Hypothalamic and hormonal control of photoperiodically induced vernal functions in the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary The effects of implantation of testosterone propionate (TP) in various sites in the hypothalamus on the photoperiodically induced vernal premigratory functions in the White-crowned Sparrows were investigated in order to assess the role of the hypothalamo-hypophysial-testicular axis in the induction of these responses.Implantation of glass capillary tubes containing TP in the basal infundibular nucleus (IN), in the median eminence, or in the pars distalis inhibited the photoperiodically induced increase in plasma levels of luteinizing hormone (LH), as measured by radioimmunoassay, and testicular growth. The effective implants significantly lowered the levels of LH in birds held on nonstimulatory short days. These TP implants apparently inhibited release from the pars distalis of both LH and follicle-stimulating hormone (FSH). It is concluded that the site of sensitivity in the negative feedback by testosterone is either the basal IN or the pars distalis, or both. The implants of TP that inhibited the increase in plasma LH and testicular growth completely did not prevent the birds from fattening.These investigations were supported, in part, by research grants from the National Institutes of Health (HD-6527) and National Science Foundation (BMS 79-13933) to Professor Donald S. Farner. This paper is based on a dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy from the University of WashingtonThe author is grateful to Professor Donald S. Farner for his guidance throughout the course of these investigations. The assistance of Mr. Philip W. Mattocks, Jr. in performing radioimmunoassays is sincerely appreciated     

Fluorescence microscopy of neurons containing primary catecholamines in the ventral hypothalamus of the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary The ventral hypothalamus of White-crowned Sparrows, Zonotrichia leucophrys gambelii, was examined for primary catecholamines with the fluorescence technique of Falck-Hillarp and Owman. Photorefractory and photosensitive birds, photoperiodically stimulated and non-stimulated, were used.Four groups of catecholaminergic fibers were demonstrated: (1) afferent fibers of extra-hypothalamic origin to the infundibular nucleus where there are extensive synaptic contacts with non-fluorescent perikarya; (2) fibers apparently extending ventrally from an area with fluorescent perikarya in the vicinity of the paraventricular organ to the infundibular nucleus via the stratum cellulare internum and lateral pathways; (3) afferent fibers to the subependymal layer of the median eminence; and (4) afferent fibers to the zona externa of the median eminence. Fluorescent fibers that pass transversely through the median eminence may be derived from any of the last three categories. It appears that the fibers of the zona externa (4) are not derived to any appreciable extent from those of the subependymal layer (3). Because fluorescent perikarya could not be demonstrated in the infundibular nucleus no conclusion can be reached with respect to this nucleus as an origin for fibers (3) and (4).Diurnal cycles in the number of fluorescent terminals observable and in the intensity of the fluorescence in the palisade layer in photosensitive birds subjected to different photoperiodic regimes, in castrates, and in photorefractory birds are described tentatively on the basis of subjective assessments. Some possible implications of the differences are discussed.This communication is based extensively on a thesis, Primary catecholamine fibers in the ventral hypothalamus of the White-crowned Sparrow, Zonotrichia leucophrys gambelii, submitted in partial fulfillment of the requirements for the degree of Master of Science, University of Washington, August 1968, by the senior author. The research on which it is based was supported extensively by Grant No. NB 06187 from the National Institutes of Health to Professor Donald S. Farner. We are grateful to Ciba Pharmaceutical Products, Inc. for a supply of Serpasil.     

Migratory sleeplessness in the white-crowned sparrow (Zonotrichia leucophrys gambelii)

Twice a year, normally diurnal songbirds engage in long-distance nocturnal migrations between their wintering and breeding grounds. If and how songbirds sleep during these periods of increased activity has remained a mystery. We used a combination of electrophysiological recording and neurobehavioral testing to characterize seasonal changes in sleep and cognition in captive white-crowned sparrows (Zonotrichia leucophrys gambelii) across nonmigratory and migratory seasons. Compared to sparrows in a nonmigratory state, migratory sparrows spent approximately two-thirds less time sleeping. Despite reducing sleep during migration, accuracy and responding on a repeated-acquisition task remained at a high level in sparrows in a migratory state. This resistance to sleep loss during the prolonged migratory season is in direct contrast to the decline in accuracy and responding observed following as little as one night of experimenter-induced sleep restriction in the same birds during the nonmigratory season. Our results suggest that despite being adversely affected by sleep loss during the nonmigratory season, songbirds exhibit an unprecedented capacity to reduce sleep during migration for long periods of time without associated deficits in cognitive function. Understanding the mechanisms that mediate migratory sleeplessness may provide insights into the etiology of changes in sleep and behavior in seasonal mood disorders, as well as into the functions of sleep itself.     

