Distribution of neuropeptide Y in the brain of the male native Thai chicken.

INTRODUCTION: Neuropeptide Y (NPY), a 36 amino acid neurotransmitter/neuromodulator, is involved in food intake and parental cares in birds. NPY is associated with the regulation of the reproductive system in the female native Thai chickens. However, the role of NPY in the male native Thai chicken has not been studied. Therefore, the objective of this study was to investigate the distributions of NPY immunoreactive (-ir) neurons and fibers in the brain of the male native Thai chickens. MATERIAL AND METHODS: The distribution of NPY-ir neurons and fibers in the hen brain was elucidated utilizing immunohistochemical technique. RESULTS: The distributions of NPY-ir neurons and fibers were located throughout the brain, predominantly in the hypothalamus. The numbers of NPY-ir neurons within the nucleus paraventricularis magnocellularis (PVN) were significantly higher than those of the nucleus septalis lateralis (SL), nucleus supraopticus (SOv), and nucleus inferioris hypothalami and nucleus infundibuli hypothalami (IH-IN). In addition, the numbers of NPY-ir neurons within the SL, SOv, and IH-IN were significantly higher than those of the tractus septomesencephalicus and nucleus dorsolateralis anterior thalami. CONCLUSIONS: These results indicated, for the first time, that the distributions of NPY-ir neurons and fibers in the brain of the male native Thai chickens were markedly observed in the hypothalamus, especially within the PVN, implicating that the NPYergic system within the PVN might be related to the regulation of feeding behavior and parental cares in this equatorial species.

Distribution of dopamine-immunoreactive neurons in the brain of the male native Thai chicken.

INTRODUCTION: Dopamine (DA) is a neurotransmitter/neuromodulator found in both central and peripheral nervous systems. It plays several physiological functions in some mammalian and avian species. DA has been indicated to be associated with the neuroendocrine regulation of the reproductive cycle and maternal behaviors in the female native Thai chickens. Indeed, male birds express parental behaviors as well. To date, there are no data describing the functional aspects of the DAergic system in the male native Thai chickens. Thus, the objective of this study was to elucidate the localization of tyrosine hydroxylase (TH; a DA marker) neuronal groups in the brain of the roosters. MATERIAL AND METHODS: The distributions of TH immunoreactivity in the brain were detected utilizing the immunohistochemical technique. RESULTS: TH immunoreactivity was located throughout the brain and extensively in the diencephalon and mesencephalon. The highest density of TH-immunoreactive (-ir) neurons and fibers was found within the nucleus intramedialis (nI) and nucleus mamillaris lateralis (ML). The numbers of TH-ir neurons within the nucleus anterior medialis hypothalami (AM), nucleus paraventricularis magnocellularis (PVN), nI, and ML were then compared and revealed that the numbers of TH-ir neurons within the nI and ML were significantly higher than those of the AM and PVN. CONCLUSIONS: These present findings suggest that the DAergic neurons within the nI and ML might play an important role in the reproductive activities of the native Thai roosters. Interestingly, the DAergic system in the nI might be involved in male reproductive activities and/or parental behaviors in this equatorial species.

Neuropeptide Y and maternal behavior in the female native Thai chicken.

Maternal care behaviors in birds include incubation and rearing behaviors. During incubating period, the hens stop laying and eating less due to food restriction as a natural fasting when compared with the rearing hens, resulting in low production of eggs and chicks. Neuropeptide Y (NPY), a neurotransmitter/neuromodulator, is very well known to be involved in food intake regulation in birds and mammals. The objective of this study is to elucidate the association between NPY and maternal behaviors in the female native Thai chicken. The distributions of NPY-immunoreactive (-ir) neurons and fibers in the brain of the incubating (INC), nest-deprived (ND), and replaced-egg-with-chicks (REC) hens at day 6 were determined utilizing immunohistochemistry technique. The results revealed that the distributions of NPY-ir neurons and fibers were observed within the septalis lateralis, nucleus rotundus, and nucleus dorsolateralis anterior thalami, with predominantly located within the the nucleus paraventricularis magnocellularis (PVN). NPY-ir fibers were located throughout the brain and the densest NPY-ir fibers were distributed in a discrete region lying close to the ventriculus tertius (third ventricle) through the hypothalamus. Changes in the number of NPY-ir neurons within the PVN of the INC, ND, and REC hens were compared at different time points (at days 6 and 14). Interestingly, the number of NPY-ir neurons within the PVN was significantly higher (P < 0.05) in the INC hens when compared with those of the ND and REC hens at day 14 but not day 6. In addition, the number of NPY-ir neurons within the PVN of the INC hens was significantly increased (P < 0.05) from day 6 to day 14 but not the ND and REC hens. These results indicated, for the first time, the asscociation between NPY and maternal behaviors in the femle native Thai chicken. Change in the number of NPY-ir neurons within the PVN during the transition from incubating to rearing behavior suggested the possible role of NPY in the regulation of the maternal behaviors in this equatorial species. In addition, the native Thai chicken might be an excellent animal model for the study of this phenomenon.

