Recovery of lordotic activity by dorsal deafferentation of the preoptic area in male and androgenized female rats

Lordotic activity was examined in male and neonatally androgenized female rats following dorsal deafferentation of the preoptic area (POA). Female pups were injected with various doses (100, 250, 500, or 1000 micrograms) of testosterone propionate (TP) on day 3 postpartum. Ten weeks after birth, all animals were castrated, then half of the castrated males and females in each group were subjected to dorsal deafferentation of the POA (anterior roof deafferentation: ARD) by using an L-shaped Halász knife in order to transect the dorsal forebrain efferents which are thought to exert an inhibitory influence on the lordosis mediating mechanism. Animals were implanted subcutaneously with Silastic tubes containing estradiol-17 beta (E2). Observations of lordosis behavior were carried out 5, 10, and 15 days after implantation of E2. Three to six hours before each behavioral test, all rats were injected with 0.5 mg progesterone. Regardless of the dose of TP given neonatally, androgenized females, as well as males, showed low levels of lordotic behavior. In contrast, males with ARD and androgenized females with ARD displayed lordosis more frequently than males without ARD, and androgenized females without ARD. Lordotic activity in the androgenized females with ARD was negatively correlated with the dose of TP given neonatally. The ARD females injected with a large dose (1000 micrograms) of TP neonatally were significantly less receptive than those injected with lower doses of TP and ARD males. These results suggest that a large dose of neonatal TP may cause permanent changes in not only the neural substrates for lordosis inhibition affected by ARD but also other structures involved in lordosis facilitation.     

Preoptic-brainstem connections and maternal behavior in rats.

This study presents evidence supporting the view that preoptic area (POA) projections through the ventral tegmental area (VTA) to lower brainstem regions are important for maternal behavior in postpartum rats. Experiment 1 demonstrated that bilateral coronal knife cuts posterior to the VTA disrupted maternal behavior, and Experiment 2 demonstrated a similar disruption when a unilateral knife cut that severed the lateral connections of the medial POA was paired with a contralateral knife cut posterior to the VTA. In a final anatomical experiment using horseradish peroxidase histochemistry, it was shown that knife cuts posterior to the VTA do sever POA efferents. However, such cuts severed other ascending and descending pathways as well, and these may also be involved in maternal behavior control.     

Medial preoptic lesions disrupt parental behavior in both male and female California mice (Peromyscus californicus)

California mice (Peromyscus californicus) are monogamous and naturally biparental, making them an ideal species in which to study the neural basis of paternal behavior. A male or female from each male-female pair was given an electrolytic or sham lesion in the medial preoptic area (MPOA), an area known to be critical for the expression of maternal behavior in rats, and retested for parental responsiveness. MPOA-lesioned males and females showed significantly longer latencies to show parental behavior and spent significantly less time near pups, sniffing pups, and licking pups than sham-lesioned mice. However, MPOA lesions did not reduce time spent hovering over pups. The results suggest that the neural mechanisms mediating paternal behavior are similar to those mediating maternal behavior in this species.     

Dopamine d1 and d2 receptor antagonism in the preoptic area produces different effects on maternal behavior in lactating rats

The preoptic area (POA) is critical for maternal behavior in rats but little is known about what neurotransmitters released here influence maternal responding. POA infusion of 10 microg (but not 2 microg) of the dopamine D1 receptor antagonist SCH-23390 greatly impaired retrieval and licking of pups but not other maternal or nonmaternal behaviors in lactating rats. In contrast, POA infusion of 10 microg (but not 2 microg) of the D2 receptor antagonist raclopride facilitated nursing but did not affect oral maternal behaviors. SCH-23390 in the medial hypothalamus tended to impair licking but not retrieval. Raclopride in the medial hypothalamus had no effects. Therefore, D1 and D2 receptor activity, particularly in the POA, is important for regulating different maternal behaviors.     

