Huperzia serrata Extract 'NSP01' With Neuroprotective Effects-Potential Synergies of Huperzine A and Polyphenols.

Huperzia serrata (Thunb.) Trevis is widely used in traditional asiatic medicine to treat many central disorders including, schizophrenia, cognitive dysfunction, and dementia. The major alkaloid, Huperzine A (HA), of H. serrata is a well-known competitive reversible inhibitor of acetylcholinesterase (AChE) with neuroprotective effects. Inspired by the tradition, we developed a green one-step method using microwave assisted extraction to generate an extract of H. serrata, called NSP01. This green extract conserves original neuropharmacological activity and chemical profile of traditional extract. The neuroprotective activity of NSP01 is based on a precise combination of three major constituents: HA and two phenolic acids, caffeic acid (CA) and ferulic acid (FA). We show that CA and FA potentiate HA-mediated neuroprotective activity. Importantly, the combination of HA with CA and FA does not potentiate the AChE inhibitory property of HA which is responsible for its adverse side effects. Collectively, these experimental findings demonstrated that NSP01, is a very promising plant extract for the prevention of Alzheimer's disease and memory deficits.

Sexually active males prevent the display of seasonal anestrus in female goats.

A well-defined season of sexual rest controlled by photoperiod is observed in female sheep and goats during spring and summer, delineating their "anestrous season"; bucks also decrease sexual activity at about the same time. Nutrition and/or socio-sexual stimuli play only secondary roles. However, the presence of sexually active males can reduce the length of seasonal anestrus. Whether it can also completely suppress anestrus has not been investigated. Here we tested this in goats in 3 experiments, using bucks rendered sexually active out of season by exposure to long days. The continuous presence of these males prevented goats to display seasonal anestrus: 12/14 females cycled the year round, vs. 0/13 and 0/11 for females with un-treated bucks or without bucks (experiment 1). When active bucks were removed, females immediately entered anestrus (7/7 stopped ovulating vs. 1/7 if maintained with active bucks; experiment 2). Finally, 7/7 anestrous does with bucks in sexual rest since 1.5months commenced cycling rapidly during mid-anestrous, when these bucks became sexually active following a treatment with artificial long days, vs. 0/7 with un-treated bucks or no bucks (experiment 3). The presence/withdrawal of active bucks had a highly significant effect in the three experiments (P≤0.002). Therefore, the presence of a mating opportunity can completely override the photoperiodic inhibition of reproduction of females throughout the anestrous season. Results suggest that we must re-evaluate the relative contributions of photoperiod vs. other external cues in controlling seasonal reproduction, thus offering new non-pharmaceutical ways for controlling out-of-season reproduction in small ruminants.

Maternal behaviour and peripartum levels of oestradiol and progesterone show little difference in Merino ewes selected for calm or nervous temperament under indoor housing conditions.

Temperament influences the expression of maternal behaviour in sheep under experimental field conditions. We investigated whether maternal behaviour between ewes selected for a calm or nervous temperament is independent from environmental conditions. In addition, the level of maternal behaviour expressed by mothers is correlated with the concentration of hormones during the peripartum period. Therefore, we investigated whether the selection for temperament had resulted in hormonal differences between the two lines with regard to the hormones that could be involved in the onset of maternal behaviour. Oestradiol, progesterone and cortisol concentrations from 4 days before parturition to 24 h after parturition were determined from blood samples collected from 10 calm and 12 nervous ewes. Behavioural interactions between ewe and lamb were also recorded for 2 h starting at parturition. Mothers of both temperament lines showed adequate maternal behaviour under the controlled conditions of the study. Therefore, the results of the study do not suggest that selection for a calm or nervous temperament has profoundly affected the intrinsic ability of mothers to display adequate maternal behaviour. Hormonal differences between the two temperament lines were generally small and their possible influence on the display of maternal behaviour in the two temperament lines would have to be demonstrated.

Four hours of daily contact with sexually active males is sufficient to induce fertile ovulation in anestrous goats.

