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PubMed2000

Prolactin and parental behavior in Adélie penguins: effects of absence from nest, incubation length, and nest failure.

Adélie penguin (Pygoscelis adeliae) males and females, nesting in Antarctica, alternate attendance at the nest with absences of many days to forage at sea. We investigated the importance of tactile input from egg and chicks on prolactin levels by observing nest attendance patterns and obtaining blood samples (1) during the first nest exchange of the incubation stage, (2) from birds whose incubation period was artificially increased or decreased by about 10 days, and (3) from birds whose nests had failed. Prolactin levels in females after 8 to 11 days of absence from the breeding colony did not differ from those in incubating males and did not change after females resumed incubation. Moving eggs between nests resulted in nests in which chicks hatched after about 26, 36 (normal), or 46 days. Duration of incubation did not affect prolactin levels in the parents measured during incubation, at the pip stage, hatch stage, or early brood stage. Adults first left their chicks unguarded on about the same calendar date, regardless of chick age. However, chicks from long incubation nests averaged 8 days younger when they were left unguarded than chicks from control or short-incubation nests. In females, there was no effect of nest failure on prolactin levels. In males, prolactin levels were slightly lower after nest failure than in males tending nests. Testosterone was significantly higher in males after nest failure than in males still tending nests. Prolactin is elevated in Adélie penguins as part of the program of cyclical hormonal changes that accompany the lengthy reproductive season and is relatively independent of tactile input. Sustained prolactin secretion is probably required for the maintenance of parental behavior in offshore feeding species that must be absent from the nest for many days at a time.

Animals↗
PubMed2000

The role of prolactin in the regulation of clutch size and onset of incubation behavior in the American kestrel.

In most bird species, the timing of incubation onset may influence the degree of hatching asynchrony, which, together with variation in clutch size, affects reproductive success. In some domesticated species that usually show no hatching asynchrony, plasma prolactin concentrations in females rise with the onset of incubation and the end of laying, and this rise enhances incubation behavior and may terminate laying. To investigate whether a rise in prolactin during laying is involved in the regulation of clutch size and incubation onset in a species with hatching asynchrony, we measured plasma concentrations of immunoreactive prolactin (ir-prolactin) in laying American kestrels, Falco sparverius, and quantified clutch size and incubation behavior. In a separate study, we administered one of three concentrations of ovine prolactin (o-prolactin) via osmotic pumps implanted in females when egg 2 of a clutch was laid. ir-Prolactin concentrations during laying were higher in small than in large clutches and increased in parallel with the development of incubation behavior. o-Prolactin treatment enhanced incubation behavior, but did not affect clutch size, possibly because the manipulation was performed after clutch size had already been determined. Consistent with studies on domesticated species that show synchronous hatching, our results indicate that rising prolactin during laying enhances the expression of incubation behavior in a species that shows hatching asynchrony. Further studies are necessary to determine whether the relationship between prolactin and clutch size in the American kestrel is one of causation or of mere association.

Animals↗
PubMed1999

Time- and concentration-dependent induction of CYP1A1 and CYP1A2 in precision-cut rat liver slices incubated in dynamic organ culture in the presence of 2,3,7,8-tetrachlorodibenzo-p-dioxin.

