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We cross-sectionally investigated prenatal ontogeny of craniofacial shape in the two subspecies of the Japanese macaque (Macaca fuscata fuscata and Macaca fuscata yakui) using a geometric morphometric technique to explore the process of morphogenetic divergence leading to the adult morphological difference between the subspecies. The sample comprised a total of 32 formalin-fixed fetal specimens of the two subspecies, in approximately the second and third trimesters. Each fetal cranium was scanned using computed tomography to generate a three-dimensional surface model, and 68 landmarks were digitized on the external and internal surface of each cranium to trace the growth-related changes in craniofacial shape of the two subspecies. The results of our study demonstrated that the two subspecies generally shared the same craniofacial growth pattern. Both crania tend to exhibit relative contraction of the neurocranium in the mediolateral and superoinferior directions, a more superiorly positioned cranial base, a more vertically oriented occipital squama, and a more anteriorly positioned viscerocranium as the cranial size increased. However, distinctive subspecific differences, for example relatively narrower orbital breadth, higher orbit, higher position of the nuchal crest, and more protrudent snout found in Macaca fuscata yakui were already present during the prenatal period. This study demonstrated that morphological differentiation in the craniofacial shape may occur at a very early stage of the fetal period even between closely related subspecies of the Japanese macaque.

Excessive preformed vitamin A (VA) intake is contraindicated during pregnancy because of teratogenic concerns. Recent studies have provided biochemical and histologic evidence of chronic hypervitaminosis A in captive Old World monkeys consuming laboratory diets containing high concentrations of retinyl acetate. To investigate the effects of maternal chronic overconsumption of preformed VA on VA storage in early fetal liver, we analyzed monkey fetal livers ranging from 35 to 93 d gestational age (comparable with mid-first to late second trimester in humans) for VA (n = 19) and retinoic acid (n = 9). Retinyl esters were identified in all fetal livers, and retinol, on a percentage basis, was more abundant in younger fetuses. Liver VA concentration increased with gestational age, ranging from 0.0011 to 0.26 micromol/g in the youngest (35 d) and oldest fetuses (93 d), respectively. Liver VA concentrations (mean +/- 1 standard deviation) were 0.023 +/- 0.008 micromol/g in early gestation and 0.19 +/- 0.06 micromol/g in midgestation fetuses. All-trans retinoic acid concentrations were higher in early gestation (99.2 +/- 57.0 pmol/g, n = 6) than in midgestation (18.2 +/- 6.1 pmol/g, n = 3) but were variable. Liver VA concentrations from midgestation fetuses were higher than those in fetal human and monkey livers from later stages of development, when growth and VA accumulation rates are assumed to be highest. Therefore, excessive intake of preformed VA by the mothers results in amplified early fetal liver retinyl ester storage.

Purpose: To characterize the spatial and temporal expression of key structural and phototransduction cascade proteins in the monkey cone outer segment (OS)., Methods: Retinas from Macaca monkeys from ages fetal day (Fd) 89 through adulthood were double labeled using immunofluorescence for short (S) or long/medium (L/M) wavelength-sensitive cone opsin and either a structural protein (peripherin) or a phototransduction cascade protein (alpha-transducin [alpha-T], phosphodiesterase [PDE], or rhodopsin kinase [RK]). The spatial and temporal patterns of expression for each protein at each age were determined and graphed as a percentage of retinal coverage., Results: In both cone types, opsins and phototransduction proteins appear first in the fovea and last at the retinal edge. Peripherin appears concomitantly with opsin in both S and L/M cones, but S cones express peripherin and opsin 1 to 3 weeks before neighboring L/M cones. Alpha-T, PDE, and RK are expressed together in the L/M cone OS shortly after L/M opsin appears. Phototransduction proteins are not expressed in S cones until 1 to 3 weeks after the appearance of S opsin and at the same time that neighboring cones are expressing both L/M opsin and phototransduction proteins., Conclusions: The concomitant appearance of opsin and peripherin strongly suggests roles in promoting the structural integrity of the developing OS. Phototransduction cascade proteins appear in the developing OS at the same time as one another, but after opsin. The significant lag between their expression and that of S cone opsin indicates that phototransduction proteins are not essential for OS formation, nor does opsin expression trigger their expression. The different temporal but similar spatial expression patterns of phototransduction proteins within S and L/M cones suggests that some local signal(s) coordinates their appearance.

