Colin A. Maxwell

Seasonal Modification of Ovine Pineal Function

Pineal β-adrenoceptor density and affinity in ewes are modified in a season-dependent manner by gonadal steroids and by the sympathetic innervation of the gland. The present study was undertaken to relate the steroidal effects on the receptors to post receptor endocrine events, and to investigate the influence of the sympathetic innervation of the pineal gland on these events. Plasma melatonin and prolactin profiles were determined during anestrus and during the normal breeding season in ewes subjected to sympathetic denervation of the pineal and/or a range of steroid-related treatments. Wherever valid comparisons could be drawn between effects of the treatments on β-adrenoceptor variables and on circulating hormone levels, similar effects were noted. Further, ganglionectomy influenced hormone profiles similarly to estradiol under all conditions tested. It appears that gonadal steroids (estradiol) and the sympathetic neurotransmitter noradrenaline have opposing actions on prolactin levels, just as they have on pineal β-adrenoceptor binding affinity. These findings suggest that steroid-mediated changes in receptor number and affinity are reflected in post receptor endocrine events. In addition, other factors (e.g. photoperiodic information transmitted via the sympathetic innervation) also play important roles in the regulation of the observed hormonal profiles. A steroid-mediated feedback regulation of pituitary prolactin release, partly direct and partly via pineal melatonin release, is suggested.

Modification of Sheep Pineal β-Adrenoceptors by Some Gonadal Steroids but Not by Melatonin

The effects of active immunization against a range of gonadal steroids on pineal beta-adrenoceptors were studied in cycling Merino ewes. In selected cases, the effects of exogenous steroids were also investigated. Immunization against progesterone had no detectable effect on pineal beta-adrenoceptor density or ligand binding affinity. Immunization against estrone or 17 beta-estradiol significantly increased binding affinity and decreased beta-receptor density. Testosterone immunization in ewes caused similar effects but to a lesser degree. In contrast, immunization against androstenedione resulted in a decreased affinity and an increase in receptor density in ewes. In subsequent experiments, some parameters of pineal function in intact cycling and ovariectomized ewes were compared; a significant decrease in beta-receptor density and an increase in binding affinity were noted in the ovariectomized animals. Androstenedione-releasing implants decreased beta-receptor density and increased beta-receptor affinity in pineals from intact ewes, but these implants had no effect on pineal parameters in ovariectomized ewes. Immunization of ewes against 17 beta-estradiol had no significant effects on basal or isoprenaline-stimulated N-acetyltransferase or adenyl cyclase activities. Immunization of ewes against melatonin, or in vitro incubation of pineal glands with the hormone had no effects on the pineal parameters studied. It is concluded that estrogens affect the pinealocyte beta-receptors in ewes, while specific androgens may act indirectly on these receptors to modify their interaction with estrogens.

Effects of pinealectomy on wool growth and wool follicle density in Merino sheep

Abstract: There is evidence to indicate that pinealectomy may enhance wool growth in the sheep. The aim of this study was to determine the effect of pinealectomy on wool growth and wool follicle density in Merino sheep.

Sheep Pineal Beta-Adrenoceptor Function – Interaction with Gamma-Aminobutyric Acid

To clarify the role and site of action of gamma-aminobutyric acid (GABA) in ovine pineal glands, we have investigated the effects of aminooxyacetic acid (AOAA), an inhibitor of GABA transaminase, on endogenous GABA content and beta-adrenoceptor mediated pineal function in Merino sheep. A significant elevation of endogenous GABA levels was noted in the glands, but no effect was observed on radioligand binding in vitro to pineal beta-adrenoceptors following in vivo administration of AOAA. Incubation of washed pineal membranes with GABA or AOAA had no effect on ligand binding to beta-adrenoceptors. Incubation of Merino pineal slices with GABA inhibited isoprenaline-stimulated but not basal serotonin N-acetyltransferase (NAT) activity. Incubation of whole pineal homogenates with GABA was without effect on either isoprenaline-stimulated or basal adenyl cyclase activity. Thus, Merino pineal glands resemble bovine pineals in that beta-adrenoceptor mediated melatonin biosynthesis in both species may be regulated in part by GABA. Our results indicate that GABA may exert its effect on Merino pineal NAT activity at a locus distal to the site of action of adenyl cyclase; however, the detailed mechanism and physiological role of this regulation remain to be elucidated.

Effect of pinealectomy and plane of nutrition on wool growth in Merino sheep

Abstract: The effects of surgical pinealectomy and plane of nutrition on wool growth and plasma prolactin concentrations in young Merino wethers were investigated. In young pinealectomized wethers maintained at a low live weight under conditions of minimal stress, the decline of conditioned clean wool production on midside patches was slowed when compared to pineal‐intact controls. This difference appeared to be due in part to the observed greater secondary wool follicle density in the pinealectomized wethers; mean fiber diameter was affected to a smaller extent, while staple length growth rate was not significantly altered. Circulating prolactin profiles showed a seasonal variation (high in summer, low in winter) in both pinealectomized and control wethers. There was no difference in wool production between pinealectomized and control wethers when the diet of the same wethers was subsequently supplemented with formaldehyde‐treated cottonseed meal. While the role of melatonin in the regulation of wool growth remains to be determined, it is suggested that the hormone may have a transient effect on wool production in young wethers under conditions of limited nutrition, but not at higher nutritional levels, and that melatonin may be involved in partitioning of nutrients to the wool follicle.