In a previous 24-h study, precision-cut rat liver slices were validated as a useful in vitro model for assessing the dose-related induction of CYP1A1 and CYP1A2 in rat liver following exposure to 2, 3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Further assessment of the utility of this model was accomplished by initially exposing rat liver slices to medium containing TCDD (0.01 nM) for 24 h and incubating the slices up to an additional 72 h in TCDD-free medium. The slices remained viable throughout the incubation period with an intracellular potassium content varying from 45.2 +/- 2.3 micromol/g at 48 h to 50.0 +/- 1.6 micromol/g at 72 h. In TCDD-exposed slices, CYP1A1 protein and its respective enzymatic activity, the O-deethylation of ethoxyresorufin (EROD), significantly increased with time over the 96-h incubation period, with EROD activity increasing from 63.6 +/- 14.2 at 24 h to 905 +/- 291 pmol/mg/min at 96 h. Under identical incubation conditions, but in the absence of TCDD, the EROD activity for the control liver slices ranged from 14. 3 +/- 4.3 to 44.9 +/- 11.9 pmol/min/mg. Conversely, the level of CYP1A2 protein and its respective activity (acetanilide hydroxylation) transiently decreased from 24 to 96 h with no significant differences observed between the control (0 nM TCDD) and treatment group (0.01 nM TCDD). The concentration-effect relationship at 96 h was characterized by incubating rat liver slices for the initial 24 h in medium containing TCDD at concentrations ranging from 0.1 pM to 10 nM. Induction of CYP1A1 protein and EROD activity was observed for all treatment groups with the 10 nM TCDD treatment group displaying greater than 100-fold induction compared to control (0 nM TCDD). Immunohistochemical localization of CYP1A1 protein within liver slices supported the time- and concentration-dependent induction of EROD activity by TCDD. The induction of CYP1A1 was initially observed to be centrilobular, with increased expression due to both elevated CYP1A1 within cells and the recruitment of additional cells expressing CYP1A1 throughout the entire liver slice. Additionally, the immunohistochemical analysis of the liver slices demonstrated the conservation of tissue architecture following up to 96 h of incubation in dynamic organ culture and provided further evidence for maintenance of tissue viability. In comparison to CYP1A1, the induction of CYP1A2 at 96 h was a less sensitive response, with significant induction of CYP1A2 protein and its respective activity occurring at a medium concentration of 0.1 nM TCDD (686 pg/g liver). In general, increasing the incubation period from 24 to 96 h markedly increased TCDD-induced expression of CYP1A1 and minimally enhanced CYP1A2 expression. Moreover, extending the incubation period to 96 h resulted in in vitro induction profiles for CYP1A1 and CYP1A2 that were qualitatively and quantitatively similar to that previously observed following in vivo exposure to TCDD (Drahushuk et al., Toxicol. Appl. Pharmacol. 140, 393-403, 1996).

Animals↗
PubMed1979

Apoproteins in association with Intralipid incubations in rat and human plasma.

Intralipid was incubated with rat and human plasma and examined for changes in lipid and aproprotein composition. Upon incubation in rat plasma, Intralipid acquired an apoprotein complement similar to that found in chylomicrons following plasma incubation or in chylomicrons after alimentary lipemia. Since the apoproteins of lipoproteins probably govern their metabolism, these results suggest that Intralipid and chylomicrons undergo similar metabolic fates. This pattern is characterized by a predominance of Apo E (the arginine-rich apoprotein) and Apo C. Incubation of Intralipid with human plasma showed the uptake of Apo A-I and Apo A-IV as well. Density fractionation of the plasma into separate lipoprotein classes identification of high density lipoprotein as the major apoprotein donor to the Intralipid. When rat lipoprotein-free plasma (delta greater than 1.21) was incubated with Intralipid, a different apoprotein pattern appeared in the particles of Sf greater than 400 depending on whether the entire Intralipid preparation or only the Sf greater than 400 fraction alone was incubated. The difference consisted of a virtual total absence of the arginine-rich protein on the Sf greater than 400 particles in whole Intralipid incubation. Density fractionation of the Sf less than 400 particles of Intralipid and recombination of these fractions with the Sf greater than 400 fraction before incubation revealed the major inhibitory fraction to be delta less than 1.006 (Sf 20-400).

Animals↗
PubMed2001

Improved yield and functionality of parathyroid cells separated by using collagenase-digestion with cold pre-incubation.

