This essay describes and analyzes of locomotion, sexual, afiliativní and agonistic behavior in a group of pigs macaque (macaca macaques) at Prague Zoo. Data collection took place in the period from December 2015 to May 2016. In the winter months (December to March) at the times from 9.00 to 16.00 hrs., And in April and May 2016 in time from 9.00 to 18.00 pm. The total observation period reached 240 hours. Data were analyzed using Statistics 12th and MS Excel 2007. Objectives of the work are three: (1) Based on the research literature to describe the behavioral ecology of macaque monkeys. (2) The detailed description and analysis of locomotion, sexual, afiliativní and agonistic behavior in a group of macaque pigs in the Prague Zoo. (3) Describe grooming, depending on time of day and social position in the group of pigs macaque in the Prague Zoo. We tested five hypotheses, which can be divided into four categories. We first investigated the hierarchy of the group. Furthermore, we investigated the behavior depending on the sex of the individual, and consequently the position in the hierarchy. In the last part we dealt with in detail during grooming. Tests confirmed the existence of a clear linear hierarchy based on agonistic interactions. The results showed that move around the enclosure depends on...

Nous possédons la faculté de reconnaître individuellement des centaines d’individus. Ceci nous permet d’évoluer dans une société complexe dont l’organisation est en partie forgée par les relations interindividuelles. La reconnaissance individuelle peut être réalisée par l'identification de divers éléments distincts, comme le visage ou la voix, qui forment chez l’Homme une seule représentation conceptuelle de l'identité de la personne. Nous avons démontré que les singes rhésus, comme les humains, reconnaissent individuellement leurs congénères familiers, mais également les individus humains connus. Ceci montre que la reconnaissance fine est une compétence partagée par un éventail d'espèces de primates pouvant servir de fondement à la vie en réseaux sociaux sophistiqués, et également que le cerveau s’adapte de façon flexible pour reconnaître les individus d'autres espèces lorsque ceux-ci ont une importance socioécologique. Par la suite, au niveau neuronal, ce projet a mis en lumière que les connaissances sociales concernant autrui sont représentées par les neurones hippocampiques ainsi que par les neurones inférotemporaux. Ainsi nous avons observé l’existence de neurones sélectifs aux visages non seulement dans le cortex inferotemporal, comme ceci a été décrit précédemment, mais également dans l’hippocampe. La comparaison des propriétés de ces neurones au sein de ces deux structures, suggère que les deux régions joueraient des rôles complémentaires au cours de la reconnaissance individuelle. Enfin, parce que l'hippocampe est une structure qui a évolué à des degrés divers chez différents mammifères pour soutenir la mémoire autobiographique et les représentations spatiales, la caractérisation des différents types de neurones et de leur connectivité a fourni un cadre commun pour comparer les fonctions de l’hippocampe à travers les espèces / Humans can individually recognize some hundreds of persons and therefore operate within a rich and complex society. Individual recognition can be achieved by identifying distinct elements such as the face or voice as belonging to one individual. In humans, those different cues are linked into one conceptual representation of individual identity. I demonstrated that rhesus monkeys, like humans, recognize familiarpeers but also familiar humans individually and that they match their voice to their corresponding memorized face. Thus it shows that fine individual recognition is a skill shared across a range of primate species, which may serve as the basis of a sophisticated social network. It also suggests that animals’ brains flexibly adapt to recognize individuals of other species when socio-ecologically relevant. Following at the neuronal level, this project put in light that social knowledge about other individuals is represented by hippocampal neurons as well as by inferotemporal neurons. For instance I observed the existence of face preferring neurons not only in the inferotemporal cortex as previously described but also in the hippocampus. Comparison of their properties across both structures, suggests that they could play complementary roles in recognition of individuals. Finally, because the hippocampus is a structure that evolved in various degrees to support autobiographical memory and spatial information in different mammals, I characterized the different subtypes of neurons and their network connectivity in the monkey hippocampus to provide a common anatomical framework to discuss hippocampal functions across species

Natal dispersal may have considerable social, ecological and evolutionary consequences. While speciesspecific dispersal strategies have received much attention, individual variation in dispersal decisions and its fitness consequences remain poorly understood. We investigated causes and consequences of natal dispersal age in rhesus macaques (Macaca mulatta), a species with male dispersal. Using long-term demographic and genetic data from a semi-free ranging population on Cayo Santiago, Puerto Rico, we analysed how the social environment such as maternal family, group and population characteristics affected the age at which males leave their natal group. While natal dispersal age was unrelated to most measures of group or population structure, our study confirmed earlier findings that sons of high-ranking mothers dispersed later than sons of low-ranking ones. Natal dispersal age did not affect males\\\'' subsequent survival, but males dispersing later were more likely to reproduce. Late dispersers were likely to start reproducing while still residing in their natal group, frequently produced extra-group offspring before natal dispersal and subsequently dispersed to the group in which they had fathered offspring more likely than expected. Hence, the timing of natal dispersal was affected by maternal rank and influenced male reproduction, which, in turn affected which group males dispersed to.

