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Muscarinic cholinergic receptors in the songbird and quail brain: a quantitative autoradiographic study.

In order to clarify the neuroanatomical basis for postulated muscarinic cholinergic control of a wide array of physiological processes in birds, the distribution of muscarinic cholinergic receptors in the brain of three avian species was investigated by quantitative autoradiography. The species consisted of two passerines (songbirds), the European starling (Sturnus vulgaris) and the song sparrow (Melospiza melodia), and one galliform, the Japanese quail (Coturnix coturnix japonica). [3H]N-methyl scopolamine (NMS), a muscarinic cholinergic antagonist was used as the ligand to label the receptors. Initial experiments demonstrated that the binding of this ligand in the three species is saturable in the nanomolar range and has a high affinity (Kd = +/- 0.6 nM). Displacement experiments revealed that three muscarinic ligands competed in an order of potency characteristic of the mammalian muscarinic receptor (i.e., atropine greater than oxotremorine greater than carbachol) for NMS binding in the avian brain. In all three species, portions of the basal ganglia, such as the parolfactory lobe and the paleostriatum augmentatum, exhibited the highest density of binding. On the other hand, the paleostriatum primitivum, the avian homologue of the mammalian globus pallidus, contained very few binding sites. Other telencephalic sites, such as the ventral and dorsal hyperstriatum, also revealed relatively high receptor density. However, the neostriatum and especially the ectostriatum showed much lower levels. In the hypothalamus, in all three species, specific binding could be observed in the ventromedial nucleus and adjacent areas. The paraventricular nucleus also showed moderate levels of binding density, especially in the two songbird taxa. At a more rostral level, the preoptic area showed low levels of binding. In the quail, the sexually dimorphic nucleus of the preoptic area was clearly outlined in the autoradiograms by the low level of binding sites compared to the surrounding areas. In the two passerine species, nuclei of the song system were identified by either high or low levels of NMS binding. High binding defined area X and the mesencephalic nucleus, intercollicularis (ICo). In contrast, the robust nucleus of the archistriatum and the magnocellular nucleus of the anterior neostriatum showed low levels of binding in comparison with the surrounding tissue. None of these nuclei were visible in the quail autoradiograms except for ICo, which appeared as in the passerines as a heavily labelled area surrounding the lightly labelled nucleus mesencephalicus lateralis pars dorsalis. In all three species, the hippocampal complex was devoid of NMS binding except for two lateral dark bands that were present along the entire rostral to caudal extent of the hippocampus.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Estrogen receptors in the avian brain: survey reveals general distribution and forebrain areas unique to songbirds.

Estrogens play an important role in the control and differentiation of species-typical behavior and in endocrine homeostasis of birds, but the distribution and evolution of cells that contain estrogen receptors in the avian brain are poorly understood. This study therefore surveys 26 species in the avian orders Anseriformes (1 species), Galliformes (2), Columbiformes (3), Psittaciformes (1), Apodiformes (2), and Passeriformes (3 suboscines, 14 oscines). Indirect immunocytochemistry with the estrogen receptor (ER) antibody H222Spy revealed a general pattern of ER-antibody-immunoreactive cells (ER-IRC) in all 26 species, with ER-IRC in consistent, well-defined locations in the limbic forebrain, the midbrain striatum, the hippocampus, the hindbrain, and especially in the preoptic area and the tuberal hypothalamus. For some species, the microdistribution of ER-IRC in some of these general areas differed, such as in the hippocampus and the anterior hypothalamus of suboscine species and in the preoptic area of the Japanese quail. Brains of oscine songbirds of both sexes, unlike brains of nonsongbirds, had ER-IRC in three specific structures of the nonlimbic forebrain: in the area surrounding the nucleus robustus archistriatalis; in the rostral forebrain; and, for all individuals, in the caudale neostriatum, including the nucleus hyperstriatalis ventrale, pars caudale (HVc). Among songbird families or subfamilies, adult males of the Estrildinae had much lower numbers of ER-IRC in HVc than did adult males of the Fringillidae, Paridae, Sturnidae, and Ploceinae. Differences occurred, too, among closely related species: the songbird canary (Serinus canaria) had an ER-IRC area in the rostral forebrain that was lacking in all other songbird species, including other cardueline finches. The cells with ER that are found only in the songbird forebrain but not in reptiles, nonpasserine birds, and nonoscine passerine birds very likely coevolved with steroid-dependent differentiation of vocal control areas. The songbird-specific expression of ER in the forebrain could be an example in which taxon-specific behavior is due to taxon specific neurochemical properties of the brain.

