Distribution of glutamine-like immunoreactivity in the cerebellum of rat and baboon (Papio anubis) with reference to the issue of metabolic compartmentation

Summary The cellular and subcellular localization of glutamine, a major glutamate precursor, was studied by means of an antiserum raised against glutaraldehydefixed glutamine. Ultrathin sections from the cerebellar cortex of rat and baboon (Papio anubis) were incubated sequentially in the primary antiserum and in a secondary antibody coupled to colloidal gold particles. The labelling intensity was quantified by computer-aided calculation of gold particle densities. High levels of immunoreactivity occurred in glial cells (Bergmann fibres, astrocytes, and oligodendrocytes), intermediate levels in cell bodies and processes of granule cells, and low levels in terminals of presumed GABAergic or glutamatergic fibres (terminals of basket and Golgi cells, and of parallel, mossy, and climbing fibres). The labelling intensity of Purkinje cells showed some variation, but never exceeded that in glial cells. Within the nerve fibre terminals, the glutamine-like immunoreactivity showed some preference for mitochondria, but was otherwise evenly distributed. The predominant glial localization of glutamine was also obvious in light microscopic preparations processed according to the postembedding peroxidase-antiperoxidase procedure. Gold particle densities over different types of profile in glutamine immunolabelled sections were compared with particle densities over the corresponding types of profiles in neighbouring sections labelled with an antiserum to glutaraldehyde-fixed glutamate. The glutamate/glutamine ratio, expressed arbitrarily by the ratio between the respective gold particle densities, varied by a factor of about 6, with the highest ratio in the putative glutamatergic mossy and parallel fibre terminals, and the lowest ratio in glial elements. The remaining tissue components displayed intermediate ratios. The present study provides direct morphological evidence for the existence in the brain of distinct compartments with differing glutamate/glutamine ratios.This paper is dedicated to Professor Fred Walberg on the occasion of his 70th birthdayOn leave of absence from Department of Anatomy, Capital Institute of Medicine, You An Men Street, Beijing, China     

N-methyl-D-aspartate receptors in the cortex and hippocampus of baboon (Papio anubis and Papio papio)

In vitro autoradiography was used to examine the N-methyl-D-aspartate receptor in the brain of a baboon species, Papio anubis, and compared to that of Papio papio which exhibits a photosensitive epilepsy. The epilepsy originates in the frontal cortex and is accompanied by an enhanced sensitivity to N-methyl-D-aspartate. In both Papio anubis and Papio papio, the density of N-methyl-D-aspartate receptors was greatest in the hippocampus, followed by associational areas including frontal cortex, and low in primary sensory areas such as the visual cortex. The receptors were concentrated in the outer cortical layers I-III, very low in layer IV except in primary visual cortex, and of intermediate density in layer V. The density of binding sites was approximately two-fold lower than previously observed in the rodent brain, whereas the affinity of the receptor for [3H]L-glutamate was greater in the primate versus the rodent brain. Glycine potentiated the binding of [3H]L-glutamate in both cortex and hippocampus. No significant differences in the properties of N-methyl-D-aspartate receptors were observed between the two baboon species, suggesting that the photosensitivity of Papio papio is not due to alterations in the binding of L-glutamate to the N-methyl-D-aspartate receptor complex.     

Profiles enriched in GABA-like immunoreactivity participate in serial synapses in the pontine nuclei of baboon (Papio anubis)

Summary Using a postembedding immunogold procedure with an antiserum against glutaraldehyde-fixed GABA, we demonstrate GABA-like immunoreactivity in two classes of synaptic profiles in the pontine nuclei of baboon. One is an axon terminal in symmetrical synaptic contact with small or medium-sized GABA-immunonegative dendrites, the other is a pale, vesicle-containing profile resembling a dendrite or dendritic process which participates in serial synaptic arrangements. These synaptic arrangements, or triads, consist of a GABA-like immunoreactive, pale vesicle-containing profile being postsynaptic to a GABA-immunonegative axon terminal, and presynaptic to a small or medium-sized GABA-immunonegative dendrite. In at least some of these triads, the GABA-immunonegative axon terminals also contact the GABA-immunonegative dendrite directly.     