Effects of steroids on β-adrenergic binding sites in sheep pineal glands

Abstract As an initial step in investigations of putative differences between central nervous system light-sensitive mechanisms in seasonally shedding and non-shedding breeds of sheep, some β-adrenoceptor characteristics of Merino sheep pineal glands were determined, using [ 3 H]dihydroalprenolol as the labelled ligand. Overall, a dissociation constant of 17.2 ± 2.6 moles/I and a daytime β-receptor density of 1.6 ± 0.3 pmoles/mg were determined at 37°. The binding sites exhibited stereospecificity, saturability and apparent homogeneity. 17β-Estradiol and progesterone implants that provided hormone concentrations in the physiological range had no significant effect on pineal β-receptors in male sheep castrated shortly after birth. Dexamethasone injections, on the other hand, in doses sufficient to loosen the attachment of wool fibres to the skin, resulted in decreased pineal β-receptor density and increased receptor affinity for dihydroalprenolol. This effect was apparently not mediated by altered plasma catecholamine concentrations, since the glucocorticoid treatment did not affect jugular venous noradrenaline, adrenaline or dopamine levels. The possible involvement of glucocorticoids in the regulation of wool growth could thus have a central neuronal component, mediated via action on pineal β-adrenoceptors in sheep; however, the existence of the putative gonadal steroid feedback on β-adrenoceptor-mediated pineal function remains to be demonstrated in this species.

Seasonal Changes in the Sensitivity of Ovine Pineal β- Adrenoceptors to Steroids

Immunoneutralization of endogenous gonadal steroids has recently been shown to modify pineal β-adrenoceptor function in intact Merino ewes. In the current study, interactions between gonadal steroids

Effect of Immunization Against Melatonin On Seasonal Fleece Growth in Feral Goats

Abstract: Four vaccination protocols were utilized to investigate the effects of immunoneutralizing circulating melatonin on the annual cashmere growth cycle and cashmere production in Australian feral goats. A fluctuating anti‐melatonin antibody response, achieved by repeated booster vaccinations, resulted in an acceleration of the growth cycle in goats which exhibited a significant immune response, compared to sham‐immunized controls. Responding goats showed two cycles of cashmere length growth in the first 16 months and increased annual cashmere production in the first year. However, in the second year, these effects were no longer apparent, suggesting either some form of desensitization to melatonin, or a diminished response due to declining antibody titre. The effects of immunization were observed in both sexes; the effect on cashmere length was greater in wethers than in does. Cashmere fibre growth in response to a continuously declining plane of specific antibody showed increased cycle frequency, albeit with a decreased amplitude; guard hair growth cycles were affected to a much lesser extent. Small transient peaks of specific immunity at the summer or winter solstice were without significant effect on cashmere growth. Immunization to provoke a persistent anti‐melatonin antibody response at the winter solstice resulted in significantly increased greasy fleece weight, % cashmere yield, and mass of cashmere produced, but no change in fibre diameter in both sexes. Thus the timing of cashmere growth cycles in goats may be, at least transiently, altered by appropriately timed immunization against melatonin. The mechanism of pineal‐mediated regulation of cashmere growth cycles may involve (i) entrainment of an endogenous rhythm by melatonin, or (ii) seasonal alteration of cashmere follicle sensitivity to the effect of melatonin.

Effects of modified circulating prolactin levels on seasonal wool growth in Romney ewes

The study investigated the effects of chronic elevation or suppression of circulating prolactin levels on seasonal wool growth in Romney sheep. Ewes (n=10/group) were housed indoors under natural lighting (latitude 34°S) and received either 50 mg bromocriptine i.m./ewe/28 days (Group B), 1 mg spiperone s.c./ewe/day (Group S) or no treatment (Group C) commencing in summer. Wool growth and liveweight were measured every 4 weeks and venous blood samples were collected for prolactin determination. Plasma prolactin levels (mean±S.E.) in Group C exhibited a gradual seasonal decline from 121±29 ng/ml to 35±9 ng/ml (.)

Effects of prolonged artificial photoperiod on circulating prolactin and melatonin levels in seasonal ewes

Crossbred ewes exposed to long days for 46 months prior to photoperiod reversal showed no alteration in the duration or amplitude of the circulating melatonin peak between 24 and 46 months of continuous long day exposure. By 3 months after photoreversal to short days, both the amplitude and duration of the peak had adapted to the new scotophase. In short day treated ewes, the melatonin peak was abolished by 46 but not 24 months of short day exposure, and was not fully restored in all ewes 3 months after photoreversal. Mean prolactin levels over 24 h remained high up to 46 months of long day treatment, and declined 3 months after short day exposure. Conversely, mean prolactin levels remained low up to 46 months of short day treatment, increasing 3 months after exposure to long days. Thus: (i) depletion of the melatonin-synthesizing capability of the ovine pineal gland by prolonged exposure to long nights is not completely reversed after 3 months of continuous long day exposure, and (ii) a nocturnal melatonin peak is not essential for maintenance of plasma prolactin levels under these conditions.