The study was conducted on two consecutive years to determine whether ovulatory activity can be induced in anovulatory goats by exposing them to sexually active bucks for 4, 8, 12 or 16 h per day during 15 consecutive days. In experiment 1, females remained continuously in the experimental pens where they were in contact with males. One group remained isolated from males (controls) and four other groups were exposed to sexually active males for 4, 8, 12 or 16 h per day. In experiment 2, females were taken away to "resting" pens free of male odours between the periods of contact with bucks. They were allocated to 5 groups as in experiment 1. Ovulations were determined by progesterone plasma levels and transrectal ultrasonography. Pregnancy was determined by abdominal ultrasonography. In both experiments, more than 90% of females exposed to the bucks had at least one ovulation during the whole experiment whereas only 11 or 0% (experiments 1 and 2, respectively) did so in the control group (P<0.001). Furthermore, the proportion of females ovulating did not differ among groups depending on duration of contact with bucks (P>0.05). In both experiments, pregnancy rates were not affected by the daily duration of contact with males (P>0.05). To conclude, 4h of daily contact with sexually active males is sufficient to stimulate ovulatory activity in anovulatory goats and this effect is not due to the presence of olfactory cues from the males remaining in the pens.

Amniotic fluid is important for the maintenance of maternal responsiveness and the establishment of maternal selectivity in sheep.

Amniotic fluid (AF) is important for the establishment of maternal behaviour in inexperienced ewes, but its role in experienced mothers remains to be studied. Here, the maintenance of post-partum maternal responsiveness and the establishment of exclusive bonding was investigated in multiparous ewes when AF was removed from the neonate or/and physical contact with the young was precluded for the first 4 h post partum. Maintenance of maternal responsiveness and establishment of exclusive bonding were measured by the proportion of mothers accepting their own lamb and alien lambs that had been either washed or not washed, and by comparing an acceptance score for each type of lamb. The acceptance score was computed by summing standardised variables of acceptance (low bleats, acceptance at udder, nursing and licking time) and subtracting standardised variables of rejection (high-pitched bleats, rejection at the udder and aggressive behaviour). Washing the neonate reduced its acceptance score, but the proportion of mothers rejecting their own lamb was reduced only when washing the neonate and prevention of physical contact for 4 h were combined (7/15 v. 0/10 in controls, P = 0.02). In addition, washing the neonate increased the acceptance score of the washed alien lamb, but not of the unwashed alien. However, washing and privation of physical contact did not increase significantly the proportion of mothers accepting an alien lamb at 4 h post partum. We conclude that AF is important in experienced ewes for the establishment of maternal responsiveness, as already found in primiparous mothers. In addition, our results indicate that AF also carries some chemosensory information facilitating exclusive bonding.

Temperament does not affect the overall establishment of mutual preference between the mother and her young in sheep measured in a choice test.

Temperament influences maternal behavior and lamb survival in Merino sheep selected for calm or nervous temperament. The impact of this selection on mother-young recognition and early expression of temperament in lambs is unknown. We tested the ability of multiparous ewes selected for calm (n = 16) or nervous (n = 18) temperament to recognize their own lambs 6 hr after parturition, the ability of the lambs to display a preference for their own mother at 18 hr, and the temperament of the lambs at 1 and 16 weeks of age. Ewes and lambs from both genotypes showed a similar preference for their familiar kin. In contrast, differences in temperament were detectable at 1 and 16 weeks of age. Nervous lambs showed higher vocal and locomotor activity than calm lambs. Thus, temperament did not affect the early process of ewe-lamb bonding but might affect the quality of the mother-young relationship under more challenging situations.

Social motivation affects the display of individual discrimination in young and adult Japanese quail (Coturnix japonica).