Hormonal, behavioral, and thermoregulatory responses to bacterial lipopolysaccharide in captive and free-living white-crowned sparrows (Zonotrichia leucophrys gambelii)

Exposing vertebrates to pathogenic organisms or inflammatory stimuli, such as bacterial lipopolysaccharide (LPS), activates the immune system and triggers the acute phase response. This response involves fever, alterations in neuroendocrine circuits, such as hypothalamo-pituitary-adrenal (HPA) and -gonadal (HPG) axes, and stereotypical sickness behaviors that include lethargy, anorexia, adipsia, and a disinterest in social activities. We investigated the hormonal, behavioral, and thermoregulatory effects of acute LPS treatment in a seasonally breeding songbird, the white-crowned sparrow (Zonotrichia leucophrys gambelii) using laboratory and field experiments. Captive male and female sparrows were housed on short (8L:16D) or long (20L:4D) day lengths and injected subcutaneously with LPS or saline (control). LPS treatment activated the HPA axis, causing a rapid increase in plasma corticosterone titers over 24 h compared to controls. Suppression of the HPG axis occurred in long-day LPS birds as measured by a decline in luteinizing hormone levels. Instead of a rise in body temperature, LPS-injected birds experienced short-term hypothermia compared to controls. Birds treated with LPS decreased activity and reduced food and water intake, resulting in weight loss. LPS males on long days experienced more weight loss than LPS males on short days, but this seasonal effect was not observed in females. These results paralleled seasonal differences in body condition, suggesting that modulation of the acute phase response is linked to energy reserves. In free-living males, LPS treatment decreased song and several measures of territorial aggression. These studies highlight immune-endocrine-behavior interrelationships that may proximately mediate life-history tradeoffs between reproduction and defense against pathogens.     

Circadian and masking control of migratory restlessness in Gambel's white-crowned sparrow (Zonotrichia leucophrys gambelii)

Avian migration is a seasonal activity that requires intricate timing on both an annual and daily basis. With increasing evidence for endogenous regulation of daily activities in migrant species, we tested whether a circadian oscillator may be involved with the expressions of daily locomotor activities and specific behaviors of the long-distance migrant, Gambel's white-crowned sparrow (Zonotrichia leucophrys gambelii). Our previous studies have identified both daytime and nighttime behavioral patterns under a photoperiod of 18L:6D. In 2 separate trials, birds in the vernal migratory life-history stage were exposed to constant dim light, (DD)dim, and constant bright light, LL, while locomotor activity and behavioral observations were collected. Under (DD)dim, the daytime behaviors that included active and quiescent components observed under 18L:6D were lost as migratory restlessness, the intense nighttime activity, persisted nonstop for 36.4 h. Furthermore, the specific behaviors of migratory restlessness that are normally confined to the dark phase of 18L:6D, beak-up and beak-up flight, were expressed also during the subjective day of (DD)dim. Birds exposed to LL retained similar patterns of activity to the 18L:6D condition for 3 days, after which they became arrhythmic. Behavioral observations of intense locomotor activity observed during the subjective night of LL revealed no beak-up and beak-up flight. Thus, the complete expression of migratory restlessness that includes beak-up and beak-up flight may not be regulated by a circadian oscillator but instigated by very low light intensity. Locomotor activity and associated daytime behaviors appear to be influenced by a circadian oscillator, given their persistent patterns under LL. Therefore, we suggest that the separate components of migratory behavior are regulated differentially by a circadian oscillator and ambient lighting conditions.     