Comparative aspects of oxytocin in baboon (Papio hamadryus anubis) and human corpora lutea

In spite of the importance of the corpus luteum in human reproduction,little is known about its formation after ovulation and duringregression in the absence of conception. This is largely dueto constraints on the availability of normal human tissue: thereforean appropriate model which could be studied and provide informationapplicable to the human was sought. The baboon (Papio), a non-humanprimate, has been determined to be one such model. Thus, inthe past several years our studies have examined the role ofluteal peptides in corpus luteum function, and, when possible,we have attempted to examine corpora lutea from the human andbaboon in parallel. Although a milk-ejection factor was recognizedto be present in luteal tissue in 1910 (Ott and Scott, Proc.Soc. Exp. Biol. Med., Vol. 8, p 49), the role of oxytocin inluteal physiology has not been easy to ascertain. This is inpart due to the methodologies employed to assess its role. Ourstudies summarized below suggest that oxytocin does not directlyaffect luteal steroidogenesis, but that it may play a role incell to cell communication involving the expression of the gapjunction proteins, the connexins. In view of the fact that oxytocin,its receptor, gap junctions and associated proteins are notunique to the human and non-human primates, the model of lutealdevelopment and demise proposed may be applicable to most species.     

Segmental morphometrics of the olive baboon (Papio anubis): a longitudinal study from birth to adulthood

The linear dimensions and inertial characteristics of the body are important in locomotion and they change considerably during the ontogeny of animals, including humans. This longitudinal and ontogenetic study has produced the largest dataset to date of segmental morphometrics in a Catarrhini species, the olive baboon. The objectives of the study were to quantify the changes in body linear and inertial dimensions and to explore their (theoretical) mechanical significance for locomotion. We took full‐body measurements of captive individuals at regular intervals. Altogether, 14 females and 16 males were followed over a 7‐year period, i.e. from infancy to adulthood. Our results show that individual patterns of growth are very consistent and follow the general growth pattern previously described in olive baboons. Furthermore, we obtained similar growth curve structures for segment lengths and masses, although the respective time scales were slightly different. The most significant changes in body morphometrics occurred during the first 2 years of life and concerned the distal parts of the body. Females and males were similar in size and shape at birth. The rate and duration of growth produced substantial size‐related differences throughout ontogeny, while body shapes remained very similar between the sexes. We also observed significant age‐related variations in limb composition, with a proximal shift of the centre of mass within the limbs, mainly due to changes in mass distribution and in the length of distal segments. Finally, we observed what we hypothesize to be ‘early biomechanical optimization’ of the limbs for quadrupedal walking. This is due to a high degree of convergence between the limbs’ natural pendular periods in infants, which may facilitate the onset of quadrupedal walking. Furthermore, the mechanical significance of the morphological changes observed in growing baboons may be related to changing functional demands with the onset of autonomous (quadrupedal) locomotion. From a wider perspective, these data provide unique insights into questions surrounding both the processes of locomotor development in primates and how these processes might evolve.     

Localization and expression of zonula occludens-1 tight junction-associated protein in baboon (Papio anubis) corpora lutea

The identification of the cell junction-forming proteins connexin-43,a gap junction protein and E-cadherin, which is a componentof adherent junctions, in the corpus luteum of both humans andbaboons suggests that cell-cell interactions and metabolic cooperationmust occur in this tissue. Occluding junctions are a third typeof junction which form a physical barrier between cells. Thus,our aims in this study were firstly to examine the presenceof the tight junction-associated protein zonula occludens-1(ZO-1) by immunohistochemistry, and secondly to determine theconcentrations of this protein in the early, mid- and late lutealphase baboon corpora lutea of the menstrual cycle by a Westernanalysis. ZO-1 was localized mainly at the periphery of theluteal cells, and the intensity of immunoreactivity varied throughthe luteal phase, with comparatively stronger immunoreactivityin the mid-luteal phase than the early and late luteal phases.Atretic corpora lutea were devoid of activity. By Western analysis,bands of immunoreactivity were observed at 225 kDa, furtherconfirming the presence of the protein. Maximum activity, asdetermined by densitometry, was observed in the mid-luteal phase.These data infer the presence of tight junctions in the corpusluteum and suggest that expression of the ZO-1 protein formingthese junctions may be hormonally regulated within this tissue.     