Age Factors↗

Expression of an onco-developmental antegen among avian species.

Rabbit antisera directed against an onco-developmental antigen on chicken red blood cells have been serologically dissected through specific adsorptions. It is now possible to detect 13 antigenic determinants with the fractionated antisera. The onco-developmental antigen referred to as chicken fetal-leukemic antigen (CFA) is fetal-specific in the white Leghorn chicken, being present on the embryonic but not adult peripheral red blood cells of non-being present on the embryonic but not adult peripheral red blood cells of non-leukemic birds. However, one or more of the onco-developmental antigenic determinants have been detected on adult peripheral red blood cells of non-Gallus avian species, as well as on red blood cells from two adult chicken varieties. For phylogenetic purposes, red blood cells from avian species were characterized for their combinations of CFA determinants. Comparisons among species revealed specific patterns of antigenic expression within phylogenetic groups. Several CFA determinants were restricted in their occurrence to species within a single family, and one determinant was found in all cases where CFA was expressed. The distribution of CFA determinants was used to determine immunological distances among four Galliform species. These distances agreed with the immunological relationships established using different serological markers.

Animals↗

Specificity of sperm-binding Wolffian duct proteins in the rooster and their persistence on spermatozoa in the female host glands.

Unlike those of mammals, chicken spermatozoa can develop their fertilizing ability before they leave the testis to pass into the Wolffian duct; moreover, chicken spermatozoa do not require a period of capacitation in the female tract. A question arises, therefore, as to the significance of secretory proteins shown to bind to the surface of chicken spermatozoa as they pass into and through the Wolffian duct. Using anti-Wolffian duct fluid IgG as a probe visualized by immunoperoxidase staining, the present investigation confirms that testicular spermatozoa of quail and turkey as well as chicken do not have any surface determinants in common with those present in Wolffian duct secretions. By contrast, those in the lower portion of the vas deferens display a strong reaction with anti-Wolffian fluid IgG over their entire surface, and immunoprecipitation studies suggest that this reflects the binding of four Wolffian duct proteins. Since a reaction to the antichicken fluid IgG is shown also by mature quail and turkey, but not duck and pigeon spermatozoa, the Wolffian components that coat spermatozoa in birds appear to have a specificity confined to the same order, in this case the Galliformes. Following vaginal or intramagnal insemination, spermatozoa present 48 hours later in the uterovaginal host glands and infundibulum glands, respectively, still reacted strongly. This finding that Wolffian duct components persist on the surface of spermatozoa in the female tract is consistent with the possibility that they have some role in sperm storage or survival in female birds.

Animals↗

The avian ceca: obligate combustion chambers or facultative afterburners? The conditioning influence of diet.

Changes in the anatomy and microbiology of the gut of many galliforme birds are influenced by the amount of crude fiber in the diet; the response is biphasic. As crude fiber in the diet increases, the bird compensates for the dilution of dietary energy content by increasing the amount it eats, which causes an increase in gizzard size. After exceeding a threshold in fiber content, where the metabolizable energy intake is limited by the sheer bulk of the food consumed relative to the ability of the bird to ingest food, there is an increase in hindgut size and a change in the microflora that allows extraction of energy from cellulose through the production of volatile fatty acids (VFAs). The result is a change in the apparent digestibility of the diet. Altering the energy demands through the application of environmental stressors, such as lowered temperature, may alter a hypothalamic or other regulatory mechanism setpoint at which hindgut hypertrophy occurs. The time frame required for such changes is between 4 and 8 weeks. Studies to determine digestibility of nutrients or the response of the gut of birds to altered diets must allow a sufficient period of time for gut adaptation prior to assessing its function. Other factors, such as age, photoperiod, and environmental factors, may affect the response of the gut.

Adaptation, Physiological↗

Water and electrolyte transport by the avian ceca.

In galliform birds with well developed ceca, these are functionally important components of the lower intestinal complex, which is capable as a whole of substantial, hormone-regulated homeostatic modification of mixed urine and intestinal luminal fluid before final excretion. Active Na+ transport drives Na-linked absorption of water and Cl- and secretion of K+; there are also lesser Na-independent components of the latter fluxes. These transport processes are appropriately enhanced by dehydration, Na depletion, or exogenous aldosterone.

Analysis of Variance↗

Dietary protein restriction stress in the domestic chicken (Gallus gallus domesticus) induces remodeling of adrenal steroidogenic tissue that supports hyperfunction.