Purpose: To determine the temporal and spatial pattern of rod opsin appearance in Macaca monkey retina., Methods: Frozen sections from fetal day (Fd) 55 to adulthood (birth = Fd168) containing the entire horizontal meridian were stained using Rho4D2 monoclonal antiserum visualized with immunofluorescent labeling. At Fd66, Fd79, and Fd89, retinal samples taken at known eccentricities were studied from the opposite eye using standard electron microscope methods., Results: Rod opsin was detected at Fd66 in or near the fovea, and a second focus appeared at Fd75 to Fd77 near the optic disc in the nasal rod ring. The earliest opsin appeared in the apical stubs, which resembled the apical connecting cilium in the electron microscope. Staining of the entire cell body membrane, including the synaptic spherule, was present 4 to 7 days later. Opsin expression had a nasal bias with rods at the nasal ora labeled at Fd140, whereas temporal ora was not labeled until Fd155. Cell body labeling disappeared by Fd132 across central retina but persisted into the first postnatal year in far peripheral retina. Outer segment (OS) length measurements showed that rods in the rod ring had the longest OS between Fd115 and postnatal week 9. Rod OS at all retinal eccentricites continued to elongate between 11 months of age and adulthood., Conclusions: Rod opsin expression follows a foveal-to-peripheral gradient beginning at Fd66 and ending near birth. Rod opsin is detected first in the connecting cilium and slightly later in the entire cell membrane, and then cell membrane labeling disappears as the heavily labeled OS elongates. Although the first OS appear on rods near the fovea, these OS still are short at birth and do not reach adult length until after 2 years of age. The longest OS at birth are found on rods at the rod ring, suggesting that this region could have higher scotopic sensitivity than central retina at birth.

Groups of fetal monkeys were studied at 50, 75, 100, 125 and 150 days gestational age, and postnatal monkeys were studied at 175 days gestational age and in adult life (full term gestation is 168 plus or minus 4 days). The ratios between the various tissues of which lung is composed were determined by light and electronmicroscopy. Characteristic changes in ratios were noted at each developmental age. Primitive bronchioles increased from 50 to 75 days, decreased as permanent bronchioles, alveolar ducts and alveoli developed, and were absent after 150 days. Mesenchymal tissue decreased progressively from 50 to 150 days. Alveoli were present at 100 days gestational age; alveolar lumens accounted for approximately 1/2 the volume of peripheral lung by 150 days, and 2/3 the volume of lung in adult animals. By 150 days alveolar epithelial cells could be identified as Type I (membranous) or Type II (granular) on the basis of the characteristic cytoplasmic lamellar inclusion bodies of the latter. The ratios of the Type II/Type I cells were .21-.23 at 150 and 175 days, and fell to approximately half that value in adult lungs.

The metabolic interrelationship between cortisol (F) and cortisone (E) was studied in four long term catheterized rhesus monkey fetuses in utero during the last third of gestation. The MCR of E (50.8 +/- 5.4 liters/day) was greater than that of F (22.4 +/- 2.1, P less than 0.005) as was the plasma concentration (187.9 +/- 5.0 vs 86.1 +/- 2.5 ng/ml, P less than 0.001). The production rate of E (9.6 +/- 1.4 mg/day) was several-fold greater than for F (1.9 +/- 0.2, P less than 0.005). Of all fetal F, 79.5 +/- 7.0% was metabolized to E, and 43.4 +/- 3.9% originated from E within the fetal circulation. A significant mass of F was infused in these experiments because of the low specific activity of [14C]F. Nevertheless, the fetus was able to maintain F concentrations in the normal range. The MCR of F was similar to that which we previously found using trace amounts of [3H]F. This indicates that the fetus regulates the amount of F in the fetal compartment, probably by decreasing fetal; adrenal secretion rate. We conclude that F in the primate fetus is extensively oxidized to E. We conclude also that E is produced and metabolized much more extensively than is F. Reduction of E back to F could be an important source of fetal F, and increasing activity of this pathway, if present, could contribute to the increase in fetal F levels observed in late gestation in the primate.