The susceptibility to praziquantel of Schistosoma haematobium in the baboon (Papio anubis) and of S. japonicum in the vervet monkey (Cercopithecus aethiops)

Summary Baboons infected with S. haematobium and vervet monkeys infected with S. japonicum were treated orally with different dosage regimens of praziquantel. The progress of the infections in the primates was followed by weekly faecal and urine egg counts before and after treatment. The response to treatment was also monitored by observing oogram changes in rectal snips. The baboons and vervet monkeys were autopsied and perfused 3–4 months after treatment.The results of the praziquantel treatments of baboons infected with S. haematobium show that a single administration of 100 mg/kg in one day was as effective as 50 mg/kg for five consecutive days in producing a complete cure. A single dose of 30 mg/kg failed to stop egg laying but retreatment with 50 mg/kg administered in one day resulted in cessation of egg laying 12 days after treatment and only one immature female worm and 33 male worms were recovered at autopsy. A baboon treated with 30 mg/kg administered at 10 mg/kg three times in one day was not cured, but when retreated with 75 mg/kg, administered in three separate doses of 25 mg/kg in one day, egg laying stopped 15 days afterwards and only three male worms were recovered at autopsy. The results suggest that a single oral dose of 75–100 mg/kg bwt is likely to be effective against S. haematobium in the baboon. An intramuscular injection of 200 mg/kg bwt was well tolerated by one animal.The results obtained with praziquantel against S. japonicum in the vervet monkey show that a complete cure was obtained in the animal given 50 mg/kg on five consecutive days, but the regimens of a single dose of 20 mg/kg followed by two separate doses of 10 mg/kg during one day, and of 10 mg/kg given three times during one day, resulted in only partial parasitological cures.The results based upon faecal egg output studies and the oograms taken after treatment suggested that praziquantel is more effective against S. japonicum than S. haematobium in the doses given, but the subsequent autopsies showed that some S. japonicum adult worms had survived treatment.A baboon naturally infected with S. mansoni, with a daily egg output of 150 eggs, was completely cured by a single oral dose of 50 mg/kg of praziquantel. This dose however failed to cure another naturally infected animal with a daily egg output of 1,800 eggs.It is considered that a predominant characteristic in the pathology of the animals given a curative dose of praziquantel was the total resolution of cellular reaction and fibrosis in the tissues containing known numbers of dead residual eggs. In the baboons with S. haematobium, the ureters and bladder had recovered their functional integrity and in the vervet monkeys infected with S. japonicum, a similar resolution of pathology in the liver and bowel was apparent.The results show that praziquantel is effective against patent S. haematobium and S. mansoni infections in baboons and against S. japonicum in vervet monkeys, in relatively low dosage regimens (100 mg/kg and < 100 mg/kg) applied in one day, as shown by suppression of egg laying and reduction or complete elimination of adult worms. Female worms of S. haematobium are apparently more susceptible to the compound than male worms. The classic hepatic shift of adult schistosomes was observed in all of the primates treated in the present series, but histopathological studies showed that numerous worms also died in situ, while only a few were found in the lungs.