The stress of dietary protein restriction in the immature domestic turkey (Meleagris gallopavo) induces adrenal steroidogenic hypofunction that is associated with an alteration in the proportion of density-dependent subpopulations of steroidogenic cells within the adrenal gland. In contrast, when imposed on immature chickens, this nutritional stressor induces long-term enhancement of adrenal steroidogenic function. However, whether this alteration in function is accompanied by a remodeling of chicken adrenal steroidogenic tissue as in the turkey is not known. To address this question, immature cockerels (2 weeks old) were fed established isocaloric synthetic diets containing either 20% (control) or 8% (restriction) soy protein for 4 weeks. Adrenal glands were processed for the isolation of defined, density-separable, adrenal steroidogenic cell subpopulations: three low-density adrenal steroidogenic cell subpopulations [LDAC-1 (rho = 1.0285-1.0430 g/ml), LDAC-2 (rho = 1. 0430-1.0485 g/ml), and LDAC-3 (rho = 1.0485-1.0500 g/ml)] and one high-density subpopulation [HDAC (rho = 1.0510-1.0840 g/ml)]. The steroidogenic function of these cell subpopulations was assessed. Protein restriction consistently, but differentially, enhanced maximal ACTH-induced corticosterone production by the subpopulations: values of LDAC-1, -2, and -3 and HDAC from protein restricted birds were, respectively, 116, 43, 33, and 20% greater than those of corresponding cell subpopulations from control birds. However, it had contrasting influences on maximal ACTH-induced aldosterone production by the cell subpopulations. Whereas the value of LDAC-1 from protein-restricted birds was 70% greater than that from control birds, the values for LDAC-2 and -3 were not different from those of the control, and the value for HDAC was 22% less than that of the control. Protein restriction also altered the cell subpopulation composition of the adrenal gland: compared to control, it increased the proportion of LDAC-1 by 46% and decreased the proportion of LDAC-3 and HDAC by 34 and 20%, respectively. Thus, dietary protein restriction increased the proportion of cells (i.e., LDAC-1) having the greatest enhancement in corticosteroid production. This pattern of remodeling of chicken adrenal steroidogenic tissue in response to dietary protein restriction contrasts sharply with the pattern that occurs in another galliform species, the domestic turkey.

Adrenal Cortex Hormones↗

Castration in Gambel's and Scaled Quail: ornate plumage and dominance persist, but courtship and threat behaviors do not.

During the breeding season, testosterone in male birds is often linked to some secondary sexual ornaments, courtship behaviors, and intrasexual aggression. I examined the effect of castration on plumage expression in Gambel's Quail (Callipepla gambelii), a species in which males are highly ornate, and in Scaled Quail (C. squamata), an unornamented species. Using male pairs, each consisting of a castrate and a control, I also assessed whether castration affected (1) the behavior of males, (2) the mating decisions of females, or (3) the outcome of male-male competition. Castration did not alter the plumage of male Gambel's or Scaled Quail. In these species, and some other members of the avian order Galliformes, production of ornate plumage appears to be independent of testosterone. In contrast, castration reduced or eliminated courtship behaviors. Females almost never preferred castrated individuals. During male-male competition, castrates also exhibited lower rates of threat behaviors, which appear to be identical to those used during courtship. Castration did not, however, influence the outcome of male-male competition. Castrates of both species exhibited overt aggression (pecks, chases, displacement) and frequently won male contests. Such results suggest that certain types of aggressive behavior may be testosterone-independent. In both Gambel's and Scaled Quail, male body size correlated positively with dominant individuals.

Agonistic Behavior↗

A molecular phylogeny of the pheasants and partridges suggests that these lineages are not monophyletic.

Cytochrome b and D-loop nucleotide sequences were used to study patterns of molecular evolution and phylogenetic relationships between the pheasants and the partridges, which are thought to form two closely related monophyletic galliform lineages. Our analyses used 34 complete cytochrome b and 22 partial D-loop sequences from the hypervariable domain I of the D-loop, representing 20 pheasant species (15 genera) and 12 partridge species (5 genera). We performed parsimony, maximum likelihood, and distance analyses to resolve these phylogenetic relationships. In this data set, transversion analyses gave results similar to those of global analyses. All of our molecular phylogenetic analyses indicated that the pheasants and partridges arose through a rapid radiation, making it difficult to establish higher level relationships. However, we were able to establish six major lineages containing pheasant and partridge taxa, including one lineage containing both pheasants and partridges (Gallus, Bambusicola and Francolinus). This result, supported by maximum likelihood tests, indicated that the pheasants and partridges do not form independent monophyletic lineages.

