Immunohistochemical localization of vasoactive intestinal polypeptide(VIP)-containing neurons in the hypothalamus of the Japanese quail,Coturnix coturnix

Summary The localization of vasoactive intestinal polypeptide (VIP) in the hypothalamus of the quail has been studied by means of light- and electron-microscopic immunohistochemistry. Numerous VIP-immunoreactive perikarya are distributed in the caudal portion of the nucleus infundibularis (n. tuberis) and nucleus mamillaris lateralis, and sparse in the preoptic area, nucleus supraopticus and nucleus paraventricularis. Dense localization of immunoreactive-VIP fibers is observed in the external layer of the median eminence, in close contact with the primary portal capillaries. The main origins of these fiber terminals are VIP-immunoreactive perikarya of the nucleus infundibularis. These neurons are spindle or bipolar and extend one process to the ventricular surface and another to the external layer of median eminence. They are CSF-contacting neurons and apparently constitute the tubero-hypophysial tract that links the third ventricle and the hypophysial portal circulation. VIP-reactive neurons in the nucleus mamillaris lateralis also project axons to the external layer of the median eminence, constituting the posterior bundle of the tuberohypophysial tract. Numerous VIP-immunoreactive perikarya occur also in the nucleus accumbens/pars posterior close to the lateral ventricle. They are also CSF-contacting neurons extending a process to the lateral ventricle. There are moderate distributions of VIP-reactive fibers in the area ventralis and in the area septalis.Ultrastructurally, the immunoreactive products against VIP are found in the elementary granules, 75–115 nm in diameter, within the nerve fibers in the median eminence.This investigation was supported by Scientific Research Grants No. 00556196, No. 56360027 and No. 56760183 from the Ministry of Education of Japan to Professor Mikami and Mr. Yamada     

Immunohistochemical localization of LH-producing cells in the adenohypophysis of the Japanese quail,Coturnix coturnix japonica

Summary The cells that produce luteinizing hormone (LH) in the adenohypophysis of the Japanese quail were identified immunohistochemically using anti-chicken LH serum and horseradish peroxidase-labeled goat anti-rabbit gamma globulin serum. The LH cells are localized in the caudal lobe of the pars distalis. They are elongate in shape and are polarized toward the sinusoids, especially in their active states. Alterations in size of LH cells are directly related to changes in circulating LH levels as induced by castration or photostimulation. The LH cells identified immunohistochemically were only stained by alcian blue with periodic acid-Schiff (PAS), alcian blue and orange G.PAS-positive gonadotropic cells in the cephalic lobe were stained immunohistochemically only slightly if at all using anti-chicken LH serum and consequently may be FSH producing cells. In the cephalic lobe another type of basophilic cell was stained with alcian blue. These cells were also stained immunohistochemically with anti-chicken LH serum. These cells may possibly be identified as TSH cells due to the characteristics of the antichicken LH serum used in this study which cross react with LH and TSH but only slightly with FSH, and also on the basis of previous light and electron microscopic studies.We express our gratitude to Professors Hideshi Kobayashi and Katsumi Wakabayashi for their valuable guidance during the experiment. We also express our cordial thanks to Professor B.K. Follett for the gift of anti-chicken LH serum and standard LH. This work was supported in part by Grants from Ministry of Education of Japan and from the Ford Foundation.     

Immunocytochemical localization of neurotensin-containing neurons in the hypothalamus of the japanese quail,Coturnix coturnix japonica

Summary The hypothalamus of Japanese quail, Coturnix coturnix japonica, has been studied by means of the peroxidase-antiperoxidase immunocytochemical method, with the use of antibodies to synthetic neurotensin (NT). A number of immunoreactive neuronal perikarya occur in the medial preoptic nucleus of the rostral hypothalamus and a few in the accessory part of paraventricular nucleus and dorsal portion of the infundibular nucleus. Some of them correspond to the parvocellular neurons previously identified tentatively as neurosecretory (Mikami et al. 1975, 1976). Large numbers of immunoreactive neuronal fibers are found in the preoptic area, which extend as a remarkable fiber tract from this area to the ventral septal area and to the subfornical organ. A few immunoreactive fibers also extend ventrocaudally to the infundibular nucleus and to the neural lobe.This investigation was supported by Scientific Research Grants No. 556196 and No. 576176 from the Ministry of Education of Japan to Professor Mikami and Mr. Yamada     

Protein polymorphism in two populations of the wild quail Coturnix coturnix japonica

To evaluate genetic variability in two populations of the wild quail Cofurnix coturnix japonica , proteins and enzymes were examined by starch gel electrophoresis.
Rare variants so far not observed in domestic quail were found in the following five enzymes; aspartate aminotransferase, acid phosphatase, pancreatic esterase, isocitrate dehydrogenase and lactate dehydrogenase. The proportion of polymorphic loci ( P _poly) and the expected average heterozygosity ( H ) in one of the two populations were estimated to be 0.484 (15/31) and 0.085, respectively. Those in another population were 0.433 (13/30) and 0.086, respectively. The genetic distance (Nei, 1975) between the two wild quail populations was D = 0.0074. D values of 0.0321 and 0.0189 were estimated between the laboratory quail population previously examined (Kimura et al., 1982) and each of these two wild populations.     