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Zusammenfassung Mit S. haematobium infizierte Paviane und mit S. japonicum infizierte Meerkatzen wurden mit unterschiedlichen oralen Dosen von Praziquantel behandelt. Der Infektionsverlauf wurde bei den Affen vor und nach der Behandlung durch wöchentliche Eizählungen im Kot und im Urin verfolgt. Die Wirkung der Behandlung wurde anhand der Oogrammveränderung in rectalen Biopsieproben untersucht. Drei bis vier Monate nach der Behandlung wurden die Tiere seziert und perfundiert.Die Behandlung S. haematobium-infizierter Paviane mit Praziquantel erbrachte folgende Ergebnisse: Die orale Verabreichung von 1×100 mg/kg oder von 50 mg/kg/die an fünf aufeinanderfolgenden Tagen führte zur parasitologischen Heilung. 1×30 mg/kg hatte keinen Einfluß auf die Eiausscheidung, diese hörte jedoch 12 Tage nach einer Nachbehandlung mit 1×50 mg/kg auf. Bei der Autopsie dieses Tieres fanden sich lediglich ein junges Weibchen und 33 männliche Würmer. Ein mit 30 mg/kg (3×10 mg/kg an einem Tag) behandelter Pavian wurde nicht geheilt. Die Nachbehandlung mit 75 mg/kg (3×25 mg/kg an einem Tag) bewirkte ein Sistieren der Eiausscheidung nach 15 Tagen. Bei der Sektion fanden sich nur drei männliche Würmer.Die Ergebnisse lassen vermuten, daß eine einmalige orale Gabe von 75–100 mg/kg KW gegen S. haematobium im Pavian wirksam ist. Eine intramuskuläre Injektion von 200 mg/kg wurde von einem Tier gut vertragen.Mit S. japonicum infizierte Meerkatzen erwiesen sich nach der Verabreichung von 50 mg/kg/die an fünf aufeinanderfolgenden Tagen als parasitologisch geheilt. Dagegen bewirkten Gaben von 1×20 mg/kg gefolgt von 2×10 mg/kg an einem Tag oder von 3×10 mg/kg/die lediglich eine Teilwirkung.Die Ergebnisse der Kotuntersuchungen und der Oogramme nach der Behandlung lassen vermuten, daß Praziquantel in den verabreichten Dosen gegen S. japonicum wirksamer ist als gegen S. haematobium. Die anschließenden Sektionen zeigten allerdings, daß einige adulte S. japonicum-Würmer die Behandlung überlebt hatten.Ein natürlich mit S. mansoni infizierter Pavian mit einer Tageseiausscheidung von 150 Eiern wurde durch einmalige orale Gabe von 50 mg/kg geheilt. Diese Dosis war bei einem anderen, ebenfalls natürlich infizierten Tier mit einer täglichen Eiausscheidung von 1.800 Eiern nicht voll wirksam.Bei den mit curativen Dosen von Praziquantel behandelten Tieren kam es zu einer bemerkenswerten Rückbildung der typischen pathologischen Organveränderungen. Die zellulären Reaktionen und die fibrösen Bildungen in den Geweben, die eine bekannte Anzahl abgestorbener Eier enthielten, waren völlig aufgelöst. Bei den mit S. haematobium infizierten Pavianen gewannen die Harnleiter und die Harnblase ihre funktionelle Integrität zurück. Auch bei den mit S. mansoni infizierten Meerkatzen war eine entsprechende Auflösung pathologischer Veränderungen in der Leber und im Darm zu erkennen.Die Ergebnisse zeigen, daß Praziquantel gegen patente Infektionen mit S. haematobium und S. mansoni im Pavian und mit S. japonicum in der Meerkatze in relativ niedrigen, an einem Tag verabreichten Dosierungen (100 mg/kg und < 100 mg/kg) wirksam ist. Die klassische liver shift der adulten Schistosomen wurde bei allen in dieser Untersuchung behandelten Tieren gesehen. Histopathologische Untersuchungen zeigten jedoch, daß zahlreiche Würmer in situ starben, während nur wenige in der Lunge gefunden wurden. Dies äußerte sich in einem Aussetzen der Eiausscheidung und einer Reduktion oder in der quantitativen Elimination der adulten Würmer.Bei S. haematobium sind die Weibchen offensichtlich gegenüber Praziquantel empfindlicher als die Männchen.

     

Compartmental distinctions in uterine Muc-1 expression during early pregnancy in cynomolgous macaque (Macaca fascicularis) and baboon (Papio anubis)

BACKGROUND: Loss of the transmembrane mucin, Muc-1, is a molecular correlate of the acquisition of uterine receptivity to embryo adhesion in most species examined. In macaques, two distinct adhesion events occur at opposite sides of the uterus. Attachment to the secondary site is delayed relative to the primary site. The aim was to determine if Muc-1 is removed at secondary sites prior to trophoblast attachment. METHODS: We examined Muc-1 expression in the uteri of cynomolgus macaque and baboon during early implantation by immunocytochemistry. RESULTS: Luminal epithelia were devoid of Muc-1 at all stages examined at both primary and secondary adhesion sites. Loss of Muc-1 in luminal epithelia was found to be maternally determined, accompanied membrane transformation in both macaque and baboon, and at secondary implantation sites, preceded trophoblast attachment. In contrast, glandular epithelia in pregnant macaques exhibited a temporal and compartmentalized gradient of Muc-1 loss confined to the implantation sites. Glandular epithelia in the pregnant baboon uterus were uniformly negative for Muc-1. CONCLUSIONS: Restriction of the Muc-1 loss in glandular epithelia to conceptual cycles may reflect the fundamental distinctions among epithelia of the various uterine compartments and the differential modulation of Muc-1 that occurs within these compartments in conceptual and non-conceptual cycles.     