A simultaneous two-choice test was used to investigate discrimination between a familiar (F) and an unfamiliar conspecific (U) in Japanese quail, selected for high (HSR) or low (LSR) social reinstatement behavior (tendency to rejoin and stay close to conspecifics). Animals were 1-week unsexed and 6-week-old male quail. One-week-old LSR quail and 6-week-old male HSR quail displayed discrimination after 24 hr of pair contact (p < 0.05), and a tendency to discriminate (p < 0.10) was found in adult male LSR quail. After 1 week of pair contact, results were similar in LSR quail chicks, but no discrimination was evidenced in adult quail of either line. Therefore, Japanese quail are able to discriminate between a familiar cagemate and an unfamiliar conspecific after only 24 hr of pair contact. Moreover, this study reveals for the first time in this species that social motivation and age both influence the display of individual discrimination.

Differential effects of undernutrition during pregnancy on the behaviour of does and their kids at parturition and on the establishment of mutual recognition.

We investigated whether undernutrition during the second half of pregnancy impaired the behaviour of does and their kids at parturition and early mutual recognition. Twenty-two control and 22 underfed mixed-breed, multiparous dairy goats were used, together with their respective kids (control, n = 31: nine singles, 16 twins and six triplets; underfed, n = 32: 11 singles, 18 twins and three triplets). Undernutrition involved limiting protein and energy intake at 70% of the nutritional requirements for maintenance and foetal growth from day 70 of pregnancy until birth. The behaviour of mothers and their two first-born kids was observed for 90 min from the birth of each kid. Maternal olfactory recognition of the kid was assessed at 4 h post partum by testing selective nursing behaviour. Non-olfactory recognition was assessed at 8 h in a two-choice test excluding olfactory cues. In kids, preference for the mother was assessed in a two-choice test at either 12 or 24 h post partum. Bodyweight of does and kids were lower in the underfed group up to 2 weeks post partum. At parturition, licking, maternal bleating frequency and latency to nursing did not differ between nutritional groups. Control kids were faster than underfed kids to stand, search for and reach the udder, but underfed kids bleated more and tended to spend more time at the udder. Both control and underfed does accepted their own kid and rejected the alien in the selectivity test at 4 h. In contrast, at 8 h post partum, only control goats showed a significant preference for their own kid in the non-olfactory recognition test. Both control and underfed kids showed a preference for their own mother at 12 and 24 h and undernutrition during pregnancy had little influence on the performance of kids. However, 12 h-old underfed kids tended to be less active than control kids and visited their own mothers less than control kids. There were no significant correlations between the behaviour of the mother or of the kid at parturition and their performance in the discrimination tests. Overall, undernutrition in the second half of pregnancy appears to be more detrimental for the behaviour of the mother than for the kid. Furthermore, it has more impact on the establishment of maternal non-olfactory recognition than on maternal care at parturition or the establishment of maternal selectivity.

Effects of restricted nursing on milk production and collection, kid growth and plasma prolactin and growth hormone concentrations in dairy goats.

The milk production of dairy goats under various regimes of mother-young contact from day 4 post partum were studied during the first 2 months of lactation, together with the prolactin (PRL) and growth hormone (GH) responses to udder stimulation. In the control group, 13 goats and their kids were left in permanent contact and did not undergo milking. In two additional groups, goats were machine milked once a day in the morning (at 0800 h) and kids were allowed 10 hours (from 1000 to 2000 h; 10H group, n = 11) or 5 h (from 1000 to 2000 h; 5H group, n = 11) of mother-young interaction per day. In the last group (MO, n = 10), mothers were permanently separated from their kids on day 4 post partum and milked once a day. Milk production during a 24-h period at 37 days post partum performed by controlled nursing and weighing of the kids (groups with kids) or by two machine milking 12 h apart (milking only group) revealed a higher production in the three groups with some mother-young contact than in the MO group. Total milk collected by milking over the 2 months of the study did not differ between the three groups that underwent milking. Kid weights at 2 months were 3.4 to 4.8 kg. lighter in the groups that underwent milking than in the control group. Hormonal profiles were significantly affected by restricted mother-young contact, with highest pre-stimulation concentrations of PRL and GH in the 5H group. Restricting mother-young contact from the first week postpartum can permit an early collection of milk without major effects on kid growth, when compared with one daily milking in goats totally separated from their young.