Genetic variation in a population of the wild quail Coturnix coturnix japonica

To investigate genetic variation and to compare it with those of domestic quail populations, enzymatic and non-enzymatic proteins in a wild quail population were examined electrophoretically.
The wild quail population had a genetic variability considerably lower than those of domestic populations. The average heterozygosity estimated for 22 loci was 0.090 in the wild quail population.     

Immunocytochemical localization of CRF in the ovine hypothalamus

A population of neuronal cell bodies and their fiber pathways have been elucidated within the ovine hypothalamus. The immunoreactive neurons were located in the anterior and dorsal hypothalamus interspersed throughout the paraventricular nucleus. These perikarya were only observed when an antiserum that was generated against the C-terminal of CRF was employed. A dense fiber projection traversed the medial-basal hypothalamus and ended within the palisade-contact zone of the median eminence and neural stem. Fibers were revealed by antisera generated against both the N-terminal and the C-terminal of CRF. Antisera pre-absorbed with synthetic CRF failed to yield immunoreactivity.     

Parenchymal fine structure of the subfornical organ in the Japanese quail,Coturnix coturnix japonica

Summary The parenchyma of the subfornical organ (SFO) of the Japanese quail was studied by light and electron microscopy. The SFO consists of ependymal, intermediate, and basal (perimeningeal) layers. In the intermediate layer, neurons, glial cells, and their processes are found. Axons containing dense core granules approximately 80 nm in diameter are numerous, some of which make synaptic contact with the neuronal perikarya or dendrites. Synaptic vesicles in some axons contain a dense dot in the interior after treatment with 5-hydroxydopamine. The activity of the SFO, which is probably concerned with elicitation of drinking by angiotensin II, may be regulated at least partly by afferent monoaminergic axons. Capillaries with a non-fenestrated endothelium are occasionally found in the parenchyma. The basal layer is occupied by glial processes abutting on the digitating layer of perivascular connective tissue of meningeal vessels. The endothelium of these vessels is occasionally fenestrated. Trypan blue injected systemically accumulated in the SFO, but not in the deeper areas of the brain. The absence of a blood-brain barrier is suggested in the SFO.     

Surface fine structure of the subfornical organ in the Japanese quail,Coturnix coturnix japonica

The surface ultrastructure of the subfornical organ (SFO) was investigated in the Japanese quail. The SFO consists of a body and a stalk. The body of the SFO can be divided into rostral and caudal parts. On the rostral part, each ependymal cell possesses a short central solitary cilium; clustered cilia are also occasionally seen. Microvilli are abundant. On the caudal part, cells with a solitary cilium are fewer in number, and clustered cilia are rarely found. Microvilli are not as abundant as on the rostral part. In addition, large bulbous protrusions, tufts of small protrusions, deep funnel-shaped hollows, small pinocytotic invaginations and possible cerebrospinal fluid-contacting axons are sporadically observed on the surface of various regions of the body. Each ependymal cell of the stalk has a wide apical surface. A central solitary cilium, microvilli and other structures are observed more rarely on the stalk than on the body, while clustered cilia are not seen on the stalk. These structures are compared with those of the mammalian SFO and further discussed in relation to the possible dipsogenic receptor function for angiotensin II.     

The acute phase response in Japanese quail (Coturnix coturnix japonica)

Experiments were conducted to determine the effect of injection of lipopolysaccharide (LPS, from S. typhimurium) or muramyl dipeptide (MDP, N-acetylmuramyl-L-ala-isoglutamine) in Japanese quail. Doses of MDP between 0.3 and 10 mg/kg body wt. had no effect on body temperature. In contrast, doses of 1.0-22.5 mg LPS/kg body wt. caused significant increases in body temperature. None of the doses of LPS or MDP resulted in mortality. The febrile response to LPS was diminished following a second injection 48 h after the first, and was absent following a third injection. Plasma zinc, an indicator of the acute phase response, was significantly reduced by either LPS or MDP after the first injection (P<0.001), but not after the second or third injection. Splenic interleukin 1-beta (IL-1beta) mRNA expression was increased after the first and last injection of LPS (P<0.001), but only after the first injection of MDP (P<0.005). Hepatic IL-1beta mRNA expression was increased after the first, but not the third injection of LPS (P<0.001), while MDP had no effect. These data indicate that Japanese quail are less sensitive to MDP than LPS, and that quail demonstrate tolerance to LPS following repeated injections.     