Targeted differential monochromatic lighting improves broiler breeder reproductive performance.

Light perception in birds is composed of the retina and extraretinal sites, located in the brain. Previous studies indicate that selective photostimulation of the eye decreased reproductive performance, whereas extraretinal photostimulation increases it. Differential photostimulation of the retina and extraretinal sites is based on the retina's sensitivity to green wavelengths and on the red wavelengths' ability to penetrate body tissues. We previously found that short-day exposure to green light within a long-day exposure to red light increases reproductive activity in female turkeys and broiler breeder hens. Furthermore, in a study conducted recently in our laboratory, we found that blue light repressed expression of green light receptor in the retina, which can further enhance reproduction activity in broiler breeders. Here, we examined the "brain activate/eye deactivate" hypothesis on gonadal axis activity and reproductive performance in a broiler breeder flock. Broiler breeder hens and roosters (ROSS 308) were divided into 5 light-treatment groups (controlled rooms with light-emitting diodes [LED] lamps): warm white (control), long-day (14 h) red (630 nm) and short-day (6 h) green (514 nm) (red-green), long-day green and short-day red (green-red), long-day red and short-day blue (456 nm) (red-blue), and long-day blue and short-day red (blue-red). Birds were reared from 20 to 55 wk of age. Eggs were collected daily. Weekly egg production calculated. All eggs were incubated for fertility and hatchability examination. Blood was drawn monthly for plasma analysis. At 35 wk of age (after peak production) and 55 wk of age (end of the experiment), 10 hens from each treatment group were euthanized, and selected tissues and glands were taken for gene expression trials. Providing long-day red light to extraretinal photoreceptors while maintaining retinal photoreceptors on short day with blue or green light significantly improved reproductive activities, manifested by elevated egg production and gonadal axis activity compared with Controls and primary breeder recommendations. Long-day green light reduced reproductive performances. We suggest that targeted photostimulation enhances reproductive and gonadal axis activities in broiler breeders.

The effects of replacing eggs with chicks on mesotocin, dopamine, and prolactin in the native Thai hen.

The mesotocinergic (MTergic) and dopaminergic (DAergic) systems have been documented to play pivotal roles in maternal behaviors in native Thai chickens. In native Thai chickens, plasma prolactin (PRL) concentrations are associated with maternal behaviors, which are also controlled by the DAergic system. However, the role of MT in conjunction with the roles of DA and PRL on the neuroendocrine regulation of the transition from incubating to rearing behavior has never been studied. Therefore, the aim of this study was to investigate the association of MT, DA, and PRL during the transition from incubating to rearing behavior in native Thai hens. Using an immunohistochemistry technique, the numbers of MT-immunoreactive (-ir) and tyrosine hydroxylase-ir (TH-ir, a DA marker) neurons were compared between incubating hens (INC; n = 6) and hens for which the incubated eggs were replaced with 3 newly hatched chicks for 3 days after 6, 10, and 14 days of incubation (REC; n = 6). Plasma PRL concentrations were determined by enzyme-linked immunosorbent assay. The results revealed that the numbers of MT-ir neurons within the nucleus supraopticus, pars ventralis (SOv), nucleus preopticus medialis (POM), and nucleus paraventricularis magnocellularis (PVN) increased in the REC hens when compared with those of the INC hens at 3 different time points (at days 9, 13, and 17). On the other hand, the number of TH-ir neurons in the nucleus intramedialis (nI) decreased in the REC13 and REC17 hens when compared with those of the INC hens. However, the number of TH-ir neurons in the nucleus mamillaris lateralis (ML) only decreased in the REC13 hens when compared with the INC13 hens. The decrease in the numbers of TH-ir neurons within the nI and ML is associated with the decrease in the levels of plasma PRL. This study suggests that the presence of either eggs or chicks is the key factor regulating the MTergic system within the SOv, POM, and PVN and the DAergic system within the nI and ML during the transition from incubating to rearing behavior in native Thai chickens. The results further indicate that these two systems play pivotal roles in the transition from incubating to rearing behavior in this equatorial species.

