlings (P < 0.05). Thus, while there were differences between hatching groups, this study showed that the hatchlings seemed capable of compensating for these as they grew.     

Development of sheep androgenetic embryos is boosted following transfer of male pronuclei into androgenetic hemizygotes

Androgenetic embryos are useful model for investigating the contribution of the paternal genome to embryonic development. Little work has been done with androgenetic embryo production in domestic animals. The aim of this study was the production of diploid androgenetic sheep embryos. In vitro matured sheep oocytes were enucleated and fertilized in vitro; parthenogenetic and normally fertilized embryos were also produced as a control. Fifteen hours after in vitro fertilization (IVF), presumptive zygotes were centrifuged and scored for the number of pronucleus. IVF, parthenogenetic, and androgenetic embryos (haploid, diploid, and triploid) were cultured in SOFaa medium with bovine serum albumin (BSA). The proportion of oocytes with polyspermic fertilization increased linearly with increasing sperm concentration. After IVF, there was no significant difference in early cleavage and morula formation rates between the groups, while there was a significant difference on blastocyst development between IVF, parthenogenetic, and androgenetic embryos, the last ones displaying poor developmental potential (IVF, parthenogenetic, and haploid, diploid, and triploid androgenetic embryos: 43%, 38%, 0%, 2%, and 2%, respectively). In order to boost androgenetic embryonic development, we produced diploid androgenetic embryos through pronuclear transfer. Single pronuclei were aspirated with a bevelled pipette from haploid or diploid embryos and transferred into the perivitelline space of other haploid embryos, and the zygotes were reconstructed by electrofusion. Fusion rates approached 100%. Pronuclear transfer significantly increased blastocyst development (IVF, parthenogenetic, androgenetic: Diploid into Haploid, and Haploid into Haploid: 42%, 42%, 19%, and 3%, respectively); intriguingly, the Haploid + Diploid group showed the highest development to blastocyst stage. The main findings of our study are: (1) sheep androgenetic embryos display poor developmental ability compared with IVF and parthenogenetic embryos; (2) diploid androgenetic embryos produced by pronuclear exchange developed in higher proportion to blastocyst stage, particularly in the Diploid-Haploid group. In conclusion, pronuclear transfer is an effective method to produce sheep androgenetic blastocysts.     

Tripronuclear human oocytes: altered cleavage patterns and subsequent karyotypic analysis of embryos

Between 1 and 4% of human oocytes fertilized in vitro are tripronuclear. It has been reported that these tripronuclear oocytes can develop to grossly normal-appearing morulae and that chromosomally, these embryos could be triploid, diploid, or severely depleted. The etiology and proportion of apparently diploid and aneuploid embryos deriving from tripronuclear human oocytes is unknown. This study provides evidence for the first time that most (18 of 29) tripronuclear human oocytes cleave directly to 3-cells at the first cleavage division. These embryos have a severely abnormal (but not triploid) chromosomal complement. Furthermore, some (4 of 29) tripronuclear human oocytes cleave to 2-cells plus an extrusion, and these embryos are diploids, whereas some (7 of 29) cleave to 2-cells, and these embryos are triploid after the first cleavage division. These findings demonstrate that most tripronuclear human oocytes have an altered cleavage pattern at the first cleavage division, that most tripronuclear human oocytes (76% in this study) do not develop into triploid embryos, and that a correlation exists between the pattern of the first cleavage division and the subsequent karyotype of these embryos. Insight into the mechanisms by which these oocytes fail to develop into triploid embryos is also provided.     

Analysis of post-implantation mouse embryos after maternal heat stress during meiotic maturation

The effects of heat stress during oocyte maturation were studied in post-implantation mouse embryos. Virgin ICR mice were exposed to 35 +/- 1 degree C and 65 +/- 3% RH for 12.5 h beginning immediately after synchronization of ovulation with PMSG and hCG. Embryos of heat-stressed dams were developmentally heterogeneous and showed significant delays in development with as much as 48 h delayed development. Nearly 6% of these embryos were triploid, and another 2% were hyper-diploid. Development of triploid embryos was delayed more than 24 h. Nine embryos with severe developmental delay had heterogeneous chromosome constitutions. Embryo mortality before and after implantation was higher in heat-stressed dams than in controls.     

