Zebra finch as a developmental model

The domesticated zebra finch (Taeniopygia guttata) is a well‐established animal model for studying vocal learning. It is also a tractable model for developmental analyses. The finch genome has been sequenced and methods for its transgenesis have been reported. Hatching and sexual maturation in this species takes only two weeks and three months, respectively. Finch colonies can be established relatively easily and its eggs are laid at a stage earlier than in other common avian experimental models, facilitating the analysis of very early avian development. Representing the Neoaves to which 95% of all bird species belong, the finch can potentially complement two existing, Galloanserae developmental models, the chick, and quail. Here, we provide a step‐by‐step guide for how to set up a finch colony in a conventional laboratory environment. Technical tips are offered to optimize hens’ productivity and ensure a constant supply of fertilized finch eggs. Methods of handling finch eggs and embryos for subsequent embryological, cellular, or molecular analyses are also discussed. We conclude by emphasizing scientific values and cost effectiveness of maintaining a finch colony for avian developmental studies. genesis 53:669–677, 2015. © 2015 Wiley Periodicals, Inc.     

Peptidergic pathways in the avian brain

The results obtained by topographical studies on the immunoreactive peptide systems in the embryonic and adult avian brain (domestic fowl, domestic mallard, pigeon, Japanese quail, and zebra finch) can be realized only by means of phylogenetical comparisons. The comparative studies mainly demonstrate a fascinating constancy of the immunological properties and the spatial distribution of the neuropeptides. Independent of the development of the neopallium, and the increasing cerebral complexity, the spatial distribution of the neuropeptides, the location of their main perikaryal accumulations which are interconnected by immunoreactive fiber projections (and thereby forming widespread but continuous peptide systems) remain nearly unchanged during vertebrate evolution. The recognition of the neuropeptides as integral parts of the central nervous system is demonstrated by the fact that neuropeptidergic structures connect sensory inputs with central nervous areas as well as with the peripheral endocrinium.     

Vaterite Deposition During Eggshell Formation in the Cormorant,Gannet and Shag,and in ‘Shell-less’ Eggs of the Domestic Fowl.

The eggshells of the cormorant (Phalacrocorax carbo), domestic fowl (Gallus domesticus), gannet (Sula bassana), guinea fowl (Numida meleagris), greater flamingo (Phoenicopterus ruber), and shag (Phalacrocorax aristotelis) have been separated into two groups on the basis of the composition of their outer stratum. In the domestic fowl, guinea fowl and greater flamingo the outer stratum is an organic cuticle while in the sea-birds it is an inorganic cover rich in vaterite. The calcareous deposits on the membranes of eggs of the domestic fowl which are shell-less at oviposition have been shown to consist essentially of the vaterite form of calcium carbonate. Reasons for the occurrence of this polymorph of calcium carbonate are discussed with relation to the physiology of the birds.     

Levels of calcium and iron in the ovaries and eggs of cod Gadus morhua L. and plaice Pleuronectes platessa L

In cod, Gadus morhua L., calcium levels expressed as percentage of tissue dry weight were (mean +/- SD) 0.0042 +/- 0.0010% of ovary before final maturation and 0.0133 +/- 0.0024% of mature eggs. In plaice Pleuronectes platessa L. these values were 0.0029 +/- 0.0006 and 0.0097 +/- 0.0001. In mature eggs of both fish species these values are at least one order of magnitude less than in the yolk of the domestic fowl Gallus domesticus L. (0.14% of dry weight). The corresponding values for iron were (2.31 +/- 0.22) X 10(-3)% of ovary and (3.07 +/- 0.29) X 10(-3)% of mature egg in cod. In plaice these values were (1.57 +/- 0.15) X 10(-3)% and (2.74 +/- 0.94) X 10(-3)%. These levels are 4-7 times less than in the yolk of the domestic fowl (0.012-0.020%). These differences between the mature pelagic eggs of marine teleosts and the eggs of other oviparous vertebrates appear to be related to the availability of these elements in sea water, the different nature of the yolk phosphoprotein and the lower levels of protein phosphorus in the eggs of these fish species.     