Distribution of mesotocin-immunoreactive neurons in the brain of the male native Thai chicken.

Mesotocin (MT), a homolog of oxytocin (OT) in mammals, is a nonapeptide neurohypophysial hormone that is mainly synthesized in specific neuronal groups within the hypothalamus and released from the posterior pituitary gland in amphibian, reptilian, and avian species. MT is associated with the neuroendocrine regulation of reproductive cycle and maternal behaviors in female native Thai chickens. Male birds exhibit parental behaviors as well. However, there are limited data regarding the role(s) of the MTergic system in males. Thus, the objective of this study was to elucidate the localization of the MT neuronal groups in the brain of male native Thai chickens. The distributions of MT-immunoreactive (-ir) neurons and fibers in the brain were studied utilizing immunohistochemistry technique. The results revealed that MT-ir neurons and fibers were distributed throughout the brain and extensively in the diencephalon. MT-ir neurons and fibers were predominantly located within the nucleus supraopticus, pars ventralis (SOv), nucleus preopticus medialis (POM), nucleus ventrolateralis thalami (VLT), nucleus paraventricularis magnocellularis (PVN), and regio lateralis hypothalami (LHy), suggesting that MT neurons in these nuclei might be involved in the reproductive activities and/or parental behavior in the male chickens. In addition, the numbers of MT-ir neurons within the SOv and POM were significantly higher than those of the VLT, PVN, and LHy. More importantly, the number of MT-ir neurons in the SOv was high in the male brain when compared with the female brain, indicating that the MTergic system in the SOv might play a significant role in male reproductive activities in this equatorial species.

Effects of nest-deprivation on hypothalamic mesotocin in incubating native Thai hens (Gallus domesticus).

Avian mesotocin (MT) is homologous to oxytocin in mammals. Native Thai chickens (Gallus domesticus) strongly express maternal behaviors including incubation and rearing. However, the role of MT during incubation behavior has never been studied. The objective of this study was to determine the physiological function(s) of the MTergic system in incubation behavior in native Thai chickens. The brains were collected from incubating (INC) and nest-deprived (ND) hens at different time points (days 3, 6, 8, 10, 14, 18, and 21; n=6). Immunohistochemistry technique was used to compare the numbers of MT-immunoreactive (-ir) neurons between the INC and ND hens within the Nucleus supraopticus, pars ventralis (SOv), Nucleus preopticus medialis (POM), and Nucleus paraventricularis magnocellularis (PVN). The results revealed that the numbers of MT-ir neurons within the SOv, POM, and PVN remained high during the incubating stage. The number of MT-ir neurons in the SOv was lower than that of the POM and PVN. Disruption of incubation behavior by nest deprivation caused the numbers of MT-ir neurons within the SOv, POM, and PVN to decrease throughout the observation periods. For the first time, this study demonstrates that the MTergic system within the SOv, POM, and PVN may be involved with incubation behavior. In addition, these results further suggest that the MTergic neurons in these nuclei are not only regulated by rearing behavior but also might have a role in the initiation and maintenance of incubation behavior in this tropical species.

Distribution of hypothalamic vasoactive intestinal peptide immunoreactive neurons in the male native Thai chicken.

Avian prolactin (PRL) secretion is under stimulatory control by the PRL-releasing factor (PRF), vasoactive intestinal peptide (VIP). The neuroendocrine regulation of the avian reproductive system has been extensively studied in females. However, there are limited data in males. The aim of this study was to elucidate the VIPergic system and its relationship to PRL and testosterone (T) in the male native Thai chicken. The distributions of VIP-immunoreactive (-ir) neurons and fibers were determined by immunohistochemistry. Changes in VIP-ir neurons within the nucleus inferioris hypothalami (IH) and nucleus infundibuli hypothalami (IN) areas were compared across the reproductive stages. Plasma levels of PRL and T were determined by enzyme-linked immunosorbent assay and then compared across the reproductive stages. The results revealed that the highest accumulations of VIP-ir neurons were concentrated only within the IH-IN, and VIP-ir neurons were not detected within other hypothalamic nuclei. Within the IH-IN, VIP-ir neurons were low in premature and aging males and markedly increased in mature males. Changes in VIP-ir neurons within the IH-IN were directly mirrored with changes in PRL and T levels across the reproductive stages. These results suggested that VIP neurons in the IH-IN play a regulatory role in year-round reproductive activity in males. The present study also provides additional evidence that VIP is the PRF in non-seasonal, continuously breeding equatorial species.