Bilateral damage to the sexually dimorphic medial preoptic area/anterior hypothalamus of male ferrets causes a female-typical preference for and a hypothalamic Fos response to male body odors

Previous studies showed that bilateral lesions of the male ferret's preoptic area/anterior hypothalamus (POA/AH), centered in the sexually dimorphic nuclei present in this region, caused subjects to seek out a same-sex male, as opposed to a female conspecific. Male subjects with POA/AH lesions (which were also castrated and given estradiol) displayed female-typical receptive behavior in response to neck gripping by a stimulus male, implying that subjects' approaches to a same-sex conspecific were sexually motivated. We asked whether the effect of POA/AH lesions on males' partner preference reflects a shift in the central processing of body odorant cues so that males come to display a female-typical preference to approach male body odorants. Sexually experienced male ferrets in which electrolytic lesions of the POA/AH caused bilateral damage to the sexually dimorphic male nucleus (MN) resembled sham-operated females by preferring to approach body odors emitted from anesthetized male as opposed to female stimulus ferrets confined in the goal boxes of a Y-maze. This lesion-induced shift in odor preference was correlated with a significant increase in the ability of soiled male bedding to induce a Fos response in the medial POA of males with bilateral damage to the MN-POA/AH. No such partner preference or neural Fos responses were seen in sham-operated males or in other groups of males with POA/AH lesions that either caused unilateral damage or no damage to the MN-POA/AH. Male-typical hypothalamic processing of conspecifics' body odorants may determine males' normal preference to seek out odors emitted by female conspecifics, leading to mating and successful reproduction.     

Cytochrome oxidase activity in the preoptic area correlates with differences in sexual behavior of intact and castrated male leopard geckos (Eublepharis macularius)

Although the utility of analyzing behavioral experience effects on neural cytochrome oxidase (CO) activity is well recognized, the behavioral correlates of endogenous differences in CO activity have rarely been explored. In male leopard geckos (Eublepharis macularius), the incubation temperature experienced during embryogenesis (IncT) and age affect CO activity in the preoptic area (POA), an area that modulates copulatory behavior. In this study, the authors assessed whether differences in POA CO activity correlate with differences in sexual behavior in intact and castrated geckos. Males with IncT- and age-dependent increases in POA CO activity mounted females with shorter latencies while intact and after castration and ejaculated more frequently after castration. The authors discuss the predictive value of CO activity and propose similar parallels in other species.     

Effects of anterior roof deafferentation on lordosis behavior and estrogen receptors in various brain regions of female rats.

This study investigated whether the estrogen receptors (ER), located at different brain areas and anterior pituitary (AP), changed after anterior roof deafferentation (ARD), and on the effects of facilitating the lordosis reflex in female rats. Female rats were ovariectomized and implanted with estradiol capsules. ARD or sham operation was performed with a Halász knife. All animals were tested for lordosis both before and after surgery. One day after the last test they were sacrificed. Cytosol and nuclear ER in the AP, medial preoptic area (MPOA), basal medial hypothalamus (BMH), amygdala (AMYG), septum (SEP), hippocampus (HPC), and cortex (CTX) were measured using an in vitro exchange assay. Rats with ARD showed significantly higher mean levels of lordosis quotient than the control and the sham groups before ARD surgery. An increase of both cytosol and nuclear ER in the BMH area compared to the control was observed, whereas the ER, in the SEP was reduced. ER in other areas were not affected by ARD. Serum estradiol and progesterone levels were not altered by the operation. These data suggest that the dorsal inhibitory pathway from the extrahypothalamus to the preoptic area and hypothalamus may modulate the estrogen receptor and the display of lordosis in female rats. Change of ER level in the BMH area may influence the hormonal sensitivity of lordosis in female rats.     