Characterization of lactate dehydrogenase enzyme in seminal plasma of Japanese quail (Coturnix coturnix japonica)

Lactate dehydrogenase enzyme present in quail seminal plasma has been characterized. Polyacrylamide gel electrophoresis and subsequently with LDH specific staining of seminal plasma revealed a single isozyme in quail semen. Studies on substrate inhibition, pH for optimum activity and inhibitor (urea) indicated the isozyme present in the quail semen has catalytic properties like LDH-1 viz. H-type. Furthermore, unlike other mammalian species, electrophoretic and kinetic investigations did not support the existence of semen specific LDH-X isozyme in quail semen. The effect of exogenous lactate and pyruvate on sperm metabolic activity was also studied. The addition of 1 mM lactate or pyruvate to quail semen increased sperm metabolic activity. Our results suggested that both pyruvate and lactate could be used by quail spermatozoa to maintain their basic functions. Since the H-type isozyme is important for conversion of lactate to pyruvate under anaerobic conditions it was postulated that exogenous lactate being converted into pyruvate via LDH present in semen may be used by sperm mitochondria to generate ATP. During conversion of lactate to pyruvate NADH is being generated that may be useful for maintaining sperm mitochondrial membrane potential.     

Cytological differentiation of the interrenal tissue of the Japanese quail,Coturnix coturnix

Summary As reported for several other avian species there are clearly distinguishable subcapsular (SCZ) and inner (IZ) zones of interrenal tissue in the Japanese quail. The SCZ contains large columnar cells (type I) with rounded nuclei, polymorphic mitochondria with shelf-like cristae, and relatively small numbers of lipid droplets. The IZ contains two and possibly three types of cells. Type II consists of large columnar cells with moderately dense cytoplasm containing large numbers of lipid droplets and many rounded mitochondria with tubular cristae. Smooth endoplasmic reticulum (SER) and Golgi apparatus are well developed; coated vesicles occur in the Golgi area and at the cell surface. Type-III cells occur in IZ and especially in its more peripheral areas. They are columnar cells with strikingly clear cytoplasm (in comparison with type II) containing mitochondria with plate-like cristae and tubular SER. Type-IV cells are sparsely distributed in IZ and occur rarely in SCZ. Type IV may be a degenerating phase of type III.After adenohypophysectomy or section of portal vessels type-I cells atrophy somewhat with a decrease in lipid droplets; type-II cells, also atrophy with conspicuous increase in size and number of lipid droplets, enlargement of mitochondria, and gradual disappearance of SER; type-III cells decrease in number whereas type-IV cells increase.After injection of ACTH, type-I cells enlarge and their mitochondria, SER and Golgi apparatus become more conspicuous; there is a decrease in lipid droplets in type-II cells and a development of SER, polysomes and Golgi apparatus; there is also a decrease in lipid droplets and a development of SER in type-III cells after injection of 2IU ACTH and an almost complete disappearance of lipid droplets after 4IU ACTH; type-IV cells increase in number.The investigation reported herein was supported by Scientific Research Grants from the Ministry of Education of Japan to Professor Mikami; and by grants from the Japan Society for the Promotion of Science, the National Science Foundation (USA), and the Graduate School Fund of the University of Washington to Professor Farner     

Genetic variants of phosvitin in egg yolk of the Japanese quail, Coturnix coturnix japonica

Summary. Phosvitin polymorphism in egg yolk of the Japanese quail was found by horizontal polyacrylamide gradient gel electrophoresis. Six phenotypes of yolk phosvitin designated A, B, C, AB, AC, and BC were observed in a population of 281 birds. Analysis of family data revealed that the phenotypic variation of quail yolk phosvitins was controlled by an autosomal Pv locus with three codominant alleles, Pv^a, Pv^b and Pv^c. The gene frequencies of Pv^a, Pv^b and Pv^c were 0.064, 0.824 and 0.112, respectively.     

