Developmental basis for telencephalon expansion in waterfowl: enlargement prior to neurogenesis

Some altricial and some precocial species of birds have evolved enlarged telencephalons compared with other birds. Previous work has shown that finches and parakeets, two species that hatch in an immature (i.e. altricial) state, enlarged their telencephalon by delaying telencephalic neurogenesis. To determine whether species that hatch in a relatively mature (i.e. precocial) state also enlarged their telencephalon by delaying telencephalic neurogenesis, we examined brain development in geese, ducks, turkeys and chickens, which are all precocial. Whereas the telencephalon occupies less than 55 per cent of the brain in chickens and turkeys, it occupies more than 65 per cent in ducks and geese. To determine how these species differences in adult brain region proportions arise during development, we examined brain maturation (i.e. neurogenesis timing) and estimated telencephalon, tectum and medulla volumes from serial Nissl-stained sections in the four species. We found that incubation time predicts the timing of neurogenesis in all major brain regions and that the telencephalon is proportionally larger in ducks and geese before telencephalic neurogenesis begins. These findings demonstrate that the expansion of the telencephalon in ducks and geese is achieved by altering development prior to neurogenesis onset. Thus, precocial and altricial species evolved different developmental strategies to expand their telencephalon.     

Genetic regulation of CFA expression in interspecific avian hybrids and somatic cell hybrids

Chicken fetal-leukemic antigen (CFA) is an oncodevelopmental antigen present on embryonic and neonatal chicken peripheral red blood cells (RBCs) but is not restricted to fetal stages of development in other avian species. Crosses between white Leghorn chickens and Japanese quail resulted in adult hybrids whose peripheral RBCs were positive for CFA. Of the four CFA determinants normally found in adult quail RBCs, only two were present on quail-chicken hybrid RBCs. Adult quail--chicken hybrid RBCs also possessed on CFA determinant associated with early development in both quail and chicken and one chicken-specific CFA determinant. Evidence is presented for the possible association of CFA-positive adult peripheral RBCs and the level of circulating reticulocytes. Crosses between pheasant and turkey (both with CFA-positive adult RBCs) resulted in hybrid adult RBCs expressing only a portion of the parental CFA determinants. Through the formation of somatic cell hybrids between adult chicken and embryonic Japanese quail RBCs, it was possible to induce the appearance of CFA determinants normally restricted to embryonic chicken RBCs. Approximately 50% of the hybrid cells showed reexpression of CFA, and this induction was both time and temperature dependent. Hybridization between RBCs of adult chicken and those of either adult Japanese quail or adult turkey failed to elicit the reexpression of chicken-specific CFA.     

Species specificity and developmental patterns of expression of the beta amyloid precursor protein (APP) gene in brain, liver and choroid plexus in birds.

1. Human APP cDNA hybridized to a 3.5 kb mRNA in liver and brain RNA from chickens, pigeons, quail and ducks as well as in RNA from choroid plexus of chicken and quail. In contrast to all other species hitherto examined a 1.6 kb mRNA hybridizing to APP cDNA was found in abundant amounts in RNA from chicken and quail livers. 2. In the chicken, before hatching, the levels of APP mRNA in total RNA from liver and choroid plexus were higher than those in RNA from liver and choroid plexus of adults. However, RNA from the rest of the brain of chicken embryos contained less APP mRNA than RNA from brain of adults. 3. In the chicken, between 10 and 40 days after hatching, APP mRNA levels in RNA from liver were higher than adult levels, APP mRNA levels in RNA from choroid plexus were similar to adult levels and APP mRNA levels in RNA from the rest of brain were below the adult levels.     

Host specificity of Cryptosporidium sp. isolated from chickens

The host specificity of Cryptosporidium sp. infecting chickens was evaluated by oral inoculation of oocysts into 6 different species of neonatal rodents, adult nude mice (athymic), neonatal conventional and gnotobiotic pigs, turkeys, muscovy ducks and bobwhite quail. Examinations of tissue sections, ileal mucosal smears, fecal flotations and stained feces failed to reveal any infections in the mammalian species examined. Oocysts were observed in the feces, and developmental stages were observed in tissue sections, of turkeys and muscovy ducks but not bobwhite quail. This study indicates that Cryptosporidium sp. infections in avian species are probably not a zoonotic threat to humans.     