Animals↗

Historical biogeography and a mitochondrial DNA phylogeny of grouse and ptarmigan.

We sequenced 2690 nucleotides of mitochondrial DNA (mtDNA) including the entire control region (CR), partial 12S and 16S ribosomal RNAs, NADH dehydrogenase subunit 2, and cytochrome b genes from representatives of all the 17 living species of grouse and ptarmigan (Aves; Galliformes; subfamily Tetraoninae). Substitution rates and phylogenetic signals were variable among genes, with the CR being more informative than protein-coding and rRNA genes. Phylogenetic trees, computed with the CR or the concatenated sequences, indicate that: (1) genus Bonasa is monophyletic and basal within the subfamily, (2) all the other currently recognized genera of Tetraoninae are monophyletic, except Dendragapus; (3) D. obscurus is related to Centrocercus urophasianus and divergent from former D. canadensis and D. falcipennis, which, accordingly, may be ascribed to the distinct genus Falcipennis; (4) Tympanuchus, Dendragapus, and Centrocercus form a clade comprising taxa distributed exclusively in North America; and (5) the North American species of Bonasa (B. umbellus) and Lagopus (L. leucurus) are basal to their Eurasian and Holarctic congeneric species. These findings, and a dispersal-vicariance analysis, support a North American origin of the subfamily and of all the genera of Tetraoninae, with the possible exception of Tetrao. Present species distributions might have been attained by at least three dispersal events from North America to Eurasia, involving the ancestors to Palearctic Bonasa, the ancestors to circumpolar Lagopus mutus/L. lagopus, and the clade leading to Tetrao/Falcipennis. According to a "standard calibration" of the mtDNA molecular clock (2% sequence divergence per million years), Bonasa split about 5-6 million years ago, the other genera diverged during the upper Pliocene, and most of the congeneric species with North American and Eurasian distributions (Bonasa, Lagopus, and Falcipennis) originated during the lower Pleistocene, well before the last interruption of the Beringian land bridge.

Animals↗

Adaptive evolution of lysozyme: changes in amino acid sequence, regulation of expression and gene number.

Adaptive evolution of lysozyme has involved remodelling of amino acid sequences and changes in patterns of gene expression and in gene number. Following an outline of the phenomena likely to be indicative of adaptive evolution and how one can assess them, this chapter focuses on four cases in which lysozyme c has been recruited as a digestive enzyme in the stomachs of creatures needing to retrieve nutrients from microorganisms in fermented food. For each case-ruminant artiodactyls, leaf-eating monkeys, a leaf-eating bird, and fruit flies-the factors likely to be of primary importance in lysozyme's adaptation are examined. Additional examples of apparent adaptation for digestion or antimicrobial defense in animals as diverse as mice, moths, and molluscs are summarized. This chapter considers also the case of three internally clustered residues which among galliform bird lysozymes c occur either as Thr 40, Ile 55, and Ser 91 (TIS) or as Ser 40, Val 55, and Thr 91 (SVT). Reconstruction and testing of six possible intermediate proteins and development of the concept of a neutral corridor of protein traits are described.

Adaptation, Physiological↗

Regressive post-hatching development of acetylcholinesterase-positive neurons in the pineal organs of Coturnix coturnix japonica and Gallus gallus.

Distribution and number of acetylcholinesterase-positive neurons were studied in the Japanese quail and the domestic fowl during the post-hatching period by means of the acetylcholinesterase method. For comparison, the development of the catecholamine-containing (sympathetic) pinealopetal fibers of the domestic fowl was demonstrated with the use of the glyoxylic acid method. The number of acetylcholinesterase-positive ganglion cells in the pineal organs of both avian species decreased rapidly after hatching, with a concentration of these elements in the basal portion (stalk) of the pineal organ. In 3-day-old chickens, perivascular catecholamine-containing nerve fibers penetrate the antero-lateral walls of the pineal organ and are found exclusively in the interfollicular and perivascular tissues. In 13-day-old and adult fowl, these fibers increase in number and terminate not only in the interfollicular space but also in the neuroepithelial parenchyma of the pineal body. The ontogenetic regression of the sensory structures paralleled by an expanding sympathetic innervation in the pineal organ of a galliform species resembles somewhat the process of phylogenetic transformation leading from pineal sense organs to pineal glands.

Acetylcholinesterase↗

Innervation of the avian pineal organ. A comparative study.