Dominance rank often determines the share of reproduction an individual male can secure in group-living animals (i.e. dominance rank-based reproductive skew). However, our knowledge of the interplay between individual and social factors in determining rank trajectories of males is still limited. The overall aim of this thesis was therefore to investigate mechanisms that underlie individual dominance rank trajectories in male crested macaques (Macaca nigra) and to highlight potential individual and social determinants of how males can achieve and maintain the highest rank possible. Data for this thesis were collected on 37 males during a field study on a natural population of crested macaques living in the Tangkoko-Batuangus Nature Reserve in Indonesia. In study 1, I validate Elo-rating as a particularly well suited method to quantify dominance hierarchies in animal species with dynamic dominance relationships. In studies 2 and 3, I suggest a personality structure for crested macaque males consisting of five distinct factors and further demonstrate that two personality factors determine whether males will rise or fall in rank. Finally, in study 4, I present results on how males utilize coalitions to increase their future rank. Together, these results shed light on how individual attributes and social environment both can impact male careers. Ultimately, in order to understand what determines rank-based reproductive skew, we need to consider the complexity and likely diversity of the mechanisms underlying rank trajectories of individual males which are likely to differ across different species.

Magister Scientiae (Medical Bioscience) - MSc(MBS) / The incidence of male infertility is increasing, with up to 50% of infertile males having “unexplained” (idiopathic) infertility. Newly developed molecular techniques have great value in detecting subtle causes of male infertility, as compared to idiopathic infertility which may be explained by standardizing and optimizing sperm functional and structural tests in non-human primate (NHP) sperm. The aim of the study was to evaluate sperm functionality utilizing the sperm of two NHP species, i.e.1) the rhesus monkey (Macaca mulatta) and 2) the vervet monkey (Chlorocebus aethiops), and further evaluate the effect of physiological media (including commonly used, and newly formulated sperm wash and sperm capacitating media) on NHP sperm functionality. Sperm functionality was evaluated by investigating the following sperm functions i.e.: sperm motility, vitality, acrosome reaction (AR), hyperactivation, and mitochondrial membrane potential (MMP). Sperm functional tests included computer-aided semen analysis (CASA), motility analysis, BrightVit staining for sperm vitality, flourescenin isothiocyanate (FITC)- conjugated peanut agglutinin (PNA) staining for sperm acrosome integrity, induction of hyperactivation by stimulants (sperm preparation media containing capacitating ingredients), and mitochondrial inhibitor (Oligomycin-A) for testing MMP. All functional and structural tests were investigated in both species, except for acrosome integrity, mitochondrial inhibition and functional tests compared over time that could not be successfully completed and investigated in the rhesus species. Motility analysis tests proved that within the vervet species, the use of different physiological media results in statistically significant differences in motility and kinematic parameters over a 1 hour time period. Hyperactivation tests proved that capacitating physiological media produced significantly higher percentages hyperactivation when compared to sperm wash media within the vervet species over a 1 hour time period. Furthermore, within both NHP species, sperm structural analysis (vitality and acrosome integrity) results showed that no significant differences are present when making use of different physiological media over a period of 1 hour incubation. The incubation of vervet sperm with different concentrations of mitochondrial inhibitor, Oligomycin-A (0 μM, 5 μM, and 25 μM), resulted in motility inhibition over a 1 hour incubation period. By the evaluation of these tests it was found that the use of different sperm wash [Human tubal fluid (HTF), Ham‟s F-10® and HD Sperm Wash Plus (HDSWP)] and sperm capacitation media [Human tubal fluid with added caffeine (HTFC) and HD Sperm Capacitating Plus (HDSCP)] resulted in significantly different results within sperm functional tests as compared to sperm structural tests. The study indicates that the composition of media, varying from simple to more complex, used for semen preparation plays an important role in determining NHP sperm functionality. Based on these findings further investigation in larger NHP sample groups and human sperm are required to evaluate the role of certain ingredients in the development of more cost-effective media producing satisfactory results in terms of sperm functionality for artificial reproductive technologies (ART).

