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The effects of neonatal exposure to testosterone on the development of behaviour in female marmoset monkeys.

Experimental investigations of sexual differentiation in primates have been mainly confined to the rhesus monkey, a highly polygamous species with marked anatomical and behavioural sexual dimorphism. The marmoset is a monogamous monkey which shows little anatomical or behavioural sexual dimorphism, and both sexes exhibit positive feedback in response to the administration of oestradiol. This monkey has a relatively short gestation period of 144 days and usually gives birth to dizygotic twins. These share a common placental circulation and hence develop as haematopoietic chimaeras. However, a female with a male co-twin is not adversely affected as a result of this and the freemartin condition does not occur. Since the newborn male experiences raised testosterone levels soon after birth it is possible that the 'critical period' for sexual marmosets were implanted with 25 mg of testosterone for 50 days after birth. After removal of the implants their genitalia remained partly masculinized and they initiated more masculine rough-and-tumble play than their controls. After puberty they exhibited a mixture of male and female sexual behaviour when presented with unfamiliar normal males and females during 15-minute behavioural trials. Four of the implanted females have also ovulated. Neonatal administration of testosterone therefore has had an organizing effect on female behaviour. Whether the marmoset is exceptional in this respect, or whether other primates which exhibit a male neonatal testosterone surge, such as man and the rhesus monkey, also undergo some postnatal behavioural differentation, remains to be seen.

Animals

Immunological interference with the maternal recognition of pregnancy in primates.

If chorionic gonadotropin is involved, as part of the luteotrophic stimulus, in the maternal recognition of pregnancy in primates, then immunization against the beta subunit of hCG should prevent a conception going beyond the length of the normal cycle. Immunization against hCG-beta suppresses fertility in marmoset monkeys, baboons and rhesus monkeys, as long as the antibody titres remained high. As titres decline, however, female marmosets experience recurrent abortions interspersed with occasional live births. Young marmosets born to mothers with low circulating levels of antibody reach apparently normal sexual maturity. Immunization against hCG-beta subunit may have potential as a possible long-term method of fertility control in humans, but its reversibility needs further study. There is still a lack of basic knowledge of the functions of chorionic gonadotropin, especially now that its restriction to pregnancy seems improbable. Immunization against hCG may be valuable as a research tool with which to study the immunological relationships between mother and fetus during early pregnancy.

Animals

Fine structural observations on hemopoiesis in the chorioallantoic placenta of the marmoset.

Hemopoietic sinuses in the chorionic trabeculae of marmoset placenta were examined electron microscopically at several stages of pregnancy. Tissues were prepared both by standard technique and following injection of exogenous peroxidase into the maternal circulation prior to autopsy, and subsequent cytochemical localization of its reaction product in the fetal placenta. Aside from their dilated size and cellular content, the sinuses resembled the allantoic capillaries of the peripheral umbilical circulation in this species; and in early and midgestational placentas, sinusal lumina, differentiating elements in the erythroid line were present in placentas at all stages. Such cells appeared to arise both by a process of proliferation of undifferentiated progenitors from the sinusal lining and by mitosis of pre-existing erythroid cells. Only the latter process was observed in the mature placenta near term. Granulopoiesis was not observed at any stage; on the other hand, cells in the thromboid line were identified in placentas believed to be near midgestation; and large intrasinusal phagocytes were present in both midgestational and late placentas. In the latter, they were seen to have endocytosed peroxidase, and, by tenuous, dendriform processes, to be related anatomically to large numbers of erythroid cells. Early in gestation, the developing erythroid elements were closely related by areas of contact, both to each other and to the delimiting endothelium of the sinuses.

Allantois

Phylogenetic implications of comparative primate growth rates.

Growth data from a number of species of Old and New World primates have been analyzed by calculating instantaneous relative growth rates. Species discussed are the New World species Saimiri sciureus and Saguinus nigricollis, and the Old World species Pan troglodytes and Macaca mulatta. The analysis of the perinatal growth data indicated that differences in relative growth rates are present during early periods of growth. More specifically, it was found that the closer taxonomically a species is to man the greater the deceleration of growth during the first postnatal year. It is suggested that this may be a general primate trend.

Age Factors

A brainstem atlas of catecholaminergic neurons and serotonergic perikarya in a pygmy primate (Cebuella pygmaea).