Induction of PGE2 by estradiol mediates developmental masculinization of sex behavior

Adult male sexual behavior in mammals requires the neuronal organizing effects of gonadal steroids during a sensitive perinatal period. During development, estradiol differentiates the rat preoptic area (POA), an essential brain region in the male copulatory circuit. Here we report that increases in prostaglandin-E(2) (PGE(2)), resulting from changes in cyclooxygenase-2 (COX-2) regulation induced by perinatal exposure to estradiol, are necessary and sufficient to organize the crucial neural substrate that mediates male sexual behavior. Briefly preventing prostaglandin synthesis in newborn males with the COX inhibitor indomethacin permanently downregulates markers of dendritic spines in the POA and severely impairs male sexual behavior. Developmental exposure to the COX inhibitor aspirin results in mild impairment of sexual behavior. Conversely, administration of PGE(2) to newborn females masculinizes the POA and leads to male sex behavior in adults, thereby highlighting the pathway of steroid-independent brain masculinization. Our findings show that PGE(2) functions as a downstream effector of estradiol to permanently masculinize the brain.     

Sex difference in the expression and regulation of nitric oxide synthase gene in the rat preoptic area

Neuronal nitric oxide synthase (nNOS) mRNA-positive cells were visualized by non-isotopic in situ hybridization histochemistry in the organum vasculosum of the lamina terminalis (OVLT) and the preoptic area (POA) in gonadectomized juvenile female and male rats. In the rostral POA (rPOA) at the level of the anteroventral periventricular nucleus, nNOS mRNA-positive cells were distributed in an inverted V-shaped area over the third ventricle and were in close proximity to cell bodies of gonadotropin-releasing hormone (GnRH)-immunoreactive neurons. In the caudal POA (cPOA) at the level of the medial preoptic nucleus, no topological association existed between GnRH and nNOS. Throughout the rPOA, both the number and the area of nNOS mRNA positive cells were significantly larger in the gonadectomized females than in the gonadectomized males. Treatment with estradiol for 2 days, followed by progesterone in the next morning, which caused an increase in serum luteinizing hormone 6 h later, induced a significant reduction of the nNOS mRNA expression in the rPOA in the female but not in the male rat at the time of sacrifice. In the OVLT and the cPOA, ovarian steroids had no effect on nNOS mRNA expression of both sexes. The results indicate that nNOS mRNA expression in the rPOA is sexually dimorphic and regulated by ovarian steroids in a sex specific manner.     

Maternal behavior in rats: evidence for the involvement of preoptic projections to the ventral tegmental area

This study provides evidence that a neural system extending from the preoptic region to the ventral tegmental area (VTA) of the midbrain is important for the normal expression of maternal behavior in lactating rats. In the first experiment, bilateral electrolytic lesions of the VTA severely disrupted the maternal behavior of postpartum rats. In the second experiment, lactating rats that received a unilateral knife cut severing the lateral connections of the medial preoptic area (MPOA) paired with a contralateral lesion of the VTA showed more severe maternal behavior deficits than females that received one of the following treatments: (a) a unilateral knife cut severing the lateral connections of the MPOA paired with an ipsilateral VTA lesion; (b) a unilateral knife cut severing the lateral connections of the MPOA paired with a contralateral lesion of the medial hypothalamus posterior to the MPOA; (c) a unilateral knife cut severing the lateral connections of the lateral preoptic area paired with a contralateral VTA lesion. The oral components of maternal behavior (retrieving and nest building) were particularly affected as a result of bilateral damage to the system extending from the preoptic area to the VTA.     

The effects of paraventricular hypothalamic lesions on maternal behavior in rats

The present study was undertaken to determine whether the disruptive effects of knife cuts which sever the lateral connections of the medial preoptic area (MPOA) on maternal behavior are mediated by interfering with the output of the paraventricular hypothalamic nucleus (PVN). Postpartum rats received one of the following: Knife cuts severing the lateral connections of the MPOA; knife cuts severing the lateral connections of the PVN; radiofrequency lesions of the PVN; sham lesions or knife cuts. Only females that received knife cuts severing the lateral connections of the MPOA showed severe deficits in maternal behavior. These results indicate that the influence of the MPOA on maternal behavior is not mediated by the output of the PVN. Since the PVN is the major source of oxytocin input to other brain regions, these results also suggest that oxytocinergic neural pathways are not critical for postpartum maternal behavior. Another important finding was that females with MPOA knife cuts that did not retrieve their young were capable of hoarding candy, suggesting that the retrieval deficit was not the result of a general oral motor deficit.     