Dopamine and prolactin involvement in the maternal care of chicks in the native Thai hen (Gallus domesticus).

The dopaminergic (DAergic) system plays a pivotal role in incubation behavior via the regulation of prolactin (PRL) secretion in birds, however the role of the DA/PRL system in rearing behavior is poorly understood. The objective of this study was to investigate the relationship between the DA/PRL system and rearing behavior in a gallinaceous bird, the native Thai chicken. Incubating native Thai hens were divided into two groups. In the first group, hens were allowed to care for their chicks (rearing hens; R). In the second group, hens were deprived of their chicks immediately after hatching (non-rearing hens; NR). In both groups, blood samples and brain sections were collected at different time points after the chicks hatched (days 4, 7, 10, 14, 17, 21, 24, and 28; 6 hens/time point/group). In this study, tyrosine hydroxylase (TH) was used as a marker for DAergic neurons. The numbers of TH-immunoreactive (-ir) neurons in the nucleus intramedialis (nI) and in the nucleus mamillaris lateralis (ML), which regulate the vasoactive intestinal peptide (VIP)/PRL system, were determined in R and NR hens utilizing immunohistochemical techniques. Plasma PRL levels were determined by enzyme-linked immunosorbent assays. The results revealed that both the number of TH-ir neurons in the nI and the plasma PRL levels were significantly higher in the R hens compared with the NR hens during the first 14 days of chick rearing (P<0.05). However, there was no significant change in the DAergic activity in the ML in either the R or NR groups throughout the 28-day rearing periods. These results suggest that the DA/PRL system is involved in early rearing behavior. The additional decline in DAergic activity and plasma PRL levels during the disruption of rearing behavior further supports their involvement in rearing behavior in this equatorial precocial species.

Differential roles of hypothalamic serotonin receptor subtypes in the regulation of prolactin secretion in the turkey hen.

In the turkey, exogenous serotonin (5-hydroxytryptamine, 5-HT) increases prolactin (PRL) secretion by acting through the dopaminergic (DAergic) system. In the present study, infusion of the 5-HT(2C) receptor agonist, (R)(-)-DOI hydrochloride (DOI), into the third ventricle stimulates PRL secretion, whereas the 5-HT(1A) receptor agonist, (+/-)-8-OH-DPAT hydrobromide (DPAT), inhibits PRL secretion. Using the immediate-early gene, c-fos, as an indicator of neuronal activity, in situ hybridization histochemistry showed preferential c-fos co-localization within tyrosine hydroxylase immunoreactive neurons (the rate limiting enzyme in DA synthesis) in the areas of the nucleus preopticus medialis (POM) and the nucleus premammillaris (PMM), in response to DPAT and DOI, respectively. To clarify the involvement of 5-HT(1A) and 5-HT(2C) receptors in PRL regulation, their mRNA expression was determined on hypothalamic tissue sections from birds in different reproductive stages. A significant difference in 5-HT1A receptor was observed, with the POM of hypoprolactinemic short day and photorefractory birds showing the highest expression. 5-HT2C receptors mRNA did not change during the reproductive cycle. The data presented support the notion that DA neurons in the PMM and POM mediate the stimulatory and inhibitory effects of 5-HT, respectively, on PRL secretion and the 5-HTergic system can both stimulate and inhibit PRL secretion.

Mesotocin and maternal care of chicks in native Thai hens (Gallus domesticus).