Résume Objet: La stimulation cérébrale profonde (SCP) est devenue une thérapie efficace dans une série de maladies cérébrales. Récemment, dans les cas des algies vasculaires de la face résistantes au traitement (intraitables), chroniques, la SCP hypothalamique a suscité un nouvel intérêt pour cette région, également bien connue pour son implication dans la régulation de la prise alimentaire et de la balance énergétique. Cependant, les risques et les problèmes connexes liés à l'implantation dans cette aire cérébrale ont soulevé plusieurs questions concernant la sûreté de cette technique chirurgicale. Dans cette étude, les auteurs ont proposé l'implantation d'une électrode intraventriculaire insérée dans le troisième ventricule au niveau de l'hypothalamus ventromedial (VMH) chez des singes macaca fascicularis non obèses dans le but de moduler la prise alimentaire et le masse corporelle des sujets. Cette méthode de SCP pourrait s'avérer être un traitement potentiel de l'obésité morbide. Méthodes: Cinq singes de macaca fascicularis adultes (4 sujets et 1 contrôle ou sham) ont été implantés de façon stéréotaxique dans le troisième ventricule. Une électrode chronique Medtronic®, habituellement utilisée dans le cadre de la SCP chez les patients atteints de la maladie de Parkinson, a été positionnées dans l'espace intraventriculaire adossée à la paroi de ce dernier au niveau du VMH. Dans la première phase de l'étude, le comportement alimentaire de chaque animal (durée du repas, quantité de nourriture avalée) et son activité motrice ont été enregistrés et analysés en fonction différents paramètres de stimulation (fréquence et intensité) après une période de jeun de 24 heures. Dans la seconde phase du protocole, trois cycles de stimulation intraventriculaire de 8 semaines chacun ont été réalisés à 130Hz, à 80Hz et à 30Hz, suivi des périodes de « washout » de 4 semaines entre les périodes« on - stimulation ». L'index de masse corporelle, le poids (masse corporelle), la « teneur « en graisse, l'épaisseur cutanée et les concentrations hormonales ont été mesurés au début de l'étude pour établir une ligne de base et après chaque session de stimulation. Résultats: Lors de la première phase du protocole réalisée sur des animaux a jeun depuis 24 heures, nous avons remarqué une diminution de la prise alimentaire comprise entre 11 et 19% chez tous les sujets stimulés à une fréquence 80 hertz. A partie de ces résultats, , une diminution de la masse corporelle et du BMI (body mass index indice de masse corporelle) ont été observés chez trois de quatre singes lors des phases de stimulation chronique à une fréquence de 80 hertz : la moyenne de perte pondérale était de 8± 4.4%. Une augmentation de 2-6 ± 2.5% et de 5 ±2,93 %de la masse corporelle a été observée respectivement chez les animaux stimulés à une fréquence de 130Hzet de 30Hz. Une diminution importante des épaisseurs sous-cutanées ( )a été observée pour chacun des quatre sujets à une fréquence de 80 hertz et dans une moindre mesure, une augmentation de cette variable ( ) a été remarquée une fréquence de 130 Hz. Tout au long de l'étude, les variables relevées sur le singe Sham sont restées stables. Sur la durée de l'étude, aucun effet potentielle ment délétère n'a été remarqués sur les animaux. Conclusion: La stimulation de la région de VMH par voie intraventriculaire pourrait s'avérer efficace pour moduler le comportement alimentaire et induire une diminution soutenue de la masse corporelle caractérisée par réduction de la masse graisseuse chez les primates non humains non obèses.

