Shape, variance and integration during craniogenesis: contrasting marsupial and placental mammals.

Studies of morphological integration can provide insight into developmental patterns, even in extinct taxa known only from skeletal remains, thus making them an important tool for studies of evolutionary development. However, interpreting patterns of integration and assessing their significance for organismal evolution requires detailed understanding of the developmental interactions that shape integration and how those interactions change through ontogeny. Thus far, relatively little comparative data have been produced for this important topic, and the data that do exist are overwhelmingly from humans and their close relatives or from laboratory models such as mice. Here, we compare data on shape, variance and integration through postnatal ontogeny for a placental mammal, the least shrew, Cryptotis parva, and a marsupial mammal, the gray short-tailed opossum, Monodelphis domestica. Cranial variance decreased dramatically from early to late ontogeny in Cryptotis, but remained stable through ontogeny in Monodelphis, potentially reflecting functional constraints related to the short gestation and early ossification of oral bones in marsupials. Both Cryptotis and Monodelphis showed significant changes in cranial integration through ontogeny, with a mixture of increased, decreased and stable levels of integration in different cranial regions. Of particular note is that Monodelphis showed an unambiguous decrease in integration of the oral region through ontogeny, potentially relating to their early ossification. Selection at different stages of development may have markedly different effects if patterns of integration change substantially through ontogeny. Our results suggest that high integration of the oral region combined with functional constraints for suckling during early postnatal ontogeny may drive the stagnant variance observed in Monodelphis and potentially other marsupials.

Cortical organization in shrews: evidence from five species.

Cortical organization was examined in five shrew species. In three species, Blarina brevicauda, Cryptotis parva, and Sorex palustris, microelectrode recordings were made in cortex to determine the organization of sensory areas. Cortical recordings were then related to flattened sections of cortex processed for cytochrome oxidase or myelin to reveal architectural borders. An additional two species (Sorex cinereus and Sorex longirostris) with visible cortical subdivisions based on histology alone were analyzed without electrophysiological mapping. A single basic plan of cortical organization was found in shrews, consisting of a few clearly defined sensory areas located caudally in cortex. Two somatosensory areas contained complete representations of the contralateral body, corresponding to primary somatosensory cortex (S1) and secondary somatosensory cortex (S2). A small primary visual cortex (V1) was located closely adjacent to S1, whereas auditory cortex (A1) was located in extreme caudolateral cortex, partially encircled by S2. Areas did not overlap and had sharp, histochemically apparent and electrophysiologically defined borders. The adjacency of these areas suggests a complete absence of intervening higher level or association areas. Based on a previous study of corticospinal connections, a presumptive primary motor cortex (M1) was identified directly rostral to S1. Apparently, in shrews, the solution to having extremely little neocortex is to have only a few small cortical subdivisions. However, the small areas remain discrete, well organized, and functional. This cortical organization in shrews is likely a derived condition, because a wide range of extant mammals have a greater number of cortical subdivisions.

The head-twitch response in the least shrew (Cryptotis parva) is a 5-HT2- and not a 5-HT1C-mediated phenomenon.

Our initial studies suggested that the 5-HT2/1C agonist (+/-)-1-(2,5-dimethoxy-4-iodophenyl-2-aminopropane [(+/-)-DOI] produces both the head-twitch response (HTR) and the ear-scratch response (ESR) in mice via stimulation of 5-HT2 receptors. However, challenge studies revealed that these behaviors are produced via two different receptors (possibly 5-HT2 and 5-HT1C). Due to a lack of selective agents one cannot designate a particular response for the activation of a specific receptor. The purpose of the present study was to investigate such behaviors in the least shrew, which is more sensitive to (+/-)-DOI than rodents. IP injection of (+/-)-DOI in shrews produced a dose-dependent (bell-shaped) and time-dependent increase in the HTR frequency. The (+/-)-DOI-induced HTR was equipotently and completely attenuated by the 5-HT2/1C antagonists ketanserin and spiperone. The 5-HT1C antagonist with 5-HT2 agonist action, lisuride, also produced the HTR in a bell-shaped dose- and time-dependent fashion. Central injections of both (+/-)-DOI (0.2 microgram) and lisuride (0.5 microgram) also induced the behavior. Both peripheral and central administration of lisuride failed to produce the ESR. (+/-)-DOI significantly induced the ESR only at the highest dose tested (2.5 mg/kg, IP). Centrally administered (+/-)-DOI (0.2 microgram) produced more ESRs relative to vehicle controls; however, the difference did not attain significance. At low doses (0.31 and 0.63 mg/kg), (+/-)-DOI had no effect on locomotor activity, but it significantly attenuated the behavior at larger doses. Both low and high doses of lisuride increased the motor activity. Spiperone dose-dependently suppressed locomotion, whereas ketanserin had no effect. The present results suggest that the HTR is a 5-HT2 receptor-mediated event and changes in locomotor activity do not affect the induced HTR.