Oxytocin (OT) is known to induce and regulate maternal behaviors in mammals via the supraoptic nucleus and paraventricular nucleus (PVN), whereas the function of mesotocin (MT; the avian homolog of OT) is poorly understood in birds. To elucidate the association of MT and the regulation of maternal behaviors in birds, we studied changes in the number of MT-immunoreactive (ir) neurons in native Thai chickens using immunohistochemistry. We observed that MT-ir neurons and fibers appeared in discrete regions located close to the third ventricle from the level of the preoptic area through the anterior hypothalamus with an abundance observed in the nucleus supraopticus, pars ventralis (SOv), nucleus preopticus medialis (POM), and PVN. The number of MT-ir neurons was low in the SOv, POM, and PVN of non-laying hens, but it increased gradually when the hens entered the laying stage, and peaked in incubating and rearing hens. We compared the number of MT-ir neurons in the SOv, POM, and PVN of native Thai hens rearing chicks (R) with that of non-rearing chicks (NR). The number of MT-ir neurons was high in the R hens, but low in the NR hens in these nuclei. For the first time, these results indicate that the association between the MT neurons and the presence of chicks might, in part, play a role in the neuroendocrine reorganization to establish and maintain maternal behaviors in native Thai chickens. MTergic activity is likely related to the contribution of rearing behavior in this equatorial precocial species.

Photoreceptive oscillators within neurons of the premammillary nucleus (PMM) and seasonal reproduction in temperate zone birds.

The pathway for light transmission regulating the reproductive neuroendocrine system in temperate zone birds remains elusive. Based on the evidence provided from our studies with female turkeys, it is suggested that the circadian clock regulating reproductive seasonality is located in putatively photosensitive dopamine-melatonin (DA-MEL) neurons residing in the premammillary nucleus (PMM) of the caudal hypothalamus. Melanopsin is expressed by these neurons; a known photopigment which mediates light information pertaining to the entrainment of the clock. Exposure to a gonad stimulatory photoperiod enhances the activity of the DAergic system within DA-MEL neurons. DAergic activity encoding the light information is transmitted to the pars tuberalis, where thyroid-stimulating hormone, beta (TSHß) cells reside, and induces the release of TSH. TSH stimulates tanycytes lining the base of the third ventricle and activates type 2 deiodinase in the ependymal which enhances triiodothyronine (T3) synthesis. T3 facilitates the release of gonadotropin-releasing hormone-I which stimulates luteinizing hormone/follicle stimulating hormone release and gonad recrudescence. These data taken together with the findings that clock genes are rhythmically expressed in the PMM where DA-MEL neurons are localized imply that endogenous oscillators containing photoreceptors within DA-MEL neurons are important in regulating the DA and MEL rhythms that drive the circadian cycle controlling seasonal reproduction.

Gonadotropin releasing hormone and brooding behavior in the native Thai hen.

Changes in the number of hypothalamic gonadotropin releasing hormone-I (GnRH-I) neurons within the Nucleus commissurae pallii (nCPa) were associated with the reproductive cycle of native Thai chickens. In order to further understand the association of GnRH-I in the regulation of brooding behavior in this bird, the native Thai chickens were divided into two groups; chick-rearing (R) and non-chick-rearing (NR) hens. Numbers of visible of GnRH-I-immunoreactive (GnRH-I-ir) neurons in the hypothalamus of R and NR hens were compared utilizing immunohistochemistry. Numbers of visible GnRH-I-ir neurons within the Nucleus anterior medialis hypothalami, Nucleus suprachaiasmaticus, pars medialis, Nucleus septalis lateralis, Nucleus paraventricularis magnocellularis, and Regio lateralis hypothalami areas were observed in both groups, but no differences were seen between R and NR hens. The number of visible GnRH-I neurons in the nCPa was higher (P<0.05) in the NR than in R hens, and increased in NR hens by day 14 after chick removal. These findings suggest, for the first time, an association of the GnRH system with brooding behavior in continuously breeding birds. Furthermore, the expression of brooding behavior of native Thai chickens might be regulated, in part, by GnRH-I neurons in the nCPa.

Neuroendocrine regulation of rearing behavior in the native Thai hen.

Vasoactive intestinal peptide (VIP) is the avian prolactin releasing factor and changes in the concentrations of plasma prolactin (PRL) are found during the avian reproductive cycle. This study investigated the changes in the VIP/PRL system of native Thai hens rearing their young as compared to hens deprived of rearing their chicks. The number of VIP-immunoreactive (VIP-ir) neurons in the Nucleus inferioris hypothalami (IH) and Nucleus infundibuli hypothalami (IN) of hens rearing chicks (R) were compared with those of non-rearing chicks (NR). Plasma PRL levels were determined by enzyme-linked immunosorbent assay. The localization and number of VIP-ir neurons were determined by immunohistochemistry. The numbers of VIP-ir neurons in the IH-IN areas were high in the R hens, whereas the number of VIP-ir neurons decreased in the NR hens as compared to their respective R hens. During the rearing period, changes in the VIP-ir neurons within the IH-IN were correlated with plasma PRL levels. The results of the present study indicate for the first time that the VIP/PRL system plays a role in neuroendocrine reorganization to establish maternal behavior in native Thai chickens. The VIP/PRL system functions not only as a well established key regulator of incubation behavior, but is also involved in the regulation of rearing behavior. It is possible that VIP and the decline in the number of VIP-ir neurons and in turn VIPergic activity and the decrease in PRL levels are related to their contribution to rearing behavior of this non-seasonal breeding, equatorial precocial species.