Preparation of cells from solid organs often induces a functional impairment due to the proteolytic cell damage by the applied digestive enzyme like collagenase, trypsin or dispase. To preserve the tissue and to enhance the yield of cells, Laue et al. reported an islet cell isolation with pre-incubation at 4 C permitting the enzyme to diffuse into the tissue and explicite activity equally throughout the whole particle. The aim of this study was to investigate whether this procedure can be applied to parathyroid glands. Therefore porcine parathyroid glands were dissected into 1 mm3 pieces. Subsequently one group of these pieces was incubated 22 h at 4 C in 2 mg/ml collagenase before activating the enzyme by elevating the temperature to 37 C for 30 min. The other group was incubated directly at 37 C for 30 min. The yield of cells and their viability was assessed by light-microscopy and staining with trypan-blue. After the cells were immobilized in barium-alginate and cultivated for 7 days, the function was tested by incubation in different calcium concentrations and PTH-measurement. Finally, the viability was assessed by histology. Using a cold pre-incubation with collagenase, a significantly higher number of isolated cells was retrieved compared with collagenase-digestion without pre-incubation. The viability was about 100% and did not differ between both groups. After immobilization and cultivation the viability decreased to less than 30%, with and without pre-incubation. In contrast to viability the PTH-secretion of the cells differed significantly between both procedures. By pre-incubation with collagenase at 4 C a gentle method is presented resulting in an enhancement of yield and function of single cells of parathyroid glands.

Animals↗
PubMed1986

Muscle cell cultures from chicken breast muscle have increased specific activities of creatine kinase when incubated at 41 degrees C compared with 37 degrees C.

Chicken muscle cell cultures were incubated at 41 degrees C, the physiological chicken body temperature, and compared with cultures incubated at 37 degrees C, the typical cell culture incubation temperature. The cultures incubated at 41 degrees C show not only an increase in creatine kinase (CK)-specific activity but also a marked increase in the percentage of adult muscle CK isozyme (MM-CK) in 7-day muscle cultures. Muscle cell cultures incubated in the presence of cytosine arabinoside (ara-C), a cell proliferation inhibitor, do not have the mononucleated cell overgrowth seen at 41 degrees C and thus exhibit a further increase in creatine kinase-specific activity compared with cultures incubated at 41 degrees C in the absence of ara-C. These results suggest that muscle cell cultures incubated at 41 degrees C are more highly differentiated than those incubated at 37 degrees C.

Animals↗
PubMed1985

The role of prolactin during incubation: comparative studies of three Diomedea albatrosses.

There is much recent evidence that prolactin is an important influence on parental and incubatory behaviour in birds. In this study prolactin was measured at various stages of the breeding cycle in three closely related albatrosses (the wandering albatross Diomedea exulans, the grey-headed albatross D. chrysostoma, and the black-browed albatross D. melanophris). Each species is monogamous, laying one egg, with the sexes sharing parental duties, including lengthy incubation shifts. In experiments where blood samples were taken daily throughout single incubation shifts (of both sexes) and every 3 hr for 36 hr, high prolactin levels were observed, but there was no indication of any changes that might suggest direct relationships between the hormone concentrations and incubatory behaviour. However, high prolactin levels were characteristic of the whole incubation period with a significant decline in concentrations towards the end of the brood-guard period. The timing of the decline in prolactin levels remained constant, even when the incubation period was artificially lengthened or shortened, as did the overall duration of the incubation-brood-guard period. Further experiments eliminated the possibility that the secretion of prolactin was a response to tactile stimulation of the brood patch by the egg. These results suggest that the incubation period is not endogenously timed but that prolactin may still affect the overall duration of the incubation-brood-guard period, although having little or no effect on the number or duration of incubation shifts.

Animals↗
PubMed1985

Stimulus requirements for prolactin and LH secretion in incubating ring doves.