Anatomic and physiologic reference values in least shrews (Cryptotis parva).

BACKGROUND AND PURPOSE: The least shrew is an established animal model for reproductive and pharmacologic research. Biologic reference data are needed to assess animal health status and provide a rationale for use of novel statistical programs to evaluate the effects of orally administered substances in toxicologic and pharmacologic studies. METHODS: Organ weights, blood biochemical and hematologic values, and food and water consumption data were collected from 50-day-old shrews after two weeks' consumption of a standard feline diet. RESULTS: In general, data correlated well with values reported for other mammalian species. Plasma phosphorus concentration was high. There was a significant difference in food and water consumption per gram of body weight between shrews at lower and upper (+/- 1 SD) weight ranges for the study. The 3.2-g animals consumed 27% more food per gram of body weight than did the 5.0-g animals. CONCLUSIONS: The high phosphorus concentration was attributed to hemolysis resulting from the axillary cut method of blood sample collection. The small size of the shrew allowed demonstration of the Kleiber effect within a +/- 1 SD weight range in a single species. The phenomenon necessitates the use of statistical methods other than the typical tests establishing the significance of the differences between the means of groups for oral toxicologic and pharmacologic studies.

The least shrew (Cryptotis parva) as a laboratory animal.

Wild least shrews, Cryptotis parva, have been maintained in the laboratory for 15 years. Animals were confined in small, open terraria with loam soil covering the floor. The shrews were fed mealworms and a mixture of ground meat and mink diet twice daily. Heterosexual pairing was the mating system most commonly employed. The gestation period was 21 to 22 days, and the mean litter size was 4.3 with a range of 1-9 offspring. The young were weaned at 18-20 days of age and were sexually mature prior to the age of 40 days. Periodontal disease was the only disease which threatened the colony, and it was controlled by adding oxytetracycline hydrochloride to the water. Maximum laboratory survival time was 889 days.

Unusual ampullary sperm crypts, and behavior and role of the cumulus oophorus, in the oviduct of the least shrew, Cryptotis parva.

The gametes of the least shrew, Cryptotis parva, were studied in regard to their maturation and structure, and with particular emphasis on their behavior in the fallopian tube, from the time of ovulation until the appearance of two-cell embryos beginning some 9 h after ovulation. Cryptotis spermatozoa are organized according to the conventional eutherian mold, with the exception of a barbed perforatorium and an unusual plasma membrane density lent by a bristly coat where it overlies the acrosome rim. In the epididymis they undergo a maturation of the capacity for motility and an -S-S-related stabilization of the nucleus and tail organelles, with the cauda housing only approximately 4-5 million spermatozoa. Mating involves penile locking and also the deposition of a modest vaginal plug that covers the cervix. The short (4-5 mm) fallopian tube has three regions-a simple isthmus, a relatively narrow ampulla populated throughout by ciliated crypts, and a crypt-free terminal infundibulum-the fertilization site. Unlike the situation in most mammals, the tubal isthmus was devoid of spermatozoa in mated females before and after ovulation, which occurred approximately 13 h post-hCG and produced a mean of 5.7 ova. However, the ampulla then housed approximately 1500 active cells in groups within the ciliated crypts, sometimes together with leukocytes but with few spermatozoa above in the infundibulum. Within about 1 h after their ovulation from approximately 400-microm follicles, eggs were penetrated while in the infundibulum despite the nonexpanded hyaluronidase-resistant state of the cumulus oophorus. However, on moving down to the ampulla by 2-4 h after ovulation, the dense cumulus around fertilized eggs appeared to proliferate and began to disperse coincidentally with secretion of a hyaluronidase-sensitive matrix in which hundreds of motile spermatozoa often became enmeshed. This cumulus change also occurred around unfertilized eggs, though more slowly, but not around fertilized or unfertilized eggs cultured in vitro. Thus, cumulus matrix production appeared to be stimulated to an important degree by factors in the oviduct, not by preovulatory gonadotropins as in many mammals. Although cumulus-invested eggs were fertilized readily in vitro, cumulus-free eggs of the same age were never fertilized, and spermatozoa bound to the zona pellucida had intact acrosomes. This and related evidence from other shrews makes it seem likely that the soricid cumulus has an essential role in fertilization and may induce the acrosome reaction.

Ultrastructure of the myocardium of the least shrew, Cryptotis parva Say.