Plasma leptin concentrations during the reproductive cycle in the native Thai chicken (Gallus domesticus).

Plasma leptin concentrations were investigated during the reproductive cycle in the native Thai chicken. The plasma leptin concentration was high during non-laying (0.69±0.15ng/ml), lowered to a minimum concentration during egg laying (0.07±0.02ng/ml), and gradually increased during egg incubation and rearing of the chicks (0.53±0.22 and 0.74±0.29ng/ml, respectively). However, the differences were not significant. Incubating chickens that were deprived of their nests for 3 weeks showed a significant decrease in plasma leptin concentrations (0.29±0.04ng/ml, P<0.05) compared to those of their corresponding incubating controls (0.77±0.08ng/ml). Similarly, plasma leptin concentration of chickens that were deprived of their chicks for 4 weeks was significantly lower (0.09±0.11ng/ml, P<0.05), when compared to those of chickens that rearing their chicks (0.71±0.18ng/ml). These findings taken together with the results that the low plasma leptin concentrations were observed in chickens having relatively greater ovary and oviduct weights led to the suggestion that circulating leptin concentrations are associated with the reproductive states of the birds, especially the ovarian activity (i.e. ovarian steroid hormone concentrations) in the native Thai chicken, a tropical and continuous breeding species.

Distribution and variation in gonadotropin releasing hormone-I (GnRH-I) immunoreactive neurons in the brain of the native Thai chicken during the reproductive cycle.

Gonadotropin releasing hormone-I (GnRH-I) is known to regulate the avian reproductive system. We investigated the roles of GnRH-I in the regulation of the reproductive system of the native Thai chicken. The distribution of GnRH-I neurons and changes in GnRH-I-immunoreactive (-ir) neurons throughout the reproductive stages and between incubating and nest-deprived hens were analyzed utilizing immunohistochemical techniques. The results revealed that GnRH-I-ir neurons were distributed in a discrete region lying close to the third ventricle from the level of preoptic area through the anterior hypothalamus, with the greatest abundance found within the nucleus commissurae pallii (nCPa). The number of GnRH-I-ir neurons in the nCPa was highest in laying hens when compared with that in the other reproductive stages. Nest deprivation caused an increase in the number of GnRH-I-ir neurons in the nCPa of nest-deprived hens when compared with incubating hens. These results indicate that GnRH-I expression is correlated with the reproductive state in the native Thai chicken and may be, in part, regulated by it. This study also confirms a pivotal role of GnRH-I in controlling avian reproduction of this non-seasonal breeding, equatorial species.

Presence of prolactin mRNA in extra-pituitary brain areas in the domestic turkey.

It is well known that prolactin plays diverse roles in vertebrate reproduction. Besides expression in the pituitary, prolactin is also found in extra-pituitary tissues. In the present study, prolactin mRNA expression was studied utilizing in situ hybridization histochemistry. Prolactin mRNA, while found throughout the turkey brain, was predominantly localized within the pituitary, confirming a pivotal role of prolactin in turkey reproduction. The expression of prolactin mRNA was also observed within extra-pituitary brain areas including the cerebellum, nucleus accumbens, lateral septum, anterior hypothalamic nucleus, lateral hypothalamus, paraventricular nucleus, ventromedial nucleus, and infundibular nuclear complex. In the hypothalamus, an abundance of prolactin mRNA-expressing cells was observed in the anterior hypothalamic nucleus, lateral hypothalamus, and ventromedial nucleus. Cells expressing the least prolactin mRNA were found in the lateral septum, paraventricular nucleus, and the infundibular nuclear complex. This study reveals, for the first time, that prolactin mRNA was expressed in extra-pituitary brain areas in birds. In addition, the diverse expression of prolactin mRNA in the brain areas suggests that prolactin plays various physiological roles in birds.

Role of vitelline envelope during fertilization in the black tiger shrimp, Penaeus monodon.