Preference of 12-h-old kids for their mother goat is impaired by pre-partum-induced anosmia in the mother.

We investigated whether kids were able to discriminate their own mother from an alien one in a two-choice test on the day of birth when they had access to acoustic, visual and olfactory cues from their mother, and whether this discrimination depended on the selective maternal behaviour of the mother (i.e. exclusive nursing of own kids). When given the choice between their own mother and an alien equivalent dam, 8-h-old kids did not show a significant preference for their dam, whereas 12- and 24-h-old kids did. When given the choice between their own and an alien mother that were both non-selective because they had been rendered peripherally anosmic by irrigation of the nostrils with zinc sulphate, 12-h-old kids did not show a significant preference for their mother. These results are similar to those reported in sheep and may suggest that the contrast of behaviour between their own and an alien mother existing in normosmic does is important for discrimination of dams by kids at this age. Finally, testing 8-h-old kids in a smaller enclosure resulted in some improvement of their performance, although they still failed to display a significant preference for their mother. On the whole, kids are able to discriminate between their own and an alien mother goat as early as previously reported in lambs. The impairment of this ability when mothers are anosmic and not selective suggests that acceptance behaviours displayed by the mother may serve as one of the cues orientating the choice of the kid when given the choice between intact mothers. Finally, the present results do not suggest the existence of fundamental differences in the establishment of a preference for the mother between lambs, which are followers, and kids, which are hiders.

Stimulation of estrous behavior in grazing female goats by continuous or discontinuous exposure to males.

Two experiments were conducted during the anestrous period to determine: (1) whether males rendered sexually active by exposure to artificial long days stimulate estrous activity of female goats under grazed conditions (Exp. 1); and (2) whether continuous presence of the buck is necessary to stimulate this estrous activity (Exp. 2). In Exp. 1, 2 groups of females (n = 20/group), one in confinement and another under grazing conditions, were exposed to 4 bucks subjected to natural photoperiod (2 males/group). Two other groups of females (n = 20/group), in confinement or grazing, were exposed to 4 males treated with artificial long days (2 males/group). All groups were exposed to males for 15 d. The percentage of does detected in estrus during these 15 d was greater (P < 0.001) in the 2 groups exposed to males sexually prepared by long days (confined, 95%; grazed, 90%) than in groups exposed to males in natural photoperiod (confined, 15%; grazed, 45%). Does in Exp. 2 were allowed to graze and were exposed continuously (n = 26) or discontinuously (from 1700 to 0900; n = 26) for 18 d to males that had been stimulated to enter the breeding season by exposure to long days. The proportion of does that displayed estrous behavior in 18 d did not differ (P = 0.55) between groups (96.2 and 92.3% for continuous and discontinuous groups, respectively). The results indicate that anestrous goats managed under grazing conditions can be stimulated to express estrus by joining with males previously exposed to artificial long days. Continuous presence of the male is not necessary for this male effect.

The role of calcium and magnesium ions in uptake of beta-amyloid peptides by microglial cells.

Amyloid peptides 1-40 and 1-42 (Abeta 1-40 and Abeta 1-42) are major components of diffuse and neuritic senile plaques present in the brain of patients with Alzheimers disease. Their interaction with microglial cells was studied using a system partly mimicking these plaques, which consisted in heat-killed yeast particles coated with either Abeta 1-40 or Abeta 1-42. Using these particles, it has been shown in our laboratory that LRP is involved mainly in the elimination of Abeta 1-42-coated heat-killed yeast particles and partly in that of Abeta 1-40-coated heat-killed yeast particles by microglial cells in culture. We show here that in the presence of calcium and magnesium ions extracellular chelators, namely EDTA (for both ions) and EGTA (for calcium ions), the internalization of coated heat-killed particles was impaired. In the presence of BAPTA-AM, an intracellular chelator of calcium ions and thapsigargin, an inhibitor of the endoplasmic reticulum calcium pump, no effect was observed on the phagocytosis of Abeta 1-40-coated heat-killed yeast particles, whereas that of Abeta 1-42-coated heat-killed yeast particles was affected. These results suggest that different signaling mechanisms are involved after the internalization of Abeta 1-40 and Abeta 1-42.