Glycine-like immunoreactivity in the cerebellum of rat and Senegalese baboon,Papio papio: a comparison with the distribution of GABA-like immunoreactivity and with [3H]glycine and [3H]GABA uptake

Summary An antiserum against conjugated glycine was characterized and applied to cerebellar sections of rats and baboons that had been perfusion-fixed with glutaraldehyde. After immunosorbent purification the serum reacted with brain protein-glutaraldehyde-glycine conjugates, but did not stain similar test conjugates prepared from other amino acids, including GABA and -alanine. In the rat cerebellum the glycine antiserum selectively labelled a subpopulation of Golgi neurons. Adjacent Vibratome sections treated with an antiserum against conjugated GABA revealed an about equally large subpopulation of immunopositive Golgi cells. A proportion of the Golgi cells that were cleaved by the plane of section contained both immunoreactivities. Additional evidence for a colocalization of glycine and GABA was obtained by postembedding staining of alternate semithin sections with the GABA antiserum and glycine antiserum, respectively. The ability of the antisera to distinguish between fixed glycine and GABA was corroborated by preincubation of the antisera with glutaraldehyde-amino acid fixation complexes: glycine complexes abolished staining with the glycine antiserum but had no effect on the GABA antiserum. The opposite effects were obtained with the GABA complexes. Matching the distributions of the respective immunoreactivities, [³H]glycine uptake was restricted to glomerulus-like structures in the granule cell layer whereas [³H]GABA uptake also occurred in punctate and fibrous profiles in the molecular layer. The baboon showed a distribution of glycine-like immunoreactivity similar to that in the rat, except that a few immunopositive neurons occurred in the molecular layer. The latter neurons were interpreted as outlying Golgi neurons; however, the possibility that they represent a subpopulation of basket cells could not be excluded. The Purkinje cells were negative in both species. Glial cells were weakly stained with the glycine antiserum but were strongly immunopositive after incubation with an antiserum raised against conjugates of the structurally similar amino acid -alanine. The present data suggest that glycine and GABA occur in about equally large subpopulations of Golgi neurons. A subpopulation of the Golgi neurons appears to contain both glycine and GABA.     

A study of germ cell morphology and duration of spermatogenic cycle in the baboon,Papio anubis

Biopsy and orchiectomy specimens were collected from two adult baboons (Papio anubis) at different intervals after intratesticular injection of H³-thymidine. Zenker-formol or Bouin's fixed materials were stained with PASWeigert-Hematoxylin and radioautographed using the H.S.R. (Harleco Synthetic Resin) coating technique. Morphological features of most germ cells appeared similar to those of other monkeys, except that the spermatids in steps 9 to 11 showed a spike-like projection of the acrosome. Also, the type A spermatogonia showed some resemblance to the human type A spermatogonia. The cell associations consisted of 12 stages and a large number of tubular cross sections showed the presence of two or more stages. In Papio anubis, the zygotene spermatocytes are formed in stage VIII, and spermatozoa are released during stages V and VI. H³-thymidine labeling of germ cells indicated that one cycle of spermatogenesis in this species takes approximately 11 days, and complete spermatogenesis occupies 3.8 cycles, or approximately 42 days. The presence of labeled B type spermatogonia 35 days after H³-thymidine injection indicated the existence of some stem cells. The presence of some labeled Ad spermatogonia in these specimens could not be explained. The data indicated that spermatogenesis in this monkey is somewhat different from that in other monkey species described.     