Animal eggs possess investments through which sperm must penetrate. The aim of the present study was to investigate the role of the egg coating, the vitelline envelope, during sperm-egg interactions in the black tiger shrimp, Penaeus monodon. The site(s) of primary binding between sperm and egg and the possible binding molecule(s) for sperm were identified. In vitro adsorption of the vitelline envelope protein onto the sperm surface showed that primary binding occurred between the sperm anterior spike of acrosome intact sperm and the vitelline envelope. Results from streptavidin blotting revealed that the component of the vitelline envelope that interacts with the sperm integral membrane protein is a 370kDa protein. In addition, it was shown that the vitelline envelope protein had no ability to induce acrosome reaction. These results suggest that the function of the vitelline envelope is as a primary binding site for sperm in shrimp, but not a sole trigger for the acrosome reaction.

Changes in vasoactive intestinal peptide and tyrosine hydroxylase immunoreactivity in the brain of nest-deprived native Thai hen.

Hyperprolactinemia is associated with incubation behavior and ovarian regression in birds. To investigate the association of prolactin (PRL), vasoactive intestinal peptide (VIP), and dopamine (DA) with the neuroendocrine regulation of incubation behavior, changes in the number of visible VIP-immunoreactive (VIP-ir) neurons in the nucleus inferioris hypothalami (IH) and nucleus infundibuli hypothalami (IN) and tyrosine hydroxylase-immunoreactive (TH-ir) neurons in the nucleus intramedialis (nI) and nucleus mamillaris lateralis (ML) of incubating native Thai hens were compared with those of nest-deprived hens. TH was used as a marker for dopaminergic (DAergic) neurons. Blood samples were collected to determine PRL levels. The localization and the number of visible VIP-ir and TH-ir neurons were determined by immunohistochemistry. Disruption of incubation behavior was accompanied by a precipitous decline in plasma PRL levels. The number of visible VIP-ir neurons in the IH-IN and TH-ir neurons in the nI and ML were high during incubation and decreased when hens were deprived of their nests. This study indicated an association between VIP neurons in the IH-IN and DA neurons in the nI and ML with the degree of hyperprolactinemia, suggesting that the expression of incubation behavior in birds might be, in part, regulated by the DAergic input from the nI and ML to VIP neurons in the IH-IN and subsequent PRL release.

Serotonin receptor subtypes influence prolactin secretion in the turkey.

Serotonin (5-HT) stimulation of prolactin (PRL) secretion is mediated through the dopaminergic (DAergic) system, with 5-HT ligands having no direct effect on pituitary PRL release. Infusion of 5-HT into the third ventricle (ICV) or electrical stimulation (ES) of the medial preoptic area (POM) or the ventromedial nucleus (VMN) induces an increase in circulating PRL in the turkey. These increases in PRL do not occur when a selective antagonist blocks the D(1) dopamine (DA) receptors in the infundibular area (INF). In this study, the ICV infusion of (R)(-)-DOI hydrochloride (DOI), a selective 5-HT(2A) eceptor agonist, caused PRL to increase. Pretreatment with Ketanserin tartrate salt (KETAN), a selective 5-HT(2A) receptor antagonist, blocked DOI-induced PRL secretion, attesting to the specificity of the response. DOI-induced PRL secretion was prevented when the D(1) DA receptors in the INF were blocked by the D(1) DA receptor antagonist, R(+)-SCH-23390 hydrochloride microinjection, suggesting that the DAergic activation of the vasoactive intestinal peptide (VIP)/PRL system is mediated by a stimulatory 5-HT(2A) receptor subtype. The DOI-induced PRL increase did not occur when (+/-)-8-OH-DPAT (DPAT) was concurrently infused. DPAT is a 5-T(1A) receptor agonist which appears to mediate the inhibitory influence of 5-HT on PRL secretion. When DPAT was microinjected directly into the VMN, it blocked the PRL release affected by ES in the POM. These data suggested that when 5-HT(2A) receptors are activated, they influence the release of DA to the INF. When 5-HT(1A) receptors are stimulated, they somehow inhibit the PRL-releasing actions of 5-HT(2A) receptors. This inhibition could take place centrally, or it could occur postsynaptically at the pituitary level. It is known that D(2) DA receptors in the pituitary antagonize PRL-releasing effect of VIP. A release of DA to the pituitary, initiated by 5-HT(1A) receptors, could effectively inhibit PRL secretion.