The heart of the least shrew, Cryptotis parva Say, is an extremely active organ, capable of achieving rates of 800-1,200 beats/minute. The general features of myocardial cell ultrastructure in this insectivore are much like those of other small mammals; no single striking feature of fine structure is present to which the physiological properties of this heart might necessarily be attributed. Still there exist in these myocardial cells a number of atypical properties. These include 1) mitochondria having a wide variety of sizes and internal configurations 2) a pleiomorphic, highly ramified, small-diameter transverse-axial tubular system (TATS) 3) numerous "labyrinths," which are proliferated components of the TATS, and 4) myofibril-free regions, located both in juxtanuclear and other myoplasmic levels and populated by a concentration of TATS elements and fibrillar structures. Features (2) and (3) are also characteristic of another fast-beating heart, that of the mouse. The sinoatrial and atrioventricular nodal regions, as well as a Purkinje system, have been identified in the least shrew heart, along with sparsely distributed atrial cells whose myofibrils contain proliferated Z-band material. A feature frequently encountered in atrial working muscle cells is the occurrence of close appositions between gap junctions and tubules of sarcoplasmic reticulum; such appositions are also present in other regions of the shrew heart, as are complexes composed of gap junctions and mitochondria.

Reproductive features of the eastern mole (Scalopus aquaticus) and star-nose mole (Condylura cristata).

Since moles are closely related to shrews, the gametes and reproductive tracts of the star-nose mole (Condylura cristata) and the eastern mole (Scalopus aquaticus) were examined to gain further insight into unusual reproductive traits of the Soricidae. Moles display many of these soricid traits, but with some important differences. The cumulus oophorus of Scalopus, ovulated about 16 h after hCG injection, was largely dispersed by hyaluronidase and, though quite dense, was nevertheless more similar to that of higher mammals than to the compact 'ball of the soricid cumulus. Within the female tract in these moles, approximately 85% of the length of the oviduct comprises a narrow ampulla with numerous differentiated crypts that, in shrews, house spermatozoa. However, in contrast to shrews, moles produce considerably larger numbers of spermatozoa, which challenges the proposal that, in shrews, oviductal sperm crypts specifically permit lower sperm production by the males. In the sperm head of these two moles, the acrosome displays the long rostrum that is typical of other Insectivora, and the perforatorium has the barbs by which soricid spermatozoa probably bind to the zona pellucida. Perhaps allied to this, immunoblots indicated that the immunoreactive acrosomal matrix of Scalopus spermatozoa is simpler than the polypeptide complex of the bovine and hamster acrosomal matrix.

Reproductive characteristics of the african pygmy hedgehog, atelerix albiventris.

To obtain further perspective on reproduction and particularly gamete function among so-called primitive mammals presently grouped in the Order Insectivora, we have examined the African hedgehog, Atelerix albiventris, in light of unusual features reported in shrews and moles. Atelerix proves to share many but not all of the characteristics seen in these other insectivores. The penis of Atelerix has a 'snail-like' form, but lacks the surface spines common in insectivores and a number of other mammals. Hedgehog spermatozoa display an eccentric insertion of the tail on the sperm head, and they manifest the barbs on the perforatorium that, in shrews, probably effect the initial binding of the sperm head to the zona pellucida. As a possible correlate, the structural matrix of the hedgehog acrosome comprises only two main components, as judged by immunoblotting, rather than the complex of peptides seen in the matrix of some higher mammals. The Fallopian tube of Atelerix is relatively simple; it displays only minor differences in width and in the arborized epithelium between the isthmus and ampulla, and shows no evidence of the unusual sperm crypts that characterize the isthmus or ampulla, depending on the species, in shrews and moles. In common with other insectivores, Atelerix appears to be an induced ovulator, as judged by the ovulation of some 6-8 eggs by about 23 h after injection of hCG. The dense cumulus oophorus appeared to have little matrix, in keeping with the modest dimensions of the tubal ampulla and, while it was not quite as discrete as that of soricids, it did show the same insensitivity to 0.5% (w/v) ovine or bovine hyaluronidase.

Longevity and the genetic determination of collagen glycoxidation kinetics in mammalian senescence.

A fundamental question in the basic biology of aging is whether there is a universal aging process. If indeed such a process exists, one would expect that it develops at a higher rate in short- versus long-lived species. We have quantitated pentosidine, a marker of glycoxidative stress in skin collagen from eight mammalian species as a function of age. A curvilinear increase was modeled for all species, and the rate of increase correlated inversely with maximum life-span. Dietary restriction, a potent intervention associated with increased life-span, markedly inhibited glycoxidation rate in the rodent. On the assumption that collagen turnover rate is primarily influenced by the crosslinking due to glycoxidation, these results suggest that there is a progressive age-related deterioration of the process that controls the collagen glycoxidation rate. Thus, the ability to withstand damage due to glycoxidation and the Maillard reaction may be under genetic control.