Early nuclear events of in vitro fertilization in the domestic fowl (Gallus domesticus)

In vitro fertilization in the domestic fowl (Gallus domesticus) was investigated by observation of the early nuclear events. Ova retrieved from the fimbria following ovulation were inseminated in vitro with 10(6)-10(7) spermatozoa in Dulbecco's modified Eagle's medium (DMEM) for 10 min and then further incubated in DMEM + albumen for 1, 2, 3, or 4 hr. These eggs were histologically examined by epifluorescent microscopy after staining with 4',6'-diamidino-2-phenylindole (DAPI). Nuclei of spermatozoa at various stages of transformation were observed in the ova incubated for 1-3 hr. Close pairing of two pronuclei, presumed to be male and female juxtaposition, was detected in ova incubated for 4 hr. These data provide direct evidence for the in vitro fertilization of fowl eggs and suggested that the early process of in vitro fertilization is comparable to that of in vivo fertilization.     

Main arteries to the female reproductive organs in the helmeted guineafowl (Numida meleagris)

The helmeted guineafowl, Numida meleagris , is a member of the Galliformes (King & McLelland, 1975). This order includes the domestic fowl, Callus domeslicus , which has undoubtedly dominated research studies in poultry production generally. Many of the anatomical studies in birds have been carried out using the domestic fowl, with comparatively little mention of other genera. Although the genera in the Galliformes have very many similarities, few anatomical differences have been shown to exist. For example, Crowe & Crowe (1979) have found some striking differences between the blood supply to the head and neck of the guineafowl and the domestic fowl.
The guineafowl has become a popular poultry bird in Nigeria, and has been said to rank second to the domestic fowl, despite its seasonal breeding pattern. The further improvement of this genus in our poultry production has necessitated increased studies into its nutritional requirements, management procedures and disease prevalence. In line with this improvement is the need for anatomical data on the female reproductive organs as a basis for the breeding and selection programme. The aim of this study is to describe the macroscopic arterial blood supply to the reproductive organs of the guineafowl.     

Intracerebral Sex Differences in the Vasotocin System in Birds: Possible Implication in Behavioral and Autonomic Functions

The brain vasotocinergic system demonstrates clear sexual dimorphism in birds investigated so far. This paper examines the evidence obtained in studies on gallinaceous (domestic fowl, Japanese quail) and passerine (canary, junco, zebra finch) birds. Vasotocin (VT)-immunoreactive parvocellular neurons are present in the nucleus of stria terminalis of males, but they are less abundant or absent in the corresponding structure of females. A similar difference has been observed in the dorsal paraventricular area of domestic fowl. Sex-related differences in VT-gene expression have been confirmed byin situhybridization. Moreover, overall brain content of VT mRNA in cockerels is about twice that of hens, suggesting that VT synthesis may also be sexually dimorphic in other brain areas where morphological sex differences have not yet been revealed. The vasotocinergic system in birds is implicated in body fluid homeostasis, and during ontogeny it starts to respond to osmotic challenges in a sexually dimorphic way. Photoperiod, aging, or castration—all associated with changes in circulating testosterone levels—affect sexually dimorphic VT pathways and cell clusters. Sexually dimorphic vasotocinergic circuits are distributed in regions containing steroid-concentrating cells and are closely intermingled with aromatase-containing neurons that may mediate activational effects of gonadal steroids on this peptidergic system. However, it remains undetermined whether the observed neuroanatomical sex differences are related to sexually dimorphic autonomic and behavioral effects induced by VT. Most likely, VT in birds has a modulatory rather than a specific regulatory function in control of male sexual behavior and vocalization.     