Rhesus monkey fetuses were surgically prosencephalectomized (Type 2) or functionally hypophysectomized (Type 1) at 75 days gestation, then returned to the uterus until elective Caesarean section on day 145--150 (term 167 days). Deprivation of fetal hypothalamic releasing factors in Type 2 and fetal pituitary tropic hormones in Type 1 significantly delayed the ontogeny and functional development of fetal endocrine tissues. Bone ossification and growth were significantly retarded in Type 1 only, not in Type 2. In Type 1 the body and all organs except the endocrine glands were about half normal weight. The adrenals, thyroids, ovaries and testes were histologically abnormal and about one-tenth normal weight. Non-endocrine organs were histologically similar to 110-130-day fetuses. Thyroxine (T4) concentrations were significantly depressed in Type 1 fetal and maternal plasma at Caesarean section but normal in Type 2. Cortisol concentrations were normal in Type 1 fetal and maternal plasma. Types 1 and 2 plasma oestradiol concentrations were significantly lower in mothers but normal in fetuses. Type 1 placentas produced significantly less progesterone in vitro than normal. Fetal endocrine autonomy is indicated (thyrotropin-releasing factor excepted). Many of the hypothalamic and anterior pituitary hormones do not pass in effective amounts from mother to fetus. Fetal endocrine autonomy is a prerequisite for the control of both development and parturition.

In the monkey lung (Macacus nemestrina) canalization of all bronchopulmonary segments begins simultaneously at the peripheral future-alveolar-duct portions of each segment and then continues progressively centralwards. Its simulataneous appearance in all segments of a human lung of the 17th postovulatory week (19th postmenstrual week) has now been confirmed (cf. Boyden, '74). The mode of canalization, as seen under the light microscope, is here demonstrated.

Stump-tailed macaque virus, a newly recognized papovavirus of the SV40 polyoma subgroup, was demonstrated in kidney cultures from each of five stump-tailed macaque fetuses in the second half of gestation and from six adult stump-tailed macaques. Such regular presence of virus in the fetus is an unusual feature for a papovavirus.

Amniotic fluid cytology is considered to reflect other organ maturity more faithfully than the L/S ratio. In order to assess the origin of amniotic fluid cells a pilot study was undertaken in the Rhesus monkey with the purpose of correlating the surface ultrastructure of amniotic fluid cells and various tissues facing, or communicating with the amniotic cavity. Three distinct cell types were found in the amniotic fluid, out of which two were obviously related to maturational events in fetal skin. The remaining cell type seemed to be derived from umbilical cord, fetal vagina and to some extent oral mucosa. No exfoliation was observed in the urinary bladder, trachea, lungs or amniotic membranes. It is concluded that correlation of cytology and tissue surface morphology in the monkey amniotic fluid compartment may give clues to the origin of amniotic fluid cells in the human.

The brain and nasal region of a fetal (124 days) female rhesus macaque (Macaca mulatta) were fixed in situ, decalcified, sectioned and treated for the immunocytochemical demonstration of LHRH. Immunoreactive neurons and fibers were found in expected sites and also in sites not reported to date in primates: there was an abundant distribution along the pathway of the nervus terminalis and clusters of neurons anterior and ventral to the olfactory bulbs and scattered in the nasal septum. Fibers extended into the epithelium lining the nasal septum. Most unexpectedly, LHRH fibers and a few neurons were observed within the optic nerve.