Incidence of chromosomal anomalies in early bovine embryos derived from in vitro fertilization

The incidence of chromosomal anomalies in early bovine embryos derived from follicular oocytes fertilized in vitro using sperm separated by Percoll density gradient centrifugation was investigated. Overall, chromosomal anomalies were observed in 13.7% (138/1005) of embryos. There were 14 haploids (1.4%), 2 hypodiploids (0.2%), 6 hyperdiploids (0.6%), 101 triploids (10.0%), 12 tetraploids (1.2%), 2 diploid/triploid mosaics (0.2%), and 1 diploid/tetraploid mosaic (0.1%). The frequency of triploidy was caused mainly by polyspermy. There was a significant difference in the frequency of embryos with abnormal chromosomes between the two bulls used (P < 0.005), but Percoll centrifugation did not affect the observed incidence of anomalies. The frequency of chromosomal anomalies in embryos at each stage increased with delay or arrest of development. These results suggest that the incidence of chromosomal anomalies depended on the conditions of in vitro fertilization and the arrest of development.     

Wasp predation and wasp-induced hatching of red-eyed treefrog eggs

Eggs often suffer high levels of predation and, compared with older animals, embryos have few options available for antipredator defence. None the less, hatchlings can escape from many predators to which eggs are vulnerable. I studied early hatching as an antipredator defence of red-eyed treefrog embryos, Agalychnis callidryas, in response to egg predation by social wasps (Polybia rejecta). Red-eyed treefrogs attach their eggs to vegetation overhanging water, where they are exposed to arboreal and aerial predators. Wasps attacked half the egg clutches and killed almost a quarter of the eggs I monitored at a natural breeding site in Panama. Hatching tadpoles fall into the water, where they face aquatic predators. As predicted from improved survival of older hatchlings with aquatic predators, most undisturbed eggs hatched relatively late. However, many younger embryos directly attacked by wasps hatched immediately. Embryos attacked by wasps hatched as much as a third younger than the peak undisturbed hatching age, and most hatching embryos escaped. Thus hatching is an effective defence against wasp predation, and plasticity in hatching stage allows embryos to balance risks from stage-specific egg and larval predators. Wasp-induced hatching is behaviourally similar to the snake-induced hatching previously described in A. callidryas, but occurs in fewer eggs at a time, congruent with the scale of the risk. Individual embryos hatch in response to wasps, which take single eggs, whereas whole clutches hatch in response to snakes, which consume entire clutches. Embryos of A. callidryas thus respond appropriately to graded variation in mortality risks. Copyright 2000 The Association for the Study of Animal Behaviour.     

Yolk partitioning in embryos of the lizard, Calotes versicolor: maximize body size or save energy for later use?

The hatchlings of Calotes versicolor and other congeners retain residual yolk for immediate post-hatching needs. Excision of 8% yolk (approximately equal residual yolk) from the eggs resulted in smaller hatchlings when compared to those emerging from sham operated eggs. However, hatchlings in both groups retained the same amount of residual yolk. The findings suggest that residual yolk in hatchlings of C. versicolor is an important part of the energy budget set aside by developing embryos and that the advantages of large hatchling size are counter balanced by selection for residual yolk.     

Development of a pheasant interspecies primordial germ cell transfer to chicken embryo: Effect of donor cell sex on chimeric semen production

This study was conducted to evaluate whether the sex of donor primordial germ cells (PGCs) influences production of chimeric semen from recipient hatchlings produced by interspecies transfer between pheasant (Phasianus colchicus) and chicken (Gallus gallus). Pheasant PGCs were retrieved from 7-d-old embryos and subsequently transferred into circulatory blood of 2.5-d-old (Stage 17) embryos. The sex of embryos was discerned 3 to 6 days after laying, and in preliminary study, overall rate of embryo survival after sexing was 74.6% with male-to-female ratio of 0.49 to 0.51. In Experiment 1, magnetic-activated cell sorting (MACS) using QCR1 antibody was effective for enriching the population of male and female PGCs in gonadal cells (9.2- to 12.5-fold and 10.8- to 19.5-fold increase, respectively). In Experiment 2, an increase in the number of hatchlings producing chimeric semen was detected after the homosexual transfer of male-to-male compared with that after the heterosexual transfer of female-to-male (68% to 88%). Significant increase was found in the frequency of chimeric semen production (0.96 to 1.68 times); production of pheasant progenies by artificial insemination using chimeric semen was also increased in the homosexual transfer (0 to 3 cases). In conclusion, the homosexual PGC transfer of male-to-male yielded better rate of generating pheasant progenies after test cross-reproduction than that of the heterosexual transfer of female-to-male, which could improve the efficiency of interspecies germ cell transfer system.     