Population expansion of the poultry fluff louse, Goniocotes gallinae (De Geer, 1778) (Ischnocera, Phthiraptera)

A study was undertaken to determine the rate of population expansion of an ischnoceran Phthiraptera, Goniocotes gallinae (infesting the domestic fowl, Gallus gallus domesticus), on the basis of in vitro and in vivo studies. Data obtained from in vitro rearing were utilized to construct the life history of the louse. The values of the gross reproductive rate (12.49 female eggs/female), net reproductive rate (8.31 female eggs/female), mean length of generation (36.91 days), precise generation time (35.65 days), finite rate of increase (1.06 female/days), and intrinsic rate of natural increase (0.059) of G. gallinae were determined. However, doubling-time values recorded during in vivo studies (14 days) was comparatively higher than those during in vitro rearing (11.73 days).     

Resource allocation across the egg laying sequence in the wild zebra finch Taeniopygia guttata

Zebra finches have played a central role in the discovery of a variety of maternal effects over the past decade, with females shown to adjust resource allocation to their eggs in response to variables such as the appearance of their partner, their own condition, and the diet on which they are maintained. In addition to being the focus of some of the most high profile individual studies that have influenced maternal effects research in birds, the multitude of zebra finch studies together provide the most comprehensive set of data to illuminate general patterns and compare different maternally derived variables. Surprisingly, to date, virtually all of this work has focused on captive populations of the zebra finch that have been domesticated for many generations, and which are typically held under relatively constant environmental and dietary conditions. Here we report the first data on resource allocation across the egg laying sequence in a free‐living wild population. Reassuringly we find that the patterns that have been found in the majority of studies of domesticated populations with respect to investment across the laying sequence were all present in the wild population. The size and mass of eggs increased through the laying sequence whilst the concentration of carotenoids significantly decreased across the laying sequence. Although there was no significant pattern with respect to testosterone across the laying sequence the first two eggs had a higher level of testosterone than the last few eggs in the clutch, which is also consistent with the findings of earlier studies in captive populations.     

Three-dimensional motion of avian spermatozoa

Observations have been made on spermatozoa from the domestic fowl, quail and pigeon (non-passerine birds) and also from the starling and zebra finch (passerine birds). In free motion, all these spermatozoa roll (spin) continuously about the progression axis, whether or not they are close to a plane surface. Furthermore, the direction of roll is consistently clockwise (as seen from ahead). The flagellar wave has been shown to be helical and dextral (as predicted) for domestic fowl sperm when they swim rapidly in low viscosity salines. Calculations have shown that their forward velocity is consistent with their induced angular velocity but that the size of the sperm head is suboptimal for progression speed under these conditions. Dextrally helical waves also occur on the distal flagellum of fowl, quail and pigeon sperm in high viscosity solutions. But in other cases, the mechanism of torque-generation is more problematical. The problem is most profound for passerine sperm, in that typically these cells spin rapidly while seeming to remain virtually straight. Because there is no evidence for a helical wave on these flagella, we have considered other possible means whereby rotation about the local flagellar axis (self-spin) might be achieved. Sometimes, passerine sperm, while maintaining their spinning motion, adopt a fixed curvature; this must be an instance of bend-transfer circumferentially around the axonemal cylinder-though the mechanism is obscure. It is suggested that the self-spin phenomenon may be occurring in non-passerine sperm that in some circumstances spin persistently, yet without expressing regular helical waves. More complex waves are apparent in non-passerine sperm swimming in high viscosity solutions: added to the small scale bends is a large scale, sinistrally helical curvature of the flagellum. It is argued that the flagellum follows this sinistrally helical path (i.e. "screws" though the fluid) because of the shape of the sperm head and the angle at which the flagellum is inserted into it. These conclusions concerning avian sperm motility are thought to have relevance to other animal groups. Also reported are relevant aspects of flagellar ultrastructure for pigeon and starling sperm.     

Oxygen uptake in fowl eggs incubated in air and pure oxygen

1. The oxygen uptake of small (bantam hen) and large (white leghorn) domestic fowl eggs was measured during incubation in air and pure O2. 2. A 2 hr exposure to pure O2 increased the O2-uptake from the 9th day in the large eggs and from the 15th day in the small eggs. 3. On the last 4 days of the incubation period, both small and large eggs increased their O2-uptake by about 22% when transferred from air to pure O2. 4. When 1 cm2 of the eggshell above the air cell was removed, the O2-uptake of the white leghorn eggs incubated in air increased by 13%.     