To study the regulation of prolactin secretion and incubation behavior during breeding, stimuli available during the prehatching period were manipulated, and plasma prolactin and LH and pituitary prolactin concentrations were assayed. Four groups were compared: (a) nest and eggs removed (b) eggs replaced with infertile eggs, (c) eggs replaced with infertile eggs and partners separated, and (d) unmanipulated controls. The increase in prolactin secretion that normally occurs at the end of incubation was reversed when the nest and eggs were removed but was unaffected by the absence of the mate. Prolactin secretion was not maintained in males sitting beyond the normal incubation period (on infertile eggs), while it was maintained in females but only in the presence of the mate. In the control birds, the increase in plasma prolactin was accompanied by an increase in pituitary content; while in the manipulated groups, an inhibition of prolactin secretion was associated with a transient increase in pituitary content. Plasma LH levels were low in control birds during the time of high prolactin secretion, and in all groups where the disruption of normal breeding stimuli caused an early fall in prolactin, there was a concomitant increase in LH secretion. In animals which had been separated from their mates and provided with a nest bowl and eggs, continuous observations of incubation behavior were made and plasma was sampled every third day. Parallel changes in prolactin and sitting behavior occurred in some of the animals, while others continued to incubate after prolactin had declined. In no instance did an animal which maintained high prolactin show a decline in incubation. This suggests that prolactin can serve to maintain incubation, while incubation does not necessarily stimulate prolactin secretion at this phase of the breeding cycle.

Animals↗
PubMed1986

Plasma concentrations of prolactin during the breeding cycle in the Cape gannet (Sula capensis): a foot incubator.

Plasma prolactin was measured in the Cape gannet (Sula capensis) which differs from most other avian species in that although both sexes share equally in incubation duties, neither sex possesses an incubation patch but rather incubates a single egg with its foot webbing. Five main categories of birds were studied: nonbreeders, unmated birds which were nevertheless defending a nest site, recently mated birds, incubators, and brooders with nestlings. Prolactin titers in the plasma were 8-9 ng/ml in nonbreeding gannets, increasing to 17.7 +/- 1.6 and 16.7 +/- 1.2 ng/ml in incubating females and males, respectively. Plasma concentrations of prolactin progressively increased in nonbreeding gannets from 5.5 +/- 0.4 ng/ml in 1-year-old birds to 8.7 +/- 1.6 ng/ml in birds defending a nest site but without an egg. Blood samples taken from both incubators and nonincubators every 3 hr over 27 hr did not reveal any significant daily cycling in prolactin titers. Plasma osmolarity did not vary significantly between nonincubators and incubators, being 300.1 +/- 3.2 and 298.2 +/- 3.0 mOsm/kg, respectively. No positive correlation was found between possible osmotic stress during incubating and plasma concentrations of prolactin. Plasma concentrations of luteinizing hormone and testosterone increased while prolactin decreased, significantly, 24 hr after clutch loss. The putative inhibition of the pituitary-gonadal axis by prolactin is discussed together with other possible roles of prolactin during the breeding season in gannets.

Aging↗
PubMed1988

Changes in plasma levels of prolactin and estradiol, nutrient intake, and time spent nesting during the incubation phase of broodiness in the Chabo hen (Japanese bantam).

The plasma concentrations of prolactin (PRL) and estradiol, time spent nesting, consumption of feed and water, body weight, and hematocrit were measured during egg laying, incubating, and brooding of the chicks in a Japanese strain of dwarf bantam hen, Chabo. Plasma levels of PRL increased before incubation, were maintained at high levels during incubation, and decreased rapidly at the onset of hatching the young. The concentration of estradiol decreased before incubation and was maintained at low levels when circulating levels of PRL were high. During incubation, hens spent greater than 95% of the day on the nest, reduced their daily intake of feed and water by 63 and 78%, respectively, and decreased their body weight by 19% by the end of incubation. Thus, Chabo hens show a mode of incubation behavior similar to that reported in larger chickens. The temporal changes between nutrient intake and plasma levels of PRL at the start and end of incubation behavior suggested that changes in nutrient intake may not cause changes in the concentration of PRL, whereas the association between increased levels of PRL and decreased levels of estradiol suggested that they may be causally associated. Plasma levels of PRL also appeared to be associated with time spent on the nest.