The innervation of the pineal organ was studied in 26 avian species under particular consideration of comparative aspects. A population of nerve cells and their pinealofugal (afferent) fiber systems were stained by means of the acetylcholinesterase method, while catecholamine-containing pinealopetal (efferent) fibers were demonstrated with the use of the glyoxylic acid method. Afferent axons were mainly found in the postero-proximal portion of the organ, and the patterns of their distribution were classified into three groups according to the characteristic densities of the reaction product. The number of acetylcholinesterase-positive neurons in the avian pineal organs examined in this study varied extremely from species to species, ranging from 0 to 362. Catecholamine-containing nerve fibers penetrating the antero-lateral walls of the pineal follicles accompanied blood vessels and were arranged more densely in the distal portion of the organ, in contrast to the distribution of the acetylcholinesterase-positive nerve fibers. Three-dimensional reconstruction of the distributional patterns of both types of neural projections was performed for the pineal organ of every avian species examined. In avian species possessing a relatively conspicuous afferent projection, such as Passeriformes, Nycticorax, and Milvus, terminals of catecholamine-containing nerve fibers were observed exclusively in the interfollicular and perivascular tissues. In Galliformes, which display only few pineal afferents, catecholamine-containing fibers terminate not only in the interfollicular space, but also in the neuroepithelial parenchyma. The regional differences in the innervation in the avian pineal organ suggest that the pinealocytes ranging from more sensory-like to more secretory-like elements are arranged in a mosaic-like pattern.

Acetylcholinesterase↗

Chromosome banding pattern conservatism in birds and nonhomology of chromosome banding patterns between birds, turtles, snakes and amphibians.

The G-banded karyotypes of 4 species of birds representing the orders Galliformes, Columbiformes and Musophagiformes were compared. Banding pattern homology between orders was limited t 5o 5 major chromosome arms and the Z chromosome. Even in these major chromosome arms pericentric and paracentric inversions produced alteration of the banding pattern sequences. Addition of constitutive heterochromatin was responsible for changes in banding patterns in the Z chromosome. The chromosome banding patterns of an emydid turtle, Terrepene carolina, 5 species of boid snakes of the genera Liasis, Acrantophis, and Sanzinia and the African clawed-frog. Xenopus muelleri, were also compared to the bird chromosome banding patterns. No homology was observed between any of these major groups: bird, snake, turtle, amphibian. However, intergroup homology was apparent. - The data obtained do not support reports of broad interordinal direct homology of the macrochromosomes of birds and refutes the idea of a primitive bird karyotype with 3 pairs of "Agroup' chromosomes and 3 pairs of "B group' chromosomes. - The major mechanisms responsible for chromosome evolution in birds appear to be centric and tandem fusions, paracentric and pericentric inversions, and addition or deletion of heterochromatin.

Animals↗

Demonstration of W chromosome-specific repetitive DNA sequences in the domestic fowl, Gallus g. domesticus.

Evidence is presented to demonstrate the presence of W chromosome-specific repetitive DNA sequences in the female White Leghorn chicken, Gallus g. domesticus, based on two different experimental approaches. First, 3H-labelled, female chicken DNA was hybridized with excess, unlabelled, mercurated, male DNA, and unhybridized single-stranded 3H-DNA (3H-SHU-DNA) was recovered by SH-Sepharose and hydroxyapatite column chromatography. Approximately 24% of the hybridizable 3H-SHU-DNA was female-specific and localized on the W chromosome. The second approach was to examine female-specific DNA fragments among the digests of chicken DNA with various restriction endonucleases. Among them, we found that digestion with XhoI produced two prominent female-specific brands of 0.60 kb (= kilobase pairs) and 1.1 kb. The 0.60 kb fragment was isolated and 3H-labelled by nick-translation. Female-specificity of the 3H-XhoI--0.60 kb DNA was judged to be at least 95% under the conditions of hybridization with membrane filter-bound DNA. Presence of amplified XhoI--0.60 kb DNA on the W chromosome seems to be limited to different lines of G.g. domesticus and no such repeat was detected in three species belonging to other genera in the order Galliformes and in three species belonging to other avian orders.

Animals↗

Occupancy of the majority of DNA in the chicken W chromosome by bent-repetitive sequences.