The CDC25B phosphatase shortens the G2 phase of neural progenitors and promotes efficient neuron production

During embryonic development, changes in cell cycle kinetics have been associated with neurogenesis. This observation suggests that specific cell cycle regulators may be recruited to modify cell cycle dynamics and influence the decision between proliferation and differentiation. In the present study, we investigate the role of core positive cell cycle regulators, the CDC25 phosphatases, in this process. We report that, in the developing chicken spinal cord, only CDC25A is expressed in domains where neural progenitors undergo proliferative self-renewing divisions, whereas the combinatorial expression of CDC25A and CDC25B correlates remarkably well with areas where neurogenesis occurs. We also establish that neural progenitors expressing both CDC25A and CDC25B have a shorter G2 phase than those expressing CDC25A alone. We examine the functional relevance of these correlations using an RNAi-based method that allows us to knock down CDC25B efficiently and specifically. Reducing CDC25B expression results in a specific lengthening of the G2 phase, whereas the S-phase length and the total cell cycle time are not significantly modified. This modification of cell cycle kinetics is associated with a reduction in neuron production that is due to the altered conversion of proliferating neural progenitor cells to post-mitotic neurons. Thus, expression of CDC25B in neural progenitors has two functions: to change cell cycle kinetics and in particular G2-phase length and also to promote neuron production, identifying new roles for this phosphatase during neurogenesis.     

A comparative study of embryonic development of some bird species with different patterns of postnatal growth

Some studies show that birds with high postnatal growth rates (e.g. altricial species) are characterized by a rapid early development of "supply" organs, such as digestive organs. Birds with low postnatal growth rates (e.g. precocial species) exhibit a slower early development of these organs and a more rapid early development of other "demand" organs, such as brain, muscles, skeleton and feathers. To test whether these differences can be traced back to early embryonic development and whether they can be associated with changes in developmental timing, i.e. heterochrony, we compared embryos of the precocial quail and the altricial fieldfare, two bird species with low and high postnatal growth rates, respectively. We used classical staging techniques that use developmental landmarks to categorize embryonic maturity as well as morphological measurements. These techniques were combined with immune detection of muscle specific proteins in the somites. Our data showed that the anlagen of the head, brain and eyes develop earlier in the quail than in the fieldfare in contrast to the gut which develops earlier in the fieldfare than in the quail. Our data also showed that the quail and the fieldfare displayed different rates of myotome formation in the somites which contribute to muscle formation in the limbs and thorax. We believe these observations are connected with important differences in neonatal characteristics, such as the size of the brain, eyes, organs for locomotion and digestion. This leads us to the conclusion that selection for late ontogenetic characteristics can alter early embryonic development and that growth rate is of fundamental importance for the patterning of avian embryonic development. It also appears that this comparative system offers excellent opportunities to test hypotheses about heterochrony.     