Histochemical identification of primordial germ cells in diandric and digynic triploid mouse embryos

Diandric and digynic triploid mouse embryos were isolated in the morning on day 10 of gestation. The embryos were separated from their extraembryonic membranes, and the latter were analysed cytogenetically by G-banding to establish the ploidy and sex chromosome constitution of these embryos. The diandric triploid embryos were produced by the technique of nuclear micromanipulation. Females were mated with male mice with a morphologically distinguishable "marker" chromosome to confirm the diandric status of these embryos. Digynic triploid and normal diploid embryos were isolated from LT/Sv strain females. These females spontaneously ovulate both primary and secondary oocytes, which are fertilisable and give rise to digynic triploid and normal diploid embryos, respectively. All the embryos were serially sectioned and processed in order to demonstrate the presence of alkaline phosphatase enzyme activity. This histochemical technique allowed primordial germ cells to be readily recognised, due to their characteristic location, cellular morphology, and staining appearance. Primordial germ cells were found in all the embryos studied, being located within the visceral yolk sac, at the base of the allantois, and/or in association with the wall or mesentery of the hindgut. The total number of germ cells present was established in nine diandric triploids and in five digynic triploids. The findings presented here represent the first demonstration that primordial germ cells can differentiate in either diandric or digynic triploid mammalian embryos.     

The development potential of ethanol-induced monosomic and trisomic conceptuses in the mouse

The development potential of fertilized embryos isolated from female mice previously given a single dose of either a dilute solution of ethanol or distilled water (controls) by mouth was studied. Exposure to ethanol occurred at various times during the cycle leading to ovulation and shortly after fertilization. The chromosome constitution of all preimplantation embryos isolated from these females was determined either at the first cleavage mitosis or at the morula stage. The incidence of aneuploidy in the ethanol-exposed groups at these times was approximately 19% and 13.5%, respectively, with a similar number of monosomic and trisomic conceptuses observed at these times. In addition, about 2% of all conceptuses examined were triploid. Further females were autopsied on the 10th or 11th day of gestation, though the chromosome constitution of only the morphologically abnormal or developmentally retarded embryos was determined. Eight embryos out of a total of 16 studied in the ethanol-exposed group were either aneuploid or triploid, whereas in the control group only one out of 11 examined proved to be aneuploid. The triploids and ethanol-induced aneuploid conceptuses appeared to be capable of surviving to the morula stage but generally failed to survive to the 10th/11th day. No monosomics were in fact observed in the postimplantation series. The present findings are briefly discussed with reference to the possible pathogenesis of spontaneous abortions in man, which often possess similar types of chromosomal anomalies.     

Development of cardiac rhythms in birds

Heart rate (HR) in avian embryos developing inside an eggshell has been measured by various means while maintaining adequate gas exchange through the eggshell. This is an important requirement in order to avoid adverse effects of impeding gas exchange on the cardiac rhythms of developing embryos. The present report is a review of our ontogenetic study on embryonic HR, which was measured with fulfillment of the above requirement and also hatchling HR measured non-invasively. Firstly, we reviewed measurements of daily changes (developmental patterns) in embryonic mean heart rate (MHR), which were determined from a short-term measurement of HR once a day, in 34 species of altricial and precocial birds. The allometric relationship between the MHR during pipping in altricial birds and their fresh egg masses was the same as that between the MHR at 80% of incubation duration and fresh egg masses in pre-cocial birds. Secondly, we presented the developmental patterns of MHR in chick embryos and hatchlings, which were determined from long-term, continuous measurement of HR before, during and after hatching. The ultradian and circadian rhythms of HR were clearly shown in embryos and hatchlings, respectively. Thirdly, we summarized instantaneous HR fluctuations: HR variability and HR irregularities, in chick embryos and hatchlings. The distinctive patterns were shown in pre-pipped and pipped embryos and newly hatched chicks, individually, which were partly related to autonomic nervous functions and physiological functions.     

Induced triploidy in carp, Cyprinus carpio L.

A method for mass production of triploid carp was developed by the cold treatment of eggs after fertilization. Triploidy was demonstrated by cytophotometry and chromosome preparation techniques. Of the haploid embryos, 100% showed morphological abnormalities and died soon after hatching. Haploid embryos of maternal and paternal origin occurring in the course of cold treatment were distinguished by using a genetic marker. Among morphologically normal larvae, the frequency of triploidy was 100%, and the viability of the triploid embryos, larvae and the juvenile fish did not differ from the diploid control. Sex differentiation started in the triploid fish, but was greatly retarded. Gonad size and the small number of growing oocytes suggest that most of the triploid individuals will be sterile. Triploid carp grew at the same rate as diploid controls in the laboratory test.     