Water loss from eggs of domestic fowl and calcium status of hatchlings

Water balance in eggs of domestic fowl was manipulated by drilling holes (each 3 mm in diameter) through the calcareous layer into the air cell on day 10 of incubation. Water loss between days 0 and 18 averaged 6 g for eggs in the control group (no hole) but increased to 8, 12, and 15 g for eggs with 1, 2, or 3 holes, respectively. Hatching success was 79–87% for eggs with 0–2 holes through the eggshell but only 43% for eggs with three holes. Live mass of hatchlings declined as the number of holes drilled in the eggshell increased, but dry mass of carcasses was unaffected by the treatments. The quantity of Ca²⁺, Mg²⁺, and phosphorus in residual yolks and yolk-free carcasses of hatchlings was not influenced by the amount of water lost from eggs. Plasma Ca²⁺ and Mg²⁺ were elevated in hatchlings from eggs with high rates of water loss. The inability to regulate plasma Ca²⁺ may be a negative consequence of excessive water loss and could contribute to increased mortality of embryos.     

Zona-induced acrosome reaction of hamster spermatozoa

It is well established that the zonae pellucidae of mature unfertilized eggs have the ability to induce the acrosome reaction of capacitated spermatozoa. To determine if this capacity of the zona is species-specific, hamster spermatozoa were allowed to attach to the zonae of homologous and heterologous eggs and examined for the acrosome reaction. The zonae of eggs from six different species were tested and the zona of hamster egg was found to have the strongest capacity to induce the acrosome reaction of hamster spermatozoa, followed by human and rat zonae. The zonae and mouse, guinea pig, and domestic fowl eggs were incapable of inducing the acrosome reaction of hamster spermatozoa. The acrosome reaction-inducing ability of the hamster zona was found to increase during maturation in the ovary. The zona of mature unfertilized hamster eggs maintained their acrosome reaction-inducing ability even after aldehyde fixation or storage in a highly concentrated solution of ammonium sulfate.     

Determination of monensin, salinomycin and narasin in muscle, liver and eggs from domestic fowl using liquid chromatography-electrospray mass spectrometry

A method is presented for the simultaneous determination of the three ionophores, monensin, salinomycin and narasin in muscle, liver and eggs from domestic fowl. Samples are homogenised in methanol and any ionophores present are extracted into toluene-hexane. The extracts are concentrated and injected into a bench-top electrospray LC-MS system. Chromatography is carried out using an end-capped reversed-phase column, with a mobile phase consisting a mixture of acetonitrile, methanol, tetrahydrofuran, trifluoroacetic acid and water. Using single-ion monitoring, the ionophores can be detected down to the 1 ng/g level.     

Sexual differentiation of the zebra finch song system parallels genetic, not gonadal, sex.

Mechanisms regulating sexual differentiation of the zebra finch song system present an intriguing puzzle. Masculine development of brain regions and behavior can be induced in genetic females by posthatching estradiol treatment. That result is consistent with the hypothesis that estradiol, converted within the brain from testicular androgen via the aromatase enzyme, masculinizes neural structure and function. In contrast, treatment during specific stages of development with the aromatase inhibitor Fadrozole has not prevented masculine development, and the presence of testicular tissue in genetic females did not induce masculine organization of neuroanatomy or singing behavior. Fadrozole treatments in those previous studies were limited, however, and most genetic females had both ovarian and testicular tissue. The present experiments were designed to provide increased aromatase inhibition and to reliably produce genetic females with only testicular tissue. Eggs received a single injection at a later age or with higher doses of Fadrozole than had been used previously. Some embryos were exposed to Fadrozole more frequently by either injecting eggs on 2 days of development or dipping them for 10-12 days in Fadrozole. Finally, in some individuals from Fadrozole-treated eggs, the left gonad was removed, leaving each genetic male and female with a single right testis. None of these treatments significantly affected development of the song system compared to appropriate control groups. These results suggest that sexual differentiation of the zebra finch song system is not regulated by embryonic aromatase activity or by gonadal secretions and instead involves events that need not be mediated by steroid hormones.     

