Effects of experimental glaucoma in macaques on the multifocal ERG

Multifocal ERGs (MERGs) of 5 adult monkeys (Macaca mulatta) with inner retinal defects caused by laser-induced glaucoma were compared to MERGs from 3 monkeys with inner retinal activity suppressed pharmacologically. MERGs were recorded with DTL fiber electrodes from anesthetized monkeys. Stimuli consisted of 103 equal size hexagons within 17° of the fovea. Stimuli at each location passed through a typical VERIS m-sequence of white (200 cd/m²) and black (12 cd/m²) presentations. In animals with laser-induced glaucoma, visual field sensitivity was assessed by static perimetry using the Humphrey C24-2 full-threshold program modified for animal behavior. Inner retinal (amacrine and ganglion cell) activity was suppressed by intravitreal injection of TTX (4.7–7.6 μM) and NMDA (1.6–5 mM). In normal eyes the first order response (1st order kernel) was larger and more complex, with more distinct oscillations (>60 Hz) in central than in peripheral locations. The 2nd order kernel also was dominated by oscillatory activity. There were naso-temporal variations in both kernels. Pharmacological suppression of inner retinal activity reduced or eliminated the oscillatory behavior, and naso-temporal variations. The 1st order kernel amplitude was increased most and was largest at the fovea. Removed inner retinal responses also were largest at the fovea. The 2nd order kernel was greatly reduced at all locations. In eyes with advanced glaucoma, the effects were similar to those produced by suppressing inner retinal activity, but the later portion of the 1st order kernel waveform was different, lacking a dip after the large positive wave. Visual sensitivity losses and MERG changes both increased over the timecourse of glaucoma, with changes in the MERG being more diffusely distributed across the visual field. We conclude that 1st and 2nd order responses of the primate MERG can be identified that originate from inner retina and are sensitive indicators of glaucomatous neuropathy. This revised version was published online in July 2006 with corrections to the Cover Date.     

Circulating concentrations of the antiprogestins CDB-2914 and mifepristone in the female rhesus monkey following various routes of administration

The overall aim of these studies was to investigate the oral and i.m. bioavailability of CDB-2914 in intact female rhesus monkeys, and to compare the serum concentrations of CDB-2914 with that of mifepristone following oral administration. In the first study, a 50 mg bolus of CDB-2914 per monkey was administered intravenously, orally or intramuscularly. The area under the serum concentration-time curve for 72 h (AUC(0-72)) following i.v. injection was 18 320 +/- 2718 ng/ml*h, and that for oral administration was 10 464 +/- 3248 ng/ml*h. Thus, the oral bioavailability of CDB-2914 equivalents was 56%. The AUC(0-168 h) following i.m. injection was 11 226 +/- 1130 ng/ml*h. Therefore, the i.m. bioavailability of CDB-2914 equivalents was 62%. In the second study, the serum concentrations of CDB-2914 and mifepristone equivalents were compared following an oral bolus dose in two different formulations. When administered at 5 mg/kg in aqueous suspending vehicle (ASV), the mean peak serum concentration (C(max)) of CDB-2914 equivalents (192 +/- 64 ng/ml) occurred at 5 +/- 1 h, whereas the C(max) of mifepristone equivalents (82 +/- 25 ng/ml) occurred at 3 +/- 1 h. Following administration in gelatin capsules (35 mg/monkey), the C(max) of CDB-2914 equivalents (129 +/- 24 ng/ml) occurred at 5 +/- 1 h, while the C(max) of mifepristone equivalents (31 +/- 8 ng/ml) occurred at 3 +/- 1 h. The serum concentration (AUC(0-120 h)) of CDB-2914 equivalents was 4.7- or 5. 3-fold greater than that of mifepristone equivalents when administered orally in ASV or gelatin capsules respectively. The serum protein binding characteristics of CDB-2914 were also studied. CDB-2914 bound to human alpha(1)-acid glycoprotein (AAG), but not with as high an affinity as mifepristone. In contrast, neither CDB-2914 nor mifepristone bound with high affinity to AAG, corticosteroid binding globulin or sex hormone binding globulin in monkey serum. Collectively, these results indicated that CDB-2914 was more efficiently absorbed than mifepristone following oral administration to female rhesus monkeys.     