Electron microscopic and experimental studies of the pineal organ in the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary The structure of the pineal organ of Zonotrichia leucophrys gambelii, as revealed by light- and electron-microscopy, resembles that of Passer domesticus (Oksche and Kirschstein, 1969; Ueck, 1970). The typical cellular element is the pinealocyte with certain basic structural features of the pineal photoreceptors of lower vertebrates (see Oksche, 1971). However, instead of the characteristic, cone-like outer segments, there are, as in other species of birds, only bulbous cilia with ectopic whorls of lamellae. This structure of the outer segment is, in a sense, contrary to the demonstration of synaptoid contacts, numerous unmyelinated, and occasional myelinated nerve fibers by electron microscopy. In Nissl preparations it was possible to demonstrate typical nerve cells. The pinealocytes of Z. l. gambelii are secretory; their Golgi complex forms granulated vesicles (800–1,400 Å in diameter) that belong to the group of granular inclusions characteristic of monoamines. Autonomie nerve fibers course within the connective tissue capsule of the pineal organ. In many pinealocytes of Z. l. gambelii, the granular endoplasmic reticulum contains extensively expanded cisternae that are filled with a flocculent material and closely associated with bundles of filaments. In a number of cases such loop-like structures are selectively stainable with aldehyde fuchsin. It was not possible to demonstrate specific secretory activity in the supporting cells. Extirpation of the pineal organ in Z. l. gambelii had no definitely detectable influence on the photoperiodic control of testicular growth.Aves, Passeriformes, Fringillidae.Supported by grants from the Deutsche Forschungsgemeinschaft to Professor Oksche and by the National Science Foundation (GB 11905) to Professor Farner. A part of this investigation was effected while Professor Kobayashi held a Visiting Professorship at the University of Giessen.     

Investigations of the butylcholinesterase-containing cells of the adenohypophysis of the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary In the adenohypophysis of Zonotrichia leucophrys gambelii two types of cells with butylcholinesterase(BuChE) activity can be demonstrated histochemically. Type I occurs in the cephalic lobe of the pars distalis and in the pars tuberalis; it consists of oval and round cells. It is a distinctive cell type that is identical with the amphophilic cells described by Matsuo, Vitums, King and Farner (1969). Whereas castration or inhibition of thyroid gland activity causes only minor changes in these cells, blocking of adrenal cortex activity, or adrenalectomy, causes conspicuous hyperplasia and hypertrophy suggesting that these cells are involved in the production or release of ACTH. The second type, which occurs in both cephalic and caudal lobes, consists mostly of irregularly formed cells. Various observations indicate that it is a composite group, consisting, at least in part, of degenerating cells.These investigations were supported by grants from the National Institutes of Health (S ROI NB 06812) and the National Science Foundation (GB 5969) to Professor Farner.     

The ACTH-immunoreactive system in the brain of the white-crowned sparrow,Zonotrichia leucophrys gambelii (Passeriformes,Emberizidae)

Summary The ACTH-immunoreactive (ir) system of the avian brain is particularly conspicuous in the male white-crowned sparrow (Zonotrichia leucophrys gambelii). The irperikaryal population is concentrated mainly within the tuberal region, projecting primarily in a dorsal direction: (i) into the striatum; (ii) into rostral diencephalic, septal, hyperstriatal, and thalamic areas; and (iii) into dorsal and ventral areas of the brain stem. Ir-fibers seemingly contact local non-immunoreactive neurons mainly in the accumbens nucleus, septum, dorsal thalamic nuclei, infundibular and interpeduncular nuclei, and in the rostral diencephalon. Neurohemal zones are not supplied by ACTH-ir terminals. Immunocytochemical problems arising from the complexity of the proopiomelanocortin molecule and its derived peptide components are discussed in relation to phylogenetically directed studies, and contradictory results.Dedicated to Professor Dr. T.H. Schiebler on the occasion of his 65th birthday     

The ACTH-immunoreactive system in the brain of the white-crowned sparrow, Zonotrichia leucophrys gambelii (Passeriformes, Emberizidae)

The ACTH-immunoreactive (ir) system of the avian brain is particularly conspicuous in the male white-crowned sparrow (Zonotrichia leucophrys gambelii). The irperikaryal population is concentrated mainly within the tuberal region, projecting primarily in a dorsal direction: (i) into the striatum; (ii) into rostral diencephalic, septal, hyperstriatal, and thalamic areas; and (iii) into dorsal and ventral areas of the brain stem. Ir-fibers seemingly contact local non-immunoreactive neurons mainly in the accumbens nucleus, septum, dorsal thalamic nuclei, infundibular and interpeduncular nuclei, and in the rostral diencephalon. Neurohemal zones are not supplied by ACTH-ir terminals. Immunocytochemical problems arising from the complexity of the proopiomelanocortin molecule and its derived peptide components are discussed in relation to phylogenetically directed studies, and contradictory results.     