The genesis of cartilage size and shape during development and evolution

How do cartilaginous elements attain their characteristic size and shape? Two intimately coupled processes underlie the patterned growth of cartilage. The first is histogenesis, which entails the production of cartilage as a discrete tissue; the second is morphogenesis, which pertains to the origins of three-dimensional form. Histogenesis relies on cues that promote the chondrogenic differentiation of mesenchymal cells, whereas morphogenesis requires information that imbues cartilage with stage-specific (e.g. embryonic versus adult), region-specific (e.g. cranial versus appendicular) and species-specific size and shape. Previous experiments indicate that early programmatic events and subsequent signaling interactions enable chondrogenic mesenchyme to undergo histogenesis and morphogenesis, but precise molecular and cellular mechanisms that generate cartilage size and shape remain unclear. In the face and jaws, neural crest-derived mesenchyme clearly plays an important role, given that this embryonic population serves as the source of chondrocytes and of species-specific patterning information. To elucidate mechanisms through which neural crest-derived mesenchyme affects cartilage size and shape, we made chimeras using quail and duck embryos, which differ markedly in their craniofacial anatomy and rates of maturation. Transplanting neural crest cells from quail to duck demonstrates that mesenchyme imparts both stage-specific and species-specific size and shape to cartilage by controlling the timing of preceding and requisite molecular and histogenic events. In particular, we find that mesenchyme regulates FGF signaling and the expression of downstream effectors such as sox9 and col2a1. The capacity of neural crest-derived mesenchyme to orchestrate spatiotemporal programs for chondrogenesis autonomously, and to implement cartilage size and shape across embryonic stages and between species simultaneously, provides a novel mechanism linking ontogeny and phylogeny.     

Effects of the organophosphate insecticides diazinon and parathion on bobwhite quail embryos: skeletal defects and acetylcholinesterase activity

Bobwhite quail eggs were injected at 48 or 72 hr of incubation with various doses of the organophosphate (OP) insecticides diazinon or parathion and the embryos were examined after an additional 48 hr of incubation by both histological and cartilage-staining methods. Bobwhite embryos did not display the notochordal folding or vascular enlargement reported for OP-injected chicken embryos. Cartilage staining of embryos injected with insecticide at 72 hr of incubation and recovered at day 12 of incubation revealed severe shortening and contortion of the vertebral axis, as well as tibiotarsal, rib, and sternum defects. Parathion was more potent in causing skeletal defects than diazinon. No type I defects (micromelia, parrot beak) were detected. Radiometric acetylcholinesterase (AChE) assays of whole embryo homogenates were performed for day 6, 9, and 12 diazinon-injected and control embryos. Diazinon effected drastic reductions in AChE activity. Although the AChE and axial skeletal responses of bobwhite embryos to OP injection are similar to those reported in the literature for other species, some major differences in the bobwhite response were noted: namely, the absence of notochordal folding in the young bobwhite embryo and the absence of type I defects at day 12. These differences suggest that further studies with the bobwhite quail would be useful in clarifying the mechanisms involved in OP-induced teratogenesis.     

Interspecies avian brain chimeras reveal that large brain size differences are influenced by cell-interdependent processes

Like humans, birds that exhibit vocal learning have relatively delayed telencephalon maturation, resulting in a disproportionately smaller brain prenatally but enlarged telencephalon in adulthood relative to vocal non-learning birds. To determine if this size difference results from evolutionary changes in cell-autonomous or cell-interdependent developmental processes, we transplanted telencephala from zebra finch donors (a vocal-learning species) into Japanese quail hosts (a vocal non-learning species) during the early neural tube stage (day 2 of incubation), and harvested the chimeras at later embryonic stages (between 9-12 days of incubation). The donor and host tissues fused well with each other, with known major fiber pathways connecting the zebra finch and quail parts of the brain. However, the overall sizes of chimeric finch telencephala were larger than non-transplanted finch telencephala at the same developmental stages, even though the proportional sizes of telencephalic subregions and fiber tracts were similar to normal finches. There were no significant changes in the size of chimeric quail host midbrains, even though they were innervated by the physically smaller zebra finch brain, including the smaller retinae of the finch eyes. Chimeric zebra finch telencephala had a decreased cell density relative to normal finches. However, cell nucleus size differences between each species were maintained as in normal birds. These results suggest that telencephalic size development is partially cell-interdependent, and that the mechanisms controlling the size of different brain regions may be functionally independent.     