The purpose of this study is to examine the influence of visitors on six species of captive primates in Shou Shan Zoo. In this article, I compared the behavior and the use of enclosure of primates with or without audiences, and I studied the correlations among the visitors¡¦ behavior, the behavior of primates, and the use of enclosure of them. Data were collected from September 2006 to March 2007 (between 09:00 to 16:00 hrs). The Shou Shan Zoo¡¦s visitors were estimated as 681 (¡Ó 323.71) individuals in non-holidays and 2824 (¡Ó 810.15) individuals in holidays. The average feeding provided by visitors was 3.65 % for 6 primate species, and this was more than four times of the feeding rate in Taipei Zoo. This phenomenon will threat the welfare and health of animals. Although resting was the most frequent behavior of these six species of primates, the proportion of abnormal behavior in Chimpanzee display site where attracted highest number of audiences and the highest proportion of animal feeding performed by visitors was relatively high. The major behavior of captive Chimpanzees were rest (53.28%), abnormal (13.75%) and food-related behavior (12.93%), and the frequency of Chimpanzees used the edge zone beyond 70%. The major behavior of Borneo Orangutan was rest (64.10%), followed by food-related behavior (20.13%) and self-directed behavior (12.18%). The frequency of Borneo Orangutan in the enclosure was highest in the back zone. The major behavior of Hamadryas Baboons was rest (49.27%), followed by affiliative (21.10%) and food-related behavior (12.68%). The frequency of Hamadryas Baboon in the enclosure was 50.27% in the other and 47.35% in the back zone. The major behavior of Celebes was rest (55.12%), followed by affiliative behavior (13.10%) and active (12.91%). The frequency of Celebes in the enclosure was highest in the back zone (41.51%), followed by the enrich (31.50%) and the edge zone (23.50%). The major behavior of the Dark-handed gibbon was rest (54.40%), active (27.10%) and food-related behavior (12.17%), and the frequency of the Dark-handed gibbon in the enclosure was 54.40% in the enrich zoon, 27.00% in the edge zoon and 10.90% in the other zoon. The major behavior of the White-handed gibbon was rest (74.28%) and active (18.07%), and the frequency of it in the enclosure was 45.58% in the enrich zoon and 39.52% in the edge zoon. The frequency of the Chimpanzees¡¦ abnormal, undesirable and self-directed behavior were influenced by audiences, and the duration of abnormal and undesirable behavior were positively correlated with the visitors¡¦ active and feeding behavior, but that were negatively correlated with the visitors¡¦ behavior (ignore and look). The duration of the Borneo Orangutan¡¦s rest, active, food-related and self-directed behavior were influenced by audiences, and the duration of the Borneo Orangutan used the back and the enrich zone were significantly correlated with the visitors¡¦ behavior (active and look). The Hamadryas Baboon¡¦s rest (time) and undesirable behavior (frequency) were influenced by audiences, and the duration of rest, food-related and undesirable behavior were significantly correlated with the visitors¡¦ behavior (active, look, feeding and ignore), and the duration of the Hamadryas Baboon used the edge and the back zone were significantly correlated with the visitors¡¦ behavior (look and feeding). The frequency of the Celebes¡¦ self-directed and agonistic behavior were influenced by audiences, and the duration of active, self-directed, food-related and affiliative behavior were significantly correlated with the visitors¡¦ feeding behavior. The duration of the Dark-handed gibbons¡¦ active and food-related behavior and the frequency of the Dark-handed gibbons¡¦ agonistic behavior were influenced by audiences, and the duration of rest, active, food-related, self-directed, undesirable and agonistic behavior were significantly correlated with the visitors¡¦ behavior (active, look, feeding and ignore). The duration of Dark-handed gibbons used the edge, the back and the enrich zoon were influenced by audiences, and the duration of the Dark-handed gibbons used these three zones were significantly correlated with the visitors¡¦ behavior (active, feeding and ignore). The frequency of the White-handed gibbon¡¦s self-directed behavior was influenced by audiences, and the duration of active, food-related, undesirable and abnormal behavior were significantly correlated with the visitors¡¦ behavior (active, look, feeding and ignore). The duration of the White-handed gibbon used the other zoon was positively correlated with the visitors¡¦ behavior (active).

