Androgen modulation of neurotransmitter receptor density was investigated in the nucleus intercol... more Androgen modulation of neurotransmitter receptor density was investigated in the nucleus intercollicularis (ICo) of male and female Japanese quail (Coturnix coturnix japonica). ICo appears to play an important role in the neural control of testosterone (T) dependent vocal behaviour. Two receptor types were investigated in this nucleus; muscarinic cholinergic receptors, labelled using [3H]N-methyl scopolamine (NMS) as the ligand, and alpha2-adrenergic receptors, labelled using [3H]p-amino-clonidine (PAC) as the ligand. Changes in receptor density were assessed using in vitro quantitative autoradiography to ensure a high degree of anatomical specificity in the identification of any steroid effects. Gonadectomy was found to reduce the density of both [3H]PAC bindings sites and [3H]NMS in specific subregions of ICo. Gonadectomized animals treated with T had levels of receptor density similar to intact birds. However, the location within ICo of the effects of T was different for each lig...
The functions of song and the contextual cues that elicit song change seasonally in parallel with... more The functions of song and the contextual cues that elicit song change seasonally in parallel with testosterone (T) concentrations in male European starlings. T is high in spring when at least one function of male song is that of immediate mate attraction, and low outside the context of ...
A partial estrogen receptor- (ER) cDNA had been previously cloned and sequenced in Japanese qua... more A partial estrogen receptor- (ER) cDNA had been previously cloned and sequenced in Japanese quail. The 3-and 5-rapid amplification of cDNA ends techniques were used here to identify a cDNA sequence of the quail ER that contains a complete open reading frame. For the first time in an avian species, this cDNA sequence and the corresponding amino acid sequence are described. They are compared with the known ER sequences previously described in mammals and with the ER␣ sequences identified in a selection of mammalian and avian species. The analysis by Northern blotting of the ER mRNA expression in the brain and kidneys revealed the presence of several transcripts. The presence of ER identified by reverse transcriptase-polymerase chain reaction demonstrated a widespread distribution quite different from the distribution of ER␣. The complete neuroanatomical distri-bution of ER mRNA as determined by in situ hybridization with 35 S-and 33 P-labeled oligoprobes is also presented. Transcripts are present in many nuclei implicated in the control of reproduction such as the medial preoptic nucleus, the nucleus striae terminalis, and the nucleus taeniae, the avian homologue of the amygdala. These data demonstrate the presence of ER in a nonmammalian species and indicate that the (neuro)anatomical distribution of this receptor type has been conserved in these two classes of vertebrates. The role of this receptor in the control of reproduction and other physiological processes should now be investigated.
Gonadotropin-releasing hormone-I (GnRH-I) cells are localized primarily to the septopreoptic area... more Gonadotropin-releasing hormone-I (GnRH-I) cells are localized primarily to the septopreoptic area (POA) and are responsible for regulating gonadotropin release from the anterior pituitary. Some songbird species exhibit dramatic seasonal variation in the number of detectable GnRH-I immunoreactive cells, with higher numbers being observed during the breeding season. Here we investigated the anatomical distribution of GnRH-I-immunoreactive cells in male starlings that varied in response to manipulations of reproductive state, social context, and gonadal condition. We housed photostimulated, intact and castrated male starlings with a female or alone. Additionally, a fifth treatment group consisted of photorefractory males (i.e., in a nonreproductive state) housed alone. All photostimulated males had significantly greater numbers of GnRH-I cells compared with photorefractory male starlings. There was a significant main effect of castration and social context. Castrated males had significantly greater numbers of GnRH-I cells compared with intact males, and males housed in male-female dyads also had greater numbers of GnRH-I cells. Furthermore, the significant main effects of castration and social context were the result of an increase in GnRH-I cell numbers specifically in the rostral and intermediate regions of the POA. These findings indicate that social context and hormonal milieu have profound effects on GnRH-I immunoreactivity in addition to the previously described effects of reproductive state. These data provide novel insight into the environmental regulation of the hypothalamopituitary axis and suggest that gonadal hormones and female presence independently regulate GnRH-I cells in specific regions of the POA in male starlings.