Adeno-Associated Virus Delivery of Anti-HIV Monoclonal Antibodies Can Drive Long-Term Virologic Suppression

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CYP2C76 deficiency is embryonic lethal in cynomolgus macaques: The potential role of CYP2C76 in early embryogenesis

Cynomolgus macaques are an important primate species for drug metabolism studies; however cynomolgus CYP2C76, an important drug-metabolizing enzyme, accounts for drug metabolism differences to humans, so that CYP2C76-null animals might show drug-metabolizing properties more similar to humans. In this study, attempts were made to produce CYP2C76-null animals by assisted reproduction technology. Oocytes and sperm collected from the heterozygotes for the null allele (c.449TG > A) were subjected to intracytoplasmic sperm injection, and the embryos produced were cultured in vitro through the blastocyst stage. Preimplantation genetic diagnosis using a biopsied portion of the blastocyst revealed that none of the 32 blastocysts analyzed were homozygotes. In contrast, 2 of the 20 embryos analyzed were homozygotes at the 8-cell stage, indicating that CYP2C76-null embryos most likely stop developing between the 8-cell and blastocyst stage. By polymerase chain reaction, expression of CYP2C76 mRNA was detected in oocytes and blastocysts, but not in 2-, 4-, 8-, or 16/32-cell stage embryos. Metabolic assays showed that CYP2C76 metabolized progesterone. These results indicated that CYP2C76 null was likely embryonic lethal, suggesting its potential role during early embryogenesis in cynomolgus macaques.     

A high frequency of Mamu-A *01 in the rhesus macaque detected by polymerase chain reaction with sequence-specific primers and direct sequencing

SIV infection of rhesus macaques is an excellent model for HIV infection of humans. Unfortunately, it is has been difficult to identify macaques expressing particular MHC class I alleles. Here we describe the use of PCR-SSP for Mamu-A *01 typing of rhesus macaques. The Mamu-A *01 allele was amplified from genomic DNA using Mamu-A *07 -specific primers and positive PCR products were directly sequenced. Our technique identified 15 Mamu-A*01-positive animals of 68 tested. We validated our molecular analysis by showing that lymphocytes from 8 Mamu-A *01 -positive animals expressed Mamu-A*01 as determined by immunoprecipitation and 1-D IEF. The technical simplicity and accuracy of this typing method should facilitate selection of Mamu-A *01 -positive rhesus macaques for AIDS virus pathogenesis and vaccine studies.     

Cognitive development in macaques: Attentional set-shifting in juvenile and adult rhesus monkeys

In humans and nonhuman primates, the structure and function of frontal cortical regions of the brain are not completely developed until early adulthood. How this cortical development affects cognitive function continues to be elucidated. To that end, this experiment tested the ability of juvenile and adult rhesus monkeys to perform a cognitive task that is dependent upon intact frontal cortical function for optimal performance. Twenty-four juvenile (mean age 2.3 years) and 16 adult (mean age 10.3 years) rhesus monkeys were tested on the Cambridge Neuropsychological Test Automated Battery intradimensional/extradimensional set-shifting (ID/ED) task. Performance on the ID/ED task has been shown to be dependent upon frontal cortical function in both humans and nonhuman primates. Compared with adults, juveniles were impaired on the reversal of simple discrimination, intradimensional shift, reversal of intradimensional shift, and the extradimensional shift stages of the task. These results indicate juveniles committed more perseverative errors and more errors on the set-formation and set-shifting components of the ID/ED task. The developmental stage of the juvenile monkeys corresponds to roughly 5 to 6-year-old children, and these results are consistent with performance of human children and adults on similar ID/ED tests and on several other tests of attentional set-shifting or attentional flexibility. Furthermore, these results are consistent with the ongoing development of frontal cortical structures relating to ongoing cognitive development in nonhuman primates.     