The purpose of this study was to investigate the post-conflict reconciliation, consolation, solicited affiliation, stress and redirection in the captive Formosan macaques¡]Macaca cyclopis¡^in the Taipei Zoo. I used the post-conflict and matched-control (PC-MC) method to calculate the conciliatory tendency (CCT) and the triadic contact tendency (TCT) in adult macaques. The mean CCT for kin (83.33%) was significantly higher than that for non-kin (0.36%), and victims initiated reconciliation toward aggressors in higher rank classes significant more than both of them were in the same rank. The mean TCTs of aggressors and the victims were similar toward different triadic contact opponents (opponent¡¦s kin, own kin, unrelated individual). The ratio of the attracted pairs of victims who reconciled with aggressors by sociosexual behavior (15.23%) was significant higher than dispersed pairs (non-exist). In addition, the ratio of attracted pairs of victims who reconciled with unrelated third party by affiliation (46.72%) was significant higher than the dispersed pairs (21.76%). The similar situation also occurred in sociosexual behavior (28.68% verse 1.75%). However, aggressors and victims had similar chance to take the initiative affiliation after conflict (P > 0.05) The frequency of self-directed behavior (SDB) of Formosan macaques was slightly higher in the first 4 minutes in PC. The frequency of SDB after reconciliation (14.6 bouts/100 min) was not significantly lower than that before reconciliation (23.2 bouts/100 min) or when affiliation behavior did not occur (22.3 bouts/100 min). When the conflict opponents were kin, the SDB frequency (16.1 bouts/100 min) was not significantly lower than non-kin (24.1 bouts/100 min). The targets of redirect aggression were mostly unrelated individuals (82.61%). The mean CCT of the victims (16.50%) did not significant differ from the mean consolation TCT of the victims (48.81%), which indicated that reconciliation and consolation played similar critical roles after conflict. The sociosexual behavior performed by victims only occurred in PC (34.85%), which indicated the purpose of sociosexual behavior in reconciliation was to prevent further attack from aggressors. The chance of victims did not involve reconciliation and consolation, but solicited affiliation with a third party in PC was 23.32%. This indicated that the solicited affiliation might function to exchange the aggressive supports from the third party in the following conflicts.

GnRH is a decapeptide hormone, which plays a major role in the process of mammalian reproduction. It is synthesized by the hypothalamus and binds to its cognate receptor on the pituitary, to bring about the release of gonadotropins LH and FSH. The gonadotropin releasing hormone receptor belongs to the family of G-protein coupled receptors that are characterized by the presence of seven putative transmembrane regions linked by extracellular and intracellular loops. It is a glycoprotein made up of 327 amino acids. During the last several years cloning of this receptor from a number of species has provided considerable insight into the molecular basis of interaction between GnRH and its receptor. The GnRH receptor has been cloned and sequenced from a large number of mammalian species such as human, sheep, cow, rat, mouse, etc. GnRH receptor is known to be unique among the G protein coupled receptors by virtue of the fact that it lacks a C terminal tail which has been implicated in coupling to G-proteins in several seven transmembrane domain receptors. Other members of this G-protein coupled receptor family such as the Luetinising hormone receptor, Follicle stimulating hormone receptor contain the characteristic cytoplasmic tail of about 68-72 amino acids, which is believed to possess a plasma membrane targeting signal sequence. Mutation studies carried out revealed that this C terminal sequence may be important in membrane trafficking in other G protein coupled receptors, since mutant forms of the receptor were not expressed on the plasma membrane. In many G-protein coupled receptors, part of the cytoplasmic tail is important for desensitization and internalization. However, the GnRH receptor is an exception in that its G protein coupling and desensitization functions are dependent on regions of the GnRH receptor other than the carboxy terminal cytoplasmic domain. It has been well established that binding of GnRH to its cognate receptor induces conformational change and it is suggested that the entire extracellular loop and transmembrane region are involved in binding and signal transduction. It is pertinent to note in this connection, that the use of both polyclonal and monoclonal antibodies has contributed significantly to the understanding of the interactions between ligands and their cognate receptors. Recent studies have established that there are several extrahypothalamic sites of production of GnRH, which include testes, lymphocytes, human placenta, mammary gland etc. Of these the production of GnRH in the human placenta has attracted attention in view of the demonstration that the placental chorionic gonadotropin production (CG) is regulated by placental GnRH. Our laboratory has been investigating the role of GnRH in regulation of Chorionic Gonadotropin (CG) using both in vitro human placental villi system and pregnant bonnet monkey as models. One important and interesting observation that has been made in our studies as well as by several others is that the affinity of the placental GnRH receptor to its ligand is quite low compared to the pituitary receptor. Available evidence indicates that the hypothalamic and the placental GnRH are similar in structure and consequently the difference in the affinity could be attributed to the differences between the pituitary and the placental GnRH receptor. Considering this, it will be ideal and of interest to compare the GnRH receptor from the pituitary and placenta of a species in which both in vitro and in vivo studies can be carried out. For obvious ethical reasons, in vivo studies cannot be carried out with humans. Since very little information is available on the GnRH receptor in non-human primates, as a first step we undertook the task of characterizing the GnRH receptor from the bonnet monkey pituitary and production of antibodies to it, since all the studies carried out so far with antibodies to GnRH receptor have employed antibodies generated to a small stretch of peptide in the extracellular region. Thus the objective of the present study is to clone and express the GnRH receptor from the pituitary of the bonnet monkey {Macaco radiata), raise antibodies and to characterize them functionally. Chapter 1 provides a general review of information currently available regarding structure of GnRH and its receptor as well as the results of studies using antibodies directed to the GnRH receptor fragments. Chapter 2 deals with the partial cloning of the GnRH receptor from the pituitary of the bonnet monkeys by the technique of RT-PCR. We were able to amplify a PCR fragment of 959bp corresponding to the almost full-length GnRH receptor sequence. Southern blot analysis using the full length human pituitary GnRH receptor cDNA as the probe revealed that the 959 bp product was able to hybridize to the probe, confirming the authenticity of the PCR product. Restriction mapping with three different restriction enzymes also gave the expected pattern. Additional evidence was obtained by cloning of this PCR product into expression vector pGEX 5X-2 and sequencing a number of clones. The sequences obtained were then subjected to homology search with other known GnRH receptor sequences available in the Genebank. The sequence was found to be 97% homologous to the human pituitary GnRH receptor sequence and also showed a high degree of homology with the GnRH receptor from other species. Although antibodies have been raised to the GnRH receptor by immunizing rabbits with synthetic peptides corresponding to extracellular regions of the receptor, most of the antibodies have a very low affinity towards the native receptor. Also results of studies using these antibodies indicated that the peptide antibodies failed to recognize the native receptor. Initially we made efforts to express the full-length receptor in E.coli BL21 cells. However, since we were not successful in our attempts to express the full length, we resorted to express a smaller fragment which corresponded to amino acids 164-266, that encompassed one extracellular, two transmembrane and one intracellular domain. Before we proceeded ahead to express this fragment, the authenticity of this fragment was established by southern hybridization, restriction mapping as well as sequencing. This monkey pituitary GnRH receptor fragment corresponding to 315 bp was cloned in the expression vector pGEX 5X-2 and the protein corresponding to this region was overexpressed as a recombinant fusion protein in E.coli. BL21 plys S strain. Overexpression of the protein was induced using IPTG and the lysate was subjected to electrophoresis on a SDS-PAGE gel A signal corresponding to 37Kda, which is in agreement with the expected size (GST portion of the fusion protein plus the peptide) was observed following induction with IPTG. The overexpressed protein was found to be localized to the inclusion bodies, and this was purified from inclusion bodies by cutting out the band corresponding to the overexpressed protein from the preparative SDS-PAGE gels and the protein was eluted out by electroelution. Sera from the rabbits, which were immunized with the overexpressed protein, were checked for the presence of antibodies by ELISA, using the purified protein as the antigen. After ascertaining the presence of high titre antibodies in the sera of immunized animals, the serum was used to detect the presence of GnRH receptor in the membrane preparations from rat pituitary, monkey pituitary and human placenta using the technique of western blotting. A signal corresponding to 68Kda was found in all the cases and the specificity of this signal was established by preabsorption of the antisemrn with pituitary and placental membrane preparations, which resulted in decrease in the intensity of the signal. . The antiserum was also used to localize the GnRH receptor in different tissues such as first trimester and term human placenta, sheep pituitary, monkey placenta, human pituitary and rat prostate by the technique of immunotlourescence using the confocal microscope. The results of the above studies are presented in Chapter 