Estradiol facilitates the expression of male sexual behavior in Japanese quail within a few minut... more Estradiol facilitates the expression of male sexual behavior in Japanese quail within a few minutes. These rapid behavioral effects of estradiol could result from rapid changes in its local production in the preoptic area by aromatase, the enzyme converting testosterone into estradiol. Alternatively, aromatase activity may remain constant but fluctuations of local estradiol production could arise from rapid changes in
Estradiol (E2) can act in the brain in a relatively fast manner (i.e., seconds to minutes) usuall... more Estradiol (E2) can act in the brain in a relatively fast manner (i.e., seconds to minutes) usually through signaling initiated at the cell membrane. Brain-derived E2 has thus been considered as another type of neurotransmitter. Recent work found that behaviors indicative of male sexual motivation are activated by estrogenic metabolites of testosterone (T) in a fast manner, while sexual performance (copulatory behavior per se) is regulated by brain E2 in a slower manner via nucleus-initiated actions. This functional division between these two types of action appears to generalize to other behavioral systems regulated by E2. We propose the dual action of estrogen hypothesis to explain this functional distinction between these two different modes of action.
European starlings (Sturnus vulgaris) exhibit seasonal changes in singing and in the volumes of t... more European starlings (Sturnus vulgaris) exhibit seasonal changes in singing and in the volumes of the neural substrate. Increases in song nuclei volume are mediated at least in part by increases in day length, which is also associated with increases in plasma testosterone (T), reproductive activity, and singing behavior in males. The correlations between photoperiod (i.e. daylength), T, reproductive state and singing hamper our ability to disentangle causal relationships. We investigated how photoperiodic-induced variation in reproductive state modulates the effects of T on singing behavior and song nuclei volumes in adult female starlings. Female starlings do not naturally produce measureable levels of circulating T but nevertheless respond to exogenous T, which induces male-like singing. We manipulated photoperiod by placing birds in a photosensitive or photorefractory state and then treated them with T-filled or empty silastic implants. We recorded morning singing behavior for 3 weeks, after which we assessed reproductive condition and measured song nuclei volumes. We found that T-treated photosensitive birds sang significantly more than all other groups including T-treated photorefractory birds. All T-treated birds had larger song nuclei volumes than with blank-treated birds (despite photorefractory T-treated birds not increasing song-rate). There was no effect of photoperiod on the song nuclei volumes of T-treated birds. These data show that the behavioral effects of exogenous T can be modulated by reproductive state in adult female songbirds. Furthermore, these data are consistent with other observations that increases in singing rate in response to T are not necessarily due to the direct effects of T on song nuclei volume.
Learned communication is a trait observed in a limited number of vertebrates such as humans but a... more Learned communication is a trait observed in a limited number of vertebrates such as humans but also songbirds (i.e. species in the suborder passeri sometimes called oscines). Robust male-biased sex-differences in song development and production have been observed in several songbird species. However, in some of these species treating adult females with testosterone (T) induces neuro-behavioral changes such that females become more male-like in brain and behavior. T-treatment in these adult females seems to stimulate sensorimotor song development to facilitate song masculinization. In male songbirds it is known that the lateral magnocellular nucleus of the anterior nidopallium (LMAN) plays a modulatory role during song development. LMAN is androgen sensitive and may be a key target of a T-induced recapitulation of a developmental process in adult females. We tested this hypothesis. Adult female canaries were given either a chemical lesion of LMAN or a control sham-surgery. Prior to ...
The Journal of comparative neurology, Jan 15, 2014
Although the existence of newborn neurons had originally been suggested, but not broadly accepted... more Although the existence of newborn neurons had originally been suggested, but not broadly accepted, based on studies in adult rodent brains, the presence of an active neurogenesis process in adult homoeothermic vertebrates was first firmly established in songbirds. Adult neurogenesis was initially studied with the tritiated thymidine technique, later replaced by the injection and detection of the marker of DNA replication 5-bromo-2'-deoxyuridine (BrdU). More recently, various endogenous markers were used to identify young neurons or cycling neuronal progenitors. We review here the respective advantages and pitfalls of these different approaches in birds, with specific reference to the microtubule-associated protein, doublecortin (DCX), that has been extensively used to identify young newly born neurons in adult brains. All these techniques of course have limitations. Exogenous markers label cells replicating their DNA only during a brief period and it is difficult to select injec...