The distribution of monoamine oxidase and acetylcholinesterase in the hypothalamus and its relation to the hypothalamo-hypophysial neurosecretory system in the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary The distribution of monoamine-oxidase and acetylcholinesterase activities in the hypothalamus of the White-crowned Sparrow has been studied in relation to the hypothalamohypophysial neurosecretory system. The enzyme activities, as revealed by the methods employed, are unaffected during photoperiodically induced testicular growth. Monoamine oxidase has a distribution distinctly different from that of the aldehyde-fuchsin positive neurosecretory material in that there is high activity in the peripheral palisade layers of both the anterior and posterior divisions of the median eminence. Intimate contact is made between these areas with the primary vessels of the hypophysial portal system. A second concentration of activity lies in a layer between the ependymal cells and the neurosecretory material of the fiber tract. In general, monoamine oxidase appears to be associated with glial elements and non-neurosecretory axons. The pars nervosa has little or no monoamine-oxidase activity. The distribution of acetylcholinesterase activity in the anterior division of the median eminence is very similar to that of the aldehyde-fuchsin positive neurosecretory neurons; however, acetylcholinesterase also occurs in the posterior division without associated neurosecretory fibers. These distribution of enzyme activities are considered in relation to possible adrenergic and cholinergic mechanisms in the median eminence.Dedicated to Professor Berta Scharrer in honor of her 60th birthday.Supported by grant NB-01353 from the National Institutes of Health to Professor Farner. This investigation was conducted while Doctor Kobayashi was a National Science Foundation Senior Foreign Scientist at Washington State University. We are indebted to Doctor Christian Da Lage, Laboratoire de Histologie, Falculté de Médecine de Paris, for the preliminary development of some of the techniques used in these investigations.     

Projection of fibers immunoreactive to an antiserum against gonadoliberin (LHRH) into the pineal stalk of the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary We have demonstrated a bundle of fibers, immunoreactive to an antiserum against gonadoliberin (luteinizing hormone-releasing hormone), that projects into the pineal stalk of photostimulated male Z. l. gambelii, a photoperiodic passerine species. These fibers have their origin in the dorsal division of the column of gonadoliberin-positive perikarya that extends caudodorsally from the ventromedial preoptic area to the level of the anterior commissure. After entering the habenula these fibers can be traced into the pineal stalk. This bundle is of interest because it apparently has not previously been reported in birds, and further, because the pineal body of this species is not known to have a role in the photoperiodic induction of increased rates of release of gonadotropins. Relationships of this bundle to other components of the gonadoliberin system are described briefly.     

Prolactin augments luteinizing hormone binding to rat Leydig cells in serum-free culture

Effect of prolactin on the testicular luteinizing hormone binding was studied in a serum-free culture system. By the collagenase digestion of decapsulated testes taken out from 25-day-old rats, Leydig cells were isolated and cultured for 7 days in DME/F12 (1:1) medium supplemented with insulin, transferrin, epidermal growth factor, and gentamicin. The cultured cells exhibited the 3β-hydroxysteroid dehydrogenase activity. Hill plots constructed from the data of competition experiment showed that the dissociation constant (K_d) was 0.33 × 10^–10M. The K_d value was approximately the same as the known value for the rat testicular homogenates. When the Leydig cells were cultured with ovine prolactin for the last 3 days of 7-day culture period, the binding of luteinizing hormone increased to 1.7-fold ofthat in the control group. From these results it is concluded that prolactin acts to up-regulate the binding of luteinizing hormone to rat testicular Leydig cells in serum-free culture     

Seasonal changes in the reproductive system of the female white-crowned sparrow,Zonotrichia leucophrys gambelii,in captivity and in the field