Formation of hematopoietic colonies of the donor type in the bone marrow of irradiated chickens after transplantation of the cells of the yolk sac and hindlimb of the quail embryo]

The ability of yolk sac and primary bone marrow cells of the quail to form hemopoietic colonies at 6 hours of incubation (i. e. before establishment of circulation) was studied in the bone marrow of 3-week sublethally irradiated chickens. The experiments were based on the possibility of differentiating between quail and chicken cells from the natural cell marker (Pheulgen-positive nucleolus). The number of hemopoietic colonies produced by cells transplanted from the primary bone marrow was three times greater than that consequent on transplantation of yolk sac cells. With the given dose of irradiation the bone marrow shows about 75% exogenous (quail) and 25% endogenous (chicken) hemopoietic colonies.     

Role of quail in the interspecies transmission of H9 influenza A viruses: molecular changes on HA that correspond to adaptation from ducks to chickens

H9 influenza viruses have become endemic in land-based domestic poultry in Asia and have sporadically crossed to pigs and humans. To understand the molecular determinants of their adaptation to land-based birds, we tested the replication and transmission of several 1970s duck H9 viruses in chickens and quail. Quail were more susceptible than chickens to these viruses, and generation of recombinant H9 viruses by reverse genetics showed that changes in the HA gene are sufficient to initiate efficient replication and transmission in quail. Seven amino acid positions on the HA molecule corresponded to adaptation to land-based birds. In quail H9 viruses, the pattern of amino acids at these seven positions is intermediate between those of duck and chicken viruses; this fact may explain the susceptibility of quail to duck H9 viruses. Our findings suggest that quail provide an environment in which the adaptation of influenza viruses from ducks generates novel variants that can cross the species barrier.     

Progranulin regulates neurogenesis in the developing vertebrate retina

We evaluated the expression and function of the microglia‐specific growth factor, Progranulin‐a (Pgrn‐a) during developmental neurogenesis in the embryonic retina of zebrafish. At 24 hpf pgrn‐a is expressed throughout the forebrain, but by 48 hpf pgrn‐a is exclusively expressed by microglia and/or microglial precursors within the brain and retina. Knockdown of Pgrn‐a does not alter the onset of neurogenic programs or increase cell death, however, in its absence, neurogenesis is significantly delayed—retinal progenitors fail to exit the cell cycle at the appropriate developmental time and postmitotic cells do not acquire markers of terminal differentiation, and microglial precursors do not colonize the retina. Given the link between Progranulin and cell cycle regulation in peripheral tissues and transformed cells, we analyzed cell cycle kinetics among retinal progenitors following Pgrn‐a knockdown. Depleting Pgrn‐a results in a significant lengthening of the cell cycle. These data suggest that Pgrn‐a plays a dual role during nervous system development by governing the rate at which progenitors progress through the cell cycle and attracting microglial progenitors into the embryonic brain and retina. Collectively, these data show that Pgrn‐a governs neurogenesis by regulating cell cycle kinetics and the transition from proliferation to cell cycle exit and differentiation. © 2017 The Authors. Developmental Neurobiology Published by Wiley Periodicals, Inc. Develop Neurobiol 77: 1114–1129, 2017     

Comparison of glutathione S-transferase activity in the rat and birds: tissue distribution and rhythmicity in chicken (Gallus domesticus) liver.

1. Mature, male chickens, Bobwhite quail, and rats differed with respect to glutathione S-transferase (GST) activity in the kidney, duodenum and testis, but species differences were not observed in the liver. 2. GST activity was present in the heart, spleen, liver, duodenum, kidney, testis, cerebral cortex, cerebellum, optic tecta, and medulla oblongata of chickens with differences in tissues and breeds. 3. Renal GST activity was higher in female chickens, whereas enzyme activity in the brain was higher in males. 4. Hepatic GST activity fluctuated about a mean of 784 nmol min-1 mg protein-1 with a 12 hr periodicity which was not a feeding phenomenon. 5. The results demonstrate that GST activity occurs in diverse tissues of the chicken and Bobwhite quail with kidney greater than liver greater than duodenum greater than testis, compared to testis greater than liver greater than duodenum greater than kidney in the rat. Hepatic GST activity exhibits an ultradian periodicity.     