Variation in environmental factors such as day length and social context greatly affects reproduc... more Variation in environmental factors such as day length and social context greatly affects reproductive behavior and the brain areas that regulate these behaviors. One such behavior is song in songbirds, which males use to attract a mate during the breeding season. In these species the absence of a potential mate leads to an increase in the number of songs produced, while the presence of a mate greatly diminishes singing. Interestingly, although long days promote song behavior, producing song itself can promote the incorporation of new neurons in brain regions controlling song output. Social context can also affect such neuroplasticity in these song control nuclei. The goal of the present study was to investigate in canaries (Serinus canaria), a songbird species, how photoperiod and social context affect song and the incorporation of new neurons, as measured by the microtubule-associated protein doublecortin (DCX) in HVC, a key vocal production brain region of the song control system....
In seasonally breeding male songbirds, both the function of song and the stimuli that elicit sing... more In seasonally breeding male songbirds, both the function of song and the stimuli that elicit singing behavior change seasonally. The catecholamine norepinephrine (NE) modulates attention and arousal across behavioral states, yet the role of NE in seasonally-appropriate vocal communication has not been well-studied. The present study explored the possibility that seasonal changes in alpha 2-noradrenergic receptors (α(2)-R) within song control regions and brain regions implicated in sexual arousal and social behavior contribute to seasonal changes in song behavior in male European starlings (Sturnus vulgaris). We quantified singing behavior in aviary housed males under spring breeding season conditions and fall conditions. α(2)-R were identified with the selective ligand [(3)H]RX821002 using autoradiographic methods. The densities of α(2)-R in song control regions (HVC and the robust nucleus of the arcopallium [RA]) and the lateral septum (LS) were lower in Spring Condition males. α(2...
Research in songbirds shows that singing behavior is regulated by both brain areas involved in vo... more Research in songbirds shows that singing behavior is regulated by both brain areas involved in vocal behavior as well as those involved in social behavior. Interestingly, the precise role of these regions in song can vary as a function of the social, environmental and breeding context. To date, little is known about the neurotransmitters underlying such context-dependent regulation of song. Dopamine (DA) modulates highly motivated, goal-directed behaviors (including sexually motivated song) and emerging data implicate DA in the context-dependent regulation of singing behavior. This study was performed to begin to examine whether differences in DA receptors may underlie, in part, context-dependent differences in song production. We used autoradiographic procedures to label D1-like and D2-like DA receptors to examine the relationship between DA receptor density and singing behavior in multiple contexts in male European starlings (Sturnus vulgaris). Within a breeding context (when test...
Songbirds produce learned vocalizations that are controlled by a specialized network of neural st... more Songbirds produce learned vocalizations that are controlled by a specialized network of neural structures, the song control system. Several nuclei in this song control system demonstrate a marked degree of adult seasonal plasticity. Nucleus volume varies seasonally based on changes in cell size or spacing, and in the case of nucleus HVC and area X on the incorporation of new neurons. Reelin, a large glycoprotein defective in reeler mice, is assumed to determine the final location of migrating neurons in the developing brain. In mammals, reelin is also expressed in the adult brain but its functions are less well characterized. We investigated the relationships between the expression of reelin and/or its receptors and the dramatic seasonal plasticity in the canary (Serinus canaria) brain. We detected a broad distribution of the reelin protein, its mRNA and the mRNAs encoding for the reelin receptors (VLDLR and ApoER2) as well as for its intracellular signaling protein, Disabled1. Thes...
Many sex differences in brain and behavior related to reproduction are thought to have evolved ba... more Many sex differences in brain and behavior related to reproduction are thought to have evolved based on sexual selection involving direct competition for mates during male-male competition and female choice. Therefore, certain aspects of brain circuitry can be viewed as secondary sexual characteristics. The study of proximate causes reveals that sex differences in the brain of mammals and birds reflect organizational and activational effects of sex steroids as articulated by Young and collaborators. However, sex differences in brain and behavior have been identified in the cognitive domain with no obvious link to reproduction. Recent views of sexual selection advocate for a broader view of how intra-sexual selection might occur including such examples as competition within female populations for resources that facilitate access to mates rather than mating competition per se. Sex differences can also come about for other reasons than sexual selection and recent work on neuroendocrine...