The present paper provides a brainstem atlas showing the distribution of catecholaminergic cells and processes, as well as serotonergic perikarya, in the pygmy marmoset. The findings revealed by the Falck and Hillarp histofluorescence method conform in essential details to what has been described in rodents. These and other comparative data indicate that a prototypical pattern of organization of aminergic systems has been retained in the evolution of primates.

Animals

Patterns of retinal terminations and laminar organization of the lateral geniculate nucleus of primates.

Autoradiographic tracing procedures have been used to study the organization of retinogeniculate axons in seven primates, i.e., four species of New World monkeys, one species of Old World monkeys and two species of prosimians. These data suggest that the basic primate pattern of geniculate lamination consists of two parvocellular layers, two magnocellular layers, and two poorly developed and highly variable superficial (S) layers which are ventrally located. Ocular input to each member of each of the three pairs differs. In the macaque, the squirrel, and the saki monkey, the parvocellular layers subdivide and interdigitate into four leaflets so as to give the appearance of four parvocellular "layers." These leaflets are much less extensive in the owl and marmoset monkeys. In some individual macaque monkeys, there is further splitting of the parvocellular leaflets into subleaflets, giving the appearance of six parvocellular "layers." The prosimians (galago and slow loris) have two additional layers that are not found in pithecoid primates, and only one superficial layer is apparent. The two additional layers are termed "koniocellular" since they consist of very small cells. Finally, New and Old World monkeys have both ipsilateral and contralateral retinal input to the interlaminar zones. We conclude that the basic pattern of lateral geniculate organization is six layers, but not the traditional six. Prosimians have evolved two additional layers, the koniocellular layers, and have possibly lost one superficial layer. Both New World and Old World monkeys have elaborated the parvocellular layers by forming leaflets to varying extents. With the possible exception of the single S layer in prosimians, layers form pairs that are similar in cell types, but different in ocular input.

Animals

Lymphoproliferative disease in a cotton-top marmoset after inoculation with infectious mononucleosis-derived Epstein-Barr virus.

Injection of concentrated EBV derived from cells of the Kaplan line of infectious mononucleosis (IM) origin resulted in malignant lymphoproliferation in one out of three cotton-top marmosets 6 weeks after inoculation. Two additional animals receiving the same isolate after incubation with an antibody-containing human serum did not develop tumors. Inoculation of concentrated virus derived from the P3HR-1 line of Burkitt origin did not lead to lymphoproliferations in five marmosets. Three of these received non-neutralized, and two received neutralized P3HR-1 virus. The tumor obtained with the Kaplan isolate revealed characteristics of a lymphosarcome. It contained EBV-specific DNA. In addition, EBV-synthesizing lymphoblastoid lines were established from a tumorous lymph-node, as well as from the spleen of the diseased marmoset. Virus recovered from these lines transformed lymphocytes derived from spleens of healthy marmosets. The tumor-bearing animal developed low levels of anti-VCA antibodies during the course of tumor growth. These data demonstrate the oncogenic potential of EBV directly derived from cells of IM origin.

Animals

Characteristics of three strains of feline fibrosarcoma virus grown in cat and marmoset monkey cells.

Two strains of feline fibrosarcoma virus (ST-FeSV and GA-FeSV) were found to induce tumors in cats and marmosets, and to transform feline and marmoset cells in vitro after primary inoculation. A third strain (SM-FeSV) failed to induce tumors or transform marmoset cells after primary inoculation; however, when SM-FeSV-injected marmoset cultures were passed 26 times in vitro, the cell cultures released infectious virus which transformed marmoset fibroblasts but still failed to induce tumors in marmosets. ST-FeSV induced mainly round-cell type transformation (r foci), GA-FeSV induced predominantly mixed round-fusiform cell type transformation (fr foci), and SM-FeSV induced r and fr type foci with a higher proportion of fusiform cells in the fr foci than seen with GA-FeSV. Transforming virus was obtained from r or mixed r/fr foci of ST-FeSV but not from fr foci; heat treatment changed the virus from producing almost exclusively r type foci to inducing an increased number of fr foci. Passage of FeSV in cat cells yielded viruses with a higher ratio of infectivity for feline vs marmoset cells, while passage of FeSV in marmoset cells yielded virus with a relatively higher infectivity ratio for marmoset cells; the three strains differed in the degree of change in the infectivity ratio. Despite the alteration of host range of SM-FeSV propagated in marmoset fibroblasts, the virus retained feline P-30 antigen by CF and FA assays. Neutralization tests did not indicate but also did not exclude an alteration of the surface antigens of ST-FeSV or SM-FeSV propagated in marmoset fibroblasts. The alteration of the relative infectivity of FeSV during passage in marmoset cells may be due to: (1) the selection of a variant present in the original heterogenous uncloned population; (2) mutation; or (3) recombination with some marmoset genetic material, possibly an as yet unidentified endogenous marmoset virus.