Moulting cycles in European quail (Coturnix coturnix coturnix) under constant photoperiodic conditions

Abstract

Male and female European quails (Coturnix coturnix coturnix) were studied whilst being maintained in LD 12:12, at 20° C, for four years from birth. The birds exhibited the following phases: end of the first phase of the post‐juvenile moult, fattening and a period of nocturnal activity. Then sexual development is observed up to complete maturation at the age of four months. The second part of post juvenile moulting occurs towards the age of 5 to 6 months. During the following 6 months an important phase of reproduction is observed, thus completing their first year of life. The same sequence is repeated during the following years: a moult which lasts for 6 months in two (sometimes three) partial phases separated by episodes of reproduction; then an important period of reproduction (6 months) without moulting. The birds are capable of reproducing at all times except during moulting. The sequence of moulting begins each year in the same months as those for natural French populations. It thus seems that the annual periodicity and the organisation itself of the phenomenon of moulting is controlled in European quail by an endogenous circannual rhythm. In contrast, the natural cyclic expression of sexuality appears to be dependent on abiotic environmental and social factors.     

Inter-individual variability during morning choruses in Japanese quail males (Coturnix coturnix japonica)

We compared the organization of dawn choruses in five groups of Japanese quail, Coturnix coturnix japonica, males (four groups of kin-related males and one group of unrelated males). We documented the structure of chorus groups: one male sang much more than the others during the dawn peak in four of the five groups, and this regardless of the number of males in the group. The dominant singers were then removed from two similar-sized groups, one of kin-related (K3) and the other of unrelated (NK) males. The quantity of song emitted by the remaining NK males was modified, but not that emitted by the K3 males.     

Oviposition patterns and photoresponsivity in Japanese quail (coturnix coturnix japonica)

Abstract

Plots of oviposition times for an unselected group of 84 female Coturnix using the circular mean time for each bird on a central index time indicated a regularity in the data for individual birds. An analysis of pooled data using a moving average showed agreement with the temporal occurrence of the photoresponsive interval reported in the literature for gonadal development. These data were used to support a modification of existing hypothesis on the control of oviposition and ovulation.     

Ultrastructural changes in the uropygial gland of the male Japanese quail,Coturnix coturnix,after testosterone treatment

Summary The ultrastructure of the uropygial gland of the male quail was compared to that of the sebaceous gland of the male rat after castration and testosterone treatment of both species. In intact animals, the differentiating cells of these glands displayed almost the same pattern as regards their smooth endoplasmic reticulum, an organelle involved in lipogenesis in both cases. Castration reduced the volume of this organelle, while testosterone administration restored cell morphology to a normal or supranormal level. Finally, this study showed that at ultrastructural level, there is a close functional analogy between the uropygial gland of quail and the sebaceous glands of rats as regards their androgen dependency. Consequently, the uropygial gland might be an attractive model for study of action of androgens on sebaceous-like glands.     

CRF-containing neurons of the rat hypothalamus

Summary The immunoreactive CRF-neurons of the rat hypothalamus have been examined immunohistochemically employing anti-rat CRF serum. These neurons are confined to the paraventricular nucleus, dorsomedial-lateral hypothalamic area, and suprachiasmatic nucleus, and are, respectively, also immunoreactive to anti-Met-enk, -alpha-MSH, and -VIP sera. Intraventricular administration of colchicine (50 g/5 l/rat) induces a dramatic enhancement of the immunostainability of the cell somata, and also accelerates the development of immunoreactivity of other stored peptides, especially in the paraventricular nucleus.The CRF-neurons respond to adrenalectomy by showing increased immunoreactivity and an increase in the number of cell bodies; in the dorsomedial-lateral area and suprachiasmatic nucleus, there is also an enhanced immunoreactivity for alpha-MSH and VIP, respectively. CRF-cells in the paraventricular nucleus become markedly hypertrophied, but do not show any enhanced immunoreactivity for Met-enk. Since the axons of the paraventricular neurons run to the median eminence, it is probable that they are involved with the endocrine control of hypophysial ACTH release. It is concluded that the CRF-containing neurons in rat hypothalamus consist of three types which are functionally and morphologically different.     

Lactate dehydrogenase isozyme patterns in the denervated quail (Coturnix coturnix japonica) muscles

Polyacrylamide gel electrophoresis of the Japanese quail (Coturnix cotunix japonica) muscle extracts revealed a single lactate dehydrogenase isozyme. A month after surgical unilateral brachiotectomy (denervation) there was significant atrophy of the triceps, biceps and radius ulnar muscles accompanied by the appearance of an additional lactate dehydrogenase isozyme band. This extra band may be the result of the synthesis of a new lactate dehydrogenase isozyme. This new isozyme exhibited a lower affinity for lactate, less sensitivity to urea denaturation and was more thermostable than the lactate dehydrogenase of normal (innervated) quail muscles. Based on these properties, it is suggested that the newly synthesised isozyme of the denervated muscles is LDH-1, (or B4/H4) type. Brachiotectomy also resulted in significant quantitative changes in the total lactate dehydrogenase activity of innervated muscles of the same animal.