History of reptile placentology, part III: Giacomini's 1891 histological monograph on lizard placentation.

By the 1890s, placental arrangements had been documented macroscopically in lizards and fishes, but placental studies on such species lagged far behind research on mammals. In 1891, the biologist Ercole Giacomini (at the University of Siena, Italy) published the first histological analysis of a reptile placenta. Focusing on a placentotrophic lizard (Chalcides chalcides) with a morphologically complex placenta, Giacomini documented the histological and cellular bases for placental nutrient transfer and gas exchange. In conjunction with a follow-up study in 1906, he demonstrated that placental structure is correlated with function and can vary dramatically between related species. Giacomini's work was highly influential in showing that placentation in lizards had converged evolutionarily on that of mammals, while establishing reptile placentology as a highly promising area for future research.

Reptilian viviparity: past research, future directions, and appropriate models.

Squamate reptiles represent an ideal group for studies of viviparity, because they have evolved this reproductive pattern frequently, relatively recently, and at low taxonomic levels. A phylogenetic approach shows particular promise in helping us interpret anatomical, physiological, and ecological diversity. This review summarizes four major categories of active investigation: (1) reproductive anatomy and physiology; (2) placental structure and function; (3) reproductive endocrinology; and (4) reproductive and physiological ecology. Evolutionary reconstructions suggest that on many occasions viviparity has evolved concomitantly with functional placentation, through reduction of the shell membrane and hormonal modifications that prolong gestation. Studies of placentotrophic clades as well as reproductively bimodal species offer great potential for explaining the evolution of viviparity and placentation. However, live-bearing squamates are reproductively diverse, and appear to have solved physiological problems associated with viviparity by a variety of mechanisms. Consequently, studies on one or a few squamate species appear increasingly unlikely to yield all-inclusive explanations. Future studies and analyses should abandon assumptions of universal physiological mechanisms and a single historical sequence, in favor of the documentation of diversity in phylogenetic and quantitative terms.

Structure, function, and evolution of the oviducts of squamate reptiles, with special reference to viviparity and placentation.

In lizards and snakes, the oviducts function in fertilization, sperm storage, egg transport, eggshell deposition, maintenance of the early embryo, and expulsion of the egg or fetus. In viviparous forms they also contribute to placentae responsible for gas exchange and nutrient provision to the fetus. Dissections of species of 30 genera coupled with data from the literature indicate that squamate oviducts vary interspecifically in seven macroscopic features, including the extent and nature of regional differentiation, vascular supply, topographic asymmetry, number of oviducts, vaginal pouches, and relationship to the cloaca. The uterus, infundibulum, and vagina differ histologically in their epithelia, glands, and myometrial layers. Season cyclicity occurs in all three oviductal regions, most prominently in the uterus, and is under endocrinological control. Regional and cytological specializations reflect the diverse functions performed by the oviduct. Definitive evidence for oviductal albumen production and egg resorption is lacking. In viviparous squamates, three uterine specializations may facilitate maternal-fetal gas exchange: an attenuated epithelium, reduced uterine glands (and a reduced shell membrane), and increased vascularization. Contrary to previous reports, pregnant uteri show no epithelial erosion or capillary exposure. Specializations for nutrient provision to the fetus include mucosal hypertrophy, enlarged glandular epithelia, and multicellular glands whose secretions are absorbed by the chorioallantois. Comparisons with other amniotes indicate that squamates inherited the oviduct as an organ with capabilities for egg uptake and transport, fertilization, eggshell deposition, and oviposition. Other features have evolved convergently among squamates: infundibular sperm receptacles, unilateral oviduct loss, uterine gestation, placentation, and specializations for placentotrophy. Cladistic analysis indicates that oviductal features associated with deposition of tertiary egg investments in reptiles reflect evolutionary convergence as well as secondary simplification, rather than a unidirectional trend towards increased specialization.

Effects of testosterone administration and castration on the forelimb musculature of male leopard frogs, Rana pipiens.

In Rana pipiens, forelimb muscles that are used by males to clasp females during amplexus are sexually dimorphic in mass, protein content, and fiber composition. This experiment examined the effects of castration and exogenous testosterone on wet mass, dry mass, and protein content of the 22 major forelimb muscles of male leopard frogs to determine whether established patterns of sexual dimorphism of the muscles are reflected in differential androgen sensitivity. Muscles ranged from highly and moderately responsive to testosterone treatment (e.g., flexors of the elbow and of the carpus; adductors of the shoulder and of the first digit) to nonresponsive to testosterone (antagonists to these muscles). The mean dry mass of the testosterone-responsive muscles ranged broadly from 28-164% over control values. Castration had little or no effect on the response to testosterone, nor did it affect muscle mass in frogs not treated with hormone, as compared to sham-operated animals. Experimental treatment did not alter water content or protein concentration of muscles. The degree of testosterone sensitivity exhibited among the muscles of males closely correlated with their degree of sexual dimorphism. We postulate that androgens influence the functional attributes of male forelimb muscles through both organizational and activational effects.

Saltationist and punctuated equilibrium models for the evolution of viviparity and placentation.