Demonstration of GABA immunoreactive cells in the inferior olive of baboons (Papio papio and Papio anubis)

The distribution of gamma-aminobutyric acid (GABA)-like immunoreactivity was studied in semithin sections through the inferior olivary complex in two baboon species. About 5% of the olivary neurons were GABA-immunoreactive. The GABA-immunoreactive neurons differed from the large majority of olivary neurons by their smaller size and their lower contents of aspartate, as judged by analysis of alternate sections labelled with an aspartate antiserum. The present observations raise the possibility that in primates the GABAergic modulation of the activity of the climbing fibre system is effected not only by the previously described input from the cerebellar nuclei and other extrinsic sources, but that intrinsic neurons also participate.     

Aspartate-like and glutamate-like immunoreactivities in the inferior olive and climbing fibre system: a light microscopic and semiquantitative electron microscopic study in rat and baboon (Papio anubis)

A post-embedding immunogold procedure was used to analyse, in a semiquantitative manner, the distributions of aspartate-like and glutamate-like immunoreactivities in the inferior olive and climbing fibre system in rats and baboons. The neurons in the inferior olive were uniformly labelled for aspartate as well as glutamate, indicating a 100% co-localization of these two amino acids in the cell bodies. The level of glutamate-like immunoreactivity in the climbing fibre terminals was similar to that in the parent cell bodies, as judged by a computer-assisted calculation of gold particle densities. In contrast, the level of aspartate-like immunoreactivity in the climbing fibre terminals was only one-seventh of that of the olivary neurons. No differences were found between the hemispheres and vermis. Nerve terminals in the inferior olive were generally moderately labelled with the aspartate antiserum, as were cell bodies of astrocytes. With a few exceptions, the results obtained in baboons were similar to those in rats. Notably, no evidence was found of an enrichment of aspartate-like immunoreactivity in climbing fibres. The present results do not support previous data suggesting that aspartate is the transmitter of the climbing fibres but indicate that glutamate or another excitatory compound should be considered as candidate for this role. Our findings show that the presence of aspartate-like immunoreactivity in cell bodies is an unreliable indicator of transmitter identity.     

Late luteal rescue in the baboon (Papio cynocephalus)

Numerous studies have used human chorionic gonadotrophin (HCG)administration to study the response of the primate ovary togonadotrophin stimulation. These studies are generally performedin the luteal phase with very few studies of the follicularphase. We have studied the effect of both HCG and gonadotrophinreleasing hormone (GnRH) agonist administered at the early follicularphase in normally cycling baboons (Papio cynocephalus). Fivebaboons were treated with increasing doses of HCG for 5 consecutivedays starting on day 1 of the cycle and three untreated baboonsserved as controls. Follicular and luteal phase lengths weredetermined and serum samples were assayed for progesterone,oestradiol and 17-OH progesterone. In a separate study, sixbaboons were treated with GnRH agonist (WY-40972) on days 2-6of the cycle and saline-treated baboons served as controls (n= 5). Mean peak progesterone concentrations (± SE) duringthe treatment interval were 3.88 ± 0.56 ng/ml in HCG-treatedbaboons compared to 0.19 ± 0.07 ng/ml in controls (P<0.001). A similar significant increase (P <0.001) inserum 17-OH progesterone concentrations was also observed (6.13± 1.12 ng/ml versus 1.13 ± 0.49 ng/ml). In associationwith the increase in luteal steroids there was also a significantprolongation of menstrual cycle length from 32.7 ± 1.2days in controls to 46.8 ± 4.9 days in HCG-treated baboons(P <0.05), which involved prolongation of the follicularphase (16.7 ± 1.2 days to 29.0 ± 4.6 days, P <0.05)with no difference in luteal phase length or progesterone concentrations.In GnRH agonist-treated baboons, mean (± SE) cycle lengthwas prolonged to 46.3 ± 1.6 days and in saline-treatedcontrols was 32.8 ± 0.8 days (P <0.001), again thiswas completely represented by the change in follicular phaselength, from 13.4 ± 0.7 days in controls to 27.2 ±2.1 days in agonist-treated baboons (P <0.001). In contrast,there was no significant difference in luteal phase length betweenthese two groups (19.4 ± 0.7 versus 19.2 ± 1.0days). The prolongation of the follicular phase was accompaniedby significant increases in both progesterone (P <0.01) andoestradiol (P <0.01) during GnRH agonist treatment abovecontrol concentrations. Luteal phase concentrations of thesehormones were not different from controls. These results demonstratethe previously unreported finding that gonadotropin stimulationwill rescue the corpus luteum in the next follicular phase.     