Animals↗
PubMed1992

Gonadal hormones determine sex differences in timing of incubation by doves.

Male and female ring doves express a sexually dimorphic pattern of incubation. The dimorphism is temporal rather than motoric. The male incubates for a block of time in the middle of the day and the female incubates the rest of the time. The present study explored the role of gonadal hormones in the control of the temporal dimorphism. Female-female pairs incubated their eggs, but it could not be predicted which of the partners would be sitting on the nest at any given time. Male-male pairs did not incubate and instead destroyed the nests that were provided and displayed aggressive behavior. Some intact males incubated when paired with gonadectomized males, although the castrates tended to ignore the nest. In contrast, when castrated birds were given heterotypical hormonal replacement therapy, they and their same-sex partners incubated the eggs that had been provided, with the gonadectomized birds sitting at a time appropriate to the hormonal state. The results indicate that gonadal hormones influence not only the expression of incubation behavior, but also its phase and duration.

Animals↗
PubMed1986

Changes of the main isoform of human apolipoprotein A-I following incubation of plasma.

This study was aimed to ascertain whether the more acidic isoforms of plasma apo A-I (A-I-1 and A-I-2) could originate in vitro from the main plasma isoform (A-I0). Apo A-I isoforms were separated by two-dimensional gel electrophoresis before and after a prolonged incubation of serum or EDTA-plasma at 37 degrees C. Incubated plasma there was a marked decrease of apo A-I0 and a concomitant linear increase of apo A-I-1 + and A-I-2. The relative content of the latter raised from 22 +/- 7% before the incubation to 60 +/- 7% after 48 h of incubation. This conversion of A-I0 was not inhibited by either pre-heating of plasma at 60 degrees C for 1 h or the addition of protease inhibitors, EDTA and p-chloromercuriphenylsulfonic acid. The conversion of apo A-I0 was not observed in isolated HDL, incubated either in the absence or in the presence of d less than 1.063 g/ml lipoproteins and lipoprotein deficient plasma, nor in plasma which had been dialyzed before being incubated at 37 degrees C. This suggests that plasma contains a low molecular weight factor capable of promoting the conversion of A-I0. The increase of the relative content of apo A-I-1 and apo A-I-2 in incubated plasma was not due to glucosylation or carbamylation of A-I0 as no radioactive glucose and urea were found to be bound to A-I. Since the conversion of apo A-I0 was prevented by the addition of an antioxidant (butylated hydroxytoluene, BHT) to the incubated plasma it is conceivable that some product of lipid peroxidation renders apo A-I0 more electronegative by reacting with some free amino groups of this peptide.

Adult↗
PubMed1996

Effects of pre-incubation egg storage on embryonic functions and growth.

The effect of pre-incubation storage on physiological functions of chick embryos during the last half of incubation and the relationship to embryonic growth were studied. In the first experiment, eggs were stored for 20 or 30 days, respectively, and the developmental patterns of oxygen consumption (MO2), heart rate (fH) and O2 pulse of individual embryos were examined. The MO2 of stored eggs increased at significantly lower rate than the control between day 12 and 17 of incubation, and the stored eggs had a significantly lower plateau MO2 between day 17 and 19. The decrease in MO2 of some stored eggs was correlated with late incubation mortality. Pre-incubation storage also resulted in significant changes in the developmental patterns of fH and O2 pulse. In the second experiment, the eggs were stored for 10 and 20 days, respectively, to also examine the effect of short-term storage on the development of MO2 in relation to embryo growth. Pre-incubation storage for 10 days had no significant effect, but 20 days storage shifted developmental patterns of MO2, wet mass, dry mass and embryo water fraction to the right of the control. Furthermore, MO2 was significantly lower than expected on the basis of embryo mass after day 17 of incubation. Prolonged pre-incubation storage caused not only a rightward shift in MO2 pattern due to retarded growth, but also severe depression of MO2 during the last stages of prenatal development.