Vertebrate viviparity (live-bearing reproduction), placentation, and placentotrophy are widely assumed to have evolved as three successive, gradualistic transformations. From empirical data and predictive tests on lizards and snakes, this paper indicates that placentae and a degree of placentotrophy have evolved repeatedly as necessary correlates of viviparity, not as subsequent modifications. In addition, information derived from studies of anatomy, physiology, biogeography and systematics is used to evaluate new saltationist and punctuated equilibrium models for the evolution of viviparity. Phylogenetic reconstruction reveals that more than 100 squamate clades have made the transition to viviparity and placentation. However, various phenotypic intermediates postulated by the gradualistic model are either scarce or unrepresented among known forms, including those in which viviparity has evolved at specific and subspecific levels. Evolution in squamates seems to have produced a dichotomy between two evolutionarily stable patterns: (i) retention of weakly shelled or shell-free eggs to term (viviparity), with development of fully functional placentae; and (ii) deposition of shelled eggs at or near the limb bud stage of development (typical oviparity). Conflicting functional demands placed on eggshell morphology may constrain establishment of prolonged, oviparous egg-retention as a viable, historically stable pattern. Alternatively, the costs of prolonged egg-retention associated with decreased female mobility or decreased fecundity may exceed the benefits in oviparous forms.

Lactation: historical patterns and potential for manipulation.

The advent of biotechnology has made data on undomesticated mammals relevant to dairy science. Such data indicate the potential of lactation for modification, reveal genetic material available for use through bioengineering, help distinguish adaptive features from historical artifacts, and clarify limits on lactational diversity that date from early evolution. Evolutionary analysis indicates that a complex degree of lactation preceded divergence of the extant mammalian lineages during the Mesozoic Era. Although aspects of monotreme lactation appear to be ancestral for extant mammals, the marsupials and eutherians exhibit divergent specializations. Evidence is consistent with the idea that protolacteal glands evolved by combining features of skin gland populations into a new functional complex. Secretions of these ancestral glands may have had antimicrobial properties that protected the eggs or hatchlings and organic components that supplemented offspring nutrition. Following development of highly nutritious milks, evolution produced diversity in milk composition and function, milk output, length of lactation, mammary gland anatomy, and contributions of lactation to offspring nutrition. Certain marsupials are specialized in terms of functional independence and temporal plasticity of mammary tissues. Mammalian diversity indicates that artificial selection and physiological manipulation of domestic artiodactyls has only modestly exploited the potential of mammary glands as a nutritional source for humans.

Obplacental giant cells of the domestic rabbit: development, morphology, and intermediate filament composition.

Obplacental giant cells are large (less than or equal to 210 microns) polyploid cells that appear in the stroma of the pregnant uterus of the rabbit following ovoimplantation. Histological examination of a complete developmental series indicates that obplacental giant cells arise from trophoblastic knobs that have traversed the uterine epithelium during early implantation. During maturation, the cells undergo a massive (approximately 6,000%) increase in volume and penetrate deeply into the uterine stroma and myometrium, where they often become associated with blood vessels and smooth muscle cells. Giant cells at mid-gestation contain one or two large nuclei with prominent nucleoli and appear to be amitotic. They are rich in Golgi complexes, RER, SER, and cortically distributed cytoplasmic filaments, and contain intracellular canaliculi lined by microvilli. Giant cells vary with respect to the occurrence of lipid droplets, phagocytotic inclusions, lysosomal structures, and electron-dense granules. Immunocytochemistry demonstrates that the giant cells exhibit intermediate filaments related to cytokeratin and vimentin, but are negative for desmin and for an endothelial cell marker, Factor VIII-related antigen. The cells are positive for cytokeratin from their inception, but only become vimentin-positive between Days 12 and 15 of pregnancy, a change seemingly related to their detachment from epithelial tissue to take on an independent existence. Our findings indicate that the giant cells originate from obplacental trophoblast and, at maturity, exhibit cytoskeletal characteristics of isolated epithelial cells, as well as a complement of organelles suggestive of synthetic activity.

Trophoblast concept as applied to therian mammals.

Available evidence provides little support for a recent proposal that the term "trophoblast" be applied solely to eutherian mammals. Arguments for such a restricted usage are based on a dichotomous interpretation of therian reproduction that underestimates the developmental, structural, and functional diversity of trophoblastic tissues occurring within the infraclass Eutherian. The occurrence of developmental patterns that are phenotypically intermediate between those of commonly studied eutherians and metatherians suggests that blastocyst development is not fundamentally different in marsupials and eutherians. The trophoblast of marsupials accomplishes most or all of the major functions of the eutherian trophoblast, including maternal-fetal physiological exchange, implantation, contribution to placental membranes, steroid metabolism, and possibly, immunological protection of the conceptus. Furthermore, application of the term "trophoblast" to marsupials is consistent with present and past usage, as well as with the original definition and etymological derivation of the term. Therefore, we recommend that the term "trophoblast" continue to be applied in a functional-morphological sense to the appropriate extraembryonic tissues of marsupials. Such use of functional (rather than taxonomic) criteria for application of this term avoids biasing interpretations of mammalian reproductive evolution.

Eutherian-like reproductive specializations in a viviparous reptile.

The viviparous Brazilian scincid lizard Mabuya heathi exhibits a suite of reproductive specializations widely believed to be confined to the eutherian mammals. This skink ovulates the smallest known reptilian egg ( approximately 1.0 mm in diameter). Placental transport accounts for >99% of the dry mass of the periparturient fetus, representing a degree of placentotrophy proportionately greater than that reported in any other non-mammalian vertebrate. Placental morphology and the timing of fetal growth implicate the chorioallantoic placenta in maternal-fetal nutrient transfer. The yolk sac placenta regresses prior to any major increase in embryonic dry mass. Precocial gonadal maturation and postponement of reproductive investment until well after ovulation enables females to become pregnant at 3-4 months of age, long before attainment of full adult body size.