Androgen modulation of neurotransmitter receptor density was investigated in the nucleus intercol... more Androgen modulation of neurotransmitter receptor density was investigated in the nucleus intercollicularis (ICo) of male and female Japanese quail (Coturnix coturnix japonica). ICo appears to play an important role in the neural control of testosterone (T) dependent vocal behaviour. Two receptor types were investigated in this nucleus; muscarinic cholinergic receptors, labelled using [3H]N-methyl scopolamine (NMS) as the ligand, and alpha2-adrenergic receptors, labelled using [3H]p-amino-clonidine (PAC) as the ligand. Changes in receptor density were assessed using in vitro quantitative autoradiography to ensure a high degree of anatomical specificity in the identification of any steroid effects. Gonadectomy was found to reduce the density of both [3H]PAC bindings sites and [3H]NMS in specific subregions of ICo. Gonadectomized animals treated with T had levels of receptor density similar to intact birds. However, the location within ICo of the effects of T was different for each lig...
The functions of song and the contextual cues that elicit song change seasonally in parallel with... more The functions of song and the contextual cues that elicit song change seasonally in parallel with testosterone (T) concentrations in male European starlings. T is high in spring when at least one function of male song is that of immediate mate attraction, and low outside the context of ...
A partial estrogen receptor- (ER) cDNA had been previously cloned and sequenced in Japanese qua... more A partial estrogen receptor- (ER) cDNA had been previously cloned and sequenced in Japanese quail. The 3-and 5-rapid amplification of cDNA ends techniques were used here to identify a cDNA sequence of the quail ER that contains a complete open reading frame. For the first time in an avian species, this cDNA sequence and the corresponding amino acid sequence are described. They are compared with the known ER sequences previously described in mammals and with the ER␣ sequences identified in a selection of mammalian and avian species. The analysis by Northern blotting of the ER mRNA expression in the brain and kidneys revealed the presence of several transcripts. The presence of ER identified by reverse transcriptase-polymerase chain reaction demonstrated a widespread distribution quite different from the distribution of ER␣. The complete neuroanatomical distri-bution of ER mRNA as determined by in situ hybridization with 35 S-and 33 P-labeled oligoprobes is also presented. Transcripts are present in many nuclei implicated in the control of reproduction such as the medial preoptic nucleus, the nucleus striae terminalis, and the nucleus taeniae, the avian homologue of the amygdala. These data demonstrate the presence of ER in a nonmammalian species and indicate that the (neuro)anatomical distribution of this receptor type has been conserved in these two classes of vertebrates. The role of this receptor in the control of reproduction and other physiological processes should now be investigated.
Gonadotropin-releasing hormone-I (GnRH-I) cells are localized primarily to the septopreoptic area... more Gonadotropin-releasing hormone-I (GnRH-I) cells are localized primarily to the septopreoptic area (POA) and are responsible for regulating gonadotropin release from the anterior pituitary. Some songbird species exhibit dramatic seasonal variation in the number of detectable GnRH-I immunoreactive cells, with higher numbers being observed during the breeding season. Here we investigated the anatomical distribution of GnRH-I-immunoreactive cells in male starlings that varied in response to manipulations of reproductive state, social context, and gonadal condition. We housed photostimulated, intact and castrated male starlings with a female or alone. Additionally, a fifth treatment group consisted of photorefractory males (i.e., in a nonreproductive state) housed alone. All photostimulated males had significantly greater numbers of GnRH-I cells compared with photorefractory male starlings. There was a significant main effect of castration and social context. Castrated males had significantly greater numbers of GnRH-I cells compared with intact males, and males housed in male-female dyads also had greater numbers of GnRH-I cells. Furthermore, the significant main effects of castration and social context were the result of an increase in GnRH-I cell numbers specifically in the rostral and intermediate regions of the POA. These findings indicate that social context and hormonal milieu have profound effects on GnRH-I immunoreactivity in addition to the previously described effects of reproductive state. These data provide novel insight into the environmental regulation of the hypothalamopituitary axis and suggest that gonadal hormones and female presence independently regulate GnRH-I cells in specific regions of the POA in male starlings.