Animals

Epstein-Barr virus: experimental infection of Callithrix jacchus marmosets.

Eight common marmoset monkeys (Callithrix jacchus) were inoculated with about 10(4) transforming units of B95-8 virus; seven of the marmosets died 50-111 days post inoculation and all seven showed microscopic and/or macroscopic lesions compatible with a diagnosis of lymphoproliferative disease. Low levels of anti-VCA antibodies were detected in plasma from six marmosets. Attempts failed to establish continuous EBV-carrying lymphoblastoid cell cultures by cultivation in vitro of circulating lymphocytes or minced lymphoid tissues obtained at necropsy.

Animals

Establishment of simian sarcoma virus, type 1 (SSV-1)-transformed non-producer marmoset cell lines.

Simian sarcoma virus, type 1 (SSV-1)-transformed non-producer cell lines were established by infection of normal marmoset fibroblast cells (HF) with limiting dilutions of SSV-1. Four focus-derived cell lines were identified as non-producers by assay of culture fluids for focus-forming activity and by the mixed culture cytopathogenicity test with XC cells. Further studies failed to detect production of type-C virus by 3H-uridine labelling, reverse transcriptase assay or electron microscopy. The non-producer cell lines, designated HF/SSV-NPI, IV, V, and VI, appeared morphologically transformed and cloned in soft agar with the same efficiency as HF/SSV-1 virus-producing transformed cells. Expression of either cytoplasmic or cell surface virus-related antigens was not detected by immunofluorescence or serum cytotoxicity tests. The presence of sarcoma genome in the transformed non-producer cell lines was demonstrated by rescue of focus-forming activity following superinfection with non-transforming helper virus or by cocultivation with helper-virus-producing cell lines. The SSV-1-transformed non-producer primate cells provide a useful tool for future studies.

Animals

Transformation of lymphocytes by Herpesvirus papio.

Cotton-topped (CT) or white-lipped (WL) marmoset lymphocytes were transformed in vitro with herpesvirus papio (HVP) into permanently growing lymphoblastoid cell lines (LCL). Five of 9 HVP-transformed CT cell lines contained cells with antigens reacting with antibodies to Epstein-Barr virus (EBV) capsid antigen (VCA) and/or to EBV-induced early antigens (EA). None of 12 WL LCL revealed such antigen-producing cells. Cells from both groups of cultures failed to react with antibodies to the EBV-specified nuclear antigen (EBNA). Exposure of baboon circulating lymphocytes to X-irradiated HVP or EBV-carring cells, or to suspensions of EBV resulted in establishment of LCL which all contained VCA and/or EA-positive, but no EBNA-positive cells. Nuclear antigens were undetectable also with anti-VCA-positive sera from baboons, chimpanzees, or other non-human primates. DNA-complementary RNA (cRNA) filter hybridization with EBV cRNA showed that with one exception transformed CT or WL marmoset cells contained at least 1-2 virus genome equivalents per cell, while at least 12-25 virus genome equivalents per cell were detected in transformed baboon cells. These data need confirmation by DNA-DNA reassociation kinetics.

Animals

Biologic and antigenic characteristics of Epstein-Barr virus-related Herpesviruses of chimpanzees and baboons.

Leukocyte-transforming agents were isolated in baboon leukocytes inoculated with oral excretions from immunosuppressed chimpanzees. The transformed lymphoblasts had B cell surface markers and harbored herpes-type virus particles; 5-10% of the cells contained cytoplasmic antigens reactive with Epstein-Barr virus (EBV)-antibody-positive chimpanzee, human and baboon sera. These sera also neutralized the transforming activity of the chimpanzee virus. Long-term lymphoid cell lines were established from circulating lymphocytes of normal baboons: two from Papio cynocephalus and three from P. hamadryas. The cells had B cell surface markers, contained herpes-type virus particles and produced virus with leukocyte-transforming activity. No virus-associated nuclear antigen was detectable with reference baboon and chimpanzee sera; however, the cells reacted with selected human sera containing antibodies to EBV nuclear antigen (EBNA). Absorption experiments confirmed the specificity of this reaction. Baboon lymphoblasts produced baboon virus-associated soluble complement-fixing (CF/S) antigen. Baboon sera had CF antibodies to viral (CF/V) antigen derived from EBV but failed to react with EBV-associated CF/S antigen. Chimpanzee and baboon herpesviruses had similar in vitro host cell ranges but were different from those of EBV. Inoculation of baboons, rhesus monkeys and cottontop marmosets failed to produce detectable illness or palpable tumors.