Cyclin dependent kinase inhibitors prevent apoptosis of postmitotic mouse motoneurons.

Recent evidence suggests that apoptosis in post-mitotic neurons involves an aborted attempt of cells to re-enter the cell cycle which is characterized by increased expression of cyclins, such as cyclin D1, prior to death. However, such cyclins activation prior to apoptotic cell death remains controversial. Many neurological disorders are characterized by neuronal loss, particularly amyotrophic lateral sclerosis (ALS). ALS is a motoneuronal degenerative condition in which motoneuron loss could be due to an inappropriate return of these cells in the cell cycle. In the present study, we observed that deprivation of neurotrophic factor in purified motoneuron cultures induces an apoptotic pathway. After neurotrophic factor withdrawal, DAPI (4,6-diamidin-2-phenylindol dichlorohydrate) staining revealed the presence of nuclear condensation, DNA fragmentation, and perinuclear apoptotic body. Similarly, release of apoptotic microparticles and activation of caspases-3 and -9 were observed within the first hours following neurotrophic factor withdrawal. Next, we tested whether inhibition of cell cycle-related cyclin-dependent kinases (cdks) can prevent motoneuronal cell death. We showed that three cdk inhibitors, olomoucine, roscovitine and flavopiridol, suppress the death of motoneurons. Finally, we observed early increases in cyclin D1 and cyclin E expression after withdrawal of neurotrophic factors. These findings support the hypothesis that after removal of trophic support, post-mitotic neuronal cells die due to an attempt to re-enter the cell cycle in an uncoordinated and inappropriate manner.

Maintaining contact with bucks does not induce refractoriness to the male effect in seasonally anestrous female goats.

A study was performed to determine if complete separation between sexes is a pre-requisite to stimulate sexual activity of anovulatory female goats by male exposure in mid-anestrus when reproductive activity is greatly inhibited. The response to the male was studied in anovulatory goats which had either been in contact with bucks before the stimulation (n = 22; CONT), or totally isolated from bucks (n = 22; ISOL). At the time of male exposure, the bucks from the CONT groups were removed and females of both groups (CONT and ISOL) were divided into two subgroups (n = 11) exposed to novel bucks (n = 1 per subgroup), either in sexual rest (sexually inactive; SI) or in which sexual activity had been induced by a photoperiodic treatment (sexually active; SA). The sexual behavior of the SA bucks during the first 5 days of stimulation was greater than that of SI bucks (P < 0.01). Regardless of whether females had been (82%) or not (91%) exposed to bucks before teasing, most goats stimulated by SA males displayed estrous behavior and ovulation in the first 15 days following stimulation, whereas no female from the two subgroups exposed to SI bucks showed estrous behavior and only one of them ovulated (P < 0.02). These results indicate that previous isolation of seasonally anovulatory goats from bucks is not required to stimulate their reproductive activity by the male effect, if sexually active bucks are used.

Xaliproden (SR57746A) induces 5-HT1A receptor-mediated MAP kinase activation in PC12 cells.