Estradiol facilitates the expression of male sexual behavior in Japanese quail within a few minut... more Estradiol facilitates the expression of male sexual behavior in Japanese quail within a few minutes. These rapid behavioral effects of estradiol could result from rapid changes in its local production in the preoptic area by aromatase, the enzyme converting testosterone into estradiol. Alternatively, aromatase activity may remain constant but fluctuations of local estradiol production could arise from rapid changes in
Estradiol (E2) can act in the brain in a relatively fast manner (i.e., seconds to minutes) usuall... more Estradiol (E2) can act in the brain in a relatively fast manner (i.e., seconds to minutes) usually through signaling initiated at the cell membrane. Brain-derived E2 has thus been considered as another type of neurotransmitter. Recent work found that behaviors indicative of male sexual motivation are activated by estrogenic metabolites of testosterone (T) in a fast manner, while sexual performance (copulatory behavior per se) is regulated by brain E2 in a slower manner via nucleus-initiated actions. This functional division between these two types of action appears to generalize to other behavioral systems regulated by E2. We propose the dual action of estrogen hypothesis to explain this functional distinction between these two different modes of action.
European starlings (Sturnus vulgaris) exhibit seasonal changes in singing and in the volumes of t... more European starlings (Sturnus vulgaris) exhibit seasonal changes in singing and in the volumes of the neural substrate. Increases in song nuclei volume are mediated at least in part by increases in day length, which is also associated with increases in plasma testosterone (T), reproductive activity, and singing behavior in males. The correlations between photoperiod (i.e. daylength), T, reproductive state and singing hamper our ability to disentangle causal relationships. We investigated how photoperiodic-induced variation in reproductive state modulates the effects of T on singing behavior and song nuclei volumes in adult female starlings. Female starlings do not naturally produce measureable levels of circulating T but nevertheless respond to exogenous T, which induces male-like singing. We manipulated photoperiod by placing birds in a photosensitive or photorefractory state and then treated them with T-filled or empty silastic implants. We recorded morning singing behavior for 3 weeks, after which we assessed reproductive condition and measured song nuclei volumes. We found that T-treated photosensitive birds sang significantly more than all other groups including T-treated photorefractory birds. All T-treated birds had larger song nuclei volumes than with blank-treated birds (despite photorefractory T-treated birds not increasing song-rate). There was no effect of photoperiod on the song nuclei volumes of T-treated birds. These data show that the behavioral effects of exogenous T can be modulated by reproductive state in adult female songbirds. Furthermore, these data are consistent with other observations that increases in singing rate in response to T are not necessarily due to the direct effects of T on song nuclei volume.
Learned communication is a trait observed in a limited number of vertebrates such as humans but a... more Learned communication is a trait observed in a limited number of vertebrates such as humans but also songbirds (i.e. species in the suborder passeri sometimes called oscines). Robust male-biased sex-differences in song development and production have been observed in several songbird species. However, in some of these species treating adult females with testosterone (T) induces neuro-behavioral changes such that females become more male-like in brain and behavior. T-treatment in these adult females seems to stimulate sensorimotor song development to facilitate song masculinization. In male songbirds it is known that the lateral magnocellular nucleus of the anterior nidopallium (LMAN) plays a modulatory role during song development. LMAN is androgen sensitive and may be a key target of a T-induced recapitulation of a developmental process in adult females. We tested this hypothesis. Adult female canaries were given either a chemical lesion of LMAN or a control sham-surgery. Prior to ...
The Journal of comparative neurology, Jan 15, 2014
Although the existence of newborn neurons had originally been suggested, but not broadly accepted... more Although the existence of newborn neurons had originally been suggested, but not broadly accepted, based on studies in adult rodent brains, the presence of an active neurogenesis process in adult homoeothermic vertebrates was first firmly established in songbirds. Adult neurogenesis was initially studied with the tritiated thymidine technique, later replaced by the injection and detection of the marker of DNA replication 5-bromo-2'-deoxyuridine (BrdU). More recently, various endogenous markers were used to identify young neurons or cycling neuronal progenitors. We review here the respective advantages and pitfalls of these different approaches in birds, with specific reference to the microtubule-associated protein, doublecortin (DCX), that has been extensively used to identify young newly born neurons in adult brains. All these techniques of course have limitations. Exogenous markers label cells replicating their DNA only during a brief period and it is difficult to select injec...