The induction by testosterone of aromatase activity in the preoptic area and activation of copulatory behavior

A series of 4 experiments was designed to study the relationships between the activity of the aromatase (AA) in the preoptic area (POA) and the activation by testosterone (T) of copulatory behavior in gonadectomized male and female Japanese quail. The induction of AA by T in the POA is dose- and time-dependent. Levels of AA seen in sexually mature males are restored in castrated birds by a treatment with 20 to 40 mm silastic T capsules which produce physiological levels of steroid in the plasma. The minimal dose of T (10 mm implant) which reliably restores copulatory behavior approximately doubles the AA in the POA. The induction of AA is significantly larger in males than in females. A significant increase in AA is observed within 16 hours after the start of the treatment with T and the induction is maximal after 48 hours. Activation of copulatory behavior follows a similar time course but occurs with a delay of 24-48 hours. These results thus suggest that, in male quail, the activity of the aromatase in the POA is a limiting factor in the activation of copulatory behavior. This idea is confirmed by direct experimentation using an aromatase inhibitor, androstatrienedione (ATD). If T-treated birds receive at the same time silastic implants filled with ATD, the activation of behavior is suppressed for at least one week. This behavioral inhibition is, as expected, accompanied and very probably caused by the inhibition of the aromatase activity in the preoptic area and anterior hypothalamus. No increase of enzyme activity over the level seen in castrates was actually detected during the first 8 days of exposure to T. A moderate increase in AA was seen on day 16 and is probably responsible for the behavioral activation which was observed at the end of the experiment.     

Synaptic changes in the hypothalamus of the prepuberal female rat administered estrogen

Subcutaneous administration of estradiol benzoate (EB) to prepuberal female rats can advance vaginal opening, phasic pituitary gland luteinizing hormone (LH) secretion, and ovulation, presumably through a neural mechanism. This study investigated whether these effects are associated with changes in synaptic profiles in the arcuate nucleus of the hypothalamus (ARC) and the preoptic area (POA). Twenty-five-day-old female rats were adminstered EB, EB followed by progesterone on day 27, or oil vehicle alone; or they received no treatment. Blood was collected by jugular venipuncture at 1600 hr, on day 27, and plasma was assayed for LH by radioimmunoassay. Rat brains were immediately perfused for electron microscopy, and the ARC and POA were dissected out. Tissue blocks from these areas were processed with phosphotungstic acid for selective staining of neural synapses. Serum LH was markedly elevated in the EB-treated rats compared with controls. In the treated groups, LH values in serum were above 1,000 ng/ml, whereas the control values were less than 50. This acute rise of serum LH was accompanied by an acute increase of synaptic volume percent, area density, and numerical density in the ARC of EB-treated rats. The numerical density of the control groups was approximately 800 million observed synapses per cubic millimeter, whereas in the EB-treated groups, there were approximately 1.8 billion synapses per cubic millimeter. We found no differences in synaptic profiles of the POA in EB-treated animals as compared to the controls. We conclude from this study that estrogens act through neural mechanisms to accelerate maturation of neuroendocrine processes that govern phasic pituitary gland LH release and that this maturation process entails synaptogenesis in the ARC.     

Telencephalic and preoptic areas integrate sexual behavior in hime salmon (landlocked red salmon, Oncorhynchus nerka): results of electrical brain stimulation experiments

Various patterns of sexual behavior were evoked in freely swimming hime salmon by electrical stimulation of specific loci in the telencephalon and the preoptic area (POA) using chronically implanted electrodes. Furthermore, co-ordinated sexual behavior corresponding to stages of the natural spawning sequence was elicited from some of these brain regions. These results suggest that (1) sexual behavior is integrated in specific parts of the telencephalon and POA, and (2) within these regions there is a hierarchy of neural systems which mediate progressively more complete components of normal sexual behavior.     