Neurotrophic growth factors are involved in cell survival. However, natural growth factors have a very limited therapeutic use because of their short half-life. In the present study, we investigated the mechanism of action of a non-peptidic neurotrophic drug, Xaliproden, a potential molecule for the treatment of motoneuron diseases, since the transduction pathways of this synthetic 5-HT1A agonist are very poorly understood. Xaliproden does not activate the Trk receptor but causes a rapid increase in the activities of the ERK1 and ERK2 isoforms of MAP kinase, which then rapidly decrease to the basal level. We demonstrate that isoforms of the SHC adapter protein are phosphorylated independently of each other and are probably not the source of the Xaliproden-induced MAP kinases activation. The inhibitor of Ras farnesylation, FPT-1, and the protein kinase C inhibitors, GF 109203X and chelerythrine, inhibited the Xaliproden-induced MAP kinase activation, suggesting p21Ras and PKC involvement. Moreover, the observations that the 5-HT1A antagonist, pindobind, and pertussis toxin abolished the Xaliproden-induced ERK stimulation suggested that Xaliproden activates the MAP kinase pathways by stimulating the G protein-coupled receptor, 5-HT1A. These results demonstrate clearly that the non-peptidic compound, Xaliproden, exerts its neurotrophic effects through a mechanism of action differing from that of neurotrophins. These findings suggest that this compound does not involve MAPK activation by TrkA receptor stimulation but acts by MAP kinase pathway by a pertussis toxin-sensitive mechanism involving 5-HT1A receptors, p21 Ras and MEK-1 and by PKC and Akt pathways.

Estradiol receptor-alpha expression in hypothalamic and limbic regions of ewes is influenced by physiological state and maternal experience.

The influence of estrus, pregnancy, parturition, and maternal experience on the expression of estrogen receptor-alpha (ERalpha) was investigated in hypothalamic and limbic regions of the sheep brain, using immunocytochemistry. Four days before parturition, previous maternal experience was associated with a higher density of ERalpha-labeled neurons in the paraventricular and supraoptic nuclei, the medial preoptic area, and the medial amygdala, but not in the mediobasal hypothalamus. Furthermore, an interaction was found between physiological state and experience in the peripartum period as the effect of experience existing 4 days prepartum was not found at parturition, when densities were lowest both in primiparous and in multiparous ewes. An additional effect of physiological state was also observed between parturition and estrus, densities being significantly lower at parturition than at estrus in the SON, PVN, and MPOA, but not in the medial amygdala. These results indicate that in sheep ERalpha expression is influenced by previous physiological and/or maternal experience at specific times of the reproductive cycle. They are also congruent with the higher ability of multiparous than nulliparous ewes to show maternal behavior several days prepartum.

MAPK activation via 5-hydroxytryptamine 1A receptor is involved in the neuroprotective effects of xaliproden.

Motoneurons require neurotrophic factors for their survival and their differentiation. Xaliproden (SR57746A) is a synthetic compound that exhibits in vivo and in vitro neurotrophic effects in several experimental studies. Here we demonstrate that neuroprotective effects of Xaliproden on motoneuron cultures are mediated by the activation of the mitogen activated protein kinase pathway. It is inhibited by PD98059, a selective and irreversible inhibitor of MEK1. The activation of this pathway seems to involve two different proteins, the protein kinase C and the Ras. Indeed, we show that Xaliproden is able to activate the MAP kinases ERK1/2 and PKC in motoneurons. In addition, the use of a 5-hydroxytryptamine 1A receptor antagonist, Pindobind and pertussis toxin, inhibits the effect of Xaliproden on motoneuron survival, suggesting the involvement of this G-protein coupled receptor. Morever, 8-OH-DPAT, an agonist of 5-hydroxytryptamine 1A receptor, increases the survival of mouse motoneurons but not by the same extent as BDNF or xaliproden. Since 8-OH-DPAT does not act synergistically with Xaliproden, it is likely that their neuroprotective properties involve a similar pathway. Taken together, these results indicate that neuroprotective effects of Xaliproden on mouse motoneurons are dependent on the mitogen-activated protein kinase activation via 5-hydroxytryptamine 1A receptor.

Olfactory regulation of maternal behavior in mammals.