Agriculture↗
PubMed1985

The release of endogenous dopamine from rat striatum during incubation in vitro.

In this study the conditions for quantitative estimation of the spontaneous and KCl-stimulated release of endogenous dopamine from rat striatum during in vitro incubation have been investigated. KCl concentrations of 15-35 mM produced increasing stimulation of dopamine release during 20 min of incubation in vitro, while KCl concentrations of 45 and 60 mM produced no further significant increase in release. In the presence of 4.7 or 20 mM KCl, dopamine release was linear throughout a 20 min period of incubation; in the presence of 25 or 60 mM KCl, however, dopamine release was not linear, reaching a maximum within 8 min of incubation. Three structurally unrelated inhibitors of monoamine uptake were shown to enhance the net spontaneous release of endogenous dopamine at low concentrations, but to inhibit release at high concentrations. The ethanol-induced enhancement of striatal dopamine release was used to demonstrate the importance of an appropriate concentration of an amine uptake inhibitor in the incubation media. It is concluded that the incubation of rat striatal tissues in vitro, coupled with amine analysis by high performance liquid chromatography with electrochemical detection, constitutes an effective technique for the quantitative estimation of endogenous dopamine release, providing appropriate concentrations of a reuptake inhibitor and a monoamine oxidase inhibitor are included in the incubation media. Where rates of KCl-stimulated amine release with time are to be determined, however, a low concentration of KCl must be used, or the tissues must be subjected to incubation for short periods of time.

Animals↗
PubMed1992

Regulation of G-PROTEIN mRNA abundancy and cAMP accumulation after long-term opioid incubation in primary cultures of astroglia from the rat cerebral cortex.

Primary astroglial cultures were incubated with delta (10(-6) M DPDPE) or kappa (10(-5) M U-50,488H) receptor agonists for 5 days. Thereafter, the acute inhibitory actions of delta or kappa receptor agonists on forskolin stimulated cAMP accumulation were assayed. The G alpha s, G alpha i-1 and G alpha i-2 mRNA levels were quantified after 5 days of either delta or kappa receptor agonist treatment using a solution hybridization, RNase protection assay. Pronounced effects were observed after 5 days of kappa receptor agonist [10(-5) M U-50,488H] incubation. This treatment resulted in an attenuation in the acute inhibitory action of delta and kappa receptor agonists. Furthermore, a decreased stimulatory action of forskolin was seen. Similar effects were also seen after delta receptor stimulation. We also investigated the effects after 24 h and 3 days of incubation with the kappa receptor agonist (10(-5) M) U-50,488H. The 24 h incubation resulted in a decreased sensitivity to the acute inhibitory action of delta and kappa receptor agonists in the astroglial cultures. This effect was further accentuated after the 3 days of incubation with 10(-5) M U-50,488H. No significant change was seen in the basal accumulation of cAMP after incubation with the kappa agonist U-50,488H. However, after 5 days of incubation with the delta agonist DPDPE, a significantly increased basal accumulation of cAMP was seen in the astroglial cultures. After 5 days of delta or kappa agonist incubation, an increase in G alpha s mRNA level and a decrease in G alpha i-2 mRNA level was seen compared with controls. No statistically significant alterations in the amount of G alpha i-1 mRNA were seen. The data obtained in the present study indicate that the effects of long-term opioid treatment alters the sensitivity of glial cell opioid receptors. Furthermore, long term opioid treatment induces alterations in glial G-protein mRNA levels.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1983

Differential labeling of sensory cell and neural populations in the organ of Corti following amino acid incubations.