Comparison of medial preoptic, amygdala, and nucleus accumbens lesions on parental behavior in California mice (Peromyscus californicus)

We have previously shown that medial preoptic area (MPOA) lesions disrupt parental behavior in both male and female California mice (P. californicus). In the present study, we compare the effects of lesions in the MPOA, with those in the basolateral amygdala (BA) and nucleus accumbens (NA) on male and female parental behaviors in the biparental California mouse. A male or multiparous female from each male-female pair was given an electrolytic or sham lesion in the MPOA, BA, or NA and tested for parental responsiveness. Since female P. californicus show postpartum estrus, they were likely pregnant during parental testing. MPOA lesions produced deficits in both male and female parental behaviors, and BA lesions disrupted male, and to a lesser extent, female parental behavior. NA lesions produced mild effects on pup-retrieval in males and no effect on parental behavior in females. However, NA lesions incompletely destroyed the NA shell, the region most relevant for maternal behavior in rats, and should be investigated further. These results support a role for the MPOA and BA in both male and female parental behaviors.     

Tyrosine hydroxylase-immunoreactivity and its relations with gonadotropin-releasing hormone and neuropeptide Y in the preoptic area of the guinea pig

The present study examines the distribution of tyrosine hydroxylase (TH) immunoreactivity and its morphological relationships with neuropeptide Y (NPY)- and gonadoliberin (GnRH)-immunoreactive (IR) structures in the preoptic area (POA) of the male guinea pig. Tyrosine hydroxylase was expressed in relatively small population of perikarya and they were mostly observed in the periventricular preoptic nucleus and medial preoptic area. The tyrosine hydroxylase-immunoreactive (TH-IR) fibers were dispersed troughout the whole POA. The highest density of these fibers was observed in the median preoptic nucleus, however, in the periventricular preoptic nucleus and medial preoptic area they were only slightly less numerous. In the lateral preoptic area, the density of TH-IR fibers was moderate. Two morphological types of TH-IR fibers were distinguished: smooth and varicose. Double immunofluorescence staining showed that TH and GnRH overlapped in the guinea pig POA but they never coexisted in the same structures. TH-IR fibers often intersected with GnRH-IR structures and many of them touched the GnRH-IR perikarya or dendrites. NPY wchich was abundantly present in the POA only in fibers showed topographical proximity with TH-IR structures. Althoug TH-IR perikarya and fibers were often touched by NPY-IR fibers, colocalization of TH and NPY in the same structures was very rare. There was only a small population of fibers which contained both NPY and TH. In conclusion, the morphological evidence of contacts between TH- and GnRH-IR nerve structures may be the basis of catecholaminergic control of GnRH release in the preoptic area of the male guinea pig. Moreover, TH-IR neurons were conatcted by NPY-IR fibers and TH and NPY colocalized in some fibers, thus NPY may regulate catecholaminergic neurons in the POA.     

Pharmacological and anatomical aspects of cholinergic activation of female sexual behavior

Forebrain infusion of cholinergic agonists activated the sexual response, lordosis, in ovariectomized female rats that had been primed with a low dose of estrogen. Carbachol, an agonist with both muscarinic and nicotinic properties, and oxotremorine, an agonist with a primarily muscarinic action, produced dose-related increases in the frequency of lordosis elicited by stimulus male rats. This facilitation of lordosis was prevented when females were pretreated systemically with atropine or scopolamine, two muscarinic receptor antagonists. These results indicate that the effect of carbachol and oxotremorine on lordosis is mediated by cholinergic muscarinic receptors. The location of these receptors within the brain has not been identified. Ventricular infusion of carbachol was as effective as infusion directly into the medial preoptic area (POA) and more effective than infusion directly into the ventromedial hypothalamus (VMH). Furthermore, when carbachol or oxotremorine was delivered to the POA through cannulae angled to avoid traversing the lateral ventricles, no facilitation of lordosis was observed. These data suggest that muscarinic receptors stimulated by central infusion of cholinergic agonists may not be located in either the POA or the VMH, two regions traditionally implicated in the regulation of lordosis.     