Another family of repetitive sequences, designated the EcoRI family, was found in the DNA of the chicken W chromosome by hybridization with the W chromosome-specific XhoI family probe under conditions of low stringency. A 1.2 kb EcoRI fragment, the major repeating unit of the family, was cloned and sequenced. The 1.2 kb unit showed an overall sequence similarity of about 68% to the 0.7 kb XhoI family repeating unit and it consisted of tandem repeats of average length 21 bp, most of which contained (A)3-5 and (T)3-4 clusters separated by 6-8 G + C-rich sequences. These features and its behavior as a strongly bent molecule in solution were very similar to those found for other W chromosome-specific repetitive sequences in the order Galliformes: XhoI family of chicken, PstI family of turkey and TaqI family of pheasant. The cloned 1.2 kb unit contained 78 CpG dinucleotide sequences and those that were in HapII, HhaI and BstUI sites were shown to be extensively methylated in the genomic DNA. Repetition frequencies of the 1.2 kb unit among the female population of chicken fell into high- and low-level classes, which accounted for about 30% and 10%, respectively, of the DNA in the W chromosome. Thus, 70% to 90% of the DNA in the chicken W chromosome was shown to be occupied by bent-repetitive sequences. The EcoRI and XhoI family sequences were not intermingled over the short range but each family formed a unique domain ranging from one to several million base pairs.

Animals↗

Intragenomic DNA sequence homologies in the chicken and other members of the class Aves: DNA re-association under reduced stringency conditions.

We have investigated the intragenomic DNA sequence homologies of twelve species of birds representing five orders, and emphasizing Galliformes. This study differs in two important ways from the classical approaches taken in constructing and evaluating phylogenies based on DNA sequence similarities. Comparisons are made on the basis of sequence homologies within genomes of related birds, rather than between genomes. DNA is reassociated at 50 degrees C in 0.5M phosphate buffer; these conditions allow formation and detection of duplexes containing more mismatch than would normally be permitted using more stringent conditions, affording an opportunity to observe more ancient sequence homologies. Thermal stability profiles of DNA duplexes formed under these conditions are the basis of comparison; three general patterns were observed. This approach emphasizes differences in sequence composition between genomes while the more traditional method of intergenomic tracer DNA hybridization at higher stringency emphasizes sequence similarities. No correlation was found between taxonomic position and intragenomic sequence composition, either within or between lineages. The thermal stability profiles of DNA duplexes formed within avian genomes did not reflect the biological similarities inferred from morphology, karyotype, and studies of interspecific hybridization. While all of the differences observed could have occurred over geological time, it was surprising that the genomes of the domestic chicken and the Red Jungle Fowl (Gallus gallus) differ in their sequence compositions. It appears that amolification/reduction events and/or positional changes occur rather often during evolution of a lineage.

Animals↗

Characterization of DNA sequences constituting the terminal heterochromatin of the chicken Z chromosome.

Two clones, pCZTH5-8 and pCZTH12-8, were isolated from a female chicken genomic library by screening with sequences obtained from genomic libraries which had been constructed from a terminal region of a single Z chromosome of chicken utilizing laser microbeam irradiation and PCR amplification. Fluorescence in situ hybridization to the mitotic Z chromosome and the lampbrush ZW bivalent of chicken demonstrated that both the cloned sequences are located in the heterochromatic region of the Z chromosome at the end opposite to the pairing region with the W chromosome. The sequences pCZTH-8 and pCZTH12-8 are distributed widely on both the telomeric bow-like loops (TBL) and the region I (short loops region) of the Z lampbrush chromosome. These clones, pCZTH12-8 particularly notably, hybridized also to the TBLs of lampbrush bivalents 1-4 of chicken. Both sequences are transcribed in the lampbrush stage oocytes on the Z chromosome and on other macrobivalents. The subfragment of pCZTH5-8 which hybridizes to the TBLs and the insert of pCZTH12-8 contain regions that are closely similar in sequence. The pCZTH-8 sequence has no internal repeats and may be part of the 24-kb macrosatellite repeating unit that is evident after Nhel digestion of the genomic DNA. A cloned 24-kb unit, pFN-1, does not show significant DNA curvature, but cytosines of its CpG dinucleotides may be highly methylated in vivo. This contrasts with the repeat sequences of the W heterochromatin which not only have highly methylated CpG but are also strongly curved. The 24-kb unit is repeated about 830 times in the diploid genome of a female chicken, suggesting that nearly the entire terminal heterochromatin on the Z chromosome consists of this macrosatellite family. Sequences of the greater part of the pCZTH-8 are restricted to the genus Gallus but the sequence of one subregion which hybridizes to TBLs is present in the genomes of the order Galliformes.

Animals↗