Immunology: Anti-endometrial lymphocytotoxicity and natural killer cell activity in baboons (Papio anubis and Papio cynocephalus) with endometriosis

This study was performed to test the hypothesis that anti-endometrial,lymphocyte-mediated cytotoxicity and natural killer (NK) activityare reduced in baboons with endometriosis when compared to animalswith a normal pelvis. Lymphocyte-mediated cytotoxicity was determinedin 28 baboons (15 with endometriosis, 13 with normal pelvis)and NK cell activity was evaluated in 42 baboons (31 with endometriosis,11 with normal pelvis). Anti-endometrial lymphocyte-mediatedcytotoxicity was determined by a 20 h assay with effector-targetratios of 50: 1 and 25: 1. The NK activity (K562 cell line astarget) was simultaneously measured in all animals during a4 h assay with effector:target ratios of 200: 1, 100: 1, 50:1, 25: 1, 12: 1, 6: 1 and 3: 1. Statistical analysis was performedusing analysis of variance, paired rank, Mann—Whitney,Kruskal—Wallis and Fisher exact tests where appropriate.Lymphocyte-mediated cytotoxicity was significantly lower (P< 0.025) in baboons with endometriosis (mean 5.9 ±8.7 %, median 0%, range 0–26%) than in animals with anormal pelvis (mean 22.9 ± 23.0 %, median 7%, range 0–78%). This difference could be explained by the absence of cytotoxicityin baboons with moderate to severe endometriosis, probably dueto high spontaneous release of ⁵¹Cr from labelled target cells.When stricter criteria were used and only animals with a labellingindex (maximal/spontaneous release) of 1.7 were analysed (n= 11), the anti-endometrial cytotoxicity was comparable betweenbaboons with and without endometriosis. NK cell activity wasalso comparable in primates with and without endometriosis.In conclusion, no difference in lymphocyte-mediated cytotoxicityand NK cell activity was observed between baboons with and withoutendometriosis.     

Ultrastructure of epithelial plaque formation and stromal cell transformation by post-ovulatory chorionic gonadotrophin treatment in the baboon (Papio anubis).

BACKGROUND: To understand factors controlling endometrial responses to pregnancy, we have established a model using the baboon and examined the effects of infused human chorionic gonadotrophin (HCG) on the preparation of the luminal epithelium and stromal cell differentiation for the establishment of pregnancy. METHODS: The ultrastructure of endometrium from normal day 10 post-ovulation animals, cycling females treated with either HCG or FSH (control), and a day 15 pregnant animal has been compared. RESULTS: In the control endometrium, the luminal epithelium was smooth and regular, with underlying spindle shaped stromal cells. In pregnancy, the luminal epithelium underwent a plaque reaction, while stromal cells enlarged and developed filament-rich cell processes. Infusion of HCG produced changes similar to those seen in pregnancy, with generalized plaque formation and stromal decidualization, while in the animal treated with FSH there was no response. CONCLUSIONS: This study indicates that infusion of HCG into the uterus can duplicate many of the responses of the endometrium to pregnancy, although in this case the plaque reaction involved the whole of the luminal epithelium, rather than only the implantation site as in pregnancy.     

Symposium: reproduction in baboons. Secretory proteins of the baboon (Papio anubis) endometrium: regulation during the menstrual cycle and early pregnancy