Pre- and postpuberal medial preoptic area lesions and maternal behavior in the rat

The ability of female rats to express maternal behavior following pre- or postpuberally-administered lesions of the medial preoptic area (MPOA) was investigated. Female Sprague-Dawley rats were electrolytically or sham-lesioned at either 28-30 days of age or 65-70 days of age and housed together in groups of 6-8 in "enriched" environments. Subsequently, the animals were mated and moved to individual cages just prior to parturition. Indices of maternal proficiency included nest ratings, pup retrieval time, percentage of pups with milk in their stomachs, and percentage of pup mortality. All animals in which lesions had substantially damaged the MPOA demonstrated significant deficits in all indices. Age at which the lesion was administered had no effect. In contrast to the recovery of male sexual behavior that has been reported for rats following prepuberally administered MPOA lesions, no recovery of maternal behaviors was seen in this study. Reasons for this lack of recovery may include the greater complexity of physiological and behavioral processes involved in maternal behavior in comparison to the rather stereotypical response patterns of male sexual behavior.     

Axon-sparing lesions of the preoptic region and substantia innominata disrupt maternal behavior in rats

In this study we investigated the effects of axon-sparing lesions of the preoptic region on the maternal behavior of postpartum rats. The lesions were produced with the excitotoxic amino acid N-methyl-D,L-aspartic acid (NMA). The first experiment determined that bilateral injections of NMA into the medial preoptic area (MPOA) of fully maternal lactating rats disrupted maternal behavior. In a second experiment, bilateral injections of NMA into the lateral preoptic area and adjoining substantia innominata (LP/SI region) also disrupted maternal behavior. A third experiment, employing horseradish peroxidase histochemistry, provided anatomical evidence that NMA destroys neuronal cell bodies while sparing fibers of passage. These findings were discussed with respect to the view that an MPOA-to-LP/SI-to-ventral tegmental area circuit underlies maternal behavior in the rat.     

Inhibitory and facilitatory neural mechanisms involved in the regulation of lordosis behavior in female rats: Effects of dual cuts in the preoptic area and hypothalamus

Two types of cuts were performed at a 4–5 week interval in the same ovariectomized rats; horizontal half-circle cut located just above the anterior commissure (ARD) and half-dome cut located anterior to the ventromedial nucleus (AD). Behavioral tests were carried out following the pretreatment with estradiol benzoate for 3 days and progesterone on the fourth day. When females received AD first (Experiment 1) the mean LQ was significantly lower than that of controls without brain surgery. Then, the AD rats were subjected to ARD or ARD sham. At the second test, the mean LQ of AD-ARD rats increased to the control level, but the LQ of AD-ARD sham rats was still low. In the experiment 2, the order of the brain surgery was just reversed. In the first test, all ARD females showed high levels of lordosis and the mean LQ was higher than that of control. Then, these ARD females receiving AD or AD sham were subjected to the second test. The mean LQ of ARD-AD rats decreased to the level of the control, but the LQ of ARD-AD sham rats was still high. Thus, dorsal neural inputs to or through the preoptic area and hypothalamus may exert inhibitory influences on a lordosis mediating system and anterolateral outputs of the medial basal hypothalamus appear to be concerned with a lordosis facilitating mechanism. These two systems seem to have a mutual correlation in regulating lordosis response. However, ARD or AD could not completely reverse the suppressive effect of AD or the facilitatory effect of ARD in the animals with dual cuts. It is suggested that the dorsal extrahypothalamic inhibitory influence and the hypothalamic facilitatory influence can regulate the display of lordosis independently in female rats.