Auditory stimulus-induced changes in immediate-early gene expression related to an inborn perceptual predisposition

Auditory-induced expression of the immediate-early gene ZENK was examined in 18 brain areas of domestic chicken and Japanese quail subjects with no previous exposure to parental vocalizations. After one 30-min exposure (approximately 120 calls) within 24 h of hatching to either the chicken or quail maternal call, paired subjects from each species showed significantly more intense levels of ZENK staining to conspecific (own species) calls. Single brain areas did not show consistent, large differences in ZENK expression across all subject pairs. Instead, a majority of brain areas exhibited a small conspecific stimulus-induced staining advantage in each species. The species differed in the quantitative distribution of ZENK responses among brain areas; qualitative patterns of call-induced staining exhibited both similarities and differences. The results suggest shared neural correlates and potential developmental/evolutionary differences in congenital brain responses to biologically significant auditory stimuli in na?ve individuals of the two species.     

Identification and characterization of tenp,a gene transiently expressed before overt cell differentiation during neurogenesis

The initiation of cellular differentiation in the vertebrate central nervous system is a cascade of regulated gene expression events involving both intrinsic and extrinsic factors. Currently, the molecular events underlying the developmental transition from the cessation of cell proliferation to the onset of cell differentiation during neurogenesis are still poorly understood. We have identified a gene, tenp, which is likely to play a role during the transition. tenp mRNA was detected in the developing retina and brain, but not in the heart, kidney, or liver. Anatomically, cells expressing tenp formed a narrow strip at the boundary between the ventricular zone (consisting of proliferating cells) and the intermediate zone (occupied by postmitotic, differentiating cells). Further analysis showed that they were bromodeoxyuridine negative and thus postmitotic, yet without apparent differentiation such as the expression of microtubule-associated protein (MAP2) and neurofilament (NF68). When expressed in chicken embryonic fibroblast cells through transfection, Tenp protein was immunodetected in membrane fractions, implying that Tenp might be a membrane protein as predicted by a computer analysis of its primary sequence. Our data suggested that tenp might be involved in an early neurogenic event that existed transiently after terminal mitosis, yet before the overt differentiation of neural precursor cells. © 1998 John Wiley & Sons, Inc. J Neurobiol 34: 319–328, 1998     

Characterization of critical factors influencing gene expression of two types of fatty acid-binding proteins (L-FABP and Lb-FABP) in the liver of birds

In avian species, two types of intracellular lipid-binding proteins are abundant in the liver, the liver fatty acid-binding protein (L-FABP) and the liver basic fatty acid-binding protein (Lb-FABP). Both FABPs are capable of forming complexes with free fatty acids and bile acids, but the functional distinction between L-FABP and Lb-FABP in avian liver is not fully understood. To gain insights into the functional distinction between L-FABP and Lb-FABP, we investigated the expression of both genes in relation to the pre- and post-hatching development, diurnal cycle and feeding state in the livers of chicken (Gallus gallus) and Japanese quail (Coturnix japonica). In chickens, the Lb-FABP mRNA was expressed only in the liver, while the L-FABP was expressed in both liver and intestinal tissues. Only small amounts of the L-FABP and Lb-FABP mRNAs were detected in the liver during chicken embryogenesis, but at the onset of hatching a dramatic increase in mRNA expression was observed for both genes, suggesting that the expression of the L-FABP and Lb-FABP genes is synchronized at developmental stages. Remarkably, the diurnal expression pattern differed between the two genes under a 16L:8D condition in sexually mature quail: L-FABP gene expression transiently increased at the end of the light cycle, whereas Lb-FABP gene expression peaked during the early part of the light cycle and gradually decreased as the dark period approached. We attempted to identify the factors regulating the diurnal gene expression pattern, and found that feeding stimulation was a critical factor inducing Lb-FABP gene expression irrespective of light condition. On the other hand, feeding stimulation only slightly stimulated expression of the L-FABP gene, and was not always its primary determinant. These results suggest that L-FABP and Lb-FABP play different roles in metabolic process during the postprandial state.