Animals

Transformation of marmoset lymphocytes in vitro with Herpesvirus ateles.

Circulating lymphocytes from three species of marmoset monkeys were transformed in vitro with Herpesvirus ateles (HVA) either by co-cultivation with lethally X-irradiated HVA-producing lymphoblastoid cells or by infection with cell-free virus: 42 transformed lymphocyte cultures were obtained from 72 transformation attempts. Attempts to transform squirrel monkey lymphocytes were unsuccessful in 29 attempts. Association of HVA with each transformed culture was demonstrated by staining of antigen-positive cells in indirect fluorescent antibody (FA) tests and recovery of HVA after co-cultivation of the transformed cells with permissive monolayer cells. Cells of most transformed cultures possessed T lymphocyte properties: i.e. formation of E rosettes, reactivity with a specific anti-marmoset T lymphocyte serum and lack of surface Ig.

Animals

Heterogeneity of Epstein-Barr virus. IV. Induction of a specific antigen by EBV from two transformed marmoset cell lines in Ramos cells.

Infection of cells of the EBV-genome-negative human B-lymphoma Ramos line with viral isolates obtained from two EBV-transformed marmoset cell lines (B95/8; Nyevu) resulted in the induction of a nuclear antigen (RAM-ag) apparently different from other EBV-associated antigen complexes. This antigen is revealed by indirect immunofluorescence and shows no detectable cross-antigenicity with EBNA or any other known EBV-associated antigen. EBV-isolates from P3HR-1 cells fail to induce a similar antigen in Ramos cells although they induce EBNA. No RAM-ag was expressed, either after infection of cells of another EBV-genome-negative human B-lymphoma line BJAB with B95-8 EBV or in a series of EBV-harbouring cell lines. Thus the antigen appears to be cell-line-specific for Ramos cells. It is also induced upon infection of either B95-8 or P3HR-1 converted Ramos sublines with EBV from B95-8 cells. All human sera with RAM-ag-reactivity revealed antibodies against VCA. However, sera from patients with acute infectious mononucleosis containing high anti-VCA-antibodies did not react with RAM-ag. Seroconversion for this antigen apparently more closely coincides with the appearance of EBNA-directed antibodies.

Animals

Biochemical identification of primate lymphoid cell-surface glycoproteins.

We have employed the galactose oxidase-tritiated sodium borohydride labelling method to examine the surface glycoproteins of cotton-topped marmoset and other primate cell lines either established from tumors or transformed in vitro by different lymphotropic herpesviruses. The labelled surface glycoproteins were separated on acrylamide gels in the presence of sodium dodecyl sulfate (SDS) and analyzed by fluorography. Our results indicate that (1) lymphocytes of the same class from different primate species are similar but can be distinguished; (2) T and B lymphocytes of the same species can be differentiated; (3) cotton-topped marmoset lymphocytes of the same class show marked similarities regardless of tumor or in vitro origin or virus used for transformation; (4) three cell lines established from different EBV-induced tumors of the same marmoset show essentially the same labelling pattern, supporting the hypothesis that they originated from a single clone.

Animals

Biological and biochemical observations on isolates of EB virus from the malignant epithelial cells of two nasopharyngeal carcinomas.

Foetal, adult seronegative, and cotton-top marmoset lymphocytes have been transformed into cell lines by EB virus from the malignant epithelial cells of two nasopharyngeal carcinomas. The nature of the cell lines was checked by karyotyping and by light and electron microscopy, and the presence of the EB virus genome was demonstrated by immunofluorescence tests. Immunofluorescence also showed the incidence of EB virus-producing cells in each line and this was checked by electron microscopy and the use of a lymphocyte transformation assay. Two foetal-derived lines did not produce virus spontaneously, could not be activated with various inducers, and were found by DNA reassociation kinetics to carry only small numbers of genome copies per cell. An adult-derived line produced virus, could be activated to produce more, and provided enough infectivity to transform marmoset cells. The resulting marmoset line made profuse transforming virus; it thus provides the first abundant source of NPC-derived infectious EB virus for comparative studies. The results are discussed in relation to the interactions with comparable target cells of EB virus from normal individuals and from patients with other diseases.

Animals