The biological function of uterine endometrial secretory proteinsin the primate remain to be elucidated. In general, during theluteal phase and under progesterone dominance, the glandularepithelial cells synthesize and secrete a number of proteins.Of these, placental protein 14 (PP₁₄; now referred to as glycodelin)and insulin-like growth factor binding protein-1 (IGFBP-1) arethe best characterized. Although induced by progesterone, theirsynthesis increases exponentially during pregnancy. In the baboon,glycodelin is immunolocalized to the mid functionalis and basalglands between days 10 and 12 post-ovulation. In response toeither exogenous or blastocyst-secreted chorionic gonadotrophin,glandular synthesis increases markedly and remains elevatedup to days 18-25 of pregnancy. The decrease in glycodelin inthe endometrium is associated with glandular regression duringthe first third of pregnancy. In contrast, IGFBP-1 is only observedin the deep basal glands during the luteal phase. Followingthe establishment of pregnancy, IGFBP-1 synthesis switches fromglandular to stromal and is correlated with the process of decidualization.IGFBP-1 synthesis continues to increase throughout gestation.We propose that glycodelin may have immunosuppressive propertiesand that IGFBP-1 may regulate trophoblast migration within theuterine endometrium.     

An immunosuppressive factor in the serum of baboons (Papio anubis) infected with Schistosoma mansoni

Serum taken from baboons infected with Schistosoma mansoni was able to suppress in vitro reactivity of normal baboon lymphocytes. The concanavalin A response was significantly suppressed by such serum, whereas the specific suppression of the phytohaemagglutinin response was minimal. Serum from S. mansoni infected donors also depressed the mixed lymphocyte reactions to xenogeneic targets, but did not affect the specific transformation of lymphocytes stimulated with a parasite Ag. Significant suppressive activity occurred in the baboon serum from 4 to 11 weeks after the initial infection. Serum from animals with a chronic infection of 6-42 months, did not suppress in vitro cell-mediated immunity. The suppressive factor was heat-stable, non-dialysable and, following ultracentrifugation of the suppressive serum, was found to be present in the high mol. wt fraction. From these studies, it is suggested that the immunosuppressive factors are immune complexes, which appear in the serum of the baboons following their infection with this blood parasite.     

Fetal infection of the baboon(Papio cynocephalus) with lymphocytic choriomeningitis virus

Summary Recent observations of LCM-induced fetal damage in humans suggested attempts to develop an animal model for studies on viral congenital malformations. We report herein viral studies on three pregnant baboons(Papio cynocephalus) inoculated subcutaneously with LCM virus strain WE3. The first animal, inoculated in the 9th week of pregnancy, aborted 9 days after a high virus dose. Inoculation of the second baboon during a later stage (23rd week) of pregnancy with a moderate virus dose, resulted in the demonstration of virus in the placenta, amniotic fluid, and cord blood. The infant showed only a slight pleocytosis of the spinal fluid, but no virus shedding and no late sequelae. The third baboon inoculated with a high virus dose during the 21st week of pregnancy delivered an underweight, icteric infant that succumbed on the 6th day. All organs of this animal that were tested contained virus. Microscopic examination of these tissues revealed multifocal necrosis, cerebral glial nodules, meningitis, and bilateral choriovasculitis. These results illustrate that fetal damage observed in the LCM-inoculated baboon resembles that seen in humans following infection with LCM virus.With 8 Figures     

Distribution of acetylcholinesterase activity in the rat embryonic heart with reference to HNK-1 immunoreactivity in the conduction tissue

Acetylcholinesterase (AChE) activity was topographically investigated in the presumptive cardiac conduction tissue regions visualized by HNK-1 immunoreactivity in rat embryos, and AChE-positive cells were examined with the electron microscope. On embryonic day (ED) 14.5, when HNK-1 was most intensely visualized, AChE activity could not be detected enzyme-histochemically in the conduction tissue regions, except in the ventricular trabeculae and part of the AV node. On ED 16.5, however, the AChE activity was clearly demonstrated in some parts of the developing conduction tissue. One exception was the AV node region, where an AChE-positive area was in close proximity to an area showing HNK-1 immunoreactivity but did not overlap. Furthermore, AChE activity was demonstrated predominantly in the ventricular trabeculae, including cardiac myocytes, but was rather weak in the atrium. With the electron microscope, AChE reaction products were observed predominantly intracellulary in both developing conduction tissue cells and developing ordinary myocytes, and no reactivity was found in neuronal components. From ED 18.5 until birth, both AChE activity and HNK-1 immunoreactivity faded away in the conduction tissue. Thus, transient AChE activity in the embryonic heart seems to be different from the developing adult form and may be related to a morphogenetic function in embryonic tissues, as proposed by other authors.