Summary The ventral apterium of free-living female White-crowned Sparrows (Zonotrichia leucophrys gambelii) becomes an incubation patch during the breeding season. At this time, it loses its feathers, increases in wet and defatted dry weight, and undergoes marked histological alterations. At times of year other than the breeding season, the apterium consists of a low squamous epidermis and a thin, poorly vascularized dermis of dense connective tissue. The dermis is separated from subcutaneous tissue by an internal elastic lamina. During the breeding season, the epidermis is a proliferative, stratified squamous eptihelium with well-defined basal, intermediate, transitional, and cornified layers; and the dermis consists of a superficial layer of collagen and a deep layer of highly vascular areolar connective tissue, noticeably edematous and mildly inflamed. Blood vessels are frequently in large groups in the center of the dermis. Edema and hypervascularity are most pronounced during incubation, but the epidermis is best developed during egg-laying. The apterium reverts to its basal state after the incubation period. Captive females, which do not breed, do not develop incubation patches.Estrogen is apparently responsible for feather loss and collagen synthesis. It and other unidentified hormones (probably prolactin and/or androgens) produce the hypervascularity, edema, and epidermal growth.This study was supported by an NIH Training Grant (GM 01276) from the National Institute of General Medical Sciences and by funds from the Faculty Research Council at Fordham University, New York, N.Y. It is a pleasure to acknowledge the assistance of Drs. Brina Kessel and George West at the University of Alaska during summer, 1968; Dr. William A. deGraw for help in the field; and Drs. Laura Englisch and James Wiggins, as well as my son, Robert, for help with the photomicrography. Wallace Moreland of the Singer-Friden Company kindly provided a desk calculator for my use. The content of this paper profitted substantially from the suggestions of Dr. Robert A. Lewis. This paper is based on part of a dissertation done to fulfill the Ph.D. requirements at Washington State University     

The effect of flight, fasting and p,p'-DDT on thyroid hormones and corticosterone in Gambel's white-crowned sparrow, Zonotrichia leucophrys gambelli

This study examined the effects of p,p'-dichlorodiphenyltrichloroethane (p,p'-DDT), fasting and flight on thyroid hormones and corticosterone in Gambel's White-crowned Sparrows (Zonotrichia leucophrys gambelli). Female sparrows were dosed daily with either 5 mg p,p'-DDT per kg body mass or corn oil vehicle over 3 days. On the fifth day the sparrows were divided into 3 groups: (1) unstressed - non-stressed control sparrows; (2) fasted - sparrows fasted for intervals ranging from 20 min to 9 h; or (3) flown - sparrows flown in a wind tunnel for intervals between 20 min and 2.5 h while fasting. Half the sparrows from each group received DDT (DDT-dosed sparrows) and the other half corn oil vehicle only (vehicle sparrows). Trunk blood plasma was analyzed for thyroxine, triiodothyronine and corticosterone using radioimmunoassay. In the flown group, corticosterone was elevated (DDT-dosed 35.52 ng/ml, P < or = 0.05), and thyroxine was depressed (DDT-dosed 4.09 ng/ml, P < or = 0.05; vehicle 4.33 ng/ml, P < or = 0.05). Elevated corticosterone likely decreased thyroid hormone production through a negative feedback mechanism originating at the hypothalamus. Mean triiodothyronine concentrations did not differ among any of the test groups. Relative to time fasted and flown, thyroxine decreased in flown birds dosed with DDT (P < 0.001) and triiodothyronine decreased in fasted birds dosed with DDT (P = 0.004). The increased rate of hormone diminution may be a result of the ability of DDT to induce microsomal enzyme production.     

Validation of an enzyme immunoassay for measurement of excreted estrogen and testosterone metabolites in the white-crowned sparrow (Zonotrichia leucophrys oriantha)

Over the past few years there has been a growing interest in hormonal analyses, despite the cost and difficulty involved in measuring hormones using radioimmunoassays. Therefore, there is a need for alternate procedures, such as enzyme immunoassays, capable of providing similar data to that provided by radioactive methods. Enzyme immunoassays are a less expensive method of measuring estrone conjugates and testosterone. With further work, the enzyme immunoassay also provides the potential to adapt the enzyme assay for use in the field. The data in this study demonstrate that an enzyme assay can accurately detect both endogenous and exogenous hormonal changes in the droppings of white-crowned sparrows. © 1995 Wiley-Liss, Inc.