3. Chapter 4 deals with the functional studies carried out using the antiserum to GnRH receptor in an in vivo system using male and female rats. As discussed earlier, all the reported studies on use of antibodies to GnRH receptor have employed a small region of the extracellular portion of the receptor for the production of antibodies. However, the antibodies in the present study have been directed towards a larger fragment, and considering this, it was of interest to evaluate the effect of these antibodies in in vivo as well as in vitro systems. Two approaches were used to evaluate the effect of antibodies, namely passive and active immunization i.e. administration of antiserum to GnRH receptor fragment raised in rabbits and also immunization with the overexpressed recombinant GnRH receptor protein. This study was carried out in both immature as well as adult male rats and also in the cycling female rats. Several parameters were monitored, which included various androgen dependent parameters in the male reproductive tissue i.e. body weight, testes weight as well as the weight of accessory sex organ-the prostate and also the fertility status. In the female rats the changes in the weight of the ovary, uterus, serum E2 and P4 were monitored. No effect on the body weight, testis weight or prostate weight was noticed in the treated animals compared to the controls. Furthermore, an indication that the hypothalamo-pituitary-gonadal axis was not compromised in the passively immunized animals was obtained from the observation that there was no decrease in the serum and testicular testosterone levels. In fact, there was a significant increase in the serum and testicular testosterone levels. This suggested the possibility that the antibodies are exerting a ßßstimulatory effect. To ascertain this possibility, two androgen dependent parameters namely the levels of mRNA for TGF ß, which is androgen repressed gene and Prostatein Cl, which is an androgen induced gene were monitored. It was observed that there was a significant increase in the steady state mRNA level of Prostatein Cl in GnRH antiserum treated animals and a corresponding decrease in TGFß mRNA levels. Active immunization study with injection of the recombinant protein was also carried out in adult male rats. All immunized animals responded to the immunization by producing high titre antibodies, the presence of which was detected by ELISA using the recombinant protein as the antigen. The results of the study revealed that there was no change in the body weight, testis weight or prostate weight. However, there was a significant increase in the serum and testicular testosterone levels compared to the control animals. Fertility studies indicated that all the animals were fertile. However, as in the case of passive immunization studies, an increase in the mRNA levels of Prostatein Cl was noted although the level of TGFß, which is an androgen repressed gene could not be monitored in this case due to the very high levels of endogenous androgens present in these animals. Thus it appears that the antibodies produced both in rabbits as well as in rats were stimulatory in nature probably indicating some specific characteristic of the region of the receptor to which the antibody has been raised. The results obtained in the present study are of significance considering the fact that studies using the antibodies to LH receptor and TSH receptor, both of which belong to the G-protein coupled family also report production of stimulatory antibodies. Active immunization studies using the GnRH receptor protein in the female rats also revealed that the antibodies were not compromising the hypothalamo-pituitary-gonadal axis. Accordingly, there was no decrease in the serum or ovarian levels of estradiol 17ß and progesterone and there was no difference in the ovarian weight. However, a significant decrease in the uterine weight and difference in the histology of the uterus of the immunized animals was observed. This is of significance, considering the fact that the presence of the GnRH receptor has been reported in the uterus also. In an attempt to develop an in vitro system to monitor the effect of GnRH receptor antibody, an in vitro incubation system with the human placental villi, which is known to produce both GnRH and hCG was standardized. Sensitive ELISA and RIA were developed for GnRH and hCG, respectively to monitor their levels.The results of the studies on the effect of addition of GnRH receptor antibody to the immunoreactive hCG levels in the placental incubation medium are presented in Chapter 5. In addition, advantage was taken of the report of the presence of the specific receptors for GnRH in the Leydig cells of the rats, to evaluate the effect of the GnRH receptor antibodies on the function of leydig cells. Results of studies in which the effect of addition of GnRH receptor antibodies on the testosterone production by purified rat Leydig cells were monitored revealed that there was no inhibitory effect. Finally in the Chapter 6, a general discussion and critical evaluation of the results obtained in the study, in light of similar studies reported in literature are presented.

Thesis (Ph. D.)--University of Washington, 2002. / Vita. Includes bibliographical references (leaves 163-232).