Ventilatory chemosensitivity of the 1-day-old chicken hatchling after embryonic hypoxia

We investigated the effects of sustained embryonic hypoxia on the neonatal ventilatory chemosensitivity. White Leghorn chicken eggs were incubated at 38 degrees C either in 21% O(2) throughout incubation (normoxia, Nx) or in 15% O(2) from embryonic day 5 (hypoxia, Hx), hatching time included. Hx embryos hatched approximately 11 h later than Nx, with similar body weights. Measurements of gaseous metabolism (oxygen consumption, Vo(2)) and pulmonary ventilation (Ve) were conducted either within the first 8 h (early) or later hours (late) of the first posthatching day. In resting conditions, Hx had similar Vo(2) and body temperature (Tb) and slightly higher Ve and ventilatory equivalent (Ve/Vo(2)) than Nx. Ventilatory chemosensitivity was evaluated from the degree of hyperpnea (increase in Ve) and of hyperventilation (increase in Ve/Vo(2)) during acute hypoxia (15 and 10% O(2), 20 min each) and acute hypercapnia (2 and 4% CO(2), 20 min each). The chemosensitivity differed between the early and late hours, and at either time the responses to hypoxia and hypercapnia were less in Hx than in Nx because of a lower increase in Ve and a lower hypoxic hypometabolism. In a second group of Nx and Hx hatchlings, the Ve response to 10% O(2) was tested in the same hatchlings at the early and late hours. The results confirmed the lower hypoxic chemosensitivity of Hx. We conclude that hypoxic incubation affected the development of respiratory control, resulting in a blunted ventilatory chemosensitivity.     

Size-dependent pigmentation-pattern formation in embryos of Alligator mississippiensis: time of initiation of pattern generation mechanism

The pigmentation pattern of Alligator mississippiensis was examined. The number of white stripes on the dorsal side of embryos (stages 21-28) and hatchlings from eggs incubated at 30 degrees C (100% females) and 33 degrees C (100% males) was recorded. Total length, nape-rump length and tail length were recorded for each embryo and hatchling. The number of white stripes was affected by incubation temperature but not sex; hatchlings incubated at 33 degrees C had two more white stripes than those at 30 degrees C, despite being the same length. Five female hatchlings produced at 33 degrees C by manipulation of the temperature, had the same number of stripes as males that developed under the same incubation temperatures. The appearance of the pigmentation was accelerated in embryos incubated at 33 degrees C, occurring eight days earlier than at 30 degrees C. At the time just before the first signs of pigment deposition, embryos from 33 degrees C were longer than those at 30 degrees C. If the stripe formation is size dependent this explains why hatchlings at 33 degrees C have more stripes than hatchlings from 30 degrees C. The mechanism that produces the stripe patterns is unknown. We describe key elements a pattern formation mechanism must possess to produce such stripes and suggest a possible mechanism, based on cell movement driven by chemotaxis. We apply the mathematical model to dorsal patterning on A. mississippiensis. We show how length at pattern formation is the prime factor in determining stripe number and how the pattern can be formed in the observed anterior-posterior sequence. We present numerical simulations and show that the qualitative behaviour is consistent with the experimental results.     

Determinants of embryonic stage at oviposition in the lizard Urosaurus ornatus

Relatively few squamate reptiles oviposit eggs with embryos at developmental stages greater than stage 30. To investigate potential proximate and ultimate bases of this phenomenon, we experimentally induced females of the lizard Urosaurus ornatus to retain their eggs past the normal time of oviposition (NTO). This procedure allowed us to determine whether the length of egg retention is fixed or facultative and to evaluate the effects of retention on embryos, hatchlings, and females. Females were able to retain eggs facultatively for at least 29 d past the NTO. However, retention resulted in arrested development of embryos; arrest occurred at stages 30-30.5, which is only slightly more advanced than that at the NTO (stage 29.5). Embryogenesis was reinitiated when eggs were removed from females and placed in incubation media. Hatching success of these eggs was high (87%), and incubation time was not affected by the number of days that development had been arrested. However, the snout-vent length and water content of hatchlings were negatively related to the length of retention, and they ran slower than hatchlings from control eggs obtained at the NTO. Retention of eggs past the NTO had no detectable effect on the body condition or running speeds of females. Developmental arrest and the adverse effects of retention on hatchling phenotype, if widespread among squamates, would account for the limited range of embryo stages at oviposition and act as major constraints on the evolution of viviparity.