Variation in environmental factors such as day length and social context greatly affects reproduc... more Variation in environmental factors such as day length and social context greatly affects reproductive behavior and the brain areas that regulate these behaviors. One such behavior is song in songbirds, which males use to attract a mate during the breeding season. In these species the absence of a potential mate leads to an increase in the number of songs produced, while the presence of a mate greatly diminishes singing. Interestingly, although long days promote song behavior, producing song itself can promote the incorporation of new neurons in brain regions controlling song output. Social context can also affect such neuroplasticity in these song control nuclei. The goal of the present study was to investigate in canaries (Serinus canaria), a songbird species, how photoperiod and social context affect song and the incorporation of new neurons, as measured by the microtubule-associated protein doublecortin (DCX) in HVC, a key vocal production brain region of the song control system....
In seasonally breeding male songbirds, both the function of song and the stimuli that elicit sing... more In seasonally breeding male songbirds, both the function of song and the stimuli that elicit singing behavior change seasonally. The catecholamine norepinephrine (NE) modulates attention and arousal across behavioral states, yet the role of NE in seasonally-appropriate vocal communication has not been well-studied. The present study explored the possibility that seasonal changes in alpha 2-noradrenergic receptors (α(2)-R) within song control regions and brain regions implicated in sexual arousal and social behavior contribute to seasonal changes in song behavior in male European starlings (Sturnus vulgaris). We quantified singing behavior in aviary housed males under spring breeding season conditions and fall conditions. α(2)-R were identified with the selective ligand [(3)H]RX821002 using autoradiographic methods. The densities of α(2)-R in song control regions (HVC and the robust nucleus of the arcopallium [RA]) and the lateral septum (LS) were lower in Spring Condition males. α(2...
Research in songbirds shows that singing behavior is regulated by both brain areas involved in vo... more Research in songbirds shows that singing behavior is regulated by both brain areas involved in vocal behavior as well as those involved in social behavior. Interestingly, the precise role of these regions in song can vary as a function of the social, environmental and breeding context. To date, little is known about the neurotransmitters underlying such context-dependent regulation of song. Dopamine (DA) modulates highly motivated, goal-directed behaviors (including sexually motivated song) and emerging data implicate DA in the context-dependent regulation of singing behavior. This study was performed to begin to examine whether differences in DA receptors may underlie, in part, context-dependent differences in song production. We used autoradiographic procedures to label D1-like and D2-like DA receptors to examine the relationship between DA receptor density and singing behavior in multiple contexts in male European starlings (Sturnus vulgaris). Within a breeding context (when test...
Songbirds produce learned vocalizations that are controlled by a specialized network of neural st... more Songbirds produce learned vocalizations that are controlled by a specialized network of neural structures, the song control system. Several nuclei in this song control system demonstrate a marked degree of adult seasonal plasticity. Nucleus volume varies seasonally based on changes in cell size or spacing, and in the case of nucleus HVC and area X on the incorporation of new neurons. Reelin, a large glycoprotein defective in reeler mice, is assumed to determine the final location of migrating neurons in the developing brain. In mammals, reelin is also expressed in the adult brain but its functions are less well characterized. We investigated the relationships between the expression of reelin and/or its receptors and the dramatic seasonal plasticity in the canary (Serinus canaria) brain. We detected a broad distribution of the reelin protein, its mRNA and the mRNAs encoding for the reelin receptors (VLDLR and ApoER2) as well as for its intracellular signaling protein, Disabled1. Thes...
Many sex differences in brain and behavior related to reproduction are thought to have evolved ba... more Many sex differences in brain and behavior related to reproduction are thought to have evolved based on sexual selection involving direct competition for mates during male-male competition and female choice. Therefore, certain aspects of brain circuitry can be viewed as secondary sexual characteristics. The study of proximate causes reveals that sex differences in the brain of mammals and birds reflect organizational and activational effects of sex steroids as articulated by Young and collaborators. However, sex differences in brain and behavior have been identified in the cognitive domain with no obvious link to reproduction. Recent views of sexual selection advocate for a broader view of how intra-sexual selection might occur including such examples as competition within female populations for resources that facilitate access to mates rather than mating competition per se. Sex differences can also come about for other reasons than sexual selection and recent work on neuroendocrine...
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