Label localization was compared by light and electron microscopic autoradiography in the organ of Corti of the gerbil, following in vivo incubation with one of eight 3H-labeled amino acids. Following incubation with GABA, efferent endings underneath the outer hair cells were heavily labeled, as were all tunnel crossing fibers, many small fibers in the inner spiral bundle and some efferents beneath the inner hair cells. This pattern was absent following incubations with the GABA analog muscimol. Following L- and especially D-aspartic acid incubations, efferent endings under the inner hair cells and small fibers of the inner spiral bundle were preferentially labeled. It is concluded that these differential labeling patterns after GABA and aspartic acid incubations reflect two populations of efferent fibers in the organ of Corti: one terminating primarily upon the outer hair cells and one underneath the inner hair cells. Cochlear hair cells showed the highest level of labeling following incubation with alanine. Inner hair cells were more heavily labeled than outer hair cells following incubations with alanine and glycine. Hair cell labeling was comparable to that of supporting cells for most amino acids. However, for alanine and glycine the labeling of inner hair cells, and for L-aspartic and glutamic acid the labeling of all hair cells, was higher than that of structural cells. Labeling of hair cells was substantially lower than that of structural cells following taurine incubations.

Amino Acids↗
PubMed2005

The impact of fine sediment accumulation on the survival of incubating salmon progeny: implications for sediment management.

This paper draws on results from a recent research programme on the impact of fine sediment transport through catchments to present a case for the development of new approaches to improving the quality of salmonid spawning and incubation habitats. To aid the development of these programmes, this paper summarises the mechanisms by which fine sediment accumulation influences the availability of oxygen (O2) to incubating salmon embryos. The results of the investigation indicate that incubation success is inhibited by: (i) the impact of fine sediment accumulation on gravel permeability and, subsequently, the rate of passage of oxygenated water through the incubation environment; (ii) reduced intragravel O2 concentrations that occur when O2 consuming material infiltrates spawning and incubation gravels; and (iii) the impact of fine particles (clay) on the exchange of O2 across the egg membrane. It is concluded that current granular measures of spawning and incubation habitat quality do not satisfactorily describe the complexity of factors influencing incubation success. Furthermore, an assessment of the trends in fine sediment infiltration indicates that only a small proportion of the total suspended sediment load infiltrates spawning and incubation gravels. This casts doubt over the ability of current catchment-based land use management strategies to adequately reduce fine sediment inputs.

Animals↗
PubMed1997

The effect of sperm-oocyte incubation time on in vitro embryo development using sperm from a tetraparental chimeric bull.

The present study was designed as 5 x 4 factorial to investigate the effects of using sperm from 5 bulls, and varied sperm-oocyte incubation times (5, 10, 15 and 20 h) on the fertilization, cleavage rates and blastocyst formation on an in vitro bovine embryo production system. The bulls included a tetraparental Chimera, its sires (Japanese Black and Limousin), its maternal grand-sires (Japanese Brown and Holstein). The proportion of polyspermy, 2-pronuclei formation, fertilization, cleavage and development to blastocyst were affected (P < 0.001) by the duration of sperm-oocyte incubation, as well as by the interaction between bulls and their corresponding sperm-oocyte incubation time. Blastocyst rate observed after 5 h in oocytes inseminated with Chimera, Japanese Black and Limousin were higher (p < 0.05) than those observed at 20 h incubation. The proportion of blastocysts from oocytes inseminated with Japanese Black observed at 10 h of incubation did not differ from that of Chimera, but both were higher (p < 0.05) than those observed for the Limousin, Japanese Brown and Holstein sires. The present study showed that there was an effect by the duration of sperm-oocyte incubation on in vitro embryo development. The optimal time of sperm-oocyte incubation for the Chimera was similar to that of its sires (Japanese Black and Limousin) but differed from its maternal grand-sires (Japanese Brown and Holstein). The fertilization rates for the sperm from the Holstein bull increased up to 15 h suggesting that this might be the only bull that would benefit from a long incubation period for insemination.

Animals↗