Recent success propagating captive beluga has resulted from combined efforts by North American zoos and aquariums to manage disparate collections as a single population. This success has provided a tremendous opportunity to increase our understanding of beluga reproductive biology. Blood samples were collected on a weekly to biweekly basis from 23 female and 12 male beluga, ranging in age from 2-15 years, for analysis of serum progesterone (P) and testosterone (T), respectively. Peri-parturient observational data, including food intake, duration and signs of labor, and nursing patterns were collected from 15 days prepartum to 30 days postpartum during 21 births. Total body lengths and weights were collected from 10 captive-born beluga. For female beluga, the mean (7SD) age, body length, and weight at first conceptions were 9.172.8 years, 318.079.1 cm, and 519784 kg. Thirty-five luteal phases and 13 conceptions were detected from January-June, and 70% of luteal phases and 80% conceptions occurred from March-May. The mean luteal phase and total estrous cycle lengths were 30.076.5 days and 48.074.6 days, respectively. For male beluga, the mean age that males sired their first calf was 13.372.6 years. Compared to younger males (o8 years of age, 0.95 ng/ml), levels of T secretion in older males (48 years of age, 5.0 ng/ml) were elevated significantly only during the interval from January-April. Highest T concentrations (6.274.9 ng/ml) were recorded from January-March, whereas nadir concentrations (1.171.0 ng/ml) were detected from August-September. The mean gestation length was 475.0720.4 days (n ¼ 9). For parturition, the mean time from the first appearance of fluke or rostrum to delivery, delivery to placental passage, and delivery to nursing were 4.472.9 hr, 7.671.8 hr, and 43745 hr, respectively. All cows had decreased food intake on the day of delivery, with 44% having zero intake. Peak 24-hr nursing activity occurred 3.972.7 days post-partum. Growth (i.e., body weight and length) as a function of age were well described by the Gompertz model (r 2 ¼ 0.91, 0.93). Based on the model, growth in body weight and length were significantly greater in males compared to females. Predicted birth weight (88.9 kg) was similar for both sexes, however, and male calves were predicted to be shorter (154.3 cm) than female calves (160.7 cm). The results provide the first descriptions of captive beluga reproductive physiology, including endocrinology, peri-parturient behavior, growth, and reproductive maturity. This knowledge is important for helping to maintain genetically diverse, self-sustaining populations of captive beluga whales.

Odontocetes have an exceptional range in body mass spanning 10(3) kg across species. Because, size influences oxygen utilization and carbon dioxide production rates in mammals, this lineage likely displays an extraordinary variation in oxygen store management compared to other marine mammal groups. To examine this, we measured changes in the partial pressures of respiratory gases ([Formula: see text], [Formula: see text]), pH, and lactate in the blood during voluntary, quiescent, submerged breath holds in Pacific white-sided dolphins (Lagenorhynchus obliquidens), bottlenose dolphins (Tursiops truncatus), and a killer whale (Orcinus orca) representing a mass range of 96-3,850 kg. These measurements provided an empirical determination of the effect of body size on the variability in blood biochemistry during breath hold and experimentally determined aerobic dive limits (ADL) within one taxonomic group (odontocetes). For the species in this study, maximum voluntary breath-hold duration was positively correlated with body mass, ranging from 3.5 min in white-sided dolphins to 13.3 min for the killer whale. Variation in breath-hold duration was associated with differences in the rate of change for [Formula: see text] throughout breath hold; [Formula: see text] decreased twice as fast for the two smaller species (-0.6 mmHg O(2) min(-1)) compared to the largest species (-0.3 mmHg O(2) min(-1)). In contrast, the rate of increase in [Formula: see text] during breath hold was similar across species. These results demonstrate that large body size in odontocetes facilitates increased aerobic breath-hold capacity as mediated by decreased mass-specific metabolic rates (rates of change in [Formula: see text] served as a proxy for oxygen utilization). Indeed the experimentally determined 5 min ADL for bottlenose dolphins was surpassed by the 13.3 min maximum breath hold of the killer whale, which did not end in a rise in lactate. Rather, breath hold ended voluntarily as respiratory gases and pH fell within a narrow range for both large and small species, likely providing cues for ventilation.

Transcription of the plasminogen activator inhibitor type-1 (PAI-1) gene appears to be growth state regulated in several cell types (e.g. , Ryan and Higgins, 1993, J Cell Physiol 155:376-384; Mu et al., 1998, J Cell Physiol 174:90-98). Transit of serum-stimulated normal rat kidney (NRK) epthelial cells through the first division cycle after release from quiescence (G(0)) provided a model system to assess the kinetics and mechanisms underlying PAI-1 expression in a growth "activated" phenotype. PAI-1 mRNA transcripts increased by more than 20-fold during the G(0)-->G(1) transition; induced expression had immediate-early response characteristics and abruptly declined prior to the onset of DNA synthesis. Transcriptional activity of the PAI-1 gene paralleled the steady-state mRNA abundance profile during this first synchronized growth cycle after release from quiescence. Although PAI-1 mRNA levels were up-regulated (approximately threefold) upon exposure to several different growth factors, neutralizing antibodies to transforming growth factor-beta1 (TGF-beta1) effectively attenuated the more than ninefold serum-associated PAI-1 inductive response by more than 70% (at both the mRNA transcript and protein levels). Similar to the metabolic requirements for serum-mediated PAI-1 transcription, PAI-1 induction upon addition of TGF-beta1 to quiescent NRK cell cultures was actinomycin D sensitive and resistant to cyclohexamide and puromycin, suggesting a primary mode of transcript control. The response to protein synthesis inhibitors, however, was complex. While cyclohexamide appeared to stabilize, or at least maintain, fetal bovine serum (FBS)- or TGF-beta1-stimulated PAI-1 mRNA levels, puromycin had no such affect. The amplitude and duration of induced PAI-1 expression were the same in either the presence or absence of puromycin. Cyclohexamide when used alone (i.e., in non-FBS- or TGF-beta1-treated cultures), moreover, effectively stimulated PAI-1 induction whereas puromycin was ineffective. Although TGF-beta1 was not a complete mitogen in the NRK cell system, incubation of quiescent renal cell cultures with TGF-beta1, prior to serum stimulation, resulted in a 10- to 12-fold increase in PAI-1 expression coincident with exit out of G(0). These data support a model in which PAI-1 gene expression is closely associated with creation of the growth-activated state and that cell cycle controls appear to be superimposed on the time course of the serum-induced expression of the PAI-1 gene.

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