Structural and physiological differences between montane and lowland avian eggs and embryos

A few avian species breed at altitudes up to 6500 m. Embryos in eggs laid at high altitudes are confronted with the problem that gases diffuse more rapidly at low barometric pressure than at sea level. Data on birds breeding up to 4500 m indicate that modifications in eggshell structure and embryonic physiology foster successful development in these groups. At moderate altitudes (up to 3600 m), shell conductance to gases (corrected to 760 torr) is decreased in approximate proportion to the reduction in barometric pressure, thus offsetting the increased tendency of gases to diffuse. At altitudes above 4000 m, the conductance is increased above levels at moderate altitudes, thus fostering improvement in oxygen availability, while increasing rates of water and CO2 losses. Above 4000 m, embryonic physiological properties become increasingly important for coping with hypoxic, hypocapnic, and dehydrated conditions inside the shell. Nothing is known about characteristics of eggshells and embryos in eggs laid between 4500 and 6500 m. Despite years of artificial selection, domestic fowl do not breed successfully much above 3000 m. Embryos of domestic fowl appear highly sensitive to the effects of hypoxia.     

Molecular genetics of avian proteins. XL Egg proteins of Gallus gallus, G. sonnerati and hybrids

By a variety of electrophoretic procedures it has been found that Gallus gallus , whether as jungle fowl or as chickens, differs consistently from Sonnerat's jungle fowl, G. sonnerati , in three egg white proteins: G₃ ovoglobulin, G₂ ovoglobulin and glutamyl peptidase. The electrophoretic patterns of egg white hybrids between G. gallus and G. sonnerati are identical to those obtained after simpiy mixing the egg whites; there is no evidence for hybrid protein zones. Although Japanese data on serum amylase can be interpreted as indicating introgressive hybridization in the origin of the domestic fowl, so far the egg protein data are negative in this respect.     

The magnum-isthmus junction of the fowl oviduct participates in the formation of the avian-type shell membrane

Avian eggs possess a shell membrane in the shape of an asymmetrical ellipsoid and with a limiting membrane that is a smooth layer of homogeneous, dense materials. We describe the role of the magnum-isthmus junction (MIJ) of the oviduct in the formation of the avian-type shell membrane in the domestic fowl Gallus domesticus. The narrow width of the lumen at the MIJ indirectly participates in the determination of the asymmetrical ellipsoid shape of eggs that are encased by the egg-white layer and subsequently by the peri-albumen layer (PL) and the shell membrane. The PL reacts with Alcian blue and exists between the egg white and the limiting membrane. It is added to the ovulating egg at the MIJ and covers the outermost surface of the egg-white layer. The function of the PL is to provide a smooth surface by covering the irregular surface of the egg-white layer. The materials of the PL consist of an Alcian blue-positive polysaccharide (or glycoprotein) of 240 kDa and five proteins of 135, 116, 72, 49, and 46 kDa. The isolated materials have an affinity to bind with the egg-white mass. An antiserum against quail PL materials stains the domestic fowl PL and secretory cells of the luminal epithelium at the MIJ, and cross-reacts with the molecules of 240, 135, and 116 kDa.     

Natural foci of ornithosis in Kazakhstan]

An investigation (clinical, immunological and epidemiological) of the workers of various fowl-farms was carried out in various regions of Kazakhstan for a number for years; the sera of domestic fowl and birds was studied as well. On the basis of examination of 6264 persons the greatest percentage of ornithosis was revealed among the workers of the slaughter house and the shop for the processing of the fowl. Examination of 4749 sera of bird blood showed that the highest incidence of ornithosis was among doves and ducks (43.2 and 11.6%), and the least--in chickens (5.3%); there proved to be a direct relationship between the infection of man and birds. Thus, it was shown that domestic fowl and also doves served as the source of ornithosis infection; foci of ornithosis were revealed in a number of Kazakhstan regions as a result of these investigations.