The Effects of an NMDA Receptor Antagonist on Delayed Visual Differentiation in Monkeys and Rearrangements of Neuron Spike Activity in the Visual and Prefrontal Areas of the Cortex

The effects of perfusion of field 17 with the glutamate receptor antagonist 2-amino-5-phosphonovaleric acid (APV) on the characteristics of visual recognition and short-term memory were studied, along with the effects of APV on the responses of neurons in the visual and prefrontal areas of the cortex in rhesus macaques. In the test for delayed visual differentiation of stimuli of different colors, behavioral data were recorded simultaneously with multichannel recordings of the spike activity of single cells in cortical field 17 (directly within the microdialysis zone) and field 8. Multifactor dispersion analysis (ANOVA) showed that APV significantly worsened the behavioral characteristics in monkeys, with significant reductions in the duration of short-term storage of information (by factors of 2–4) and significant increases in the motor response times. These changes in cognitive characteristics induced by APV were accompanied by changes in the spike activity of neurons in the visual and prefrontal areas of the cortex during the sensory analysis and delay stages; changes in spike activity consisted of significant desynchronization. These results show that cognitive dysfunctions consisting of worsening of short-term remembering of information and increases in the duration of motor responses during exposure to APV may be caused by desynchronization of neuron activity in various areas of the cortex, these being involved in neuron ensembles responsible for the mechanisms of short-term memory, in which glutamatergic structures play an important role.     

Effects of progesterone and estradiol on sexual attractivity of female rhesus monkeys.

The mechanism whereby progesterone (P) reduces sexual attractivity was studied in female rhesus monkeys following ovariectomy and SC implantation of silastic capsules containing estradiol (E₂). Daily IM administration of 10 mg P to such females caused a significant decline in mounting rate and ejaculation by male partners. Females' willingness to accept male mounts (receptivity) was not affected, and the display of sexual invitations by females (proceptivity) actually increased after treatment with P, which produced plasma levels of P similar to those present during the luteal phase of the menstrual cycle and in pregnancy. Similar behavioral effects were obtained by instilling 250 μg P daily directly into the vagina, which did not significantly increase plasma P. Daily IM treatment with 250 μg P caused a significant increase in plasma P without affecting sexual interaction. Removal of E₂ capsules caused a significant decline in mounting rate and ejaculation without affecting the females' receptivity or proceptivity. This effect of E₂ withdrawal was not accentuated by perineal application just prior to testing of vaginal secretions collected from donor females treated with E₂ and P. It is concluded that P probably disrupts sexual interaction in monkeys by antagonizing the facilitatory effect of E₂ on production of a vaginal factor which otherwise enhances sexual attractivity. Ovariectomized females displayed significantly more sexual invitations following combined treatment with E₂ and P than with P alone, suggesting that E₂ may directly affect neural tissues controlling proceptivity.     

Sexual receptivity of bilaterally adrenalectomised female rhesus monkeys

Bilateral adrenalectomy (with cortisol replacement) of ovariectomised, oestrogentreated rhesus monkeys greatly decreased their sexual receptivity: they presented less to and refused the mounting attempts of males with whom they were paired. As a consequence the levels of the latter's sexual activity was low. Two hundred or 400 μg androstenedione/day increased the receptivity of these females, but dehydroepiandrosterone (up to 5,000 μg/day) was ineffective. Vaginal cornification, sexual skin colour and the size of the clitoris were measured and showed no significant change during the experiment. It is concluded that adrenal androgens are necessary to maintain sexual receptivity in the female rhesus monkey: the site of action of these androgens is discussed.