In mammals, olfactory cues are extensively used in many aspects of maternal care to ensure the coordination of mother-infant interactions and consequently the normal development of the offspring. Outside the period of parturition and lactation, when the young are not a behavioral priority, olfactory cues play an inhibitory role on maternal responsiveness since in most mammalian species studied so far, nonpregnant females find the odor of young aversive. On the contrary at the time of parturition, a shift in the hedonic value of infantile odors occurs so that the young now become a very potent stimulus and this sensorial processing constitutes an important part of the maternal motivational system. Moreover, infants' odors provide a basis for individual recognition by their mothers and some species (ungulates) have developed highly specialized mechanisms for processing of the infant signals. Perception of the smell of the young also regulates various aspects of maternal behavior. Dodecyl propionate, a compound released by of pup's preputial glands, has been shown to influence anogenital licking behavior, a fundamental pattern of maternal behavior in rodents. While there is no functional specificity of either the main or the accessory olfactory systems in the development of maternal behavior amongst species, it appears that only the main olfactory system is implicated when individual odor discrimination of the young is required. Neural structures, such as the main olfactory bulb, undergo profound changes when exposed to offspring odors at parturition. These changes in synaptic circuitry contribute both to maternal responsiveness to these odors, to their memorization, and to effects of long-term maternal experience.

Quantification of neurotrophin mRNA expression in PMN mouse: modulation by xaliproden.

Compounds possessing neurotrophic properties may represent a possible treatment for neurodegenerative disorders such as amyotrophic lateral sclerosis. Xaliproden (SR57746A), an orally-active non-peptide compound, which has been found to exhibit neurotrophic effects in vitro and in vivo, increased the lifespan and delayed the progression of the motor neuron degeneration in PMN mice. We have used a quantitative reverse transcription/polymerase chain reaction amplification technique to study the regulation of neurotrophin mRNA and trk mRNA expression in PMN mice. NGF and NT-3 mRNA are downregulated in PMN mice. These deficiencies can be overcome by a treatment with xaliproden. Such an effect could contribute to neurotrophic effects of xaliproden in vivo and in vitro.

Induction of sexual activity in lactating anovulatory female goats using male goats treated only with artificially long days.

Two experiments were conducted to determine the response of Creole male goats treated with long days and melatonin implants, and the response of the anovulatory does to male effect using males treated only with artificially long days. All animals were allocated to open sheds. In Exp. 1, one group of males was under natural photoperiod (CG; n = 7); the second group was submitted to 2.5 mo of long days followed by the insertion of two s.c. melatonin implants (LD+MEL; n = 7); the third group was subjected only to 2.5 mo of long days (LD; n = 7). Testicular weight was measured every 2 wk. Plasma testosterone concentrations were determined weekly. A treatment x time interaction was detected (P < 0.001) for testicular weight and plasma testosterone concentration. In the LD+MEL and LD groups, testicular size and plasma testosterone levels varied in a similar way, but differed from those observed in CG (P < 0.001). In this latter group, testicular weight displayed seasonal variations and peaked in June, whereas in treated groups this peak occurred in March. In CG, testosterone varied in a seasonal manner and plasma concentrations increased in June and remained elevated throughout the study. In experimental groups, testosterone increased in February and peaked in March. In Exp. 2, one group of males was left under natural photoperiod (CG, n = 5) and the other one was submitted to 2.5 mo of artificially long days (LD, n = 4). On March 16, two control and two treated males were put in contact with 20 and 19 females, respectively. Sexual behavior of the bucks was observed during the 5 d following male introduction. Progesterone assays and estrous behavior were used to determine ovarian and behavioral responses of the females to teasing. The anogenital sniffing, nudging, and mount instances registered in LD-treated males were greater than those observed in CG (P < 0.05). Of the does exposed to CG, none ovulated and only two of 20 females displayed estrous behavior. All does in contact with LD-treated males ovulated and showed at least one estrous behavior during the 15 d following joining (P < 0.001). These results indicate that the sexual activity of male goats from subtropical latitudes can be induced using only artificially long days. In addition, males treated in this way are capable of stimulating sexual activity in anovulatory females by the male effect.